Immunocytochemical Localization of Substance P Receptor in Hypothalamic Oxytocin-containing Neurons of C57 Mice

Brain Research. Sep, 2002  |  Pubmed ID: 12383972

With the use of double immunofluorescence, we have examined the distribution of oxytocin-containing neurons that express substance P receptor (SPR) in the hypothalamus of C57 mice. The distribution of oxytocin-like immunoreactive neurons overlapped with that of SPR-like immunoreactive neurons in the paraventricular nucleus and supraoptic nucleus of the hypothalamus. Neurons showing both oxytocin- and SPR-like immunoreactivities were predominantly found in both nuclei. A few neurons that were double-labeled with oxytocin- and SPR-like immunoreactivities were also scattered in the hypothalamic periventricular and preoptic regions. Semi-quantitative analysis indicated that about 94% of the oxytocin-like neurons displayed SPR-like immunoreactivity. These double-labeled cells constituted about 91% of the SPR-like neurons in the aforementioned regions. The present study provides morphological evidence for tachykinin-induced modulation of oxytocin-containing neurons as mediated by substance P receptor in the hypothalamus of mammals.

Leporin B: a Novel Hexokinase II Gene Inducing Agent from an Unidentified Fungus

Bioorganic & Medicinal Chemistry Letters. Apr, 2003  |  Pubmed ID: 12668006

Leporin B (1), a novel demethylated analogue of leporin A (2), was isolated from a taxonomically unidentified fungal strain as part of an effort to discover compounds with the ability to increase expression levels of the enzyme hexokinase II. The structure was determined by spectral methods, including 1D and 2D NMR, and HRMS. The relative stereochemistry was assigned by NOESY experiments and coupling constants.

[Operative Treatment of Displaced Talar Neck Fractures with Absorbable Lag Screw]

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery. Sep, 2003  |  Pubmed ID: 14551931

To study a new kind of operation for displaced talar neck fractures.

[Cloning of the Complete Coding Sequence of Mouse Oxytocin Receptor Gene and Its Eukaryotic Expression]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi = Chinese Journal of Cellular and Molecular Immunology. Jul, 2004  |  Pubmed ID: 15207101

To clone the complete coding sequence of mouse oxytocin receptor (mOTR) gene and to express it in mammalian cells.

Identification of Cell Surface and Secreted Proteins Essential for Tumor Cell Survival Using a Genetic Suppressor Element Screen

Oncogene. Oct, 2004  |  Pubmed ID: 15361835

Survival factors play critical roles in regulating cell growth in normal and cancer cells. We designed a genetic screen to identify survival factors which protect tumor cells from apoptosis. A retroviral expression library of random cDNA fragments was constructed from cancer cells and used to transduce the colon carcinoma cell line HCT116. Recipient cells were functionally selected for induction of caspase 3-mediated apoptosis. Analyses of over 10,000 putative genetic suppression elements (GSEs) sequences revealed cognate gene candidates that are implicated in apoptosis. We further analysed 26 genes encoding cell surface and secreted proteins that can potentially serve as targets for therapeutic antibodies. Tetracycline-inducible GSEs from several gene candidates induced apoptosis in stable HCT 116 cell lines. Similar phenotypes were caused by RNAi derived from the same genes. Our data suggest requirement for the cell surface targets IGF2R, L1CAM and SLC31A1 in tumor cell growth in vitro, and suggests that IGF2R is required for xenograft tumor growth in a mouse model.

Nicotinic Receptor Alpha Subunits in Magnocellular Neurons of Rat Hypothalamus

Neuroreport. Oct, 2004  |  Pubmed ID: 15640750

Mab35 is a monoclonal antibody against one specific immunogenic region in alpha1, alpha3, alpha5 subunits of nicotinic acetylcholine receptors (N-AChR) of a variety of species. It has previously been claimed that N-AChR-like immunoreactivity (-LI) identified by mab35 is present in vasopressin-containing magnocellular neurons. However, we show here by double immunofluorescence labelling that mab35 immunoreactivity is predominantly localized to oxytocinergic rather than vasopressinergic magnocellular neurons. We further infer that mab35 predominantly stained the alpha3 and/or alpha5 subunits in rat oxytocinergic neurons, and suggest that the unbalanced distribution of these subunits may contribute to some specific physiological properties of oxytocinergic neurons.

