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Other Publications (199)

Articles by Min Wei in JoVE

 JoVE Biology

Studying Age-dependent Genomic Instability using the S. cerevisiae Chronological Lifespan Model

1Andrus Gerontology Center, Department of Biological Sciences, Department of Molecular and Computational Biology, University of Southern California, Los Angeles


JoVE 3030

Here we describe a set of DNA mutation assays that can be combined with the yeast chronological life span model to study the genes/pathways that regulate or contribute to genomic DNA instability during aging.

Other articles by Min Wei on PubMed

Altered Gene Expression in Rat Colonic Adenocarcinomas Induced in an Azoxymethane Plus 2-amino-1-methyl-6-phenylimidazo[4,5-b]- Pyridine Initiation-promotion Model

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), the most abundant food-derived mutagenic/carcinogenic heterocyclic amine (HCA), has attracted particular attention as a probable human colon carcinogen. Some studies have shown that PhIP administered in the post-initiation phase is able to enhance rat colon carcinogenesis remarkably. To determine whether this genotoxicant leaves a DNA footprint in colon carcinogenesis, 6-week-old male F344 rats were first subcutaneously injected with azoxymethane (AOM) and then continuously treated with various doses (0-200 ppm) of PhIP added to their diet. Animals were killed at week 36 for histopathological examination, and colonic adenocarcinomas derived from animals receiving 0, 50 and 200 ppm PhIP were subjected to a novel three-dimensional (3D)-microarray and real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis. A total of five candidate genes were identified in adenocarcinomas following 200 ppm of PhIP and AOM initiation, with a dose-dependent increment. Among them, Stat1 (signal transducer and activator of transcription 1) and VEGFc (vascular endothelial growth factor c) demonstrated statistically significant upregulation by real-time RT-PCR. In addition, HSP90 (heat shock protein 90) and VEGFa showed a non-significant tendency to increase. In summary, overexpression of Stat1, VEGF and other genes could be involved in PhIP-enhanced colon tumorigenesis in the post-initiation phase.

[Catastrophic Antiphospholipid Syndrome in Children]

[Clinical and Molecular Genetic Study on Two Patients of the Juvenile Form of Pompe Disease in China]

Glycogen-storage disease type II (GSD II, Pompe's disease) is an autosomal recessive disorder caused by a functional deficiency of acid alpha-glucosidase (GAA) that leads to glycogen accumulation within lysosomes in most tissues. The GAA gene is located to human chromosome 17q25 and contains 20 exons, 19 of which are coding. Clinically, patients with the severe infantile form of GSD II have muscle weakness and cardiomyopathy eventually leading to death before the age of two years. Patients with the juvenile or the adult form of GSD II present with myopathy with a slow progression over several years or decades. A broad genetic heterogeneity has been described in GSD II in Europe, South Africa, USA, Japan and Korea, however, the investigation has not been performed in the patients from the mainland of China. In this study, clinical analysis and mutation detection were done on Chinese patients.

[Roles of Integrin-linked Kinase in Renal Diseases]

[Preparation and Application of Oligo Microarrays for Hepatitis Virus Detection and Genotyping]

To prepare oligo microarrays for hepatitis virus detection and genotyping.

Swiveling Domain Mechanism in Pyruvate Phosphate Dikinase

Pyruvate phosphate dikinase (PPDK) catalyzes the reversible conversion of phosphoenolpyruvate (PEP), AMP, and Pi to pyruvate and ATP. The enzyme contains two remotely located reaction centers: the nucleotide partial reaction takes place at the N-terminal domain, and the PEP/pyruvate partial reaction takes place at the C-terminal domain. A central domain, tethered to the N- and C-terminal domains by two closely associated linkers, contains a phosphorylatable histidine residue (His455). The molecular architecture suggests a swiveling domain mechanism that shuttles a phosphoryl group between the two reaction centers. In an early structure of PPDK from Clostridium symbiosum, the His445-containing domain (His domain) was positioned close to the nucleotide binding domain and did not contact the PEP/pyruvate-binding domain. Here, we present the crystal structure of a second conformational state of C. symbiosum PPDK with the His domain adjacent to the PEP-binding domain. The structure was obtained by producing a three-residue mutant protein (R219E/E271R/S262D) that introduces repulsion between the His and nucleotide-binding domains but preserves viable interactions with the PEP/pyruvate-binding domain. Accordingly, the mutant enzyme is competent in catalyzing the PEP/pyruvate half-reaction but the overall activity is abolished. The new structure confirms the swivel motion of the His domain. In addition, upon detachment from the His domain, the two nucleotide-binding subdomains undergo a hinge motion that opens the active-site cleft. A similar hinge motion is expected to accompany nucleotide binding (cleft closure) and release (cleft opening). A model of the coupled swivel and cleft opening motions was generated by interpolation between two end conformations, each with His455 positioned for phosphoryl group transfer from/to one of the substrates. The trajectory of the His domain avoids major clashes with the partner domains while preserving the association of the two linker segments.

[Surgical Correction of Craniofacial Dysostosis with Midface Distraction Osteogenesis]

To investigate the effect of distraction osteogenesis on correction of craniofacial dysostosis.

[Clinical Study of Terminating Biochemical Pregnancy and Early Clinical Pregnancy with Mifepristone and Misoprostol]

To explore the efficacy and safety of terminating biochemical pregnancy (the stage in which intrauterine or ectopic pregnancy cannot be confirmed) with mifepristone and misoprostol.

Significant and Systematic Expression Differentiation in Long-lived Yeast Strains

Recent studies suggest that the regulation of longevity may be partially conserved in many eukaryotes ranging from yeast to mammals. The three yeast mutants sch9Delta, ras2Delta, tor1Delta show extended chronological life span up to three folds. Our aim is to dissect the mechanisms that lead to the yeast life span extension.

[Nephropathy Associated with Vertically Transmitted Virus Infection]

[Analysis of Influencing Factors on the Sub-health Status in Guangdong Province]

[Research and Clinical Application of Computer-aided Design of Craniofacial Prosthesis Using Mirror Technique]

To explore the feasibility of creating a 3D-CAD model of craniofacial prostheses through mirror technique to repair the unilateral craniofacial defects and restore craniofacial symmetry.

[Diagnosis and Treatment of Pulmonary Arterial Hypertension Associated Connective Tissue Diseases in Children]

[Clinical Analysis of Pulmonary Arterial Hypertension Secondary to Connective Tissue Disease in Children]

To study the epidemiology, clinical characteristics and prognosis of children with pulmonary arterial hypertension (PAH) secondary to connective tissue disease (CTD).

Label-free Biomedical Imaging with High Sensitivity by Stimulated Raman Scattering Microscopy

Label-free chemical contrast is highly desirable in biomedical imaging. Spontaneous Raman microscopy provides specific vibrational signatures of chemical bonds, but is often hindered by low sensitivity. Here we report a three-dimensional multiphoton vibrational imaging technique based on stimulated Raman scattering (SRS). The sensitivity of SRS imaging is significantly greater than that of spontaneous Raman microscopy, which is achieved by implementing high-frequency (megahertz) phase-sensitive detection. SRS microscopy has a major advantage over previous coherent Raman techniques in that it offers background-free and readily interpretable chemical contrast. We show a variety of biomedical applications, such as differentiating distributions of omega-3 fatty acids and saturated lipids in living cells, imaging of brain and skin tissues based on intrinsic lipid contrast, and monitoring drug delivery through the epidermis.

[Impacted Cancellous Autograft for Reconstructing Bone Defects of Tibial Plateau in Total Knee Arthroplasty]

To observe the effects of a newly developed technique to transform the dish-like bone defect into contained bone defect and adopt the cancellous bone obtained from bone cut to make parvula impacted bone graft to repair the bone defect of tibial plateau in total knee arthroplasty (TKA) for osteoarthritis (OA) patients with severe varus or valgus.

[Arthroscope Monitored Solution of Adult Intramuscular Injection Associated Gluteal Muscle Contracture by Radiofrequency]

To evaluate the result of releasing adult intramuscular injection associated gluteal muscle contracture under the monitor of arthroscope by radiofrequency probe.

Heterogeneous Ultrathin Films Fabricated by Alternate Assembly of Exfoliated Layered Double Hydroxides and Polyanion

Transparent heterogeneous ultrathin films of exfoliated layered double hydroxide (LDHs) nanosheets, fabricated alternately with polyanion, have been obtained via a layer-by-layer electrostatic self-assembly which yields a series of novel LDH films with potential multifunctionality.

[Monobloc Distraction Osteogenesis and Cranial Vault Remodeling in a Pediatric Patient with Severe Crouzon's Syndrome]

To report the treatment of a case of severe Crouzon's syndrome using monobloc distraction osteogenesis and cranial vault remodeling.

[Signal Pathway in Apoptosis of K562 Cells Induced by STI571]

This study was aimed to investigate the effect of tyrosine kinase inhibitor (STI571) on growth and proliferation of K562 cells by using microarray method, the changes of gene expression in the process of K562 cell apoptosis induced by STI571 and the mechanism of K562 cell apoptosis. The gene microarray probes were prepared by RD-PCR technique, then the microarray of gene expression map was constructed; the morphologic changes of K562 cells were observed under phase-contrast microscopy before and after treatment with STI571; the apoptosis of K562 cells treated with STI571 was assayed by MTT method; the expression level of genes was analyzed by self-made microarray. The results indicated that after the treatment of STI571 for 24 hours, in K562 cells appeared major morphological changes, which included nuclear shrinkage, membrane bleb and scattered apoptotic bodies. DNA gel electrophoresis also showed that the typical "DNA ladder" phenomena existed in the treated group. After hybridization, detection and analysis with microarray method, expression of 9 genes significantly down-regulated and expression of 4 genes up-regulated. These differentially expressed genes included cell cycle related genes, cell metabolizing pathway related genes, signal transduction and transcription regulation related genes and antiapoptosis genes. It is concluded that STI571 can effectively inhibit the K562 cell growth and induce K562 cell apoptosis. The genes screened from this microarray offer new information for exploration of pathogenesis of K562 cell malignant transformation and shows abundant potential targets for the treatment of CML.

Inability of Human Immunodeficiency Virus Type 1 Produced in Murine Cells to Selectively Incorporate Primer Formula

Attempts to use the mouse as a model system for studying AIDS are stymied by the multiple blocks to human immunodeficiency virus type 1 (HIV-1) replication that exist in mouse cells at the levels of viral entry, transcription, and Gag assembly and processing. In this report, we describe an additional block in the selective packaging of tRNA(3Lys) into HIV-1 produced in murine cells. HIV-1 and murine leukemia virus (MuLV) use tRNA(3Lys) and tRNA(Pro), respectively, as primers for reverse transcription. Selective packaging of tRNA(3Lys) into HIV-1 produced in human cells is much stronger than that for tRNA(Pro) incorporation into MuLV produced in murine cells, and different packaging mechanisms are used. Thus, both lysyl-tRNA synthetase and GagPol are required for tRNA(3Lys) packaging into HIV-1, but neither prolyl-tRNA synthetase nor GagPol is required for tRNA(Pro) packaging into MuLV. In this report, we show that when HIV-1 is produced in murine cells, the virus switches from an HIV-1-like incorporation of tRNA(3Lys) to an MuLV-like packaging of tRNA(Pro). The primer binding site in viral RNA remains complementary to tRNA(3Lys), resulting in a significant decrease in reverse transcription and infectivity. Reduction in tRNA(3Lys) incorporation occurs even though both murine lysyl-tRNA synthetase and HIV-1 GagPol are packaged into the HIV-1 produced in murine cells. Nevertheless, the murine cell is able to support the select incorporation of tRNA(3Lys) into another retrovirus that uses tRNA(3Lys) as a primer, the mouse mammary tumor virus.

[Stress Distribution on Periodontium of Separated Removable Partial Denture Assembly]

The objects of the present study were to construct a computer model and to evaluate the load and stress of denture by means of three dimensional finite element analysis in order to provide evidence for clinic application.

TGF-beta Utilizes SMAD3 to Inhibit CD16-mediated IFN-gamma Production and Antibody-dependent Cellular Cytotoxicity in Human NK Cells

TGF-beta can be a potent suppressor of lymphocyte effector cell functions and can mediate these effects via distinct molecular pathways. The role of TGF-beta in regulating CD16-mediated NK cell IFN-gamma production and antibody-dependent cellular cytotoxicity (ADCC) is unclear, as are the signaling pathways that may be utilized. Treatment of primary human NK cells with TGF-beta inhibited IFN-gamma production induced by CD16 activation with or without IL-12 or IL-2, and it did so without affecting the phosphorylation/activation of MAP kinases ERK and p38, as well as STAT4. TGF-beta treatment induced SMAD3 phosphorylation, and ectopic overexpression of SMAD3 resulted in a significant decrease in IFN-gamma gene expression following CD16 activation with or without IL-12 or IL-2. Likewise, NK cells obtained from smad3(-/-) mice produced more IFN-gamma in response to CD16 activation plus IL-12 when compared with NK cells obtained from wild-type mice. Coactivation of human NK cells via CD16 and IL-12 induced expression of T-BET, the positive regulator of IFN-gamma, and T-BET was suppressed by TGF-beta and by SMAD3 overexpression. An extended treatment of primary NK cells with TGF-beta was required to inhibit ADCC, and it did so by inhibiting granzyme A and granzyme B expression. This effect was accentuated in cells overexpressing SMAD3. Collectively, our results indicate that TGF-beta inhibits CD16-mediated human NK cell IFN-gamma production and ADCC, and these effects are mediated via SMAD3.

A Novel Riboflavin Gastro-mucoadhesive Delivery System Based on Ion-exchange Fiber

A novel gastro-mucoadhesive delivery system based on ion-exchange fiber has been developed. Riboflavin-5'-phosphate sodium salt (RF5P), which is site-specifically absorbed from the upper gastrointestinal tract, was used as model drug. A modified dissolution system, which can also be called 'flow through diffusion cell' (FTDC), was used to study the drug release from the drug fibers. Gastrointestinal transit studies of the RF5P fiber complexes in rats and gamma imaging study in volunteer was carried out to evaluate the gastro-retentive behavior of the fiber. The pharmacokinetic profile and parameters of riboflavin via analysis of urinary excretion of riboflavin on man were measured. Study on rat and man provide evidence for the validity of the hypothesis that the drug fiber provided good mucoadhesive properties in vivo and should therefore be of considerable interest for the development of future mucoadhesive oral drug delivery dosage forms.

Inhibitory Effects of Baicalin on Ultraviolet B-induced Photo-damage in Keratinocyte Cell Line

Baicalin, one kind of Chinese herbal medicine with anti-inflammatory and anti-oxidant property, has been commonly used as a clinical medicine. However, little has been known about the effects of Baicalin on ultraviolet (UV) induced photo-aging and photo-carcinogenesis. The photoproduct is critical to the initial event of UV-induced photo-carcinogenesis. The purpose of the present study was to investigate whether Baicalin, in immortalized human keratinocyte HaCaT cells, could inhibit ultraviolet-B (UVB) induced skin damage and its possible underlying mechanisms, such as inhibiting UVB-induced cytotoxicity and apoptosis, cyclobutane pyrimidine dimers (CPDs), down-regulating the expression of regulatory proteins which are related to cell apoptosis and DNA damage/repair. Our study revealed that Baicalin treatment could inhibit the UVB-induced cytotoxicity, apoptosis and CPD level. It also decreased the mRNA expression of apoptosis-regulatory genes (p53-p21 and c-fos), the protein levels of p53, proliferating cell nuclear antigen (PCNA) and repair protein A (RPA), and the secretion of cytokines [interleukin(IL)-6 and tumor necrosis factor (TNF-alpha)]. These results suggested that Baicalin may have an inhibitory effect on the UVB-induced photo-damage by blocking the relevant cytokine secretion and expression of p53-p21, c-fos, PCNA and RPA genes.

[Diagnosis of Prader-Willi Syndrome by Methylation-specific PCR]

Prader-Willi syndrome (PWS) is a complex, multisystem disorder, which is difficult to be diagnosed based on clinical symptoms and the purpose of this study is to establish methylation-specific PCR (MS-PCR) assay for the diagnosis of PWS, and evaluate its use in clinical cases. MS-PCR assay has been developed abroad for 10 years, and it is efficient, fast, specific and sensitive but it has not yet been used in clinical diagnosis in our country.

[Cloning and Secretory Expression of Islet Neogenesis-associated Protein in Pichia Pastoris]

To clone the recombinant human islet neogenesis-associated protein (rhINGAP) gene for its secretory expression in Pichia pastoris.

[Effects of Lanthanum on the Plant Growth and Leaf Anti-oxidative Enzyme Activities of Cucumber Seedlings Under Nitrate Stress]

The study with water culture showed that nitrate stress had significant inhibitory effects on the plant growth, especially the shoot growth of cucumber seedlings. On the 7th day under nitrate stress, the shoot fresh mass per plant decreased by 12.77 g, leaf SOD, POD and CAT activities increased, while leaf APX, DHAR and GR activities decreased significantly, compared with the control. The addition of low concentration (0.05 mmol L(-1)) LaCl3 increased the shoot fresh mass per plant by 35%, enhanced the leaf thermally stable protein content and SOD, POD, CAT, APX, DHAR and GR activities, and decreased the leaf MDA content and electrolytic leakage, resulting in a definite alleviation of the inhibitory effects of nitrate. However, an addition of high concentration (0.5 mmol L(-1)) LaCl3 had no obvious alleviation effect. It was concluded that the addition of certain concentration LaCl3 could alleviate the nitrate stress on cucumber via increasing anti-oxidative enzymes activities and thermally stable protein content.

In Situ Polymerization of the 4-vinylbenzenesulfonic Anion in Ni-Al-layered Double Hydroxide and Its Molecular Dynamic Simulation

This paper describes a systematic study on the thermal polymerization of both pristine 4-vinylbenzenesulfonic anion (VBS) and intercalated VBS in the two-dimensional (2D) gallery of Ni-Al layered double hydroxide (VBS/Ni-Al-LDH), by virtue of combining experimental and theoretical investigations. In situ FT-IR, in situ high-temperature X-ray diffraction (HT-XRD), UV-vis absorption spectroscopy, TG-DTA and elemental analysis were used to study the polymerization process, and it was found that the polymerization of VBS/Ni-Al-LDH occurs at ca. 150-170 degrees C, at least 40 degrees C lower than that of the pristine VBS, indicating that the layered structure of LDH is favorable for thermal polymerization of VBS. Therefore, this layered inorganic material may have potential application as a "molecular reactor" for enhancing the efficiency of polymerization reaction. Furthermore, the sheet-like polymerization product was obtained with the LDHs lamella as template. For better understanding the structure and arrangement of intercalated VBS and the polymerization product between the layers of Ni-Al-LDH, molecular dynamics (MD) simulation method was employed. The simulation results of hydration energies show that there are two relatively stable stages upon the increase of the number of interlayer water molecules. VBS molecules exhibit a tendency from tilted to vertical orientation with respect to the layers as the interlayer water content increases. Compared with the experimental results, the calculated interlayer spacing is more severely affected by interlayer water content. Finally, a typical tetramer product of VBS intercalated LDH was studied and the simulated equilibrium interlayer spacing is consistent with the experimental result of in situ HT-XRD. Based on the combination of experimental and theoretical studies on the interlayer polymerization system, the aim of this work is to deeply investigate the differences in thermal polymerization process between pristine monomers and intercalated ones in the gallery of LDHs, and to give detailed information of the arrangement and swelling behavior of guest molecules confined between the sheets of host layers.

[Stressors, Coping Strategies and Psychosocial State of Children with Chronic Illness]

Identifying the stressors, coping strategies, and psychosocial state of children with chronic illness would be very useful to help them to adapt to chronic medical conditions. This study aimed to investigate the stressors, coping strategies, and psychosocial state of Chinese children with chronic illness.

Soybean Seed Extracts Preferentially Express Genomic Loci of Bradyrhizobium Japonicum in the Initial Interaction with Soybean, Glycine Max (L.) Merr

Initial interaction between rhizobia and legumes actually starts via encounters of both partners in the rhizosphere. In this study, the global expression profiles of Bradyrhizobium japonicum USDA 110 in response to soybean (Glycine max) seed extracts (SSE) and genistein, a major soybean-released isoflavone for nod genes induction of B. japonicum, were compared. SSE induced many genomic loci as compared with genistein (5.0 microM), nevertheless SSE-supplemented medium contained 4.7 microM genistein. SSE markedly induced four predominant genomic regions within a large symbiosis island (681 kb), which include tts genes (type III secretion system) and various nod genes. In addition, SSE-treated cells expressed many genomic loci containing genes for polygalacturonase (cell-wall degradation), exopolysaccharide synthesis, 1-aminocyclopropane-1-carboxylate deaminase, ribosome proteins family and energy metabolism even outside symbiosis island. On the other hand, genistein-treated cells exclusively showed one expression cluster including common nod gene operon within symbiosis island and six expression loci including multidrug resistance, which were shared with SSE-treated cells. Twelve putatively regulated genes were indeed validated by quantitative RT-PCR. Several SSE-induced genomic loci likely participate in the initial interaction with legumes. Thus, these results can provide a basic knowledge for screening novel genes relevant to the B. japonicum- soybean symbiosis.

[Arthroscopically Assisted Radiofrequency Probe to Treat Achilles Tendinitis]

To evaluate the effectiveness of micro-tenotomy using a radiofrequency (RF) probe to treat chronicity achilles tendinitis.

Cytotoxicity of Combinations of Arsenicals on Rat Urinary Bladder Urothelial Cells in Vitro

Based on epidemiological data, chronic exposure to high levels of inorganic arsenic in the drinking water is carcinogenic to the urinary bladder of humans. The highly reactive trivalent organic arsenicals dimethylarsinous acid (DMA(III)) and monomethylarsonous acid (MMA(III)) are formed during the metabolism of inorganic arsenic in vivo in addition to the corresponding mono-, di- and trimethylated pentavalent arsenicals. The objective of this study was to determine if combining arsenicals was additive or synergistic toward inducing cytotoxicity in a rat urothelial cell line. The MYP3 cell line, an immortalized but not transformed rat urinary bladder epithelial cell line, was seeded into appropriate culture wells. Treatment with the arsenicals was begun 24 h after seeding and continued for 3 days. Combinations of arsenicals used were DMA(III) with arsenite, dimethylarsinic acid (DMA(V)) or trimethylarsine oxide (TMAO). Combinations of concentrations used were the LC50, one-quarter or one-half the LC50 of one arsenical with one-half or one-quarter the LC50 of the other arsenical. To determine if MYP3 cells metabolize arsenicals, cells were treated with arsenate, arsenite and MMA(V) as described above and the medium was analyzed by HPLC-ICPMS to determine species and quantity of arsenicals present. When cells were treated with one-quarter or one-half the LC50 concentration of both arsenicals, the cytotoxicity was approximately the same as when cells were treated with half the LC50 concentration or the LC50 concentration, respectively, of either arsenical. Treatment with one-quarter the LC50 concentration of one arsenical plus the LC50 concentration of a second arsenical had similar cytotoxicity as treatment with the LC50 concentration of either of the arsenicals. Quantitation and speciation of arsenicals in the cell culture medium showed that MYP3 cells have some reductase activity but the cells do not methylate arsenicals. The effect on the cytotoxicity of arsenicals in combination was additive rather than synergistic toward a rat urothelial cell line.

[Movement Capture of Knot-tying Manipulation in Microsurgeries]

The surgical dexterous suturing and knotting movements are photographed with binocular stereo vision and the movements' locus is captured through identifying the RGB values in a series of key-frames. Then a normal movement path of knot-tying is obtained through curve-fitting and smoothing. And meanwhile, the workspace of knot-tying manipulation in microsurgeries is got. And thus, a good foundation is laid for the design of an automatic knotting mechanism and for the establishment of a virtual surgery training system.

Starvation-dependent Differential Stress Resistance Protects Normal but Not Cancer Cells Against High-dose Chemotherapy

Strategies to treat cancer have focused primarily on the killing of tumor cells. Here, we describe a differential stress resistance (DSR) method that focuses instead on protecting the organism but not cancer cells against chemotherapy. Short-term starved S. cerevisiae or cells lacking proto-oncogene homologs were up to 1,000 times better protected against oxidative stress or chemotherapy drugs than cells expressing the oncogene homolog Ras2(val19). Low-glucose or low-serum media also protected primary glial cells but not six different rat and human glioma and neuroblastoma cancer cell lines against hydrogen peroxide or the chemotherapy drug/pro-oxidant cyclophosphamide. Finally, short-term starvation provided complete protection to mice but not to injected neuroblastoma cells against a high dose of the chemotherapy drug/pro-oxidant etoposide. These studies describe a starvation-based DSR strategy to enhance the efficacy of chemotherapy and suggest that specific agents among those that promote oxidative stress and DNA damage have the potential to maximize the differential toxicity to normal and cancer cells.

Lack of Mutagenic and Toxic Effects of Low Dose Potassium Bromate on Kidneys in the Big Blue Rat

Potassium bromate (KBrO3) has been classified as a genotoxic carcinogen based on positive results in the Ames test, and chromosome aberration and micronucleus tests. The purpose of the present study was to investigate the dose-response relationship for in vivo mutagenic and toxic effects of KBrO3 in the kidneys of Big Blue rats. In experiment 1, male Big Blue rats were divided into 8 groups. KBrO3 was dissolved in tap water and administered to groups 1-8 at concentrations of 0, 0.02, 0.2, 2, 8, 30, 125 and 500 ppm, respectively, for 16 weeks. Experiment 2 was performed to investigate the effects of KBrO3 at the 0.002 ppm dose approximately contained in the tap water on rat kidneys. Ten Big Blue rats were divided into 2 groups and given distilled water and tap water, respectively, for 16 weeks. In experiment 1, treatment with 500 ppm KBrO3 significantly increased the mutant and total mutation frequencies and frequency of GC to TA transversion of the lacI gene in the kidney compared to non-treatment control group, but 125 ppm and lower doses of KBrO3 had no effects. Histopathologically, renal toxic changes were observed in groups administered KBrO3 at 30 ppm or higher in a dose-dependent manner. PCNA positive cell indices in renal tubular cells were significantly increased in the kidney at doses of 125 and 500 ppm, but not at 30 ppm or lower doses, as compared to the control group. Furthermore, 8-hydroxy-2'-deoxyguanosine formation, a marker of oxidative stress, was significantly increased at 500 ppm. In experiment 2, there were no differences in any parameter between the distilled water and tap water groups. These results suggest the existence of no-effect levels for in vivo mutagenic and toxic effects, proliferation stimulus, and oxidative stress of KBrO3 in rat kidneys.

[A Study on Gene Expression Profiles of HepG2 Cell Transfected with Hepatitis B Virus Using Oligonucleotide Microarray]

Life Span Extension by Calorie Restriction Depends on Rim15 and Transcription Factors Downstream of Ras/PKA, Tor, and Sch9

Calorie restriction (CR), the only non-genetic intervention known to slow aging and extend life span in organisms ranging from yeast to mice, has been linked to the down-regulation of Tor, Akt, and Ras signaling. In this study, we demonstrate that the serine/threonine kinase Rim15 is required for yeast chronological life span extension caused by deficiencies in Ras2, Tor1, and Sch9, and by calorie restriction. Deletion of stress resistance transcription factors Gis1 and Msn2/4, which are positively regulated by Rim15, also caused a major although not complete reversion of the effect of calorie restriction on life span. The deletion of both RAS2 and the Akt and S6 kinase homolog SCH9 in combination with calorie restriction caused a remarkable 10-fold life span extension, which, surprisingly, was only partially reversed by the lack of Rim15. These results indicate that the Ras/cAMP/PKA/Rim15/Msn2/4 and the Tor/Sch9/Rim15/Gis1 pathways are major mediators of the calorie restriction-dependent stress resistance and life span extension, although additional mediators are involved. Notably, the anti-aging effect caused by the inactivation of both pathways is much more potent than that caused by CR.

Longevity Mutation in SCH9 Prevents Recombination Errors and Premature Genomic Instability in a Werner/Bloom Model System

Werner and Bloom syndromes are human diseases characterized by premature age-related defects including elevated cancer incidence. Using a novel Saccharomyces cerevisiae model system for aging and cancer, we show that cells lacking the RecQ helicase SGS1 (WRN and BLM homologue) undergo premature age-related changes, including reduced life span under stress and calorie restriction (CR), G1 arrest defects, dedifferentiation, elevated recombination errors, and age-dependent increase in DNA mutations. Lack of SGS1 results in a 110-fold increase in gross chromosomal rearrangement frequency during aging of nondividing cells compared with that generated during the initial population expansion. This underscores the central role of aging in genomic instability. The deletion of SCH9 (homologous to AKT and S6K), but not CR, protects against the age-dependent defects in sgs1Delta by inhibiting error-prone recombination and preventing DNA damage and dedifferentiation. The conserved function of Akt/S6k homologues in lifespan regulation raises the possibility that modulation of the IGF-I-Akt-56K pathway can protect against premature aging syndromes in mammals.

Single-crystalline ZnGa2O4 Spinel Phosphor Via a Single-source Inorganic Precursor Route

The synthesis of single-crystalline ZnGa 2O 4 spinel phosphor with intense ultraviolet-emitting properties through a novel single-source inorganic precursor route is reported. This synthetic approach involves the calcination of a Zn-Ga layered double hydroxide precursor followed by selective leaching of the self-generated zinc oxide. Material characterization has been presented by chemical analysis, X-ray diffraction analysis, thermogravimetric-differential thermal analysis, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, electron paramagnetic resonance, nuclear magnetic resonance, extended X-ray absorption fine structure analysis, UV-vis, and photoluminescence measurements. The results indicate that a single-crystalline ZnGa 2O 4 spinel with an average particle size of around 150 nm has been obtained at a lower calcination temperature and shorter calcination time compared with that with the high-temperature solid-state reaction method, based on the fact that the large amount of highly dispersed ZnO particles generated during the high-temperature calcination of the single-source inorganic precursor has a remarkable segregation and inhibition effect on the growth of ZnGa 2O 4 spinel. Furthermore, it has been confirmed that that Ga (3+) ions locate not only on the octahedral sites but also on the tetrahedral sites in the matrix of the ZnGa 2O 4 spinel structure, and the Ga-O coordination environment has a great influence on the photoluminescence of ZnGa 2O 4 phosphors.

Two-dimensional Reaction Free Energy Surfaces of Catalytic Reaction: Effects of Protein Conformational Dynamics on Enzyme Catalysis

We introduce a two-dimensional (2D) multisurface reaction free energy description of the catalytic cycle that explicitly connects the recently observed multi-time-scale conformational dynamics as well as dispersed enzymatic kinetics to the classical Michaelis-Menten equation. A slow conformational motion on a collective enzyme coordinate Q facilitates the catalytic reaction along the intrinsic reaction coordinate X, providing a dynamic realization of Pauling's well-known idea of transition-state stabilization. The catalytic cycle is modeled as transitions between multiple displaced harmonic wells in the XQ space representing different states of the cycle, which is constructed according to the free energy driving force of the cycle. Subsequent to substrate association with the enzyme, the enzyme-substrate complex under strain exhibits a nonequilibrium relaxation toward a new conformation that lowers the activation energy of the reaction, as first proposed by Haldane. The chemical reaction in X is thus enslaved to the down hill slow motion on the Q surface. One consequence of the present theory is that, in spite of the existence of dispersive kinetics, the Michaelis-Menten expression of the catalysis rate remains valid under certain conditions, as observed in recent single-molecule experiments. This dynamic theory builds the relationship between the protein conformational dynamics and the enzymatic reaction kinetics and offers a unified description of enzyme fluctuation-assisted catalysis.

Isothiazolidinone Inhibitors of PTP1B Containing Imidazoles and Imidazolines

The structure-based design and synthesis of isothiazolidinone (IZD) inhibitors of PTP1B containing imidazoles and imidazolines and their modification to interact with the B site of PTP1B are described here. The X-ray crystal structures of 3I and 4I complexed with PTP1B were solved and revealed the inhibitors are interacting extensively with the B site of the enzyme.

[The Expression Status of 11 Genes, Located at the Commonly Deleted Region 3p21.3, in Non-small Cell Lung Cancer.]

The deletion of allele located at 3p21.3 region is found in 50%-80% of nonsmall cell lung cancer (NSCLC), suggesting that this region might harbor some tumor suppressor genes (TSGs). It is anticipated that some TSGs closely related to NSCLC carcinogenesis might be found by screening the expression of genes, located at the commonly deleted region 3p21.3, in NSCLC.

[Effects of Exogenous NO on the Growth and Antioxidant Enzyme Activities of Cucumber Seedlings Under NO3- Stress]

In this study, cucumber seedlings were cultivated in nutrient solution added with different concentration (0.05, 0.1, 0.2, and 0.3 mmol x L(-1)) sodium nitroprusside (SNP) as nitric oxide (NO) donor to study the effects of exogenous NO on the growth of cucumber seedlings and the activities of antioxidant enzymes in cucumber leaves under NO3- stress. Under the stress of 140 mmol x L(-1) NO3-, treating with 0.1 mmol x L(-1) of SNP for 1 d or 7 d increased the leaf soluble protein content and superoxidase (SOD), catalase (CAT), and ascorbic acid peroxidase (APX) activities, and decreased the leaf malondialdehyde (MDA) content significantly, suggesting that exogenous NO could enhance the capacity of cucumber seedlings in scavenging active oxygen species, protect the seedlings from the peroxidation of membrane lipids, and promote the seedlings growth and increase their resistance to high concentration NO3- stress. After the cucumber seedlings grew in 0.3 mmol x L(-1) of SNP for 7 d, the activities of leaf SOD, POD, and CAT decreased, and the MDA content increased, resulting in the injury of cucumber seedlings. It was indicated that certain concentration (0.1-0.2 mmol x L(-1)) exogenous NO could alleviate the NO3- stress to cucumber seedlings.

[Effects of Rotation and Fallowing on the Microbial Communities and Enzyme Activities in a Solar Greenhouse Soil Under Continuous Cucumber Cropping]

A pot experiment was conducted to study the effects of rotation and fallowing on the microbial communities and enzyme activities in a greenhouse soil continuously cropped with cucumber and on the growth and yield of followed cucumber. Comparing with continuous cropping, rotation improved the components of soil microbial communities, which was manifested in the increase of bacteria and actinomycetes and the decrease of fungi. Rotation and fallowing enhanced the activities of soil invertase, urease, catalase, and polyphenol oxidase significantly. The quantities of soil bacteria and actinomycetes and the activity of soil invertase increased at the fruiting stage of cucumber plants, being the maximum at harvest stage, but decreased thereafter. In contrast, the quantity of soil fungi had a linear increase, and the activities of soil urease, catalase, and polyphenol oxidase decreased gradually during fruit development. Welsh onion and waxy maize promoted the growth and fruiting of the followed cucumber plants significantly, being the optimal rotation crops for cucumber.

Fasting and Cancer Treatment in Humans: A Case Series Report

Short-term fasting (48 hours) was shown to be effective in protecting normal cells and mice but not cancer cells against high dose chemotherapy, termed Differential Stress Resistance (DSR), but the feasibility and effect of fasting in cancer patients undergoing chemotherapy is unknown. Here we describe 10 cases in which patients diagnosed with a variety of malignancies had voluntarily fasted prior to (48-140 hours) and/or following (5-56 hours) chemotherapy. None of these patients, who received an average of 4 cycles of various chemotherapy drugs in combination with fasting, reported significant side effects caused by the fasting itself other than hunger and lightheadedness. Chemotherapy associated toxicity was graded according to the Common Terminology Criteria for Adverse Events (CTCAE) of the National Cancer Institute (NCI). The six patients who underwent chemotherapy with or without fasting reported a reduction in fatigue, weakness, and gastrointestinal side effects while fasting. In those patients whose cancer progression could be assessed, fasting did not prevent the chemotherapy-induced reduction of tumor volume or tumor markers. Although the 10 cases presented here suggest that fasting in combination with chemotherapy is feasible, safe, and has the potential to ameliorate side effects caused by chemotherapies, they are not meant to establish practice guidelines for patients undergoing chemotherapy. Only controlled-randomized clinical trials will determine the effect of fasting on clinical outcomes including quality of life and therapeutic index.

[Relapsing Polychondritis in Childhood: Report of Three Cases and Review of the Literature]

To analyze the clinical features, diagnosis, treatment and prognosis of relapsing polychondritis (RP) in childhood.

Compared Study of Asian Reduction Malarplasty: Wedge-section Osteotomy Versus Conventional Procedures

The zygomatic body and arch are usually prominent in Asians; therefore, malar reduction is one of the most popular procedures for aesthetic facial contouring. The purpose of this study was to establish a new option for reduction malarplasty and analyze the merits and demerits of conventional operative techniques, thus searched for more effective and reliable surgical procedures.

[Clinical Characteristics and Follow-up of 12 Cases with Severe Chronic Active Epstein-Barr Virus Infection]

There are two different types of chronic active Epstein-Barr virus (CAEBV) infection: chronic EBV (CEBV) having persistent infectious mononucleosis (IM)-like illness with relatively good prognosis, and severe CAEBV (SCAEBV)infection that has rather severe manifestations and generally poor prognosis with many life-threatening complications, such as EBV-associated malignant lymphoma and hemophagocytic syndrome (HPS). The purpose of this study was to clarify the clinical and prognostic characteristics in 12 cases with SCAEBV infection.

[RIGIDfix Tibial and Femur Cross Pin System Used for Hamstring Grafted Anterior Cruciate Ligament Reconstruction]

To evaluate the curative effect of RIGIDfix tibial and femur cross pin system used for hamstring grafted reconstruction of anterior cruciate ligament (ACL) in arthroscopy.

[Characteristics of Peripheral Blood Lymphocyte Immune Subsets in Patients with Chronic Active Epstein-Barr Virus Infection]

To study the characteristics of the peripheral blood lymphocyte subsets in pediatric patients with chronic active EBV (CAEBV) infection.

Biotemplated Hierarchical Nanostructure of Layered Double Hydroxides with Improved Photocatalysis Performance

We report a biomorphic hierarchical mixed metal oxide (MMO) framework through a biotemplated synthesis method. A uniform Al(2)O(3) coating was deposited on the surface of the biotemplate with an atomic layer deposition (ALD) process, and the film of ZnAl-layered double hydroxide (ZnAl-LDH), which faithfully inherits the surface structure of the biotemplate, was prepared by an in situ growth technique. Subsequently, a polycrystal ZnAl-MMO framework obtained by calcination of the LDH precursor has been demonstrated as an effective and recyclable photocatalyst for the decomposition of dyes in water, owing to its rather high specific surface area and hierarchical distribution of pore size. Therefore, the new strategy reported in this work can be used to fabricate a variety of biomorphic LDHs as well as MMO frameworks through replication of complicated and hierarchical biological structures for the purpose of catalysis, adsorbents, and other potential applications.

Imaging Chromophores with Undetectable Fluorescence by Stimulated Emission Microscopy

Fluorescence, that is, spontaneous emission, is generally more sensitive than absorption measurement, and is widely used in optical imaging. However, many chromophores, such as haemoglobin and cytochromes, absorb but have undetectable fluorescence because the spontaneous emission is dominated by their fast non-radiative decay. Yet the detection of their absorption is difficult under a microscope. Here we use stimulated emission, which competes effectively with the nonradiative decay, to make the chromophores detectable, and report a new contrast mechanism for optical microscopy. In a pump-probe experiment, on photoexcitation by a pump pulse, the sample is stimulated down to the ground state by a time-delayed probe pulse, the intensity of which is concurrently increased. We extract the miniscule intensity increase with shot-noise-limited sensitivity by using a lock-in amplifier and intensity modulation of the pump beam at a high megahertz frequency. The signal is generated only at the laser foci owing to the nonlinear dependence on the input intensities, providing intrinsic three-dimensional optical sectioning capability. In contrast, conventional one-beam absorption measurement exhibits low sensitivity, lack of three-dimensional sectioning capability, and complication by linear scattering of heterogeneous samples. We demonstrate a variety of applications of stimulated emission microscopy, such as visualizing chromoproteins, non-fluorescent variants of the green fluorescent protein, monitoring lacZ gene expression with a chromogenic reporter, mapping transdermal drug distributions without histological sectioning, and label-free microvascular imaging based on endogenous contrast of haemoglobin. For all these applications, sensitivity is orders of magnitude higher than for spontaneous emission or absorption contrast, permitting nonfluorescent reporters for molecular imaging.

Layer-by-layer Assembly of Ruthenium(II) Complex Anion/layered Double Hydroxide Ordered Ultrathin Films with Polarized Luminescence

Ordered ultrathin films of tris(1,10-phenanthroline-4,7-diphenylsulfonate)ruthenium(ii) anions and cationic layered double hydroxide monolayers have been fabricated by the layer-by-layer assembly technique and show well-defined polarized red luminescence.

Potassium Bromate Enhances N-ethyl-N-hydroxyethylnitrosamine-induced Kidney Carcinogenesis Only at High Doses in Wistar Rats: Indication of the Existence of an Enhancement Threshold

As susceptibility to carcinogens varies considerably among different strains of experimental animals, evaluation of dose-response relationships for genotoxic carcinogen in different strains is indispensable for risk assessment. Potassium bromate (KBrO(3)) is a genotoxic carcinogen inducing kidney cancers at high doses in male F344 rats, but little is known about its carcinogenic effects in other strains of rats. The purpose of the present study was to determine dose-response relationships for carcinogenic effects of KBrO(3) on N-ethyl-N-hydroxyethylnitrosamine (EHEN)-induced kidney carcinogenesis in male Wistar rats. We found that KBrO(3) showed significant enhancement effects on EHEN-induced kidney carcinogenesis at above 250 ppm but not at doses of 125 ppm and below when evaluated in terms of induction of either preneoplastic lesions or tumors in male Wistar rats. Furthermore, KBrO(3) significantly increased the formation of oxidative DNA damage at doses of 125 and above but not at doses of 30 ppm and below in kidneys. These results demonstrated that low doses of KBrO(3) exert no effects on development of EHEN-initiated kidney lesions and induction of oxidative DNA damage. Taking account of previous similar findings in male F344 rats, it is strongly suggested that a threshold dose exists for enhancement effects of KBrO(3) on kidney carcinogenesis in rats.

A Combined Study Based on Experiment and Molecular Dynamics: Perylene Tetracarboxylate Intercalated in a Layered Double Hydroxide Matrix

This paper describes a combined experimental and theoretical simulation investigation on the photophysical properties, thermolysis, and orientation of 3,4,9,10-perylene tetracarboxylate (PTCB) intercalated Mg-Al-layered double hydroxide (PTCB/Mg-Al-LDH). UV-vis absorption and fluorescence spectroscopy show the existence of PTCB aggregates within the gallery of LDH, indicative of H-type (blue-shifted absorption band) and J-type dimers (red-shifted absorption band). In situ high-temperature X-ray diffraction (HT-XRD), thermogravimetry and differential thermal analysis (TG-DTA) and elemental analysis were used to study the thermal decomposition properties of PTCB/Mg-Al-LDH, and it was found that the decomposition temperature of the intercalated PTCB is lower than its pristine form (460 vs 565 degrees C), indicating that the strong pi-pi interaction among PTCB was weakened by the positively charged LDH host layers. Molecular dynamics (MD) calculations were employed to simulate the molecular arrangement and aggregation behavior of intercalated PTCB in the gallery of Mg-Al-LDH. The simulation results show that the intercalated PTCB anions exhibit a tendency from tilted to vertical orientation with respect to the layers as the interlayer water content increases, furthermore, the H and J-type dimer species are most likely populated under low and high hydration conditions, respectively. Moreover, the distribution of interlayer water molecules are also discussed.

CD94 Defines Phenotypically and Functionally Distinct Mouse NK Cell Subsets

Understanding of heterogeneous NK subsets is important for the study of NK cell biology and development, and for the application of NK cell-based therapies in the treatment of disease. Here we demonstrate that the surface expression of CD94 can distinctively divide mouse NK cells into two approximately even CD94(low) and CD94(high) subsets in all tested organs and tissues. The CD94(high) NK subset has significantly greater capacity to proliferate, produce IFN-gamma, and lyse target cells than does the CD94(low) subset. The CD94(high) subset has exclusive expression of NKG2A/C/E, higher expression of CD117 and CD69, and lower expression of Ly49D (activating) and Ly49G2 (inhibitory). In vivo, purified mouse CD94(low) NK cells become CD94(high) NK cells, but not vice versa. Collectively, our data suggest that CD94 is an Ag that can be used to identify functionally distinct NK cell subsets in mice and could also be relevant to late-stage mouse NK cell development.

Super-resolution Laser Scanning Microscopy Through Spatiotemporal Modulation

Super-resolution optical microscopy has attracted great interest among researchers in many fields, especially in biology where the scale of physical structures and molecular processes fall below the diffraction limit of resolution for light. As one of the emerging techniques, structured illumination microscopy can double the resolution by shifting unresolvable spatial frequencies into the pass-band of the microscope through spatial frequency mixing with a wide-field structured illumination pattern. However, such a wide-field scheme typically can only image optically thin samples and is incompatible with multiphoton processes such as two-photon fluorescence, which require point scanning with a focused laser beam. Here, we propose two new super-resolution schemes for laser scanning microscopy by generalizing the concept of a spatially nonuniform imaging system. One scheme, scanning patterned illumination (SPIN) microscopy, employs modulation of the excitation combined with temporally cumulative imaging by a nondescanned array detector. The other scheme, scanning patterned detection (SPADE) microscopy, utilizes detection modulation together with spatially cumulative imaging, in this case by a nondescanned single-element detector. When combined with multiphoton excitation, both schemes can image thick samples with three-dimensional optical sectioning and much improved resolution.