Active Pixel Sensor Array As a Detector for Electron Microscopy

Ultramicroscopy. Sep, 2005  |  Pubmed ID: 15890445

A new high-resolution recording device for transmission electron microscopy (TEM) is urgently needed. Neither film nor CCD cameras are systems that allow for efficient 3-D high-resolution particle reconstruction. We tested an active pixel sensor (APS) array as a replacement device at 200, 300, and 400 keV using a JEOL JEM-2000 FX II and a JEM-4000 EX electron microscope. For this experiment, we used an APS prototype with an area of 64 x 64 pixels of 20 microm x 20 microm pixel pitch. Single-electron events were measured by using very low beam intensity. The histogram of the incident electron energy deposited in the sensor shows a Landau distribution at low energies, as well as unexpected events at higher absorbed energies. After careful study, we concluded that backscattering in the silicon substrate and re-entering the sensitive epitaxial layer a second time with much lower speed caused the unexpected events. Exhaustive simulation experiments confirmed the existence of these back-scattered electrons. For the APS to be usable, the back-scattered electron events must be eliminated, perhaps by thinning the substrate to less than 30 microm. By using experimental data taken with an APS chip with a standard silicon substrate (300 microm) and adjusting the results to take into account the effect of a thinned silicon substrate (30 microm), we found an estimate of the signal-to-noise ratio for a back-thinned detector in the energy range of 200-400 keV was about 10:1 and an estimate for the spatial resolution was about 10 microm.

Evaluation of the Bioequivalence and Pharmacokinetics of Two Formulations of Rizatriptan After Single Oral Administration in Healthy Volunteers

Arzneimittel-Forschung. 2005  |  Pubmed ID: 16080273

The pharmacokinetic parameters of two oral formulations of rizatriptan (CAS 144034-80-0, a capsule preparation as test and rizatriptan tablet as reference), given at a single dose of 10 mg each, were compared in an open-label, randomized, single oral dose, two-period cross-over design in 20 healthy volunteers under fasting conditions. Plasma concentrations of rizatriptan were measured by a validated HPLC assay. The parametric 90% confidence intervals of the geometric mean values of the test/reference ratios were 91.9% to 101.9% (point estimate: 97.3%) for AUC(0-infinity), 93.0% to 102.2% (point estimate: 96.5%) for AUC(0-t), 90.1% to 100.0% (point estimate: 95.4%) for Cmax, being within the acceptance criteria for bioequivalence (80%-125%). Tmax values were analyzed by the nonparametric Wilcoxon test and the difference was not statistically significant. Therefore, it is concluded that the test and reference rizatriptan formulations are bioequivalent with regard to both the extent and the rate of absorption.

Caspase Family Proteases and Apoptosis

Acta Biochimica Et Biophysica Sinica. Nov, 2005  |  Pubmed ID: 16270150

Apoptosis, or programmed cell death, is an essential physiological process that plays a critical role in development and tissue homeostasis. The progress of apoptosis is regulated in an orderly way by a series of signal cascades under certain circumstances. The caspase-cascade system plays vital roles in the induction, transduction and amplification of intracellular apoptotic signals. Caspases, closely associated with apoptosis, are aspartate-specific cysteine proteases and members of the interleukin-1beta-converting enzyme family. The activation and function of caspases, involved in the delicate caspase-cascade system, are regulated by various kinds of molecules, such as the inhibitor of apoptosis protein, Bcl-2 family proteins, calpain, and Ca2+. Based on the latest research, the members of the caspase family, caspase-cascade system and caspase-regulating molecules involved in apoptosis are reviewed.

[Clinical Application of Maxillary Sinus Lifting, Bone Graft and Simultaneous Placement of Implant]

Shanghai Kou Qiang Yi Xue = Shanghai Journal of Stomatology. Oct, 2005  |  Pubmed ID: 16288340

To evaluate the results of two kinds of sinus lifting techniques with simultaneous implant placement.