Role of Conformational Dynamics in Kinetics of an Enzymatic Cycle in a Nonequilibrium Steady State

Enzyme is a dynamic entity with diverse time scales, ranging from picoseconds to seconds or even longer. Here we develop a rate theory for enzyme catalysis that includes conformational dynamics as cycling on a two-dimensional (2D) reaction free energy surface involving an intrinsic reaction coordinate (X) and an enzyme conformational coordinate (Q). The validity of Michaelis-Menten (MM) equation, i.e., substrate concentration dependence of enzymatic velocity, is examined under a nonequilibrium steady state. Under certain conditions, the classic MM equation holds but with generalized microscopic interpretations of kinetic parameters. However, under other conditions, our rate theory predicts either positive (sigmoidal-like) or negative (biphasic-like) kinetic cooperativity due to the modified effective 2D reaction pathway on X-Q surface, which can explain non-MM dependence previously observed on many monomeric enzymes that involve slow or hysteretic conformational transitions. Furthermore, we find that a slow conformational relaxation during product release could retain the enzyme in a favorable configuration, such that enzymatic turnover is dynamically accelerated at high substrate concentrations. The effect of such conformation retainment in a nonequilibrium steady state is evaluated.

Oncogene Homologue Sch9 Promotes Age-dependent Mutations by a Superoxide and Rev1/Polzeta-dependent Mechanism

Oncogenes contribute to tumorigenesis by promoting growth and inhibiting apoptosis. Here we examine the function of Sch9, the Saccharomyces cerevisiae homologue of the mammalian Akt and S6 kinase, in DNA damage and genomic instability during aging in nondividing cells. Attenuation of age-dependent increases in base substitutions, small DNA insertions/deletions, and gross chromosomal rearrangements (GCRs) in sch9Delta mutants is associated with increased mitochondrial superoxide dismutase (MnSOD) expression, decreased DNA oxidation, reduced REV1 expression and translesion synthesis, and elevated resistance to oxidative stress-induced mutagenesis. Deletion of REV1, the lack of components of the error-prone Polzeta, or the overexpression of SOD1 or SOD2 is sufficient to reduce age-dependent point mutations in SCH9 overexpressors, but REV1 deficiency causes a major increase in GCRs. These results suggest that the proto-oncogene homologue Sch9 promotes the accumulation of superoxide-dependent DNA damage in nondividing cells, which induces error-prone DNA repair that generates point mutations to avoid GCRs and cell death during the first round of replication.

Urinary Bladder Carcinogenesis Induced by Chronic Exposure to Persistent Low-dose Ionizing Radiation After Chernobyl Accident

Urinary bladder urothelium as well as cells in the microenvironment of lamina propria (endothelial elements, fibroblasts and lymphocytes) demonstrate a number of responses to chronic persistent long-term, low-dose ionizing radiation (IR). Thus, oxidative stress occurs, accompanied by up-regulation of at least two signaling pathways (p38 mitogen-activated protein kinase and nuclear factor-kappaB cascades) and activation of growth factor receptors, in the bladder urothelium of people living in Cesium 137-contaminated areas of Ukraine, resulting in chronic inflammation and the development of proliferative atypical cystitis, so-called Chernobyl cystitis, which is considered a possible pre-neoplastic condition in humans. Furthermore, significant alterations in regulation of cell cycle transitions are associated with increased cell proliferation, along with up-regulated ubiquitination and sumoylation processes as well as inefficient DNA repair (base and nucleotide excision repair pathways) in the affected urothelium. The microenvironmental changes induced by chronic long-term, low-dose IR also appear to promote angiogenesis and remodeling of the extracellular matrix that could facilitate invasion as well as progression of pre-existing initiated cells to malignancy. Based on the available findings, new strategies have been developed for predicting and treatment of Chernobyl cystitis-a first step in urinary bladder carcinogenesis in humans.

Modified Heparins Inhibit Integrin Alpha(IIb)beta(3) Mediated Adhesion of Melanoma Cells to Platelets in Vitro and in Vivo

The adhesion of tumor cells with platelets is important in the process of tumor metastasis. A huge work has indicated that anti-adhesion is an effective strategy for metastasis inhibition. In this article, we assess the role of platelet integrin alpha(IIb)beta(3) in adhesion of melanoma cells to platelets and the effects of heparin and modified heparins on the adhesion in vitro and in vivo. We show that platelet integrin alpha(IIb)beta(3) is involved in the interaction of human melanoma A375 cells with platelets, and the high affinity epitope resides on the alpha(IIb) subunit rather than beta(3) subunit. Heparin sulfate-like proteoglycans on tumor cell surface are implicated in the adhesion of A375 cells to integrin alpha(IIb)beta(3). We also show that RO-heparin, CR-heparin, N-2,3-DS-heparin and 2,3-O-DS-heparin can significantly inhibit A375 cells binding to the CHO cells expressing integrin alpha(IIb)beta(3) under static and flow conditions, and remarkably inhibit the adhesion of A375 cells to the immobilized platelet layers under flow conditions. We find that A375 cells and B16F10 cells are arrested in the pulmonary vessels and adhered to platelets, and the initial interaction of tumor cells with platelets in lung vessel and long-term establishment of metastatic foci can be inhibited by heparin as well as CR-heparin and N-2,3-DS-heparin. These data suggest that modified heparins can inhibit tumor cell-platelet interaction mediated by platelet integrin alpha(IIb)beta(3) and modified heparins may be a potential substitute for heparin in inhibiting tumor metastasis.

[Clinical and Pathological Features of Glycogen Storage Disease Type III]

To summarize the clinical and pathological features of glycogen storage disease (GSD) type III.

[Carvedilol Therapy in Pediatric Patients with Takayasu's Arteritis Complicated with Congestive Heart Failure: a Case Report]

[Clinical Manifestations, Laboratory Test, Treatment and Outcome of Neuropsychiatric Involvement in Pediatric Systemic Lupus Erythematosus Patients]

To investigate the clinical manifestations, laboratory test, treatment and outcome of neuropsychiatric (NP) involvement in pediatric systemic lupus erythematosus (SLE) patients.

Response Linearity of Alert Monkey Non-eye Movement Vestibular Nucleus Neurons During Sinusoidal Yaw Rotation

Vestibular afferents display linear responses over a range of amplitudes and frequencies, but comparable data for central vestibular neurons are lacking. To examine the effect of stimulus frequency and magnitude on the response sensitivity and linearity of non-eye movement central vestibular neurons, we recorded from the vestibular nuclei in awake rhesus macaques during sinusoidal yaw rotation at frequencies between 0.1 and 2 Hz and between 7.5 and 210 degrees/s peak velocity. The dynamics of the neurons' responses across frequencies, while holding peak velocity constant, was consistent with previous studies. However, as the peak velocity was varied, while holding the frequency constant, neurons demonstrated lower sensitivities with increasing peak velocity, even at the lowest peak velocities tested. With increasing peak velocity, the proportion of neurons that silenced during a portion of the response increased. However, the decrease in sensitivity of these neurons with higher peak velocities of rotation was not due to increased silencing during the inhibitory portion of the cycle. Rather the neurons displayed peak firing rates that did not increase in proportion to head velocity as the peak velocity of rotation increased. These data suggest that, unlike vestibular afferents, the central vestibular neurons without eye movement sensitivity examined in this study do not follow linear systems principles even at low velocities.

Boron-doped Diamond Nanograss Array for Electrochemical Sensors

A novel BDD nanograss array has been prepared simply on a heavily doped BDD film by reactive ion etching for use as an electrochemical sensor, which improved the reactive site, promoted the electrocatalytic activity, accelerated the electron transfer, and enhanced the selectivity.

A Novel Method Developed for Acetylcholine Detection in Royal Jelly by Using Capillary Electrophoresis Coupled with Electrogenerated Chemiluminescence Based on a Simple Reaction

A novel method for highly sensitive detection of acetylcholine in royal jelly was proposed by using CE coupled with electrogenerated chemiluminescence (ECL). Acetylcholine, which could not react with Tris(2,2'-bipyridine)ruthenium(II) to strengthen its ECL signals, decomposed into trimethylamine and strengthened the ECL signals sharply when it was heated to its melting point. This reaction needed no additional reagent and it was mild, simple, stable and rapid, without any side reaction. By combining the above process with CE separation technique, trimethylamine in royal jelly was completely separated from interfering substances and was successfully detected within 4 min. The limit of detection for acetylcholine was found to be 6.3 x 10(-8) g/mL with a signal-to-noise ratio of 3:1. Acetylcholine in the royal jelly was detected to be 912+/-58 microg/g. The recoveries of acetylcholine chloride in the sample were in the ranges of 92-106%. The coefficients of variation for intra-day and inter-day reproducibility were equal to or less than 4.9 and 6.8%, respectively.

Microarray Analysis Identifies Differentially Expressed Genes Induced by Human Papillomavirus Type 18 E6 Silencing RNA

The oncoprotein E6 of high-risk human papillomavirus (HPV) types promotes cell proliferation and contributes to carcinogenesis of HPV-positive cervical cancer cells. In this study, we used small interfering RNA (siRNA) technology to silence the E6 gene in HPV-18-transformed human cervical cell line HeLa and determined the effects of E6 gene knockdown on the cell by using microarray-based gene expression profiling coupled with gene functional classification with bioinformatics methods. Silencing RNA prepared by siRNA expression cassettes against HPV-18 E6 gene could significantly inhibit E6 gene expression and induce HeLa cells to apoptosis. The microarray analysis identified 359 differentially expressed genes containing 307 up-regulated and 52 down-regulated genes. We analyzed the gene functions and cellular pathways in detail, including cell cycle-related genes, CCNG1 and p21; apoptosis-related genes, CASP4, CASP6, IGFBP3, and DFFA; ubiquitin proteolysis pathway-related genes, UBE3A and UBE2C; keratinocyte differentiation-related genes, KRT4, KRT6E, and KRT18; and antioncogenes, RECK and VEL. In addition, it can be concluded that cellular apoptosis induced by HPV-18 E6 siRNA mainly depends on the P53 and ubiquitin proteolysis pathway to regulate gene expression, consequently inhibiting cell proliferation and promoting cell apoptosis. Meanwhile, activation of antioncogene and upper regulation of immunization-related genes signified the degression of the malignant extent of tumor cells after E6 inhibition. Our approach, which combines the use of siRNA-mediated gene silencing, microarray screening, and functional classification of differential genes, can be used in functional genomics study to elucidate the role of E6 oncogene in the carcinogenesis of HPV-18 and provide some possible targets for clinical treatment and drug development of cervical cancer.

Discovery of Potent Competitive Inhibitors of Indoleamine 2,3-dioxygenase with in Vivo Pharmacodynamic Activity and Efficacy in a Mouse Melanoma Model

A hydroxyamidine chemotype has been discovered as a key pharmacophore in novel inhibitors of indoleamine 2,3-dioxygenase (IDO). Optimization led to the identification of 5l, which is a potent (HeLa IC(50) = 19 nM) competitive inhibitor of IDO. Testing of 5l in mice demonstrated pharmacodynamic inhibition of IDO, as measured by decreased kynurenine levels (>50%) in plasma and dose dependent efficacy in mice bearing GM-CSF-secreting B16 melanoma tumors.

Near-degenerate Four-wave-mixing Microscopy

Fluorescence microscopy has been widely used to explore the nanoscale world because of its superb sensitivity, but it is limited to fluorescent samples. Hence, various spectroscopic contrasts have been explored for imaging nonfluorescent species. Here we report a multiphoton microscopy based on single-beam near-degenerate four wave mixing (ND-FWM), by detecting a coherent signal generated by the sample at frequencies close to the "edge" of the spectrally "truncated" incident femtosecond pulses. ND-FWM microscopy allows label-free biomedical imaging with high sensitivity and spatial resolution. In particular, by achieving a nearly perfect phase matching condition, ND-FWM generates almost the highest nonlinear coherent signal in a bulk medium and provides a contrast mechanism different from other nonlinear imaging techniques. More importantly, we developed an electronic resonant version of ND-FWM for absorbing but nonfluorescent molecules. Ultrasensitive chromophore detection (approximately 50 molecules) and hemoglobin imaging are demonstrated, by harnessing a fully (triply) resonant enhancement of the nonlinear polarization and using optical heterodyne detection.

Tor1/Sch9-regulated Carbon Source Substitution is As Effective As Calorie Restriction in Life Span Extension

The effect of calorie restriction (CR) on life span extension, demonstrated in organisms ranging from yeast to mice, may involve the down-regulation of pathways, including Tor, Akt, and Ras. Here, we present data suggesting that yeast Tor1 and Sch9 (a homolog of the mammalian kinases Akt and S6K) is a central component of a network that controls a common set of genes implicated in a metabolic switch from the TCA cycle and respiration to glycolysis and glycerol biosynthesis. During chronological survival, mutants lacking SCH9 depleted extracellular ethanol and reduced stored lipids, but synthesized and released glycerol. Deletion of the glycerol biosynthesis genes GPD1, GPD2, or RHR2, among the most up-regulated in long-lived sch9Delta, tor1Delta, and ras2Delta mutants, was sufficient to reverse chronological life span extension in sch9Delta mutants, suggesting that glycerol production, in addition to the regulation of stress resistance systems, optimizes life span extension. Glycerol, unlike glucose or ethanol, did not adversely affect the life span extension induced by calorie restriction or starvation, suggesting that carbon source substitution may represent an alternative to calorie restriction as a strategy to delay aging.

Cytokeratin 8/18 Overexpression and Complex Formation As an Indicator of GST-P Positive Foci Transformation into Hepatocellular Carcinomas

Screening of the proteome of microdissected glutathione S-transferase placental form (GST-P) positive foci and normal-appearing liver on anionic (Q10), and cationic (CM10) surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) ProteinChip arrays demonstrated significant overexpression of cytokeratin 8 (CK8; m/z 54,020), cytokeratin 18 (CK18; m/z 47,760), microsomal cytochrome 5A (m/z 15,224) and histone type 2 H2aa3 (m/z 15,964) in the livers of rats initiated with diethylnitrosamine (DEN) followed by 10 weeks on phenobarbital (PB) at a dose of 500 ppm. Furthermore, formation of CK8 and CK18 complexes due to CK8 phosphorylation at Ser73 and Ser431 was found to be strongly associated with promotion of hepatocarcinogenesis by PB and the development of hepatocellular carcinomas. The data were confirmed by immunohistochemistry and real-time Q-PCR and profound overexpression of CK8 and CK18 (CK8/18) proteins and mRNAs were detected in several large size GST-P positive foci and liver tumors. A strong correlation between CK8/18 positive foci development and multiplicity of hepatocellular carcinomas was further observed. Moreover, elevation of CK8/18 was strongly associated with induction of cell proliferation in GST-P positive foci and tumors. In conclusion, our data imply that CK8/18 overexpression, those two cytokeratins complex formation associated with histone type 2 H2aa3 up-regulation and intermediate filament reorganization may drive neoplastic transformation of GST-P positive foci during rat hepatocarcinogenesis leading to the formation of hepatocellular carcinomas.

Theoretical Study on the Structural Properties and Relative Stability of M(II)-Al Layered Double Hydroxides Based on a Cluster Model

The [M2Al(OH2)9(OH)4]3+ clusters (M = divalent cation Mg2+, Ca2+, Mn2+, Fe2+, Co2+, Ni2+, Cu2+, Zn2+, or Cd2+), which include the basic information of layered double hydroxides (LDHs) lattice structure with the most economical size, have been investigated by density functional theory (DFT) to shed light on the structural properties and relative stability of M(II)-Al binary LDHs layers with a M2+/Al3+ ratio of 2. The geometric parameters (bond distance and bond angle), natural bond orbitals (NBO), stretching vibration frequencies of three-centered bridging OH groups (nu(O3-H)), as well as binding energy of the cluster model were systematically studied. It was found that the geometries and the nu(O3-H) frequency for the calculated clusters are remarkably influenced by the electronic structure of the divalent cations, such as valence electronic configuration, natural bond orbitals, natural charge transfer, and bond order. The calculated binding energies are in good agreement with the relative stability of the experimental results for the corresponding LDHs. The calculation results reveal that the 2Ni-Al cluster shows the highest stability among the open-shelled cation-containing clusters, while the stability of the 2Cu-Al cluster is the weakest; the 2Mg-Al and 2Zn-Al clusters are the most stable ones among the closed-shelled cation-containing clusters. These findings are in high accordance with the experimental results. Therefore, this work provides a detailed understanding of how the electronic structure of cations plays a more significant role in the structural properties and relative stability of the corresponding LDHs layers rather than ionic size.

Segmental Osteotomy in the Treatment of Obsolete Orbitozygomatic Fractures

Different kinds of orbitozygomatic fractures lead to different levels of orbital structural destructions. Although the magnitude of the management of orbitozygomatic fractures varies considerably, an unsolved and important question remains regarding how to select the best surgical modality according to different kinds of fractures. Among 26 cases of unilateral noncomminuted orbitozygomatic fractures, a segmental osteotomy technique was used to repair the displaced orbitozygomatic complex. The fragment contributing to the orbital structure was mobilized and reduced, whereas the fragment without contribution to the orbital structure was not mobilized and left in situ. Next, the cuneate fragment was imbricated with the orbital osteotomy sites unilaterally or bilaterally. Miniplates and screws were used to span the osteotomy sites and provided fixation once the alignment of the orbit was achieved. The mean difference in volume between fractured orbits preoperatively and postoperatively was 2.47 mL. The mean difference in ocular globe projection was 2.76 mm. When postoperative results were compared with those of the preoperative site, statistically significant difference was noted. At follow-up 1 year postoperatively, the management of orbitozygomatic fractures using segmental osteotomy reduces complications and attains aesthetically satisfying results. Subjective assessment of the patients' globe position found that 88.5% of the patients were satisfied with the outcome, and 11.5% of the patients found it unacceptable. Compared with the traditional method, segmental osteotomy is a simple technique requiring less dissection and can reconstruct the orbital anatomic structure and restore globe position effectively.

Oriental Surgical Protocol in Orbital Hypertelorism

To summarize and analyze the diagnosis, treatment protocol, and therapeutic effect of orbital hypertelorism in the past 30 years in our center.

Ultralong Pedicled Superficial Temporal Fascia Island Flaps for Lower Nasal Defect

To explore the method of repairing nose defects of the apex, ala, septum, and even opposite ala nasi with ultralong pedicled superficial temporal fascia (STF) island flaps.

TSC-22 Contributes to Hematopoietic Precursor Cell Proliferation and Repopulation and is Epigenetically Silenced in Large Granular Lymphocyte Leukemia

Aberrant methylation of tumor suppressor genes can lead to their silencing in many cancers. TSC-22 is a gene silenced in several solid tumors, but its function and the mechanism(s) responsible for its silencing are largely unknown. Here we demonstrate that the TSC-22 promoter is methylated in primary mouse T or natural killer (NK) large granular lymphocyte (LGL) leukemia and this is associated with down-regulation or silencing of TSC-22 expression. The TSC-22 deregulation was reversed in vivo by a 5-aza-2'-deoxycytidine therapy of T or NK LGL leukemia, which significantly increased survival of the mice bearing this disease. Ectopic expression of TSC-22 in mouse leukemia or lymphoma cell lines resulted in delayed in vivo tumor formation. Targeted disruption of TSC-22 in wild-type mice enhanced proliferation and in vivo repopulation efficiency of hematopoietic precursor cells (HPCs). Collectively, our data suggest that TSC-22 normally contributes to the regulation of HPC function and is a putative tumor suppressor gene that is hypermethylated and silenced in T or NK LGL leukemia.

Platelets and Fibrinogen Facilitate Each Other in Protecting Tumor Cells from Natural Killer Cytotoxicity

The functions of platelets and fibrinogen in protecting tumor cells from natural killer cytotoxicity have been discussed for more than 20 years. However, their exact roles and relationships in the process are still not clear. In this study, we show that tumor cells prefer to adhere to fibrinogen than to platelets, and fibrinogen can enhance the adhesion of tumor cells to platelets. Beta3 integrin plays an important role in the adhesion of B16F10 to platelets enhanced by fibrinogen. In the presence of thrombin, fibrinogen forms dense fibrin(ogen) layers around tumor cells. Tumor cells can induce platelets to aggregate and form thrombin. Platelets, as well as thrombin, can help fibrinogen protect tumor cells from lethal contact with natural killer cells and natural killer cytotoxicity. Hirudin, a specific inhibitor of thrombin, can reverse the effect of platelets on fibrinogen in blocking natural killer cytotoxicity. Our results suggest that fibrinogen helps platelets to adhere to tumor cells, and platelets in turn promote more fibrinogen to aggregate around tumor cells by forming thrombin. They facilitate each other in protecting tumor cells from natural killer cytotoxicity.

Ordered Poly(p-phenylene)/layered Double Hydroxide Ultrathin Films with Blue Luminescence by Layer-by-layer Assembly

Lavender layers: A poly(p-phenylene) anionic derivate and exfoliated Mg-Al layered double hydroxide monolayers were assembled into ultrathin films with well-defined blue fluorescence (see picture; the numbers indicate the number of bilayers), long-range order, and high photostability. These films work as multiple quantum-well structures for valence electrons.

Ultralong Pedicled Superficial Temporal Fascia Island Flaps for Lower Nasal Defect

To explore the method of repairing nose defects of apex, ala, septum, and even opposite ala nasi with ultralong pedicled superficial temporal fascia (STF) island flaps. There were 29 cases with defects of apex nasi, ala nasi, and nasal columella, of which 12 cases were repaired with frontal-branched STF island flaps, 14 cases with apical-branched STF postauricular island flaps, and 3 cases with prefabricated apical-branched STF postauricular island flaps. The flap areas were arranged from 1.2 x 2.3 to 2.0 x 2.8 cm2; the length was more than 15 cm on average. Liners were reconstructed at the stage of prefabricating flaps with free skin graft in the cases of ala nasi defects. The surfaces of wound after flap prefabrications were covered by skin graft as well. Twenty-seven cases were successfully taken without blood circulation blocks; the color, texture, and figure were good, and the outcomes were satisfying. Seven nonprefabricated flap cases have epidermis necrosis because of the lack of artery perfusion pressure and venous return handicap, and the epidermis fell off after 1 month, 2 cases of which required secondary surgery because of partial necrosis. Ultralong pedicled STF island flap is an available way to repair defects of apex nasi, ala nasi, and nasal columella. Prefabricated flaps are with benefits of good blood circulation, primary-made liner, and minute injury of the donor site. It is a good method to repair defects of apex nasi, ala nasi, nasal septum, and opposite ala nasi simultaneously.

Ethanol Does Not Promote MeIQx-initiated Rat Colon Carcinogenesis Based on Evidence from Analysis of a Colon Cancer Surrogate Marker

Epidemiological studies suggest that alcohol consumption increases the risk of developing colorectal cancer. However, the data are confounded by numerous cosegregating variables. To cast further light on the relationships between alcohol intake and colon cancer development, 21-day-old male F344/DuCrj rats were fed 200 ppm 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) in their diet for 8 weeks and doses of 0, 0.1, 0.3, 1, 3, 10 and 20% of ethanol in their drinking water ad libitum for 16 weeks thereafter. The rats were sacrificed after 24 weeks of experiment, and aberrant crypt foci (ACF), surrogate lesions for colon cancer, were examined under a light microscope at low magnification. Ethanol was found not to affect the ACF formation at any dose compared with the initiated-controls. Furthermore, ethanol did not alter colon epithelial cell proliferation. These data, obtained by analysis of a colon cancer surrogate marker lesion, indicate that ethanol lacks promotion activity for MeIQx-initiated rat colon carcinogenesis.

A Transposable Class I Composite Transposon Carrying Mph (methyl Parathion Hydrolase) from Pseudomonas Sp. Strain WBC-3

Pseudomonas sp. strain WBC-3 utilizes methyl parathion (O,O-dimethyl O-p-nitrophenol phosphorothioate) or para-nitrophenol as the sole source of carbon, nitrogen and energy. A gene encoding methyl parathion hydrolase (MPH) had been characterized previously and found to be located on a typical class I composite transposon that comprised IS6100 (Tnmph). In this study, the transposability of this transposon was confirmed by transposition assays in two distinct mating-out systems. Tnmph was demonstrated to transpose efficiently in a random manner in Pseudomonas putida PaW340 by Southern blot and in Ralstonia sp. U2 by sequence analysis of the Tnmph insertion sites, both exhibiting MPH activity. The linkage of the mph-like gene with IS6100, together with the transposability of Tnmph, as well as its capability to transpose in other phylogenetically divergent bacterial species, suggest that Tnmph may contribute to the wide distribution of mph-like genes and the adaptation of bacteria to organophosphorus compounds.

Increased Susceptibility to Spontaneous Lung Cancer in Mice Lacking LIM-domain Only 7

LIM-domain only (LMO) 7 is a multifunctional protein that is predicted to regulate the actin cytoskeleton, assembly of adherens junctions in epithelial cells, and gene expression. LMO7 was highly expressed in the mouse lung and predominantly localized to the apical membrane domain of bronchiolar epithelial cells. Although mice lacking LMO7 were viable and fertile in specific pathogen-free conditions, they developed protruding epithelial lesions in the terminal and respiratory bronchioles and alveolar ducts at 14-15 weeks of age. Furthermore, they tended to develop spontaneous adenocarcinoma in the lung at over 90 weeks of age. The cumulative incidence ratios of lung cancer were 22% in LMO7(-/-) mice and 13% in LMO7(+/-) mice whereas no primary lung cancer was observed in wild-type mice. Ex vivo analyses of the cancer cells showed numerical chromosome abnormalities and tumorigenicity in nude mice. These results suggest that LMO7 can act as a tumor suppressor whose deficiency confers a genetic predisposition to naturally occurring lung cancer.

The C-terminal Region of Activation-induced Cytidine Deaminase is Responsible for a Recombination Function Other Than DNA Cleavage in Class Switch Recombination

Activation-induced cytidine deaminase (AID) is an essential factor for the class switch recombination (CSR) and somatic hypermutation (SHM) of Ig genes. CSR and SHM are initiated by AID-induced DNA breaks in the S and V regions, respectively. Because truncation or frame-shift mutations at the carboxyl (C)-terminus of AID abolishes CSR but not SHM, the C-terminal region of AID likely is required for the targeting of DNA breaks in the S region. To test this hypothesis, we determined the precise location and relative amounts of AID-induced DNA cleavage using an in situ DNA end-labeling method. We established CH12F3-2 cell transfectants expressing the estrogen receptor (ER) fused with wild-type (WT) AID or a deletion mutant lacking the C-terminal 16 aa, JP8Bdel. We found that AID-ER, but not JP8Bdel-ER, caused a CSR to IgA from the addition of 4-hydroxy tamoxifen. In contrast, both WT AID and JP8Bdel induced DNA breaks in both the V and S regions. In addition, JP8Bdel enhanced c-myc/IgH translocations. Our findings indicate that the C-terminal domain of AID is not required for S-region DNA breaks but is required for S-region recombination after DNA cleavage. Therefore, AID does not distinguish between the V and S regions for cleavage, but carries another function specific to CSR.

[Caroli's Syndrome]

OBJECTIVE: Caroli's syndrome is a rare autosomal recessive hereditary disease. Here a case of Caroli's syndrome associated with medullary sponge kidney was reported. The patient was a 2-years and 10 months-old boy. He presented with hepatosplenomegaly. Fever, abdominal pain or jaundice was not found. The imaging examination showed intrahepatic bile duct dilation, splenomegaly, medullary sponge kidney and nephrocalcinosis. After introduction of the case, this paper reviewed the clinical characteristics, diagnosis and treatment of Caroli's syndrome.

Sulforhodamine B Intercalated Layered Double Hydroxide Thin Film with Polarized Photoluminescence

Sulforhodamine B (SRB) and dodecylbenzenesulfonate (DBS) with different molar ratios cointercalated into the interlayer region of Mg-Al-layered double hydroxide (SRB-DBS/LDH) were prepared. The structure and chemical composition of the composites were characterized by X-ray diffraction, elemental analysis, thermogravimetry, and differential thermal analysis (TG-DTA). Fluorescence spectra demonstrate that the sample with 4.76% SRB molar percentage, with respect to the total organic material, exhibits the optimal luminous intensity. The fluorescence lifetime of SRB in SRB-DBS/LDH is enhanced significantly compared with that of pristine SRB in solution (4.14 vs 2.05 ns). SRB-DBS/LDH thin films on the quartz substrates were constructed by the solvent evaporation method. Steady-state polarization photoemission spectra show that the luminescence anisotropy of SRB-DBS/LDH thin films (r=0.10) was enhanced remarkably compared with that of a powder sample (r=0+/-0.01) at ambient temperature, whereas the anisotropy of both film and powder samples was largely improved at low temperature (77 K). Furthermore, the SRB-DBS/LDH thin films exhibit a luminescence anisotropy decay effect in the range of their fluorescence lifetime. These results demonstrate that the SRB-DBS/LDH thin films could be used as a good candidate for the immobilization of laser dyes and polarized luminescence materials.

Triple-resonance Coherent Anti-stokes Raman Scattering Microspectroscopy

Fluorescence-free microscopy: A new nonlinear optical microspectroscopy technique, femtosecond (fs) triple-resonance coherent anti-Stokes Raman scattering, in which the amplitude and phase of input fs laser pulses are optimally shaped to be in triple resonance with the molecular electronic and vibrational transitions, generates a coherent nonlinear signal beam at a new color with a highest possible efficiency (see figure).

Stem Cell Factor and Interleukin-2/15 Combine to Enhance MAPK-mediated Proliferation of Human Natural Killer Cells

Stem cell factor (SCF) promotes synergistic cellular proliferation in combination with several growth factors, and appears important for normal natural killer (NK)-cell development. CD34(+) hematopoietic precursor cells (HPCs) require interleukin-15 (IL-15) for differentiation into human NK cells, and this effect can be mimicked by IL-2. Culture of CD34(+) HPCs or some primary human NK cells in IL-2/15 and SCF results in enhanced growth compared with either cytokine alone. The molecular mechanisms responsible for this are unknown and were investigated in the present work. Activation of NK cells by IL-2/15 increases expression of c-kit whose kinase activity is required for synergy with IL-2/15 signaling. Mitogen-activated protein kinase (MAPK) signaling intermediaries that are activated both by SCF and IL-2/15 are enhanced in combination to facilitate earlier cell-cycle entry. The effect results at least in part via enhanced MAPK-mediated modulation of p27 and CDK4. Collectively the data reveal a novel mechanism by which SCF enhances cellular proliferation in combination with IL-2/15 in primary human NK cells.

Enhancement of Preneoplastic Lesion Yield by Chios Mastic Gum in a Rat Liver Medium-term Carcinogenesis Bioassay

The mastic (Pistacia lentiscus var. chia) tree is native throughout the Mediterranean region and has long proved a source of food additives and medical treatments. To investigate the modifying effects of Chios Mastic Gum on rat liver carcinogenesis, 6-week-old male F344 rats were subjected to the established rat liver medium-term carcinogenesis bioassay (Ito-test). At the commencement, rats (groups 1-4) were intraperitoneally injected with 200 mg/kg body weight of diethylnitrosamine (DEN). After two weeks, mastic was added to CRF (Charles River Formula)-1 powdered basal diet at doses of 0, 0.01, 0.1 and 1% in groups 1-4, respectively. At week 3, all rats were underwent two-thirds partial hepatectomy. The experiment was terminated at week 8. As results show, liver weights were significantly increased in a mastic dose-dependent manner among groups 1-4. The numbers (/cm(2)) and the areas (mm(2)/cm(2)) of glutathione S-transferase placental form (GST-P)-positive cell foci (>or=0.2 mm in diameter) were significantly increased in the DEN-1% group compared to the DEN-alone group, along with the average areas per foci and larger-sized foci (>or=0.4 mm). 5-Bromo-2'-deoxyuridine (BrdU)+GST-P double-immunohistochemistry showed the highest BrdU-labeling indices within GST-P foci in the DEN-1% group. 8-hydroxydeoxyguanosine (8-OHdG) levels in liver DNA did not vary, while real-time quantitative polymerase chain reaction (PCR) analysis of livers revealed many up- or down-regulated genes in the DEN-1% group. In conclusion, this is the first report to display a promotion potential of Chios Mastic Gum on the formation of preneoplastic lesions in the established rat liver medium-term carcinogenesis bioassay.

Progesterone Influence on Neurite Outgrowth Involves Microglia

Progesterone (P4) antagonizes estradiol (E2) in synaptic remodeling in the hippocampus during the rat estrous cycle. To further understand how P4 modulates synaptic plasticity, we used entorhinal cortex lesions, which induce E2-dependent neurite sprouting in the hippocampus. In young ovariectomized rats, the E2-dependent entorhinal cortex lesion-induced sprouting was attenuated by concurrent treatment with P4 and E2. Microglial activation also showed the E2-P4 antagonism. These findings extend reports on the estrous cycle synaptic remodeling without lesions by showing the P4-E2 antagonism during simultaneous treatment with both E2 and P4. Glial mechanisms were analyzed with the wounding-in-a-dish model of cocultured glia and embryonic d-18 cortical neurons from rat. In cocultures of mixed glia (astrocytes plus 30% microglia), P4 antagonized the E2-dependent neurite outgrowth (number and length) and neuron viability in the presence of E2, as observed in vivo. However, removal of microglia (astrocyte-neuron coculture) abolished the antagonism of E2 by P4 on neuron sprouting. The P4 receptor antagonists ORG-31710 and RU-486 blocked the antagonism of P4 on E2-dependent sprouting. These findings suggest a new role for microglia in P4 antagonism of E2 in neuronal plasticity and show its dependence on progesterone receptors. These findings are also relevant to the inclusion of progestins in hormone therapy, which is controversial in relation to cognitive declines during aging and in Alzheimer's disease.

Promoter Polymorphism of Interleukin-18 in Angiographically Proven Coronary Artery Disease

Interleukin 18 (IL-18) is a pro-atherogenic cytokine associated with the occurrence of various cardiac complications. The IL-18 gene has a functional -137 G/C polymorphism (rs187238) in the promoter region. Using the ligase detection reaction-polymerase chain reaction, we genotyped a cohort of patients in Chinese Han population in Xiangfan region. Case patients of coronary artery disease and control patients were identified by coronary angiography. The plasma IL-18 concentrations were measured by ELISA. A significant increase of G allele or GG-genotype was observed in 241 case patients compared to 145 control individuals (frequency of G allele = 0.90 vs 0.83, p=0.004; frequency of GG-genotype = 0.81 vs 0.68, p = 0.005). In case patients, G allele carriers in multi-vessel disease patients had a higher occurrence rate when compared to single-vessel disease patients, but no significant difference was detected (frequency of G allele = 0.92 vs 0.88, p=0.107; frequency of GG-genotype = 0.84 vs 0.75, p = 0.089). IL-18 protein concentration of the -137GG genotype was much higher than concentration of the CG and CC genotype (case patients: 229.1+/-131.5 vs 122.7+/-73.6 pg/ml, P < 0.001; control patients: 65.9+/-31.6 vs 42.4+/-19.5 pg/ml, P < 0.001). To conclude, IL-18 promoter -137G/C polymorphism influences IL-18 levels and the occurrence of coronary artery disease, suggesting that IL-18 is causally involved in the development of atherosclerosis.

Evaluation of the Subchronic Toxicity of Dietary Administered Equisetum Arvense in F344 Rats

Equisetum arvense, commonly known as the field horsetail, has potential as a new functional food ingredient. However, little information is available on its side effects, and the general toxicity of Equisetum arvense has yet to be examined in detail. In the present study, we evaluated the influence of administration in diet at doses of 0, 0.3, 1 and 3% for 13 weeks in male and female F344 rats. No toxicity was detected with reference to clinical signs, body weight, urinalysis, hematology and serum biochemistry data and organ weights. Microscopic examination revealed no histopathological lesions associated with treatment. In conclusion, the no-observed-adverse-effect level (NOAEL) for Equisetum arvense was determined to be greater than 3% in both sexes of F344 rat (males and females: >1.79 g/kg BW/day and >1.85 g/kg BW/day, respectively) under the conditions of the present study.

[Arthroscopic Reconstruction of Anterior Cruciate Ligament for Single Bundle Rupture with Hamstring Autograft]

To investigate the effectiveness of anterior cruciate ligament (ACL) reconstruction for single bundle rupture using hamstring autograft with preservation of the left bundle.

Reversibly Thermochromic, Fluorescent Ultrathin Films with a Supramolecular Architecture

[Classification and Arthroscopic Surgery of Chronic Achilles Tendinitis]

To investigate the clinical classification of chronic achilles tendinitis and analyze the surgical technique and efficacy of arthroscopic surgery.

[Mechanism of Telomere Shortening in Photoaging Model Induced by 8-methoxypsoralen and Ultraviolet A]

To investigate the mechanism of telomere shortening through 8-methoxypsoralen (8-MOP) and subsequent ultraviolet A (UVA) irradiation-induced photoaging model in human dermal fibroblasts (HDFs).

Thin Film of Sulfonated Zinc Phthalocyanine/layered Double Hydroxide for Achieving Multiple Quantum Well Structure and Polarized Luminescence

Ordered thin films based on alternate assembly of zinc phthalocyanine complex and layered double hydroxide monolayers have been fabricated, which exhibit multiple quantum well structure and well-defined polarized luminescence.

Benzocarbazole Anions Intercalated Layered Double Hydroxide and Its Tunable Fluorescence

Luminescent benzocarbazole anions (BCZC) intercalated into the interlayer region of Mg-Al-layered double hydroxides (BCZC/LDH) with different layered charge densities (LCD) were prepared. The structure and chemical composition of the composites were characterized by X-ray diffraction, elemental analysis, thermogravimetry and differential thermal analysis (TG-DTA), infrared spectra (FT-IR), UV-vis absorption and fluorescence spectroscopy. The photoemission behavior of BCZC in the LDH matrix with high (Mg/Al ratio = 1.801) and low (Mg/Al ratio = 3.132) LCD is similar to that of BCZC solid and aqueous solution states respectively, indicating that the luminescence performances of the intercalated dye anions can be tuned by adjusting the LCD of the LDH layer. Moreover, the thermal stability and stacking order of BCZC are largely improved upon intercalation, and the BCZC/LDH thin film exhibits well polarized luminescence with the luminescent anisotropy of 0.15-0.20. In addition, molecular dynamics (MD) simulation was employed to calculate the basal spacing and molecular arrangement of the intercalated BCZC within the LDH matrix. The simulation results show that the distribution of BCZC anions is much broader in the gallery of Mg-Al-LDH with high LCD, while BCZC anions exhibit a more ordered arrangement in LDH with low LCD. Furthermore, the radial distribution functions of interlayer water molecules were also studied. Based on the combination of experiment and theoretical simulation, this work provides a detailed understanding of the tunable photoluminescence, orientation and diffusion behavior of the luminescent molecules confined within the gallery of a 2D inorganic matrix.

ZnO Nanotube-based Dye-sensitized Solar Cell and Its Application in Self-powered Devices

High-density vertically aligned ZnO nanotube arrays were fabricated on FTO substrates by a simple and facile chemical etching process from electrodeposited ZnO nanorods. The nanotube formation was rationalized in terms of selective dissolution of the (001) polar face. The morphology of the nanotubes can be readily controlled by electrodeposition parameters for the nanorod precursor. By employing the 5.1 microm-length nanotubes as the photoanode for a dye-sensitized solar cell (DSSC), a full-sun conversion efficiency of 1.18% was achieved. Furthermore, we show that the DSSC unit can serve as a robust power source to drive a humidity sensor, with a potential for self-powered devices.

The Synergy of 6-O-sulfation and N- or 3-O-sulfation of Chitosan is Required for Efficient Inhibition of P-selectin-mediated Human Melanoma A375 Cell Adhesion

We prepared chitosan sulfated derivatives to address the common structural requirement of the sulfate pattern to block P-selectin-mediated tumor cell adhesion. Our results indicate that 6-O-sulfation of chitosan is indispensable for inhibition of P-selectin binding to human melanoma A375 cells. Furthermore, additional N-sulfation or 3-O-sulfation dramatically enhanced the inhibitory activity of 6-O-sulfated chitosan, suggesting that efficient anti-P-selectin adhesion activity of sulfated saccharides requires the synergy of 6-O-sulation and N- or 3-O-sulfation in glucosamine units.

Layer-by-layer Assembly of Bi-protein/layered Double Hydroxide Ultrathin Film and Its Electrocatalytic Behavior for Catechol

This paper reports the fabrication of a bi-protein/layered double hydroxide (LDH) ultrathin film in which hemoglobin (HB) and horseradish peroxidase (HRP) molecules were assembled alternately with LDH nanosheets via the layer-by-layer (LBL) deposition technique, and its electrocatalytic performances for oxidation of catechol were demonstrated. The results of XRD indicate that the HB-HRP/LDH ultrathin film possesses a long range stacking order in the normal direction of the substrate, with the two proteins accommodated in the LDH gallery respectively as monolayer arrangement. SEM images show that the film surface exhibits a continuous and uniform morphology, and AFM reveals the Root-Mean-Square (RMS) roughness of ∼10.2 nm for the film. A stable direct electrochemical redox behavior of the proteins was successfully obtained for the HB-HRP/LDH film modified electrode. In addition, it exhibits remarkable electrocatalytic activity towards oxidation of catechol, based on the synergistic effect of the two proteins. The catechol biosensor in this work displays a wide linear response range (6-170 μM, r=0.999), low detection limit (5 μM), high sensitivity and good reproducibility.

Genome-wide Screen in Saccharomyces Cerevisiae Identifies Vacuolar Protein Sorting, Autophagy, Biosynthetic, and TRNA Methylation Genes Involved in Life Span Regulation

The study of the chronological life span of Saccharomyces cerevisiae, which measures the survival of populations of non-dividing yeast, has resulted in the identification of homologous genes and pathways that promote aging in organisms ranging from yeast to mammals. Using a competitive genome-wide approach, we performed a screen of a complete set of approximately 4,800 viable deletion mutants to identify genes that either increase or decrease chronological life span. Half of the putative short-/long-lived mutants retested from the primary screen were confirmed, demonstrating the utility of our approach. Deletion of genes involved in vacuolar protein sorting, autophagy, and mitochondrial function shortened life span, confirming that respiration and degradation processes are essential for long-term survival. Among the genes whose deletion significantly extended life span are ACB1, CKA2, and TRM9, implicated in fatty acid transport and biosynthesis, cell signaling, and tRNA methylation, respectively. Deletion of these genes conferred heat-shock resistance, supporting the link between life span extension and cellular protection observed in several model organisms. The high degree of conservation of these novel yeast longevity determinants in other species raises the possibility that their role in senescence might be conserved.

[Effect of Arthroscopic Debridement for Adolescent Ankylosing Spondylitis with Early Hip-joint Disease]

To study the effect of arthroscopy debridement for adolescent ankylosing spondylitis (AS) with early hip-joint disease.

Modulators of Multidrug Resistance Associated Proteins in the Management of Anticancer and Antimicrobial Drug Resistance and the Treatment of Inflammatory Diseases

Human multidrug resistance protein (MRP/ABCC) family contains 9 members (MRP1-9) which transport a structurally diverse array of anticancer and antimicrobial drugs and several important endogenous substances including prostaglandins (PGs) and leukotrienes (LTs) with different substrate specificity. MRP1-5 can collectively confer resistance to natural product anticancer drugs and their conjugated metabolites, platinum compounds, folate antimetabolites, nucleoside and nucleotide analogs, and alkylating agents. MRP1-3 are often associated with tumor resistance which is often caused by an increased efflux and decreased intracellular accumulation of natural product anticancer drugs and other anticancer agents. Both PGE(1) and PGE(2) are known high-affinity substrates of MRP4, but not MRP1, MRP2, MRP3 and MRP5. LTC(4) is a substrate of MRP1, MRP2, MRP3, and MRP6-8. MRP2 is also able to transport LTD(4) and LTE(4). Experimental studies in Abcc1-difficient mice have demonstrated a role of MRP1 in inflammation process in vivo. Abcc3-difficient mice have normal bile salt transport, however they have decreased blood bilirubin glucuronide levels. Abcc6-difficient mice show remarkable mineraliztion of the connective tissues, including skin, arterial blood vessels, and retina. Most MRP/ABCC transporters are subject to inhibition by a variety of compounds. Drug targeting of these transporters to overcome MRP/ABCC-mediated multidrug resistance may play a role in cancer and infection (e.g. HIV infection) chemotherapy. Some modulators of MRPs have shown reversing effects on MDR phenotype in preliminary clinical studies and some modulators of MRPs may modify the inflammatory process and consequently ameliorate the inflammatory symptoms. A better understanding of the interactions of these modulators with MRPs has important implications in development of novel drugs for treatment of cancer, infection and inflammation.

A Strategy to the Ordered Assembly of Functional Small Cations with Layered Double Hydroxides for Luminescent Ultra-thin Films

A cationic functional molecule (BNMA) was assembled with a positively-charged LDH monolayer through a polyanion (PVS) as the intermediary. The approach allows fine-tuning and ordered assembly of functional cations with LDH monolayers for designing and achieving novel organic-inorganic ultra-thin films.

Plane-wave Density Functional Theory Study on the Structural and Energetic Properties of Cation-disordered Mg-Al Layered Double Hydroxides

Periodic solid models have been employed to investigate the structural and energetic properties of cation-disordered Mg-Al layered double hydroxides (LDHs), including composition, ordering, and stacking pattern of layers. The geometry, lattice energy and density of states (DOS) of the periodic models were computed using the plane-wave pseudopotential implementation of density functional theory (DFT) with the virtual crystal approximation (VCA). The calculation results for the MgAl-Cl-LDH model show that a pure LDH phase is difficult to sustain when the Mg/Al ratio (R) is R > or = 4. The stability of the cation-disordered MgAl-Cl-LDH unit cell increases upon increasing R, owing to the decrease in the number of Al(III)-O-Al(III) linkages in the LDH sheets. The highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) of MgAl-Cl-LDH receive contributions from the 3s and 3p orbitals of Mg and Al and the 2p orbitals of O and Cl, respectively, implying significant host-guest interactions. The increase of R leads to the decrease in electron density of 3s and 3p of metal cation and that of 2p in Cl. Consequently, the HOMO-LUMO energy gap as well as the systematical stability increase upon increasing R. From this point of view, it has been demonstrated that the cations distribute in an ordered arrangement with the absence of Al(III)-O-Al(III) linkages if R is in the low range (R = 2-3). It was also found that the 3R polytype is the most stable stacking pattern with the same Mg/Al ratio, owing to the fact that the conductive band energy levels decline with the increase of electron density of 3s and 3p of the metal cation in the 3R-stacking system. These findings agree well with the experimental results and provide a profound understanding of critical factors influencing the structure of LDHs, including metal composition and ordering, the stacking sequence, as well as the host-guest interactions.