[Simulation of Greenhouse Tomato Dry Matter Partitioning and Yield Prediction]

Ying Yong Sheng Tai Xue Bao = The Journal of Applied Ecology / Zhongguo Sheng Tai Xue Xue Hui, Zhongguo Ke Xue Yuan Shenyang Ying Yong Sheng Tai Yan Jiu Suo Zhu Ban. May, 2006  |  Pubmed ID: 16883806

Based on the relationships between dry matter partitioning index, harvest index, and product of thermal effectiveness and PAR, a simulation model for greenhouse tomato dry matter partitioning and yield prediction was built, and validated by independent experimental data of different cultivars, substrates and locations. The coefficient of determination (R2) between simulated and measured shoot, root, stem, leaf and fruit dry matter weight based on 1:1 line was 0.95, 0.57, 0.82, 0.79 and 0.93, the root mean squared error (RMSE) between them was 647.0, 78.1, 279.0, 496.9 and 381.8 kg x hm(-2), and the R2 and RMSE between predicted and measured yield based on 1:1 line were 0.88 and 5 828.5 kg x hm(-2), respectively. Compared to 'source-sink' theory, the model developed in this study could give satisfactory prediction of the dry weight of leaf, stem, fruit and yield, with fewer parameters that could be easily obtained in practice.

Identification of Simple Sequence Repeat (SSR) Markers for Acid Detergent Fiber in Rice Straw by Bulked Segregant Analysis

Journal of Agricultural and Food Chemistry. Oct, 2006  |  Pubmed ID: 17002430

Rice straw is a significant energy source for ruminant animals. The acid detergent fiber (ADF) content of rice straw is negatively related to intake potential of forages. Therefore, improving the digestibility of rice straw by reducing ADF content is a necessary goal in breeding programs. In the present study, simple sequence repeat (SSR) markers and the bulked segregant analysis (BSA) approach were used to identify molecular markers associated with ADF. A total of 121 BC1F1 plants derived from the cross of JX974 (a cultivar with high ADF, 36.6%) and Dongxiang wild rice (a wild rice with low ADF, 31.3%), with JX974 as a recurrent parent, were used to conduct BSA. Phenotypic analysis showed that ADF displayed a normal distribution in BC1F1 population, indicating the involvement of polygenes. A SSR marker, RM566 on chromosome 9, was identified for ADF. A small linkage map consisting of five markers was constructed by adding four other markers, and a quantitative trait locus (QTL) controlling ADF was mapped at the RM321-RM566 interval, with a distance of 3.9 cM to RM566. This QTL explained 12% of the total phenotypic variation of ADF, and its additive effect was 3%. This study is the first step to map QTL for ADF, one of the plant cell wall components in rice.

Deferiprone Protects the Isolated Atria from Cardiotoxicity Induced by Doxorubicin

Acta Pharmacologica Sinica. Oct, 2006  |  Pubmed ID: 17007740

To investigate the effects of deferiprone on doxorubicin-induced cardiotoxicity and determine its protection on cardiac contractility in vivo at tissue level.

Long-lasting Specific Antibodies Against P277 Induced by Mucosal Administration of P277 Repeat Sequences Carried by Hsp65 in the Absence of Adjuvants

Vaccine. Mar, 2007  |  Pubmed ID: 17224213

To improve the weak immunogenicity of peptide P277, the recombinant expression plasmid pET28-Hsp65-6 x P277 was constructed by inserting 5 x P277 which was amplified by PCR into the vector pET28-Hsp65-P277. It was transformed into Escherichia coli BL21 (DE3) and the fusion protein (Hsp65-6 x P277) was expressed effectively as soluble protein after inducing by lactose. The fusion protein was purified by means of cell disruption, ammonium sulfate precipitation, double-distilled H2O dialysis, DEAE52-cellulose column chromatography, and then used to immunize female NOD mice with three i.n. inoculations in the absence of adjuvants. Serum samples from the immunized mice were collected at 3 weeks interval. Antibodies against P277 and HSP65 were detected in immunized mice sera by enzyme-linked immunosorbent assay (ELISA) and Western blot. Specific antibodies were successfully induced and lasted for more than 20 weeks in animals immunized with the fusion protein via intranasal route even in the absence of adjuvants.