Enhancement of the Nitrogen Fixation Efficiency of Genetically-engineered Rhizobium with High Catalase Activity

The vktA catalase gene, which had been cloned from Vibrio rumoiensis S-1T having extraordinarily high catalase activity, was introduced into the root nodule bacterium, Rhizobium leguminosarum bv. phaseoli USDA 2676. The catalase activity of the vktA-transformed R. leguminosarum cells (free-living) was three orders in magnitude higher than that of the parent cells and this transformant could grow in a higher concentration of exogenous hydrogen peroxide (H2O2). The vktA-transformant was inoculated to the host plant (Phaseolus vulgaris L.) and the nodulation efficiency was evaluated. The results showed that the nitrogen-fixing activity of nodules was increased 1.7 to 2.3 times as compared to the parent. The levels of H2O2 in nodules formed by the vktA-transformant were decreased by around 73%, while those of leghemoglobins (Lba and Lbb) were increased by 1.2 (Lba) and 2.1 (Lbb) times compared with the parent. These results indicated that the increase of catalase activity in rhizobia could be useful to improve the nitrogen-fixing efficiency of nodules by the reduction of H2O2 content concomitantly with the enhancement of leghemoglobins contents.

[Effectiveness of Educational Interventions in Children with Chronic Diseases and Their Parents]

To evaluate the effectiveness of educational interventions in children with chronic illness and their parents.

A Novel Plant Homeodomain Finger 10-mediated Antiapoptotic Mechanism Involving Repression of Caspase-3 in Gastric Cancer Cells

The mechanisms governing tumorigenesis of gastric cancer have been an area of intense investigation. Currently, plant homeodomain (PHD) finger (PHF) proteins have been implicated in both tumor suppression and progression. However, the function of PHF10 has not been well characterized. Here, we show that various levels of PHF10 protein were observed in gastric cancer cell lines. Alteration of PHF10 expression, which is associated with tumor cell growth, may result in apoptosis in gastric cancer cells both in vitro and in vivo. Knockdown of PHF10 expression in gastric cancer cells led to significant induction of caspase-3 expression at both the RNA and protein levels and thus induced alteration of caspase-3 substrates in a time-dependent manner. Moreover, results from luciferase assays indicated that PHF10 acted as a transcriptional repressor when the two PHD domains contained in PHF10 were intact. Combined with previous findings, our data suggest that PHF10 transcriptionally regulates the expression of caspase-3. Finally, by using systematic reporter deletion and chromatin immunoprecipitation assays, we localized a region between nucleotides -270 and -170 in the caspase-3 promoter that was required for the efficient inhibition of caspase-3 promoter activity by PHF10. Collectively, our findings show that PHF10 repressed caspase-3 expression and impaired the programmed cell death pathway in human gastric cancer at the transcriptional level.

Layer-by-layer Ultrathin Films of Azobenzene-containing Polymer/layered Double Hydroxides with Reversible Photoresponsive Behavior

We report the preparation of a reversible photoresponsive ultrathin film containing a photoactive azobenzene polymer poly{1-4[4-(3-carboxy-4-hydroxyphenylazo)benzenesulfonamido]-1,2-ethanediyl sodium salt} (PAZO) and exfoliated layered double hydroxide (LDH) nanosheets using a layer-by-layer (LBL) self-assembly technique. Alternate irradiation with UV and visible light (lambda > 450 nm) results in a reversible switching between the trans isomer and the cis-rich photostationary state of the azobenzene group with concomitant significant changes in film color. Fluorescence polarization measurements indicated that the orientation of the azobenzene chromophores in the LDH matrix undergoes a reversible realignment during the photoisomerization processes. Photoisomerization induced by alternate UV and visible light irradiation was accompanied by reversible morphological changes of the LBL film, observable by atomic force microscopy (AFM). Molecular dynamics (MD) studies demonstrated that the LDH monolayers allow sufficient free space for the PAZO to undergo isomerization of its azobenzene chromophores. The reversible photoalignment of PAZO was also followed by MD simulations, and the results are in reasonable agreement with the experimental findings, further validating the configurational and orientational changes proposed for PAZO during the reversible photoprocess. It has been demonstrated that the host (LDH nanosheet)-guest (PAZO) interactions are key factors in determining the reversible photoresponsive performances of the film, since the comparative pristine PAZO film shows no such properties. Therefore, the incorporation of a photoactive moiety within the inorganic nanosheets of LDH by the LBL technique provides an attractive and feasible methodology for creating novel light-sensitive materials and devices with potential read-write capabilities.

Hierarchical Films of Layered Double Hydroxides by Using a Sol-gel Process and Their High Adaptability in Water Treatment

Layered double hydroxides (LDHs) films with hierarchical morphologies have been fabricated on different templates via the sol-gel nanocopying and in situ growth process, which exhibit desirable mechanical properties and high adaptability in water treatment.

[Clinical Manifestations and Mutation Study in 16 Chinese Patients with Fabry Disease]

To investigate the clinical manifestations and to characterize mutations of the GLA gene in Chinese patients with Fabry disease so to enhance the diagnosis of Fabry disease.

Temperature-dependent Expression of Type III Secretion System Genes and Its Regulation in Bradyrhizobium Japonicum

The genome-wide expression profiles of Bradyrhizobium japonicum in response to soybean (Glycine max (L.) Merr.) seed extract (SSE) and genistein were monitored with time at a low temperature (15 degrees C). A comparison with the expression profiles of the B. japonicum genome previously captured at the common growth temperature (30 degrees C) revealed that the expression of SSE preferentially induced genomic loci, including a large gene cluster encoding the type III secretion system (T3SS), were considerably delayed at 15 degrees C, whereas most nodulation (nod) gene loci, including nodD1 and nodW, were rapidly and strongly induced by both SSE and genistein. Induction of the T3SS genes was progressively activated upon the elevation of temperature to 30 degrees C and positively responded to culture population density. In addition, genes nolA and nodD2 were dramatically induced by SSE, concomitantly with the expression of T3SS genes. However, the deletion mutation of nodD2 but not nolA led to elimination of the T3SS genes expression. These results indicate that the expression of the T3SS gene cluster is tightly regulated with integration of environmental cues such as temperature and that NodD2 may be involved in its efficient induction in B. japonicum.

Para-Nitrophenol 4-monooxygenase and Hydroxyquinol 1,2-dioxygenase Catalyze Sequential Transformation of 4-nitrocatechol in Pseudomonas Sp. Strain WBC-3

Pseudomonas sp. strain WBC-3 utilizes para-nitrophenol (PNP) as a sole source of carbon, nitrogen and energy. PnpA (PNP 4-monooxygenase) and PnpB (para-benzoquinone reductase) were shown to be involved in the initial steps of PNP catabolism via hydroquinone. We demonstrated here that PnpA also catalyzed monooxygenation of 4-nitrocatechol (4-NC) to hydroxyquinol, probably via hydroxyquinone. It was the first time that a single-component PNP monooxygenase has been shown to catalyze this conversion. PnpG encoded by a gene located in the PNP degradation cluster was purified as a His-tagged protein and identified as a hydroxyquinol dioxygenase catalyzing a ring-cleavage reaction of hydroxyquinol. Although all the genes necessary for 4-NC metabolism seemed to be present in the PNP degradation cluster in strain WBC-3, it was unable to grow on 4-NC as a sole source of carbon, nitrogen and energy. This was apparently due to the substrate's inability to trigger the expression of genes involved in degradation. Nevertheless, strain WBC-3 could completely degrade both PNP and 4-NC when PNP was used as the inducer, demonstrating its potential in bioremediation of the environment polluted by both 4-NC and PNP.

[Establishment of a Cell Model System of Herpes Simplex Virus Type II Latent Infection and Reactivation in SH-SY5Y Cells]

To establish an experimental culture system of herpes simplex virus type II (HSV-2) latent infection and reactivation in SH-SY5Y cells.

Complex Kinetics of Fluctuating Enzymes: Phase Diagram Characterization of a Minimal Kinetic Scheme

Enzyme molecules are dynamic entities with stochastic fluctuation in both protein conformation and enzymatic activity. However, such a notion of fluctuating enzymes, best characterized by recent single-molecule experiments, was not considered in the classic Michaelis-Menten (MM) kinetic scheme. Here we incorporate the fluctuation concept into the reversible MM scheme, and solve analytically all the possible kinetics (i.e., substrate concentration dependent enzymatic velocity) for a minimal model of fluctuating enzymes. Such a minimal model is found to display a variety of distinct kinetic behaviors (phases) in addition to the classic MM kinetics; excess substrate inhibition, sigmoidal kinetics, and concave biphasic kinetics. We find that all these kinetic phases are interrelated and unified under the framework of fluctuating enzymes and can be adequately described by a phase diagram that consists of two master parameters. Functionally, substrate inhibition, sigmoidal kinetics, and convex biphasic phases exhibit positive cooperativity, whereas concave biphasic phases display negative cooperativity. Remarkably, all these complex kinetics are produced by fluctuating enzymes with single substrate binding site, but the two conformations are, therefore, fundamentally different from the classic MWC and KNF models that require multiple subunit or binding sites. This model also suggests that, for a given enzyme/substrate pair, the non-MM behaviors could undergo transitions among different kinetic phases induced by varying product concentrations, owing to the fundamental Haldane symmetry in the reversible MM scheme.

Erasable Nanoporous Antireflection Coatings Based on the Reconstruction Effect of Layered Double Hydroxides

Reduced Levels of IGF-I Mediate Differential Protection of Normal and Cancer Cells in Response to Fasting and Improve Chemotherapeutic Index

Inhibitors of the insulin-like growth factor-I (IGF-I) receptor have been widely studied for their ability to enhance the killing of a variety of malignant cells, but whether IGF-I signaling differentially protects the host and cancer cells against chemotherapy is unknown. Starvation can protect mice, but not cancer cells, against high-dose chemotherapy [differential stress resistance (DSR)]. Here, we offer evidence that IGF-I reduction mediates part of the starvation-dependent DSR. A 72-hour fast in mice reduced circulating IGF-I by 70% and increased the level of the IGF-I inhibitor IGFBP-1 by 11-fold. LID mice, with a 70% to 80% reduction in circulating IGF-I levels, were protected against three of four chemotherapy drugs tested. Restoration of IGF-I was sufficient to reverse the protective effect of fasting. Sixty percent of melanoma-bearing LID mice treated with doxorubicin achieved long-term survival whereas all control mice died of either metastases or chemotherapy toxicity. Reducing IGF-I/IGF-I signaling protected primary glia, but not glioma cells, against cyclophosphamide and protected mouse embryonic fibroblasts against doxorubicin. Further, S. cerevisiae lacking homologs of IGF-I signaling proteins were protected against chemotherapy-dependent DNA damage in a manner that could be reversed by expressing a constitutively active form of Ras. We conclude that normal cells and mice can be protected against chemotherapy-dependent damage by reducing circulating IGF-I levels and by a mechanism that involves downregulation of proto-oncogene signals.

Differential Effect of Three Mitogen-activated Protein Kinases on Lipoprotein (a)-induced Human Mesangial Cell Proliferation

Mesangial hypercellularity is a critical early histopathological finding in human and experimental glomerular diseases. Hyperlipidemia and the glomerular deposition of lipoproteins are commonly associated with mesangial hypercellularity and play an important pathobiological role in the development of glomerular diseases. The activated cytoplasmic mitogen-activated protein kinase (MAPK), including mainly extracellular-signal regulated protein kinase (ERK), c-Jun amino-terminal kinase (JNK), and p38, has been thought to translocate into the nucleus and activate various transcription factors and protooncogenes associated with cell growth and proliferation. Lipoprotein (a) (Lp(a)) has been shown to stimulate proliferation of mesangial cells, but the events of Lp(a) signaling have not yet been characterized. The purpose of this study was to investigate the signal transduction pathways involved in Lp(a)-induced cell proliferation and provide an evidence for the participation of Lp(a) in intracellular signaling pathways for mesangial cell proliferation.

[Inhibitory Effect of SiRNA Targeting Gene Aqp1 on K562 Cells]

The objective of this study was to establish an RNAi approach that can specifically target aqp1 gene sequence in vitro, and to assess the inhibitory effect of this siRNA on K562 cells. The siRNA targeting aqp1-mRNA was designed and transfected into K562 cells by using Lipofectamine(TM) 2000 reagent. Phase-contrast microscopy was used to analyze morphology changes of K562 cells. Cell viability was determined by MTT assay, and flow cytometry and DNA ladder analysis were carried out to identify siRNA-induced apoptosis. The expression levels of aqp1-mRNA at different transfection time were detected by RT-PCR. The results showed that the siRNA was successful by established. The transfected K562 cells displayed the significant apoptosis. The aqp1-siRNAs could obviously inhibit the activity of K562 cells. Cellular DNA fragmentation was observed in the siRNA group after transfection for 48 hours, the apoptosis rates at 24, 48 and 72 hours after transfection were 24.2%, 36.1% and 42.9% respectively. The aqp1-mRNA expression in the cells treated by aqp1-siRNA for 24, 48 and 72 hours were significantly reduced by 33%, 46% and 57% respectively. It is concluded that the aqp1-siRNA can efficiently and specifically inhibited the proliferation and inducing apoptosis of K562 cells. Gene aqp1 can be a potential target point for therapy of malignant tumor.

[Survey of the Evolutionary Characteristics of Influenza H1N1 Hemagglutinin Gene HA1 in 2000-2009]

To study the global evolutionary characteristics of hemagglutinin gene HA1 of influenza H1N1 infecting different species during 2000-2009.

Anionic Poly(p-phenylenevinylene)/layered Double Hydroxide Ordered Ultrathin Films with Multiple Quantum Well Structure: a Combined Experimental and Theoretical Study

The sulfonated phenylenevinylene polyanion derivate (APPV) and exfoliated Mg-Al-layered double hydroxide (LDH) monolayers were alternatively assembled into ordered ultrathin films (UTFs) employing a layer-by-layer method, which shows uniform yellow luminescence. UV-vis absorption and fluorescence spectroscopy present a stepwise and regular growth of the UTFs upon increasing deposited cycles. X-ray diffraction, atomic force microscopy, and scanning electron microscopy demonstrate that the UTFs are orderly periodical layered structure with a thickness of 3.3-3.5 nm per bilayer. The APPV/LDH UTFs exhibit well-defined polarized photoemission characteristic with the maximum luminescence anisotropy of approximately 0.3. Moreover, the UTF exhibit longer fluorescence lifetime (3-3.85-fold) and higher photostability than the drop-casting APPV film under UV irradiation, suggesting that the existence of a LDH monolayer enhances the optical performance of the APPV polyanion. A combination study of electrochemistry and periodic density functional theory was used to investigate the electronic structure of the APPV/LDH system, illustrating that the APPV/LDH UTF is a kind of organic-inorganic hybrid multiple quantum well (MQW) structure with a low band energy of 1.7-1.8 eV, where the valence electrons of APPV can be confined into the energy wells formed by the LDH monolayers effectively. Therefore, this work not only gives a feasible method for fabricating a luminescence ultrathin film but also provides a detailed understanding of the geometric and electronic structures of photoactive polyanions confined between the LDH monolayers.

Profiling Non-lysyl TRNAs in HIV-1

During its assembly, human HIV-1 selectively packages the tRNA(Lys) isoacceptors, including tRNA(Lys3), the primer for the reverse transcriptase. However, other low molecular weight RNA species are also seen in the virus. We profiled the tRNAs packaged into HIV-1 using microarray analysis and validated our results by two-dimensional gel electrophoresis and RT-PCR. In addition to tRNA(Lys) isoacceptors, tRNA(Asn) and the rare isoacceptor of tRNA(Ile) are also selectively packaged. In Gag viral-like particles missing the GagPol protein, overall tRNA incorporation is reduced by >80%. This reduction is significantly greater than can be accounted for by the reduction in tRNA(Lys) isoacceptors, tRNA(Asn) and tRNA(Ile), suggesting that incorporation of other tRNAs may also require the GagPol protein. These results demonstrate selective incorporation of non-lysyl tRNAs into HIV-1 and highlight the application of microarrays as a novel method to study tRNA incorporation into viruses.

Repair of Canine Mandibular Bone Defects with Bone Marrow Stromal Cells and Coral

Tissue engineering has become a new approach for repairing bone defects. Previous studies indicated that coral scaffolds had been utilized with bone marrow stromal cells (BMSCs) in a variety of approaches for bony reconstruction. In these applications, the degradation rate of the material did not match the rate at which bone was regenerated. In this study, a previously established 30 mm long mandibular segmental defect was repaired with engineered bone using green fluorescent protein-labeled osteogenic BMSCs seeded on porous coral (n = 12). Defects treated with coral alone (n = 12) were used as an experimental control. In the BMSCs/coral group, new bone formation was observed from 4 weeks postoperation, and bony-union was achieved after 32 postoperative weeks. The residual coral volume of the BMSCs/coral grafts at 12 weeks (20-30%) was significantly higher than that at 32 weeks (10-15%, p < 0.05), which was detected by microcomputed tomography and histological examination. The engineered bone with BMSCs/coral achieved satisfactory biomechanical properties at 32 weeks postoperation, which was very close to that of the contralateral edentulous mandible. More importantly, immunostaining demonstrated that the implanted BMSCs differentiated into osteoblast-like cells. In contrast, minimal bone formation with almost solely fibrous connection was observed in the group treated with coral alone. Based on these results, we conclude that engineered bone from osteogenically induced BMSCs and biodegradable coral can successfully repair the critical-sized segmental mandibular defects in canines and the seeding cells could be used for bony restoration.

Suppressive Effect of Diazepam on IFN-gamma Production by Human T Cells

Many studies showed that benzodiazepines could modulate immune responses through interaction with peripheral benzodiazepine receptors (PBRs) in immune cells but most of the studies were focused on monocytes and macrophages. In the present study, we revealed that diazepam, a mixed-type benzodiazepine, inhibited IFN-gamma production by human peripheral blood mononuclear cells (PBMCs) induced by anti-CD3 in dose-dependent manner. Flow cytometry analysis demonstrated that diazepam could inhibit the frequency of IFN-gamma-producing CD4(+) and CD8(+) T cells. The inhibitory effect of diazepam on IFN-gamma production is similar to that of R(0)5-4864, a selective PBRs ligand. However, D8555, a selective ligand for PBRs in microglia in the central nervous system, is a much weak inhibitor compared with R(0)5-4864 or diazepam. The inhibitory effect of R(0)5-4864 could be antagonized by PK11195, which is recognized as selective PBRs antagonist, and suppressive effect of diazepam on T cells is partially antagonized by PK11195. Collectively, these results suggested that diazepam suppressed human T cell function through PBRs.

CD94 Surface Density Identifies a Functional Intermediary Between the CD56bright and CD56dim Human NK-cell Subsets

Human CD56(bright) natural killer (NK) cells possess little or no killer immunoglobulin-like receptors (KIRs), high interferon-gamma (IFN-gamma) production, but little cytotoxicity. CD56(dim) NK cells have high KIR expression, produce little IFN-gamma, yet display high cytotoxicity. We hypothesized that, if human NK maturation progresses from a CD56(bright) to a CD56(dim) phenotype, an intermediary NK cell must exist, which demonstrates more functional overlap than these 2 subsets, and we used CD94 expression to test our hypothesis. CD94(high)CD56(dim) NK cells express CD62L, CD2, and KIR at levels between CD56(bright) and CD94(low)CD56(dim) NK cells. CD94(high)CD56(dim) NK cells produce less monokine-induced IFN-gamma than CD56(bright) NK cells but much more than CD94(low)CD56(dim) NK cells because of differential interleukin-12-mediated STAT4 phosphorylation. CD94(high)CD56(dim) NK cells possess a higher level of granzyme B and perforin expression and CD94-mediated redirected killing than CD56(bright) NK cells but lower than CD94(low)CD56(dim) NK cells. Collectively, our data suggest that the density of CD94 surface expression on CD56(dim) NK cells identifies a functional and likely developmental intermediary between CD56(bright) and CD94(low)CD56(dim) NK cells. This supports the notion that, in vivo, human CD56(bright) NK cells progress through a continuum of differentiation that ends with a CD94(low)CD56(dim) phenotype.

Comparative Analyses of Time-course Gene Expression Profiles of the Long-lived Sch9Delta Mutant

In an attempt to elucidate the underlying longevity-promoting mechanisms of mutants lacking SCH9, which live three times as long as wild type chronologically, we measured their time-course gene expression profiles. We interpreted their expression time differences by statistical inferences based on prior biological knowledge, and identified the following significant changes: (i) between 12 and 24 h, stress response genes were up-regulated by larger fold changes and ribosomal RNA (rRNA) processing genes were down-regulated more dramatically; (ii) mitochondrial ribosomal protein genes were not up-regulated between 12 and 60 h as wild type were; (iii) electron transport, oxidative phosphorylation and TCA genes were down-regulated early; (iv) the up-regulation of TCA and electron transport was accompanied by deep down-regulation of rRNA processing over time; and (v) rRNA processing genes were more volatile over time, and three associated cis-regulatory elements [rRNA processing element (rRPE), polymerase A and C (PAC) and glucose response element (GRE)] were identified. Deletion of AZF1, which encodes the transcriptional factor that binds to the GRE element, reversed the lifespan extension of sch9Delta. The significant alterations in these time-dependent expression profiles imply that the lack of SCH9 turns on the longevity programme that extends the lifespan through changes in metabolic pathways and protection mechanisms, particularly, the regulation of aerobic respiration and rRNA processing.

Effects of Pioglitazone, a Peroxisome Proliferator-activated Receptor Gamma Agonist, on the Urine and Urothelium of the Rat

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors, which belong to the nuclear receptor superfamily. Some PPARgamma agonists, such as pioglitazone, and dual PPARgamma/PPARalpha agonists, such as muraglitazar, induced urothelial bladder tumors in rats but not in mice. In this study, we investigated the early effects in the urine and bladder of rats treated with pioglitazone to evaluate the possible relation between urinary solids formation and urothelial cytotoxicity and regenerative proliferation. In a 4-week experiment, treatment of rats with 16 mg/kg pioglitazone induced cytotoxicity and necrosis of the urothelial superficial layer, with increased cell proliferation measured by bromodeoxyuridine labeling index and hyperplasia by histology. It also produced alterations in urinary solid formation, especially calcium-containing crystals and calculi. PPARgamma agonists (pioglitazone and troglitazone) in vitro reduced rat urothelial cell proliferation and induced uroplakin synthesis, a specific differentiation marker in urothelial cells. Our data support the hypothesis that the bladder tumors produced in rats by pioglitazone are related to the formation of urinary solids. This strongly supports the previous conclusion in studies with muraglitazar that this is a rat-specific phenomenon and does not pose a urinary bladder cancer risk to humans treated with these agents.

Cytokeratin 8/18 As a New Marker of Mouse Liver Preneoplastic Lesions

To search for a reliable biomarker of preneoplastic lesions arising early in mouse hepatocarcinogenesis the proteomes of microdissected basophilic foci, hepatocellular adenomas (HCAs), carcinomas (HCCs) and normal-appearing liver of B6C3F1 mice initiated with diethylnitrosamine (DEN) were analysed on anionic (Q10) surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) ProteinChip arrays. Significant overexpression of cytokeratin 8 (CK8; m/z 54, 565), cytokeratin 18 (CK18; m/z 47,538) proteins was found in basophilic foci as well as in HCAs and HCCs. Furthermore, immunohistochemistry demonstrated profound overexpression of CK8 and CK18 proteins (CK8/18) in all basophilic foci, mixed cell type foci, HCAs and HCCs in B6C3F1 and C57BL/6J mice initiated with DEN. A strong correlation between CK8/18-positive foci development and multiplicity of liver tumors in B6C3F1 and C57Bl/6J mice was further observed. Moreover, formation of CK8 and CK18 complexes due to CK8 phosphorylation at Ser73 and Ser431 was found to be strongly associated with neoplastic transformation of mice liver basophilic foci. Elevation of CK8/18 was strongly correlated with induction of cell proliferation in basophilic foci and tumors. In conclusion, our data imply that CK8/18 is a novel reliable marker of preneoplastic lesions arising during mouse hepatocarcinogenesis which might be used for prediction of tumor development and evaluation of environmental agents as well as drugs and food additives using mouse liver tests.

A Thermochromic Thin Film Based on Host-guest Interactions in a Layered Double Hydroxide

Optically transparent thin films with thermochromic properties have been fabricated by means of cointercalation of different molar ratios of 4-(4-anilinophenylazo)benzenesulfonate (AO5) and sodium dodecylbenzene sulfonate (SDS) into the galleries of a ZnAl layered double hydroxide (LDH). The X-ray diffraction (XRD) patterns of these thin films show that they are assembled in a highly c-oriented manner, and the basal spacing ranges from 2.95 to 2.63 nm with varying AO5/SDS molar ratio. The preferential orientation of AO5 in the galleries of 10% AO5-LDH (AO5/SDS = 10:90, molar percentage) was evaluated by the fluorescence polarization technique; the results show that AO5 anions are accommodated between sheets of ZnAl-LDH as monomeric units with a tilt angle Psi (defined as the angle between the transition dipole moment of the AO5 anion with respect to the normal to the LDH layer) of 74 degrees. It was found that the composite film exhibits marked thermochromic behavior (light yellow <==> reddish-orange) in the temperature range of 35-65 degrees C, which is reversible over a number of heating-cooling cycles. It has been demonstrated that the thermochromic behavior results from tautomerism of interlayer AO5 and furthermore that both the host-guest and guest-guest interactions are key factors, since pristine AO5 shows no thermochromic performance. The 10% AO5-LDH film shows the highest thermochromic efficiency of all the films examined. Furthermore, a reversible contraction and expansion of the LDH basal spacing was also observed for this thin film over the same temperature range.

Chemopreventive Effects of 13alpha,14alpha-epoxy-3beta-methoxyserratan-21beta-ol (PJJ-34), a Serratane-type Triterpenoid, in a Rat Multi-organ Carcinogenesis Bioassay

A novel serratane-type triterpenoid, 13alpha,14alpha-epoxy-3beta-methoxyserratan-21beta-ol (PJJ-34) derived from cuticles of Picea jezoensis Carr. var. jezoensis, has proved to be highly effective at suppressing carcinogenesis both in vitro and in vivo. To investigate possible anti-carcinogenic efficacy at the whole-body level, male Fischer 344 rats were subjected to an established rat multi-organ carcinogenesis bioassay (DMBDD model). After initiation with five carcinogens, groups 1-3 (20 in each) were intragastrically (i.g.) administered PJJ-34 dissolved in 1 ml of 0.5% CMC (5 times/week) at doses of 0, 5 and 10mg/kg body weight (b.w.), respectively, until the end of week 30. PJJ-34 did not show apparent toxicity. Incidences of adenomas (100-->75%) and carcinomas (63-->30%) in the lung were significantly decreased in the 5mg/kg b.w. group, and multiplicity of alveolar hyperplasias and total lung tumors (adenomas+carcinomas) were significantly reduced by both 5 and 10mg/kg. The incidence of colorectal tumors was also significantly decreased in the 10mg/kg group (63-->28%) along with the multiplicity. Rat liver pre-neoplastic lesions, glutathione S-transferase placental form (GST-P) foci, and tumor development in the other organs were not affected. Immunohistochemical indices for proliferating cell nuclear antigen (PCNA) and cyclin D1 in normal alveolar epithelium of the lung were significantly suppressed at both doses. In conclusion, PJJ-34 is chemopreventive against lung and colon carcinogenesis without exerting apparent toxicity, and suppression of cell proliferation could play a key role in the underlying mechanisms.

[Clinical Characteristics and Long-term Follow-up Analysis of Three Cases with Newborn Aristolochic Acid Nephropathy]

To summarize the clinical characteristics and prognosis of newborn aristolochic acid nephropathy induced by akebia.

Enhanced Urinary Bladder, Liver and Colon Carcinogenesis in Zucker Diabetic Fatty Rats in a Multiorgan Carcinogenesis Bioassay: Evidence for Mechanisms Involving Activation of PI3K Signaling and Impairment of P53 on Urinary Bladder Carcinogenesis

In the present study, modifying effects of diabetes on carcinogenesis induced in type 2 diabetes mellitus model Zucker diabetic fatty (ZDF) rats were investigated using a multiorgan carcinogenesis bioassay. Our re sults demonstrated enhancement of urinary bladder, colon and liver carcinogenesis in ZDF rats treated with five types of carcinogens (DMBDD). Elevated insulin and leptin and decreased adiponectin levels in the serum may be responsible for the high susceptibility of type 2 diabetes mellitus model rats to carcinogenesis in these organs. Possible mechanisms of increased susceptibility of diabetic rats to bladder carcinogenesis could be activation of the PI3K pathway and suppression of p53 in the urothelium in consequence of the above serum protein alterations.

Conserved Role of Medium Acidification in Chronological Senescence of Yeast and Mammalian Cells

Proteome Analysis of Laser Microdissected Glomeruli from Formalin-fixed Paraffin-embedded Kidneys of Autopsies of Diabetic Patients: Nephronectin is Associated with the Development of Diabetic Glomerulosclerosis

BACKGROUND: To date, little proteomic information has been available from the glomeruli of diabetic patients, possibly due to the clinical limitations of renal biopsy in diabetic patients and insufficient quantities of such specimens for proteome analysis. The purpose of the present study was to identify altered protein expression profiles in diabetic glomeruli using formalin-fixed paraffin-embedded (FFPE) kidney tissues from diabetic patients.METHODS: Glomeruli were laser microdissected from FFPE autopsy kidney tissues from 10 patients with diabetic nephropathy and 10 non-diabetic control patients and underwent proteome analysis using QSTAR Elite liquid chromatography with tandem mass spectrometry and iTRAQ technology. Immunohistochemical analysis was performed on 93 autopsy samples from diabetic patients with and without nephropathy (n = 45 and n = 48, respectively).RESULTS: Thirty-one renal and urological disease-related proteins displayed a differential abundance in glomerular samples from patients with diabetic nephropathy compared with non-diabetic control patients. Among them, we found that nephronectin, which functions in the assembly of extracellular matrix, showed clearly positive immunoreactivity in diabetic glomeruli. The numerical fraction of nephronectin-positive glomerular cross sections was increased significantly in diabetic patients with nephropathy compared to those without nephropathy (32.1 ± 31.5 versus 4.14 ± 5.65%, P < 0.0001). Furthermore, there was a significant positive correlation between this numerical fraction of nephronectin-positive glomerular cross sections and the glomerular sclerosis index (ρ = 0.881, P < 0.0001, n = 93).CONCLUSION: The present study demonstrated, for the first time, that nephronectin may be associated with the development of diabetic glomerulosclerosis and that proteome analysis with FFPE kidney tissues from diabetic patients with nephropathy is useful in understanding diabetic nephropathy.

[Significance of Detecting the EBV-DNA Level in Peripheral Blood Mononuclear Cells and the EBV-infected Cell Type in Patients with Chronic Active EBV Infection]

To study the difference in the EBV-DNA level in peripheral blood mononuclear cells (PBMC) and the type of Epstein-Barr virus (EBV)-infected cells in pediatric patients with chronic active EBV (CAEBV) infection, acute EBV infection (AEBV) and healthy children, and to analyze the relationship between the above difference and the clinical manifestation of CAEBV.

A Cocrystal Strategy to Tune the Luminescent Properties of Stilbene-type Organic Solid-state Materials

The one- and two-photon luminescence of stilbene-type solid-state materials can be tuned and controlled from blue to yellow color by a supramolecular cocrystal method.

Label-free Optical Imaging of Nonfluorescent Molecules by Stimulated Radiation

Imaging contrasts other than fluorescence are highly desirable for label-free detection and interrogation of nonfluorescent molecular species inside live cells, tissues, and organisms. The recently developed stimulated Raman scattering (SRS) and stimulated emission microscopy techniques provide sensitive and specific contrast mechanisms for nonfluorescent species, by employing the light amplification aspect of stimulated radiation. Compared to their spontaneous counterparts, stimulated radiation can enhance the imaging performance significantly, making the previously 'dark' molecules observable. Here we review and summarize the underlying principles of this emerging class of molecular imaging techniques.

[Outcome Evaluation of Arthroscopy-assisted Ankle Arthrodesis]

To evaluate the methods and results of arthroscopy-assisted ankle arthrodesis.

Targeted Proteomics of Isolated Glomeruli from the Kidneys of Diabetic Rats: Sorbin and SH3 Domain Containing 2 is a Novel Protein Associated with Diabetic Nephropathy

To evaluate proteins associated with the development of diabetic nephropathy, a major cause of the end-stage renal disease, we analyzed protein expression in isolated glomeruli from spontaneous type 2 diabetic (OLETF) rats and their age-matched control littermates (LETO) in the early and proteinuric stages of diabetic nephropathy using QSTAR Elite LC-MS/MS. Among the 191 and 218 proteins that were altered significantly in the OLETF rats, twenty-four were actin cytoskeleton-associated proteins implicated in the formation of stress fibers, and the impairment of actin polymerization, intermediate filaments and microtubules. Importantly, sorbin and SH3 domain containing 2 (SORBS2), which is involved in the formation of stress fibers, was significantly upregulated in both stages of diabetic nephropathy (1.49- and 1.97-fold, resp.). Immunohistochemical and quantitative-PCR analyses revealed upregulation of SORBS2 in podocytes of glomeruli of OLETF rats. Our findings suggested that SORBS2 may be associated with the development of diabetic nephropathy possibility by reorganization of actin filaments.

A Family of Visible-light Responsive Photocatalysts Obtained by Dispersing CrO6 Octahedra into a Hydrotalcite Matrix

Self-assembled Multivalent DNA Nanostructures for Noninvasive Intracellular Delivery of Immunostimulatory CpG Oligonucleotides

Designed oligonucleotides can self-assemble into DNA nanostructures with well-defined structures and uniform sizes, which provide unprecedented opportunities for biosensing, molecular imaging, and drug delivery. In this work, we have developed functional, multivalent DNA nanostructures by appending unmethylated CpG motifs to three-dimensional DNA tetrahedra. These small-sized functional nanostructures are compact, mechanically stable, and noncytotoxic. We have demonstrated that DNA nanostructures are resistant to nuclease degradation and remain substantially intact in fetal bovine serum and in cells for at least several hours. Significantly, these functional nanostructures can noninvasively and efficiently enter macrophage-like RAW264.7 cells without the aid of transfection agents. After they are uptaken by cells, CpG motifs are recognized by the Toll-like receptor 9 (TLR9) that activates downstream pathways to induce immunostimulatory effects, producing high-level secretion of various pro-inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-12. We also show that multivalent CpG motifs greatly enhance the immunostimulatory effect of the nanostructures. Given the high efficacy of these functional nanostructures and their noncytotoxic nature, we expect that DNA nanostructures will become a promising tool for targeted drug delivery.

Influence of the Maxillary Sinus Exposure in Reduction Malarplasty with an L-shaped Osteotomy

The zygomatic bone forms the prominence of the midface and plays an important role in determining the human facial attractiveness. The Oriental face is mesocephalic, with a prominent zygomatic body and wide zygomatic arch. Nevertheless, as an oval face is considered ideal, reduction malarplasty is always requested by Oriental people. Currently, L-shaped zygomatic osteotomy is often reported in literature yet no previous study has documented the exact outcome of this approach after exposure of the maxillary sinus. The objective of the study was to investigate the outcome and complications of intraoral L-shaped zygomatic osteotomy with the opening of maxillary sinus.

Does Orbital Proprioception Contribute to Gaze Stability During Translation?

Translational motion induces retinal image slip which varies with object distance. The brain must know binocular eye position in real time in order to scale eye movements so as to minimize retinal slip. Two potential sources of eye position information are orbital proprioception and an internal representation of eye position derived from central ocular motor signals. To examine the role of orbital proprioceptive information, the position of the left eye was perturbed by microstimulation of the left abducens nerve during translational motion to the right or left along the interaural axis in two rhesus macaques. Microstimulation rotated the eye laterally, activating eye muscle proprioceptors, while keeping central motor commands undisturbed. We found that microstimulation-induced eye position changes did not affect the translational VOR in the abductive (lateral rectus) direction, but it did influence the responses in the adductive (medial rectus) direction. Our findings demonstrate that proprioceptive inputs appear to be involved in the TVOR responses at least during ipsilateral head movements and proprioceptive influences on the TVOR may involve vergence-related signals to the oculomotor nucleus. However, internal representation of eye position, derived from central ocular motor signals, likely plays the dominant role in providing eye position information for scaling eye movements during translational motion, particularly in the abducent direction.

Label-free Imaging of Lipid Dynamics Using Coherent Anti-stokes Raman Scattering (CARS) and Stimulated Raman Scattering (SRS) Microscopy

The recently developed Coherent Anti-stokes Raman Scattering (CARS) microscopy and Stimulated Raman Scattering (SRS) microscopy have provided new methods to visualize the localization and regulation of biological molecules without the use of invasive and potentially perturbative labels. They allow rapid imaging of specific molecules with high resolution and sensitivity. These tools have been effectively applied to the study of lipid metabolism using Caenorhabditis elegans as a genetic model, unraveling new lipid storage phenotypes and their regulatory mechanisms. Here we review the underlying principle of CARS and SRS microscopy, as well as their recent applications in lipid biology research in C. elegans.

[Arthroscopic Reduction and Fixation of Tibial Intercondylar Eminence Avulsion Fractures Using Nonabsorbable Suture with Neckwear Knot Loop Ligature]

To investigate the effectiveness of percutaneous reduction by leverage and fixation using nonabsorbable suture with neckwear knot loop ligature to treat tibial intercondylar eminence avulsion fractures under the arthroscope.

Enhancement of Visible Light Photocatalysis by Grafting ZnO Nanoplatelets with Exposed (0001) Facets Onto a Hierarchical Substrate

A ZnO nanocatalyst with a high percentage exposure of (0001) facets embedded on a hierarchical flower-like matrix has been prepared by an in situ topotactic transformation of a layered double hydroxide precursor, and exhibits significantly higher visible light photocatalytic performance than other ZnO nanomaterials with fewer exposed (0001) facets.

[Advances of Treatment for Extranodal NK/T-cell Lymphoma --- Review]

Extranodal NK/T-cell lymphoma is a rare pathological type, incidence of which is 2% - 10% of the primary NHL cases, showing a broad morphologic spectrum with frequent necrosis. Extranodal NK/T-cell lymphoma occurs more frequently in Asian population, especially in Southern China and Southeast Asian population. It is reported that the incidence of ENKL among lymphomas (1314 cases) in Asian countries was 4 times as much as the Western countries (22% vs. 5%) by the International Peripheral T-cell Lymphoma Project. Nasal NK/T-cell lymphoma most commonly occurred in the nasal cavity or other parts of the upper aerodigestive tract, and is highly associated with EBV (Epstein-Barr virus) infection, which is highly aggressive and its prognosis generally is poor. The mean survival time is about 12-38 months. It is important to accurately assess the patients prognosis for an optimal treatment. Localized disease (stage I and II) often has a relatively good prognosis through local radiation or combined therapy. There is a paucity of data to guide therapy in advanced disease. Generally, combining therapy is the most commonly selected approach for advanced disease. The conditions of the patients with advanced, relapsed or refractory diseases have been improved by high-intensity chemotherapy combined with radiation, and a regimen containing L-asparaginase L-Asp. Recently some studies have demonstrated promising outcomes in the selected cases by high-dose chemotherapy supplemented with auto-or allo-HSCT. Targeting therapy is also developing quickly. This current review mainly focuses on the advance of treatment for extranodal NK/T-cell lymphoma.

Application of Tyrosine Kinase Inhibitors As a Promising Targeting Treatment for Myeloproliferative Neoplasms --- Review

As well as playing vital roles in main cellular processes, such as abnormal proliferation, differentiation, survival, apoptosis, and a lot of tyrosine kinases (TK) are involved in oncogenesis. TK or components of their signal pathways have been found abnormal in many hematological malignancies. Therefore, tyrosine kinase inhibitors (TKI) have been provided a great deal of enthusiasm for development of therapy in myeloproliferative neoplasms (MPN). Representatively, the treatment of chronic myelogenous leukemia (CML) was revolutionary for the design of imatinib mesylate (IM), which is a BCR/ABL TKI. Subsequently, because of need for the resistance or intolerance, novel agents are being explored and imatinib has now been extended to eosinophilia-associated myeloid neoplasms with PDGFRA, PDGFRB or FGFR1 gene mutations. Recently, JAK2 inhibitor drugs are currently being tested in clinical trials. Here, the current review mainly focuses on the role of TK in classic MPN including CML, polycythemia vera (PV), primary myelofibrosis (PMF), essential thrombocythemia (ET), and advances of targeting these abnormalities with small molecule inhibitors.

Ordered Blue Luminescent Ultrathin Films by the Effective Coassembly of Tris(8-hydroxyquinolate-5-sulfonate)aluminum and Polyanions with Layered Double Hydroxides

This article reports a novel method to assemble a small anion with exfoliated Mg-Al-layered double hydroxide (LDH) nanosheets into ordered ultrathin films (UTFs) by employing the layer-by-layer assembly technique. The premixing solution of tris(8-hydroxyquinolate-5-sulfonate)aluminum(III) (AQS(3-)) with three kinds of polyanions-poly(acrylic acid), ((C(3)H(4)O(2))(n), PAA), poly(styrene 4-sulfonate) ([CH(2)CH(C(6)H(4))SO(3)](m), PSS), and poly[5-methoxy-2-(3-sulfopropoxy)-1,4-phenylene vinylene] (C(12)H(13)O(5)S)(n), PPV)-has been used as building blocks to assemble alternatively with LDH nanosheets. The UV-vis absorption and fluorescence spectroscopy of (AQS-polyanion/LDH)(n) UTFs presents stepwise growth upon increasing deposited cycles in comparison with the (AQS/LDH)(n) film under the same experimental process. (AQS-PPV/LDH)(n) UTF displays complex fluorescence originating from AQS and PPV. The (AQS/LDH)(n) and (AQS-polyanion/LDH)(n) UTFs exhibit higher blue-polarized photoemission character with a luminescence anisotropy (r) of ca. 0.12-0.20 and a longer fluorescence lifetime than that of the Na(3)AQS film with r = 0.04. X-ray diffraction, scanning electron microscopy, and atomic force microscopy demonstrated that the UTFs were orderly periodically layered structures with a thickness of ca. 3.0 nm per bilayer. Therefore, this work gives a feasible method for immobilizing small anions into the gallery of LDHs.

An Optical Sensor Based on H-acid/layered Double Hydroxide Composite Film for the Selective Detection of Mercury Ion

A novel optical chemosensor was fabricated based on 1-amino-8-naphthol-3,6-disulfonic acid sodium (H-acid) intercalated layered double hydroxide (LDH) film via the electrophoretic deposition (EPD) method. The film of H-acid/LDH with the thickness of 1 μm possesses a well c-orientation of the LDH microcrystals confirmed by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The fluorescence detection for Hg(II) in aqueous solution was performed by using the H-acid/LDH film sensor at pH 7.0, with a linear response range in 1.0 × 10(-7) to 1.0 × 10(-5) mol L(-1) and a detection limit of 6.3 × 10(-8) mol L(-1). Furthermore, it exhibits excellent selectivity for Hg(II) over a large number of competitive cations including alkali, alkaline earth, heavy metal and transitional metals. The specific fluorescence response of the optical sensor is attributed to the coordination between Hg(II) and sulfonic group in the H-acid immobilized in the LDH matrix, which was verified by NMR spectroscopy and UV-vis spectra. In addition, density functional theory (DFT) calculation further confirms that the coordination occurs between one Hg(2+) and two O atoms in the sulfonic group, which is responsible for the significant fluorescence quenching of the H-acid/LDH film. The results indicate that the H-acid/LDH composite film can be potentially used as a chemosensor for the detection of Hg(2+) in the environmental and biomedical field.

Knockdown of Metallopanstimulin-1 Inhibits NF-κB Signaling at Different Levels: The Role of Apoptosis Induction of Gastric Cancer Cells

The ribosomal protein S27 (metallopanstimulin-1, MPS-1) has been reported to be a multifunctional protein, with increased expression in a number of cancers. We reported previously that MPS-1 was highly expressed in human gastric cancer. Knockdown of MPS-1 led to spontaneous apoptosis and repressed proliferation of human gastric cancer cells in vitro and in vivo. However, how does MPS-1 regulate these processes is unclear. Here we performed microarray and pathway analyses to investigate possible pathways involved in MPS-1 knockdown-induced apoptosis in gastric cancer cells. Our results showed that knockdown of MPS-1 inhibited NF-κB activity by reducing phosphorylation of p65 at Ser536 and IκBα at Ser32, inhibiting NF-κB nuclear translocation, and down-regulating its DNA binding activity. Furthermore, data-mining the Gene-Regulatory-Network revealed that growth arrest DNA damage inducible gene 45β (Gadd45β), a direct NF-κB target gene, played a critical role in MPS-1 knockdown-induced apoptosis. Over-expression of Gadd45β inhibited MPS-1 knockdown-induced apoptosis via inhibition of JNK phosphorylation. Taken together, these data revealed a novel pathway, the MPS-1/NF-κB/Gadd45β signal pathway, played an important role in MPS-1 knockdown-induced apoptosis of gastric cancer cells. This study sheds new light on the role of MPS-1/NF-κB in apoptosis and the possible use of MPS-1 targeting strategy in the treatment of gastric cancer.