Liver Cancer: Increased Microwave Delivery to Ablation Zone with Cooled-shaft Antenna--experimental and Clinical Studies

Radiology. Mar, 2007  |  Pubmed ID: 17229876

To prospectively investigate whether the ablation zone induced with microwaves could be increased by delivering greater energy with a cooled-shaft antenna.

Future Directions for Camera Systems in Electron Microscopy

Methods in Cell Biology. 2007  |  Pubmed ID: 17327181

Peroxisome Proliferator-activated Receptor-gamma Agonists Induce Neuroprotection Following Transient Focal Ischemia in Normotensive, Normoglycemic As Well As Hypertensive and Type-2 Diabetic Rodents

Journal of Neurochemistry. Apr, 2007  |  Pubmed ID: 17394460

Thiazolidinediones (TZDs) are synthetic agonists of the ligand-activated transcription factor peroxisome proliferator-activated receptor-gamma (PPARgamma). TZDs are known to curtail inflammation associated with peripheral organ ischemia. As inflammation precipitates the neuronal death after stroke, we tested the efficacy of TZDs in preventing brain damage following transient middle cerebral artery occlusion (MCAO) in adult rodents. As hypertension and diabetes complicate the stroke outcome, we also evaluated the efficacy of TZDs in hypertensive rats and type-2 diabetic mice subjected to transient MCAO. Pre-treatment as well as post-treatment with TZDs rosiglitazone and pioglitazone significantly decreased the infarct volume and neurological deficits in normotensive, normoglycemic, hypertensive and hyperglycemic rodents. Rosiglitazone neuroprotection was not enhanced by retinoic acid x receptor agonist 9-cis-retinoic acid, but was prevented by PPARgamma antagonist GW9662. Rosiglitazone significantly decreased the post-ischemic intercellular adhesion molecule-1 expression and extravasation of macrophages and neutrophils into brain. Rosiglitazone treatment curtailed the post-ischemic expression of the pro-inflammatory genes interleukin-1beta, interleukin-6, macrophage inflammatory protein-1alpha, monocyte chemoattractant protein-1, cyclooxygenase-2, inducible nitric oxide synthase, early growth response-1, CCAAT/enhancer binding protein-beta and nuclear factor-kappa B, and increased the expression of the anti-oxidant enzymes catalase and copper/zinc-superoxide dismutase. Rosiglitazone also increased the expression of the anti-inflammatory gene suppressor of cytokine signaling-3 and prevented the phosphorylation of the transcription factor signal transducer and activator of transcription-3 after focal ischemia. Thus, PPARgamma activation with TZDs might be a potent therapeutic option for preventing inflammation and neuronal damage after stroke with promise in diabetic and hypertensive subjects.

Clinical Evaluation of a Computer-aided Diagnosis (CAD) Prototype for the Detection of Pulmonary Embolism

Academic Radiology. Jun, 2007  |  Pubmed ID: 17502254

To evaluate the performance of a prototype computer-aided diagnosis (CAD) tool using artificial intelligence techniques for the detection of pulmonary embolism (PE) and the possible benefit for general radiologists.

[A Comparison of Therapeutic Outcomes in Liver Fibrosis of Murine Schistosomiasis Japonica with Interferon-alpha and Interferon-gamma]

Zhonghua Gan Zang Bing Za Zhi = Zhonghua Ganzangbing Zazhi = Chinese Journal of Hepatology. Jun, 2007  |  Pubmed ID: 17594819

Genome Size and Sequence Composition of Moso Bamboo: a Comparative Study

Science in China. Series C, Life Sciences / Chinese Academy of Sciences. Oct, 2007  |  Pubmed ID: 17879070