Molecular Characterization of a Cucumber Nitrate Reductase (CsNR) Gene Under NO(3) (-) Stress

Nitrate reductase is a key enzyme in the overall process of nitrate assimilation by plants. A full-length cDNA clone encoding nitrate reductase (NR; EC 1.6.6.1) was isolated from cucumber (Cucumis sativus L.) by RT-PCR and RACE techniques. The NR of cucumber (CsNR), a full-length cDNA sequence of 3032 bp contains an open reading frame of 2748 bp encoding 915 amino acids. The deduced 915 amino acid sequence showed high identities with NR from other plants. Quantitative real-time PCR analysis indicated that CsNR expression was different in root, stem, leaf, flower and mature fruit tissues. CsNR transcript level and nitrate reductase activity (NRA) was down-regulated and the change in NO(3) (-) concentration showed a negative trend with NRA in leaves when subjected to the 182 mM NO(3) (-) treatment. However, the CsNR transcript level was up-regulated in roots by 182 mM NO(3) (-) treatment. Furthermore, NRA in roots lagged behind CsNR expression and there was no obvious lag of NRA in leaves. This study found that in roots, there was no obvious relationship between NRA and NO(3) (-) content. These results indicated that NRA was not only controlled by the level of CsNR mRNA and there was an obvious negative relationship between NO(3) (-) content and NRA in leaves but not in roots.

Effect of Low-energy Shock Waves in Microfracture Holes in the Repair of Articular Cartilage Defects in a Rabbit Model

Microfracture is a type of bone marrow stimulation in arthroscopic cartilage repair. However, the overall concentration of the mesenchymal stem cells is quite low and declines with age, and in the end the lesion is filled by fibrocartilage. The aim of this research was to investigate a novel method of enhancing microfracture by determining whether low-energy shock waves in microfracture holes would facilitate cartilage repair in a rabbit model.

Electrochemical and Spectroscopic Study on the Interaction Between Isoprenaline and DNA Using Multivariate Curve Resolution-alternating Least Squares

Interaction of isoprenaline (ISO) with calf-thymus DNA was studied by spectroscopic and electrochemical methods. The behavior of ISO was investigated at a glassy carbon electrode (GCE) by cyclic voltammetry (CV) and differential pulse stripping voltammetry (DPSV); ISO was oxidized and an irreversible oxidation peak was observed. The binding constant K and the stoichiometric coefficient m of ISO with DNA were evaluated. Also, with the addition of DNA, hyperchromicity of the UV-vis absorption spectra of ISO was noted, while the fluorescence intensity decreased significantly. Multivariate curve resolution-alternating least squares (MCR-ALS) chemometrics method was applied to resolve the combined spectroscopic data matrix, which was obtained by the UV-vis and fluorescence methods. Pure spectra of ISO, DNA and ISO-DNA complex, and their concentration profiles were then successfully obtained. The results indicated that the ISO molecule intercalated into the base-pairs of DNA, and the complex of ISO-DNA was formed.

Layered Host-guest Materials with Reversible Piezochromic Luminescence

Magnetic-field-assisted Assembly of Layered Double Hydroxide/metal Porphyrin Ultrathin Films and Their Application for Glucose Sensors

The ordered ultrathin films (UTFs) based on CoFe-LDH (layered double hydroxide) nanoplatelets and manganese porphyrin (Mn-TPPS) have been fabricated on ITO substrates via a magnetic-field-assisted (MFA) layer-by-layer (LBL) method and were demonstrated as an electrochemical sensor for glucose. The XRD pattern for the film indicates a long-range stacking order in the normal direction of the substrate. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) images of the MFA LDH/Mn-TPPS UTFs reveal a continuous and uniform surface morphology. Cyclic voltammetry, impedance spectroscopy, and chronoamperometry were used to evaluate the electrochemical performance of the film, and the results show that the MFA-0.5 (0.5 T magnetic field) CoFe-LDH/Mn-TPPS-modified electrode displays the strongest redox current peaks and fastest electron transfer process compared with those of MFA-0 (without magnetic-field) and MFA-0.15 (0.15 T magnetic field). Furthermore, the MFA-0.5 CoFe-LDH/Mn-TPPS exhibits remarkable electrocatalytic activity toward the oxidation of glucose with a linear response range (0.1-15 mM; R(2) = 0.999), low detection limit (0.79 μM) and high sensitivity (66.3 μA mM(-1) cm(-2)). In addition, the glucose sensor prepared by the MFA LBL method also shows good selectivity and reproducibility as well as resistance to poisoning in a chloride ion solution. Therefore, the novel strategy in this work creates new opportunities for the fabrication of nonenzyme sensors with prospective applications in practical detection.

Layer-by-layer Assembly of Electroactive Dye/inorganic Matrix Film and Its Application As Sensor for Ascorbic Acid

A novel inorganic-organic composite ultrathin film was fabricated by layer-by-layer assembly of naphthol green B (NGB) and layered double hydroxides (LDHs) nanoplatelets, which shows remarkable electrocatalytic behavior for oxidation of ascorbic acid. LDHs nanoplatelets were prepared using a method involving separate nucleation and aging steps (particle size: 25±5 nm; aspect ratio: 2-4) and used as building blocks for alternate deposition with NGB on indium tin oxide (ITO) substrates. UV-vis absorption spectroscopy and XRD display regular and uniform growth of the NGB/LDHs ultrathin film with extremely c-orientation of LDHs nanoplatelets (ab plane of microcrystals parallel to substrates). A continuous and uniform surface morphology was observed by SEM and AFM image. The film modified electrode displays a couple of well-defined reversible redox peaks attributed to Fe(2+)/Fe(3+) in NGB (ΔE(p)=68 mV and I(a)/I(c)=1.1). Moreover, the modified electrode shows a high electrocatalytic activity towards ascorbic acid in the range 1.2-55.2 μM with a detection limit of 0.51 μM (S/N=3). The Michaelis-Menten constant was calculated to be K(M)(app)=67.5 μM.

[Mutation Analysis and Prenatal Diagnosis of 2 Cases with Mucopolysaccharidosis Type I]

Mucopolysaccharidosis type I (MPS I; MIM# 252800) is an autosomal recessive disease that results from the deficiency in the lysosomal enzyme α-L-iduronidase(IDUA). IDUA is one of the enzymes involved in degradation of glycosaminoglycans heparan sulphate and dermatan sulphate. The deficiency of IDUA leads to widespread accumulation of partially degraded mucopolysaccharides inside lysosomes, resulting in progressive cellular and multiorgan dysfunction. Up to now there is no definitely effective treatment for this disorder, therefore it is important to provide an accurate genetic diagnosis and prenatal diagnosis for the MPSI families. This study was conducted to detect IDUA gene mutation in patients with MPSIand make a definite diagnosis of homozygote or heterozygote and make first trimester prenatal diagnosis.

Role of Th-2 Cytokines in the Development of Barrett's Esophagus in Rats

Barrett's esophagus is characterized by a distinct Th-2-predominant cytokine profile, unlike the pro-inflammatory nature of reflux esophagitis. The aim of this study was to examine the role of Th-2 cytokines during the development of Barrett's esophagus, using a rat model.

[The Relationship Between Hepatocellular Carcinoma Recurrence and Hepatitis B Virus Recurrence After Liver Transplantation]

To investigate the relationship between hepatocellular carcinoma (HCC) recurrence and hepatitis B virus (HBV) recurrence.

[Mutation in the SLC37A4 Gene of Glycogen Storage Disease Type Ib in 15 Families of the Mainland of China]

Glycogen storage disease type Ib (GSDIb, MIM: 232220) is an autosomal recessive inborn error of metabolism caused by deficiency of the glucose-6-phosphate translocase. The clinical manifestations include symptoms and signs of both the typical GSDIa, including hepatomegaly, fasting hypoglycemia, lactic acidemia and hyperlipidemia, and the dysfunction of neutrophils of recurrent infection and neutropenia. More than 84 mutations have been identified since the discovery of the SLC37A4 gene as the disease causing gene. Up to date, 5 mutations in 4 Chinese patients were reported from Hong Kang and Taiwan. In order to see the spectrum of the SLC37A4 gene mutations and the correlation between genotype and phenotype in patients with GSDIb of the mainland of China, the authors investigated 17 GSDIb patients from 15 families in this study.

Controllable Photoluminescence Properties of an Anion-dye-intercalated Layered Double Hydroxide by Adjusting the Confined Environment

This article reports a novel method to tune the photoluminance properties of ammonium 1-anilinonaphthalene-8-sulfonate (ANS) in a 2D matrix of layered double hydroxide (LDH) by changing the interlayer microenvironment. ANS and a series of surfactants with different alkyl chain lengths (pentanesulphonate (PES), hexanesulphonate (HES), heptanesulphonate (HPS), decanesulphonate (DES), and dodecylsulphonate (DDS)) were respectively cointercalated into the galleries of ZnAl-LDH by the anion exchange method. Thin films of ANS/surfactant-LDHs obtained by the solvent evaporation method possess good c orientation as revealed by XRD and SEM. It was found that the ANS/HPS-LDH film showed the maximum fluorescence efficiency and the longest intensity-average lifetime among these ANS/surfactant-LDH composites owing to the "size-matching" rule between the organic dye and surfactant. Moreover, the fluorescence properties can be tuned by changing the relative molar ratio of ANS/HPS, and the film containing 20% ANS (molar percentage, expressed as ANS(20%)/HPS-LDH) exhibits the maximum fluorescence efficiency, the longest average lifetime, and significantly enhanced photo and thermal stability. In addition, the composite films show fluorescence anisotropy, attributed to the preferential orientation of ANS in the LDH gallery. Therefore, this work demonstrates a feasible approach to tuning the photoluminescence properties of a dye confined in an inorganic 2D matrix via changing the interlayer microenvironment, which may be considered to be a good candidate for solid photoluminescence materials, nonlinear optics, and polarized luminescence materials.

Knocking Down Cyclin D1b Inhibits Breast Cancer Cell Growth and Suppresses Tumor Development in a Breast Cancer Model

Cyclin D1 is aberrantly expressed in many types of cancers, including breast cancer. High levels of cyclin D1b, the truncated isoform of cyclin D1, have been reported to be associated with a poor prognosis for breast cancer patients. In the present study, we used siRNA to target cyclin D1b overexpression and assessed its ability to suppress breast cancer growth in nude mice. Cyclin D1b siRNA effectively inhibited overexpression of cyclin D1b. Depletion of cyclin D1b promoted apoptosis of cyclin D1b-overexpressing cells and blocked their proliferation and transformation phenotypes. Notably, cyclin D1b overexpression is correlated with triple-negative basal-like breast cancers, which lack specific therapeutic targets. Administration of cyclin D1b siRNA inhibited breast tumor growth in nude mice and cyclin D1b siRNA synergistically enhanced the cell killing effects of doxorubicin in cell culture, with this combination significantly suppressing tumor growth in the mouse model. In conclusion, the results indicate that cyclin D1b, which is overexpressed in breast cancer, may serve as a novel and effective therapeutic target. More importantly, the present study clearly demonstrated a very promising therapeutic potential for cyclin D1b siRNA in the treatment of cyclin D1b-overexpressing breast cancers, including the very malignant triple-negative breast cancers.

Optical Heterodyne-detected Raman-induced Kerr Effect (OHD-RIKE) Microscopy

Label-free microscopy based on Raman scattering has been increasingly used in biomedical research to image samples that cannot be labeled or stained. Stimulated Raman scattering (SRS) microscopy allows signal amplification of the weak Raman signal for fast imaging speeds without introducing the nonresonant background and coherent image artifacts that are present in coherent anti-Stokes Raman scattering (CARS) microscopy. Here we present the Raman-induced Kerr effect (RIKE) as a contrast for label-free microscopy. RIKE allows us to measure different elements of the nonlinear susceptibility tensor, both the real and imaginary parts, by optical heterodyne detection (OHD-RIKE). OHD-RIKE microscopy provides information similar to polarization CARS (P-CARS) and interferometric CARS (I-CARS) microscopy, with a simple modification of the two-beam SRS microscopy setup. We show that, while OHD-RIKE microspectroscopy can be in principle more sensitive than SRS, it does not supersede SRS microscopy of heterogeneous biological samples, such as mouse skin tissue, because it is complicated by variations of linear birefringence across the sample.

Three-dimensional WO3 Nanostructures on Carbon Paper: Photoelectrochemical Property and Visible Light Driven Photocatalysis

Three-dimensional (3D) WO(3) nanostructures were grown on carbon paper by a catalyst-free high temperature reactive vapor deposition process, which exhibit a good photoelectrochemical property and visible light driven photocatalytic performance.

[Mutation Analysis of 11 Chinese Patients with Attenuated Mucopolysaccharidosis Type]

Mucopolysaccharidosis type I (MPS I) is an autosomal recessive disease resulting from the deficiency in the lysosomal enzyme alpha-L-iduronidase (IDUA). The present study was conducted to identify IDUA gene mutations in attenuated (MPS I H/S and MPS I S) patients with MPS I in northern China.

Biomimetic Design and Assembly of Organic-inorganic Composite Films with Simultaneously Enhanced Strength and Toughness

Inorganic nanoplatelet reinforced polymer films were fabricated via alternate layer-by-layer assembly of layered double hydroxide (LDH) nanoplatelets with poly(vinyl alcohol) (PVA), which showed largely enhanced strength and good ductility simultaneously.

Coherent Nonlinear Optical Imaging: Beyond Fluorescence Microscopy

The quest for ultrahigh detection sensitivity with spectroscopic contrasts other than fluorescence has led to various novel approaches to optical microscopy of biological systems. Coherent nonlinear optical imaging, especially the recently developed nonlinear dissipation microscopy (including stimulated Raman scattering and two-photon absorption) and pump-probe microscopy (including excited-state absorption, stimulated emission, and ground-state depletion), provides new image contrasts for nonfluorescent species. Thanks to the high-frequency modulation transfer scheme, these imaging techniques exhibit superb detection sensitivity. By directly interrogating vibrational and/or electronic energy levels of molecules, they offer high molecular specificity. Here we review the underlying principles and excitation and detection schemes, as well as exemplary biomedical applications of this emerging class of molecular imaging techniques.

P27(Kip1), Regulated by Glycogen Synthase Kinase-3β, Results in HMBA-induced Differentiation of Human Gastric Cancer Cells

Gastric cancer is the second most common cause of global cancer-related mortality. Although dedifferentiation predicts poor prognosis in gastric cancer, the molecular mechanism underlying dedifferentiation, which could provide fundamental insights into tumor development and progression, has yet to be elucidated. Furthermore, the molecular mechanism underlying the effects of hexamethylene bisacetamide (HMBA), a recently discovered differentiation inducer, requires investigation and there are no reported studies concerning the effect of HMBA on gastric cancer.

[Inhibitory Effect of Chailing Decoction on Renal Tubular Epithelial Phenotype Transformation in Rats with Cyclosporine A-induced Nephropathy]

To observe the effect of Chailing Decoction (CLD) on alpha-smooth muscular actin (alpha-SMA, a marker of renal tubular epithelial phenotype transformation) in rats with cyclosporine A (CsA)-induced nephropathy for investigate its mechanism of action in inhibiting tubulo-interstitial fibrosis.

[Arthroscopic Treatment for Tibial Eminence Avulsion Fracture Using Absorbable Double Suture Anchors]

To investigate the outcomes of arthroscopic reduction and internal fixation of tibial eminence avulsion fracture using absorbable double suture anchors.

Protective Effects of Astragaloside Against Ultraviolet A-induced Photoaging in Human Fibroblasts

In this study, we aim to investigate the protective effects of astragaloside on ultraviolet A (UVA)-induced photoaging in human fibroblasts and its possible mechanisms.

[Treatment of Adult Congenital Muscular Torticollis with Radiofrequency Carbonation Under Arthroscope]

To evaluate the efficacy of sternocleidomastoideus radiofrequency carbonation with local anesthesia under arthroscope for the treatment of congenital muscular torticollis in adults.

NKp46 Identifies an NKT Cell Subset Susceptible to Leukemic Transformation in Mouse and Human

IL-15 may have a role in the development of T cell large granular lymphocyte (T-LGL) or NKT leukemias. However, the mechanisms of action and the identity of the cell subset that undergoes leukemic transformation remain elusive. Here we show that in both mice and humans, NKp46 expression marks a minute population of WT NKT cells with higher activity and potency to become leukemic. Virtually 100% of T-LGL leukemias in IL-15 transgenic mice expressed NKp46, as did a majority of human T-LGL leukemias. The minute NKp46+ NKT population, but not the NKp46⁻ NKT population, was selectively expanded by overexpression of endogenous IL-15. Importantly, IL-15 transgenic NKp46⁻ NKT cells did not become NKp46+ in vivo, suggesting that NKp46+ T-LGL leukemia cells were the malignant counterpart of the minute WT NKp46+ NKT population. Mechanistically, NKp46+ NKT cells possessed higher responsiveness to IL-15 in vitro and in vivo compared with that of their NKp46⁻ NKT counterparts. Furthermore, interruption of IL-15 signaling using a neutralizing antibody could prevent LGL leukemia in IL-15 transgenic mice. Collectively, our data demonstrate that NKp46 identifies a functionally distinct NKT subset in mice and humans that appears to be directly susceptible to leukemic transformation when IL-15 is overexpressed. Thus, IL-15 signaling and NKp46 may be useful targets in the treatment of patients with T-LGL or NKT leukemia.

[Therapy and Prognosis Analysis of 47 Patients with Extranodal NK/T-cell Lymphoma]

This study was purposed to explore the clinical characteristics, therapy and prognosis of patients with extranodal NK/T cell lymphoma (ENKL). 47 patients with ENKL from October 1995 to December 2008 in our hospital were analyzed retrospectively. The survival of patients was analyzed by using Kaplan-Meier methods, the prognosis of patients was evaluated by multivariate analysis using COX regression model. The clinical parameters used included CD56, Ann Arbor stage, international prognostic index (IPI) and B symptom. The results showed that the 2-year and 5-year overall survival (OS) rates were 91%, 71% respectively. Multivariant analysis by COX regression showed the CD56 and Ann Arbor stage were independent prognostic factors. Single factor analysis with staging in CHOP chemotherapy group indicated that more than stage IIIE (including IIIE) was a prognostic factor. Single factor analysis with B symptom showed that B symptom also was a prognostic factor. The cumulative survival rate of patients received radiotherapy alone was higher than that of patients got chemotherapy alone, its difference had a statistical significance, but there was no statistical significance between radiotherapy group and chemoradiotherapy group. It is concluded that CD56, Ann Arbor stage and B symptoms are prognostic factors. Radiotherapy alone is better than chemotherapy alone in therapy of ENKL. Chemotherapy combined with radiotherapy can not improve the survival. Improving the status of patients can make prognosis better.

[Differential Gene Expressions in Kidney Yang Deficiency in Individuals with Sub-health Status]

To study the gene expression profiles in kidney Yang deficiency in individuals with sub-health status.

Growth Hormone Receptor Deficiency is Associated with a Major Reduction in Pro-aging Signaling, Cancer, and Diabetes in Humans

Mutations in growth signaling pathways extend life span, as well as protect against age-dependent DNA damage in yeast and decrease insulin resistance and cancer in mice. To test their effect in humans, we monitored for 22 years Ecuadorian individuals who carry mutations in the growth hormone receptor (GHR) gene that lead to severe GHR and IGF-1 (insulin-like growth factor-1) deficiencies. We combined this information with surveys to identify the cause and age of death for individuals in this community who died before this period. The individuals with GHR deficiency exhibited only one nonlethal malignancy and no cases of diabetes, in contrast to a prevalence of 17% for cancer and 5% for diabetes in control subjects. A possible explanation for the very low incidence of cancer was suggested by in vitro studies: Serum from subjects with GHR deficiency reduced DNA breaks but increased apoptosis in human mammary epithelial cells treated with hydrogen peroxide. Serum from GHR-deficient subjects also caused reduced expression of RAS, PKA (protein kinase A), and TOR (target of rapamycin) and up-regulation of SOD2 (superoxide dismutase 2) in treated cells, changes that promote cellular protection and life-span extension in model organisms. We also observed reduced insulin concentrations (1.4 μU/ml versus 4.4 μU/ml in unaffected relatives) and a very low HOMA-IR (homeostatic model assessment-insulin resistance) index (0.34 versus 0.96 in unaffected relatives) in individuals with GHR deficiency, indicating higher insulin sensitivity, which could explain the absence of diabetes in these subjects. These results provide evidence for a role of evolutionarily conserved pathways in the control of aging and disease burden in humans.

Magnetic-field-assisted Assembly of CoFe Layered Double Hydroxide Ultrathin Films with Enhanced Electrochemical Behavior and Magnetic Anisotropy

Magnetic films based on CoFe LDH nanoplatelets and porphyrin anions were fabricated by the layer-by-layer assembly technique with an assistance of an external magnetic field, which show enhanced electrochemical behavior and magnetic anisotropy.

Mice Carrying a Knock-in Mutation of Aicda Resulting in a Defect in Somatic Hypermutation Have Impaired Gut Homeostasis and Compromised Mucosal Defense

To elucidate the specific role of somatic hypermutation (SHM) in mucosal immunity, we generated mice carrying a knock-in point mutation in Aicda, which encodes activation-induced cytidine deaminase (AID), an enzyme essential to SHM and class-switch recombination (CSR). These mutant AID(G23S) mice had much less SHM but had normal amounts of immunoglobulin in both serum and intestinal secretions. AID(G23S) mice developed hyperplasia of germinal center B cells in gut-associated lymphoid tissues, accompanied by expansion of microflora in the small intestine. Moreover, AID(G23S) mice had more translocation of Yersinia enterocolitica into mesenteric lymph nodes and were more susceptible than wild-type mice to oral challenge with cholera toxin. Together our results indicate that SHM is critical in maintaining intestinal homeostasis and efficient mucosal defense.