Moso bamboo (Phyllostachys pubescens) is one of the world's most important bamboo species. It has the largest area of all planted bamboo--over two-thirds of the total bamboo forest area--and the highest economic value in China. Moso bamboo is a tetraploid (4x=48) and a special member of the grasses family. Although several genomes have been sequenced or are being sequenced in the grasses family, we know little about the genome of the bambusoids (bamboos). In this study, the moso bamboo genome size was estimated to be about 2034 Mb by flow cytometry (FCM), using maize (cv. B73) and rice (cv. Nipponbare) as internal references. The rice genome has been sequenced and the maize genome is being sequenced. We found that the size of the moso bamboo genome was similar to that of maize but significantly larger than that of rice. To determine whether the bamboo genome had a high proportion of repeat elements, similar to that of the maize genome, approximately 1000 genome survey sequences (GSS) were generated. Sequence analysis showed that the proportion of repeat elements was 23.3% for the bamboo genome, which is significantly lower than that of the maize genome (65.7%). The bamboo repeat elements were mainly Gypsy/DIRS1 and Ty1/Copia LTR retrotransposons (14.7%), with a few DNA transposons. However, more genomic sequences are needed to confirm the above results due to several factors, such as the limitation of our GSS data. This study is the first to investigate sequence composition of the bamboo genome. Our results are valuable for future genome research of moso and other bamboos.

[Diagnostic Performance of 64-channel Multislice Computed Tomography in Assessment of Significant Coronary Artery Disease in Subjects Suspected As Acute Coronary Syndrome and Stable Angina.]

Beijing Da Xue Xue Bao. Yi Xue Ban = Journal of Peking University. Health Sciences. Dec, 2007  |  Pubmed ID: 18087560

To evaluate the diagnostic accuracy of 64-channel multislice spiral computed tomography (MSCT) in subjects with presentations suggestive of stable angina or acute coronary syndrome.

A Th1-recognized Peptide P277, when Tandemly Repeated, Enhances a Th2 Immune Response Toward Effective Vaccines Against Autoimmune Diabetes in Nonobese Diabetic Mice

Journal of Immunology (Baltimore, Md. : 1950). Jan, 2008  |  Pubmed ID: 18097004

Subunit immunogens containing tandemly repeated copies of T and B cell epitopes have been shown to be more immunogenic than the respective immunogen containing only a single copy of the sequence. To investigate whether the increased copies of the Th2-activated peptide sequence will enhance the Th2-like immune response, we compared the cytokine secreted in mice that inoculated with two immunogens containing one or six tandemly repeated copies of a Th2-activated peptide sequence P277. Immunization of mice with a 6xP277 fusion protein elicited much higher levels of Th2-type cytokines and lower Th1-type cytokines than with a fusion protein with one P277 peptide. The data of tandemly repeated peptide P277 potentiate the anti-inflammatory in NOD mice, most likely associated with a Th1 to Th2 cytokine shift specific for the autoimmune T cells, which suggested that application of multiple tandem repeats of a Th2-activated epitope is an efficient method to enhance the anti-inflammatory immune response by shifting the immune response from Th1-like to Th2-like. The subunit immunogens containing tandemly repeated copies of peptide P277 might be effective vaccines against autoimmune diabetes in NOD mice.

Starch Physicochemical Properties and Their Associations with Microsatellite Alleles of Starch-synthesizing Genes in a Rice RIL Population

Journal of Agricultural and Food Chemistry. Mar, 2008  |  Pubmed ID: 18254594

The physicochemical properties of starch, such as apparent amylose content, gelatinization temperature, and pasting viscosities, determine the eating, cooking, and processing qualities of various products of rice. A recombinant inbred line (RIL) population derived from the reciprocal cross of Lemont (a premium high-quality tropical japonica rice) and Jiayu 293 (a high-yield but low-quality indica rice) was used to test the association of microsatellite markers of starch-synthesizing genes with starch quality parameters. The results confirmed the association of Wx and starch synthase I (SSI) alleles with various starch properties measured in rice flour. However, the starch properties were not associated with the starch branching enzyme 1 (SBE 1) gene alleles.

Effects of Aliskiren on Blood Pressure, Albuminuria, and (pro)renin Receptor Expression in Diabetic TG(mRen-2)27 Rats

Hypertension. Jul, 2008  |  Pubmed ID: 18490518

The aim of this study was to explore the effects of the renin inhibitor aliskiren in streptozotocin-diabetic TG(mRen-2)27 rats. Furthermore, we investigated in vitro the effect of aliskiren on the interactions between renin and the (pro)renin receptor and between aliskiren and prorenin. Aliskiren distributed extensively to the kidneys of normotensive (non)diabetic rats, localizing in the glomeruli and vessel walls after 2 hours exposure. In diabetic TG(mRen-2)27 rats, aliskiren (10 or 30 mg/kg per day, 10 weeks) lowered blood pressure, prevented albuminuria, and suppressed renal transforming growth factor-beta and collagen I expression versus vehicle. Aliskiren reduced (pro)renin receptor expression in glomeruli, tubules, and cortical vessels compared to vehicle (in situ hybridization). In human mesangial cells, aliskiren (0.1 micromol/L to 10 micromol/L) did not inhibit binding of (125)I-renin to the (pro)renin receptor, nor did it alter the activation of extracellular signal-regulated kinase 1/2 by renin (20 nmol/L) preincubated with aliskiren (100 nmol/L) or affect gene expression of the (pro)renin receptor. Evidence was obtained that aliskiren binds to the active site of prorenin. The above results demonstrate the antihypertensive and renoprotective effects of aliskiren in experimental diabetic nephropathy. The evidence that aliskiren can reduce in vivo gene expression for the (pro)renin receptor and that it may block prorenin-induced angiotensin generation supports the need for additional work to reveal the mechanism of the observed renoprotection by this renin inhibitor.

[Improved Efficacy of P277 Fused to Heat Shock Protein 65 of Mycobacterium Tuberculosis Against Diabetes in Nonobese Diabetic Mice]

Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology. Apr, 2008  |  Pubmed ID: 18616176

To improve the efficacy of peptide P277 in preventing autoimmune diabetes, heat shock protein 65 kD (HSP65) of Mycobacterium tuberculosis var. bovis was fused with linear polypeptide epitope of P277 and expressed as soluble protein in Escherichia coli. The fusion protein HSP65-P277 was purified by anion exchange column chromatography and then used to immunize prediabetic NOD mice with three ip inoculations in absence of adjuvants. Serum samples from the immunized mice were collected monthly and the concentration of blood glucose was measured. The study showed that administration of HSP65-P277 to NOD mice could prevent the development of diabetes more efficiently than the peptide P277 itself or HSP65. Fused to heat shock protein 65 of Mycobacterium tuberculosis could improve the efficacy of diabetes prevention of P277 in nonobese diabetic mice. The results suggest the fusion protein of HSP65-P277 would be useful for treating insulin-dependent diabetes mellitus.

Applications of Direct Detection Device in Transmission Electron Microscopy

Journal of Structural Biology. Mar, 2008  |  Pubmed ID: 18054249

A prototype direct detection device (DDD) camera system has shown great promise in improving both the spatial resolution and the signal to noise ratio for electron microscopy at 120-400 keV beam energies (Xuong et al., 2007. Methods in Cell Biology, 79, 721-739). Without the need for a resolution-limiting scintillation screen as in the charge coupled device (CCD), the DDD camera can outperform CCD based systems in terms of spatial resolution, due to its small pixel size (5 microm). In this paper, the modulation transfer function (MTF) of the DDD prototype is measured and compared with the specifications of commercial scientific CCD camera systems. Combining the fast speed of the DDD with image mosaic techniques, fast wide-area imaging is now possible. In this paper, the first large area mosaic image and the first tomography dataset from the DDD camera are presented, along with an image processing algorithm to correct the specimen drift utilizing the fast readout of the DDD system.

[Analysis of BRCA1 Gene Mutations in Patients with Early-onset Breast Cancer and Their Affected Relatives in Guangdong Province]

Nan Fang Yi Ke Da Xue Xue Bao = Journal of Southern Medical University. Feb, 2009  |  Pubmed ID: 19246281

To study the BRCA1 mutations in patients with early-onset breast cancer and their affected relatives in Guangdong province and explore the relationship between BRCA1 mutation and the expressions of estrogen receptor(ER), progesterone receptor(PR), HER2 and ALN.