Tumor-associated MUC5AC Stimulates in Vivo Tumorigenicity of Human Pancreatic Cancer

MUC5AC, a high molecular weight glycoprotein, is overexpressed in the ductal region of human pancreatic cancer but is not detectable in the normal pancreas, suggesting its association with disease development. In the present study, we investigated the in vitro and in vivo effects of MUC5AC knockdown by short interfering RNA (siRNA) in the MUC5AC-overexpressing SW1990 and BxPC3 human pancreatic cancer cell lines in order to clarify its function. Significant decreases in the expression levels of MUC5AC mRNA and protein were observed in SW1990 and BxPC3 cells that had been stably transfected with a MUC5AC siRNA expression vector (SW1990/si-MUC5AC and BxPC3/si-MUC5AC cells) compared to those in cells transfected with an si-mock vector (SW1990/si-mock and BxPC3/si-mock cells). In in vitro studies, neither type of MUC5AC-knockdown cell showed any difference in cell survival, proliferation, or morphology from the si-mock cells or parental cells. However, in vivo xenograft studies demonstrated that MUC5AC knockdown significantly reduced the tumorigenicity and suppressed the tumor growth of si-MUC5AC cells compared to those of the si-mock cells. Immunohistochemical analysis revealed that CD45R/B220+ and Gr-1+ cells had infiltrated into the tumor tissue of the SW1990/si-MUC5AC cells. Furthermore, cancer-associated antigen specific antibodies were detected at high levels in the sera from the SW1990/si-MUC5AC cell-bearing mice. These results suggest that tumor-associated MUC5AC expressed on the surface of pancreatic cancer cells supports the escape of pancreatic cancer cells from immunosurveillance. The present findings highlight a new dimension of MUC5AC as a functional immunosuppressive agent and its important role in pancreatic cancer progression.

RNAi Screening for Fat Regulatory Genes with SRS Microscopy

Identification of genes regulating fat accumulation is important for basic and medical research; genetic screening for those genes in Caenorhabditis elegans, a widely used model organism, requires in vivo quantification of lipids. We demonstrated RNA interference screening based on quantitative imaging of lipids with label-free stimulated Raman scattering (SRS) microscopy, which overcomes major limitations of coherent anti-Stokes Raman scattering microscopy. Our screening yielded eight new genetic regulators of fat storage.

Reversibly Thermochromic, Fluorescent Ultrathin Films with a Supramolecular Architecture

IκBζ Augments IL-12- and IL-18-mediated IFN-γ Production in Human NK Cells

Interferon-γ (IFN-γ) production by natural killer (NK) cells and cytotoxic lymphocytes is a key component of innate and adaptive immune responses. Because inhibitor of κB-ζ (IκBζ), a Toll-like receptor (TLR)/interleukin-1 receptor (IL-1R) inducible transcription factor, regulates IFN-γ production in KG-1 cells, we tested IκBζ's role in the classic lymphocyte pathway of IL-12/IL-18-induced IFN-γ. Upon stimulation with IL-12/IL-18, monocyte-depleted human peripheral blood lymphocytes expressed the 79-kDa form of IκBζ and released IFN-γ. CD56(+) NK cells were shown to be the IκBζ-producing lymphocyte subpopulation, which also released abundant IFN-γ in response to IL-12/IL-18. Importantly, IκBζ was undetectable in CD56(-) lymphocytes where IFN-γ release was 10-fold lower. In addition, small interfering RNA knockdown of IκBζ suppressed IFN-γ expression in CD56(+) cells. The association of IκBζ with the IFN-γ promoter was documented by chromatin immunoprecipitation. IFN-γ promoter activity from IκBζ overexpression was confirmed by luciferase reporter assay. Finally, IκBζ coprecipitated with p65 and p50 NF-κB in NK cells in response to IL-12/IL-18, suggesting that IκBζ's effects on IFN-γ promoter activity are coregulated by NF-κB. These results suggest that IκBζ functions as an important regulator of IFN-γ in human NK cells, further expanding the class of IκBζ-modulated genes.

Non-genotoxic Mode of Action and Possible Threshold for Hepatocarcinogenicity of Kojic Acid in F344 Rats

Kojic acid (KA), a naturally occurring compound, is contained in traditional Japanese fermented foods and is used as a food additive, preservative and a dermatological skin-lightening agent. In the present experiment, initiation (experiment 1) and promotion (experiment 2) effects of KA-induced hepatocarcinogenesis were studied by rat medium-term bioassay for carcinogenicity. Male F344 rats were administered a diet containing 0-2% KA. Experiment 1 demonstrated that KA had no effect on induction of liver preneoplastic lesions or glutathione S-transferase placental form (GST-P) positive foci, in either number or area. In experiment 2, 2% KA treatment significantly increased the number and area of GST-P positive foci, but concentrations less than 0.5% did not. Moreover, 2% KA treatment significantly increased 8-OHdG levels and PCNA positive hepatocytes. The results indicated that low concentrations of KA do not have initiation effects on rat hepatocarcinogenesis, while higher concentrations of KA do promote hepatocarcinogenesis in rats. Thus, the results indicate that KA is a non-genotoxic hepatocarcinogen, showing the possible existence of a perfect threshold.

Low-dose Carcinogenicity of 2-amino-3-methylimidazo[4,5-f ]quinoline in Rats: Evidence for the Existence of No-effect Levels and a Mechanism Involving P21(Cip / WAF1)

The carcinogenicity of the low amounts of genotoxic carcinogens present in food is of pressing concern. The purpose of the present study was to determine the carcinogenicity of low doses of the dietary genotoxic carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and to investigate mechanisms by which IQ exerts its carcinogenic effects. A total of 1595 male F344 rats were divided into seven groups and administered with IQ at doses of 0, 0.001, 0.01, 0.1, 1, 10 and 100 p.p.m. in the diet for 16 weeks. We found that IQ doses of 1 p.p.m. and below did not induce preneoplastic lesions in either the liver or the colon, while IQ doses of 10 and 100 p.p.m. induced preneoplastic lesions in both of these organs. These results demonstrate the presence of no-effect levels of IQ for both liver and colon carcinogenicity in rats. The finding that p21(Cip/WAF1) was significantly induced in the liver at doses well below those required for IQ mediated carcinogenic effects suggests that induction of p21(Cip/WAF1) is one of the mechanisms responsible for the observed no-effect of low doses of IQ. Furthermore, IQ administration caused significant induction of CYP1A2 at doses of 0.01-10 p.p.m., but administration of 100 p.p.m. IQ induced CYP1A1 rather than CYP1A2. This result indicates the importance of dosage when interpreting data on the carcinogenicity and metabolic activation of IQ. Overall, our results suggest the existence of no-effect levels for the carcinogenicity of this genotoxic compound.

Mitochondrial Prohibitins and Septin 9 Are Implicated in the Onset of Rat Hepatocarcinogenesis

In the present study, protein lysates from microdissected glutathione S-transferase placental-form-positive (GST-P(+)) foci and hepatocellular carcinomas from livers of rats treated with N-diethylnitrosamine followed by phenobarbital at doses of 0 and 500 ppm in the diet for 10 and 33 weeks were analyzed using QSTAR Elite liquid chromatography with tandem mass spectrometry and iTRAQ technology. Among 75 proteins, a total of 27 and 50 proteins displaying significant quantitative changes comparing with adjacent normal-appearing liver tissue were identified in GST-P(+) foci of initiation control and promotion groups, respectively, which are related to transcription, protein folding, cytoskeleton filaments reorganization, cell cycle control, nuclear factor (erythroid-derived 2)-like 2 (NRF2)-mediated oxidative stress responses, lipid metabolism, glutathione metabolism, oxidative phosphorylation, and signal transduction. Furthermore, Ingenuity Pathway and bioinformatic analyses revealed that expression changes of genes encoding proteins with altered expression detected in GST-P(+) foci are likely to be controlled by c-myc, NRF2, aryl hydrocarbon receptor, nuclear factor kappa B, and hepatocyte nuclear factor 4 transcriptional factors. Coordinated overexpression of mitochondrial chaperons prohibitin (PHB) and prohibitin 2 (PHB2), septin 9 (SEPT9), neurabin 1, and other cytoskeletal and functional proteins in areas of GST-P(+) foci during initiation and/or promotion stages of rat hepatocarcinogenesis was associated with induction of cell proliferation and might be responsible for the neoplastic transformation of rat liver preneoplastic lesions. Newly discovered elevation of PHB, PHB2, and SEPT9 in GST-P(+) foci and tumors, imply that they might play important role in the onset of liver cancer and be of potential values in the studies of hepatocarcinogenesis.

Ascl1 Lineage Cells Contribute to Ischemia-induced Neurogenesis and Oligodendrogenesis

Neural and oligodendrocyte progenitor cells in the adult brain express Ascl1 (also known as Mash1), a basic helix-loop-helix transcription factor. We examined the progeny and fate of this progenitor population in adult male Ascl1-CreER(TM);R26R-stop-yellow fluorescent protein mice subjected to right middle cerebral occlusion over 60 days after stroke using inducible Cre recombination to label Ascl1-expressing cells at poststroke days 2 to 6 in vivo. Seven days after stroke, a substantial increase in Ascl1 lineage cells was detected in the ipsilateral subventricular zone (SVZ), striatum, and corpus callosum. These cells exhibited proliferating progenitor cell phenotypes (Sox2(+), BrdU(+), and Ki67(+)). Although Ascl1 lineage cells in the ipsilateral SVZ gradually decreased during 14 to 60 days after stroke, Ascl1 lineage cells in the ischemic striatum revealed a remarkable increase during this period. Thirty and sixty days after stroke, Ascl1 lineage cells in the ischemic striatum gave rise to GABAergic neurons and mature oligodendrocytes. In contrast, none of the Ascl1 lineage cells in the contralateral striatum exhibited neuronal and oligodendrocyte phenotypes. Moreover, Ascl1 lineage cells in the corpus callosum were only fated to become mature oligodendrocytes. Our data suggest that Ascl1 lineage cells contribute to stroke-induced neurogenesis and oligodendrogenesis in the adult ischemic brain.

Protein-flexibility Mediated Coupling Between Photoswitching Kinetics and Surrounding Viscosity of a Photochromic Fluorescent Protein

Recent advances in fluorescent proteins (FPs) have generated a remarkable family of optical highlighters with special light responses. Among them, Dronpa exhibits a unique capability of reversible light-regulated on-off switching. However, the environmental dependence of this photochromism is largely unexplored. Herein we report that the photoswitching kinetics of the chromophore inside Dronpa is actually slowed down by increasing medium viscosity outside Dronpa. This finding is a special example of an FP where the environment can exert a hydrodynamic effect on the internal chromophore. We attribute this effect to protein-flexibility mediated coupling where the chromophore's cis-trans isomerization during photoswitching is accompanied by conformational motion of a part of the protein β-barrel whose dynamics should be hindered by medium friction. Consistent with this mechanism, the photoswitching kinetics of Dronpa-3, a structurally more flexible mutant, is found to exhibit a more pronounced viscosity dependence. Furthermore, we mapped out spatial distributions of microviscosity in live cells experienced by a histone protein using the photoswitching kinetics of Dronpa-3 fusion as a contrast mechanism. This unique reporter should provide protein-specific information about the crowded intracellular environments by offering a genetically encoded microviscosity probe, which did not exist with normal FPs before.

Fasting Cycles Retard Growth of Tumors and Sensitize a Range of Cancer Cell Types to Chemotherapy

Short-term starvation (or fasting) protects normal cells, mice, and potentially humans from the harmful side effects of a variety of chemotherapy drugs. Here we show that treatment with starvation conditions sensitized yeast cells (S. cerevisiae) expressing the oncogene-like RAS2(val19) to oxidative stress and 15 of 17 mammalian cancer cell lines to chemotherapeutic agents. Cycles of starvation (fasting) were as effective as chemotherapeutic agents in delaying progression of specific tumors and increased the effectiveness of these drugs against melanoma, glioma, and breast cancer cells. In mouse models of neuroblastoma, fasting cycles plus chemotherapy drugs-but not either treatment alone-resulted in long-term cancer-free survival. In 4T1 breast cancer cells, short-term starvation resulted in increased phosphorylation of the stress-sensitizing AKT and S6 kinases, increased oxidative stress, caspase-3 cleavage, DNA damage and apoptosis. These studies suggest that multiple cycles of fasting promote differential stress sensitization in a wide range of tumors and could potentially replace or augment the efficacy of certain toxic chemotherapy drugs in the treatment of various cancers.

The HDAC Inhibitor Valproic Acid Lessens NK Cell Action Against Oncolytic Virus-infected Glioblastoma Cells with Inhibition of STAT5/T-BET Signaling and IFNγ Generation

Tumor virotherapy has and continues to be used in clinical trials. One barrier to effective viral oncolysis, consisting of the interferon response induced by viral infection, is inhibited by valproic acid (VPA) and other histone deacetylase inhibitors (HDACi). Innate immune cell recruitment and activation have been shown to be deleterious to the efficacy of oncolytic HSV (oHSV) and in this study we demonstrate that VPA limits this deleterious response. VPA, administered prior to oHSV inoculation into an orthotopic glioblastoma mouse model, resulted in a decline in NK and macrophage recruitment into tumor bearing brains at 6 and 24 hours post-oHSV. Interestingly, there was a robust rebound of recruitment of these cells at 72 hours-post-oHSV. The observed initial decline in immune cell recruitment was accompanied by a reduction in their activation status. VPA was also found to have a profound immunosuppressive effect on human NK cells in vitro. NK cytotoxicity was abrogated following exposure to VPA, consistent with down-modulation of cytotoxic gene expression of granzyme B and perforin at the mRNA and protein levels. In addition, suppression of IFN-γ production by VPA was associated with decreased STAT5 phosphorylation and dampened T-BET expression. Despite VPA mediated immune suppression, mice were not at significantly increased risk for HSV encephalitis. These findings indicate that one of the avenues in which VPA enhances oHSV efficacy is through initial suppression of immune cell recruitment and inhibition of inflammatory cell pathways within NK cells.

The Prostate Cancer-up-regulated Long Noncoding RNA PlncRNA-1 Modulates Apoptosis and Proliferation Through Reciprocal Regulation of Androgen Receptor

OBJECTIVE: Emerging evidences implicate long noncoding RNAs (lncRNAs) are deregulated in cancer development. The purpose of the current study is to investigate the role of new lncRNA, named PlncRNA-1, in prostate cancer (CaP) pathogenesis. MATERIALS AND METHODS: In this study, real-time q-PCR was used to demonstrate the expression of PlncRNA-1 in 16 pairs CaP tissues and matched normal tissues, 14 pairs CaP tissues and BPH tissues, 4 CaP cell lines, including LNCaP, LNCaP-AI, PC3, and C4-2, and 2 normal prostate epithelial cell lines RWPE-1 and PWR-1E. After PlncRNA-1 was suppressed by siRNA in LNCaP and LNCaP-AI cell lines, cell proliferation and apoptosis were assessed using CCK-8 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). After PlncRNA-1 and AR was suppressed by siRNA in LNCaP and LNCaP-AI cell lines, real-time q-PCR and Western blotting were used to measure reciprocal regulation of PlncRNA-1 and AR. RESULTS: We showed that expression PlncRNA-1, was significantly higher in CaP cells relative to normal prostate epithelial cells, as well as higher in human CaPs compared with normal tissues and benign prostatic hyperplasia (BPH). Silencing of PlncRNA-1 significantly reduced cell proliferation and induced apoptosis in CaP cell lines LNCaP and LNCaP-AI. Mechanistically, PlncRNA-1 suppression by siRNA resulted in a decrease of androgen receptor (AR) mRNA, protein and AR downstream target. Of note, blockade of AR signaling with siRNA also resulted in a suppression of PlncRNA-1 expression in CaP cell lines. CONCLUSIONS: Our study suggests reciprocal regulation of PlncRNA-1 and androgen receptor contribute to CaP pathogenesis and that PlncRNA-1 is a potential therapy target.

Hormonally Active Doses of Isoflavone Aglycones Promote Mammary and Endometrial Carcinogenesis and Alter the Molecular Tumor Environment in Donryu Rats

Our research is focused on modifying effects of an isoflavone aglycones (IA)-rich extract at a hormonally active dose of 150 mg/kg b.w./day, on mammary and endometrial carcinogenesis in female Donryu rats. IA administered for 2 weeks in a phytoestrogen-low diet exerted estrogenic activity and induced cell proliferation in the uterus of ovariectomized rats. Furthermore, administration for 4 weeks resulted in elevation of cell proliferation in the mammary glands of 7,12-dimethylbenz[a]anthracene (DMBA) treated animals. Forty weeks of postpubertal administration of IA to 5-week-old rats after initiation of mammary and endometrial carcinogenesis with DMBA and N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) caused significant increase of incidence, and multiplicity of mammary adenocarcinoma, multiplicities of endometrial atypical hyperplasia, adenomatous polyps, and a trend for increase in uterine adenocarcinomas. LC-MS/MS and immunohistochemical analyses revealed significant elevation of tumorigenesis-related proteins such as S100 calcium binding protein A8, kininogen 1 and annexins 1 and 2 in mammary adenocarcinomas, and cadherin EGF LAG seven-pass G-type receptor 2, DEAD box polypeptide 1 and cysteine and glycine-rich protein 1 in uterine proliferative lesions of IA-treated animals. Those changes are likely to be related to modulation of estrogen receptor, AP1, NF-kB and actin signaling pathways. Our results indicate that postpubertal exposure of Donryu rats to IA at an estrogenic dose resulted in promotion of mammary and uterine carcinogenesis induced by DMBA and ENNG, which might be related to the activation of ER-dependent signaling, and alteration of the molecular tumor environment in the mammary gland and endometrium.

Preparation of Fe(3)O(4)@SiO(2)@Layered Double Hydroxide Core-Shell Microspheres for Magnetic Separation of Proteins

Three-component microspheres containing an SiO(2)-coated Fe(3)O(4) magnetite core and a layered double hydroxide (LDH) nanoplatelet shell have been synthesized via an in situ growth method. The resulting Fe(3)O(4)@SiO(2)@NiAl-LDH microspheres display three-dimensional core-shell architecture with flowerlike morphology, large surface area (83 m(2)/g), and uniform mesochannels (4.3 nm). The Ni(2+) cations in the NiAl-LDH shell provide docking sites for histidine and the materials exhibit excellent performance in the separation of a histidine (His)-tagged green fluorescent protein, with a binding capacity as high as 239 μg/mg. The microspheres show highly selective adsorption of the His-tagged protein from Escherichia coli lysate, demonstrating their practical applicability. Moreover, the microspheres possess superparamagnetism and high saturation magnetization (36.8 emu/g), which allows them to be easily separated from solution by means of an external magnetic field and subsequently reused. The high stability and selectivity of the Fe(3)O(4)@SiO(2)@NiAl-LDH microspheres for the His-tagged protein were retained over several separation cycles. Therefore, this work provides a promising approach for the design and synthesis of multifunctional LDH microspheres, which can be used for the practical purification of recombinant proteins, as well as having other potential applications in a variety of biomedical fields including drug delivery and biosensors.

Polyvalent Immunostimulatory Nanoagents with Self-assembled CpG Oligonucleotide-conjugated Gold Nanoparticles

Delivery vehicles: Gold nanoparticles (AuNPs) have been used as a nanocarrier to non-invasively deliver synthetic cytosine-phosphate-guanosine (CpG) oligodeoxynucleotides (ODNs) into cells. Compared to unconjugated single-stranded CpG ODNs, self-assembled polyvalent CpG-AuNP conjugates enhance the efficiency of cellular uptake and stimulate secretion of cytokines.

Changes in the Prevalence of Visual Impairment Due to Blinding Trachoma in Sichuan Province, China: a Comparative Study Between 1987 and 2006

Purpose: The aim of this study was to assess the changes in the prevalence of visual impairment due to blinding trachoma in Sichuan, China between 1987 and 2006. Methods: Over 125,000 residents of Sichuan province participated in the National Sample Survey on Disabilities, conducted in both 1987 and 2006. Changes in the prevalence of visual impairment due to blinding trachoma were analyzed while accounting for other factors including gender, age and residence (urban vs. rural). Results: In 1987, blinding trachoma ranked second among the eight major visual impairment causes assessed with a prevalence of 172.9 (95% CI: 152.2-196.6) per 100,000, while in 2006 it ranked eighth with a prevalence of 58.1 (95% CI: 45.5-73.0) per 100,000. The prevalence in 2006 decreased significantly compared to 1987 in both rural and urban areas, as well as in both genders. Furthermore, in both 1987 and 2006, visual impairment due to blinding trachoma tended to be concentrated among individuals over 40, although it was also seen in young people. Conclusion: In the past 20 years, the prevalence and percentage of visual impairment due to blinding trachoma was significantly reduced in Sichuan province, and we anticipate a further decrease in the future.

2-Amino-3-Methylimidazo[4,5-f]Quinoline (IQ) Promotes Mouse Hepatocarcinogenesis by Activating Transforming Growth Factor-β and Wnt/β-Catenin Signaling Pathways

The purposes of the present study were to investigate the modifying effects of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), a genotoxic carcinogen produced during cooking of protein-rich foods, and elucidate underlying mechanisms in a two-stage hepatocarcinogenesis mice model. Six-week-old B6C3F1 mice were subjected to two-thirds partial hepatectomy at the beginning of the study, followed by an intraperitoneal injection of diethylnitrosamine on day 1. Starting 1 week later, they were fed diets containing IQ at doses of 30, 100, or 300 ppm for 39 weeks. A dose-dependent trend for increase in eosinophilic altered foci as well as eosinophilic hepatocellular adenomas was observed, along with significant elevation in the incidence of hepatocellular carcinomas in the 100- and 300-ppm IQ groups as compared with initiation control group. Furthermore, IQ elevated the protein expression levels of Wnt1, transforming growth factor-β (TGF-β), TGF-β receptors 1 and 2 (TβR1 and TβR2), and phosphorylated c-Jun (p-c-Jun), while suppressing those of E-cadherin and p21(WAF1/Cip1). Moreover, translocation of β-catenin to the nuclei as well as upregulated nuclear expression of c-Myc and cyclin D1, which are downstream targets of β-catenin and p-c-Jun, were detected at 100 and 300 ppm. These findings suggest that IQ exerts dose-dependent promoting effects on mice hepatocarcinogenesis by activating TGF-β and Wnt/β-catenin signaling pathways and inhibiting cell adhesion.

Anti-angiogenic Genistein Inhibits VEGF-induced Endothelial Cell Activation by Decreasing PTK Activity and MAPK Activation

Genistein (Gen), a soy isoflavone, is considered to exert potent antitumor effect partially through its anti-angiogenesis property. However, the precise molecular mechanism is still unknown. Our previous investigations have demonstrated that genistein down-regulates expression of pro-angiogenic factors via inhibiting protein tyrosine kinase (PTK) activity both in breast cancer cells and in xenograft tumors. In the present experiment, we chose cultured human umbilical vein endothelial cells (HUVECs), which have a considerable role in tumor angiogenesis formation, to explore the influence of genistein on VEGF-stimulated endothelial cell activation and the underlying mechanism. Stimulation of human primary HUVECs by VEGF not only increased endothelial cell protein tyrosine kinase (PTK) activity but also augmented matrix metalloproteinase-2 (MMP-2), -9 secretions and increased MMP-2, -9 activities. Treatment of ECs with genistein induced VEGF-loaded endothelial apoptosis by inhibiting production and activity of matrix metalloproteinases (MMPs). In addition, exposure to genistein decreased activation of JNK and p38, not ERK-1/2, induced by VEGF. Collectively, our findings suggested that the inhibition of PTK activity and MAPK activation and the decrease in MMPs production and activity by genistein interrupt VEGF-stimulated endothelial cell activation, which thereby may represent a mechanism that would explain the anti-angiogenesis effect of genistein and its cancer-protective function.

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