Ultrahigh Spacing Tissue Arrays for Screening Hybridoma by Immunohistochemistry

Journal of Immunological Methods. Apr, 2009  |  Pubmed ID: 19374005

We present a technology to make ultrahigh spacing tissue arrays (USTA) which facilitates screening hybridoma by immunohistochemistry, results in easier performance of staining and image analysis, minimal sample mixed-up and less bookkeeping efforts. 32 array cores are designated a 4 x 8 microarray, leaving 4 mm spacing between two adjacent array elements. After separated each array spot with PAP pen, one USTA is sufficient for testing of supernatants from 32 different hybridomas.

A Th2 Immune Shift to Heat Shock Protein 65 Fails to Arrest Atherosclerosis: Proatherogenic Role of Th2-deviated Autoantibodies

Autoimmunity. Sep, 2009  |  Pubmed ID: 19418313

Many reports regarding the cytotoxicity of antibodies to heat shock protein (HSP) 65/60 have implied the potential disadvantage and risk of HSP65/60-specific Th2 shifting strategy in arresting atherosclerosis. In this study, experiments were specifically designed to investigate the effect of a HSP65-specifc Th1 to Th2 immune shift accompanied with high-titer antibodies on atherosclerosis and explore the proatherogenic cytotoxicity of Th2-deviated anti-HSP65 antibodies to endothelial cells. Rabbits were nasally immunized with a fusion protein HSP65-6 x P277 10 times every other day. Immunologic results, including the repressed T-cell proliferation, increased interleukin-10 production and IgG1-predominated isotype of antibodies, revealed a significant Th1 to Th2 shift of response to HSP65. However, rabbits showed no reduction in atherosclerotic lesions. As a control, HSP65 immunization, which induced no antibodies, obviously attenuated atherosclerosis. Further studies on endothelial cells showed that the Th2-deviated anti-HSP65 antibodies could cross-react with HSP60 highly expressed in stressed cells and mediate damage to cells in the presence of complement. In conclusion, the Th2-deviated antibodies to HSP65 that were induced by over-regulated Th2 shift are cytotoxic to endothelial cells. This proatherogenic effect, in contradiction to the positive impact of Th1 suppression, can eventually invalidate the efficacy of Th2 shift in arresting atherosclerosis.

STAMENLESS 1, Encoding a Single C2H2 Zinc Finger Protein, Regulates Floral Organ Identity in Rice

The Plant Journal : for Cell and Molecular Biology. Sep, 2009  |  Pubmed ID: 19453444

Floral organ identity is defined by organ homoetic genes whose coordinated expression is crucial with respect to the time and place of floral organ formation. Here, we report molecular cloning and characterization of the rice STAMENLESS 1 (SL1) gene that is involved in floral development. The sl1 mutant largely resembles the rice B-class gene mutant spw1; both exhibit homeotic conversions of lodicules and stamens to palea/lemma-like organs and carpels. Additionally, sl1 produces flowers with varied numbers of inner floral organs, and amorphous tissues without floral organ identity were frequently formed in whorls 3 and 4. We also show that SL1 specifies lodicule and stamen identities through positive transcriptional regulation of SPW1/OsMADS16 expression. SL1 encodes a member of the C2H2 family of zinc finger proteins, closely related to JAG of Arabidopsis. The functional divergence between SL1 and JAG implies that SL1 was co-opted for its distinctive roles in specification of floral organ identity in rice after the lineage split from Arabidopsis.

Specific Antibodies Elicited by a Novel DNA Vaccine Targeting Gastrin-releasing Peptide Inhibit Murine Melanoma Growth in Vivo

Clinical and Vaccine Immunology : CVI. Jul, 2009  |  Pubmed ID: 19458203

The elevated expression and receptor binding of gastrin-releasing peptide (GRP) in various types of cancer, especially in malignant melanoma of the skin, suggest that GRP might be a putative target for immunotherapy in neoplastic diseases. We have therefore constructed a novel DNA vaccine coding for six tandem repeats of a fragment of GRP from amino acids 18 to 27 (GRP6) flanked by helper T-cell epitopes for increased immunogenicity, including HSP65, a tetanus toxoid fragment from amino acids 830 to 844 (T), pan-HLA-DR-binding epitope (PADRE) (P), and two repeats of a mycobacterial HSP70 fragment from amino acids 407 to 426 (M). The anti-GRP DNA vaccine (pCR3.1-VS-HSP65-TP-GRP6-M2) was constructed on a backbone of a pCR3.1 plasmid vector with eight 5'-GACGTT-3' CpG motifs and the VEGF183 signal peptide (VS). Intramuscular (IM) injections of anti-GRP vaccine in mice stimulated the production of high titers of specific antibodies against GRP and suppressed the growth of subcutaneous tumors of B16-F10 melanoma cells. Parallel results were obtained in vitro, showing inhibition of B16-F10 cell proliferation by GRP antisera. IM injections of the DNA vaccine also significantly attenuated tumor-induced angiogenesis associated with intradermal tumors of B16-F10 cells. In addition, lung invasion of intravenously injected cells was highly diminished, suggesting potent antimetastatic activity of the DNA vaccine. These findings support the highly immunogenic and potent antitumorigenic activity of specific anti-GRP antibodies elicited by the anti-GRP DNA vaccine.

Nasal Immunization with Heat Shock Protein 65 Attenuates Atherosclerosis and Reduces Serum Lipids in Cholesterol-fed Wild-type Rabbits Probably Through Different Mechanisms

Immunology Letters. Jun, 2009  |  Pubmed ID: 19505506

In recent years atherosclerosis has proved to be associated with microbial infection, inflammation and autoimmunity. Conservative heat shock protein (HSP) 65/60 is a major autoantigen of atherosclerosis. In the current study, experiments were specifically designed to investigate whether a nasal immunization with HSP65 could attenuate atherosclerosis in a cholesterol-fed wild-type animal model and explore its influence on serum lipids. Wild-type rabbits were nasally treated with HSP65 10 times on alternate days. At the end of the experiment, the rabbits showed remarkably lightened lesions in aortas. The suppression of T cell proliferation, increase of IL-10 production and absence of related antibodies implied that a tolerance to HSP65 was successfully established. Simultaneously, the serum lipid levels were down-regulated significantly in this group. Further results of another group immunized with conjugated protein HSP65+CTB-P277 showed that the lipid reduction could also be achieved by an immunization without inducing tolerance. But this simple reduction of lipids could not eventually alleviate atherosclerosis. In conclusion, nasal administration of HSP65 can effectively attenuate atherosclerosis in cholesterol-fed wild-type rabbits primarily by inducing an unresponsive state of tolerance. The accompanying reduction of lipids, which probably results from a different immune mechanism other than tolerance, cannot ultimately prevent the development of atherosclerotic lesions alone.

Improved Efficacy of DNA Vaccination Against Prostate Carcinoma by Boosting with Recombinant Protein Vaccine and by Introduction of a Novel Adjuvant Epitope

Vaccine. Aug, 2009  |  Pubmed ID: 19616501

DNA vaccine represents an attractive approach for cancer treatment by inducing active immune-deprivation of gastrin-releasing peptide (GRP) from tumor cells, the growth of which is dependent on the stimulation of GRP. In this study, we developed a DNA vaccine using a plasmid vector to deliver the immunogen of six copies of the B cell epitope GRP(18-27) (GRP6). In order to increase the potency of this DNA vaccine, multiple strategies have been applied including DNA-prime protein-boost immunization and introduction of a foreign T-helper epitope into DNA vaccine. Mice vaccinated DNA vaccine boosting with HSP65-GRP6 protein induced high titer and relatively high avidity of anti-GRP antibodies as well as inhibition effect on the growth of murine prostate carcinoma, superior to the treatment using DNA alone or BCG priming HSP65-GRP6 protein boosting. Furthermore, the introduction of a novel foreign T-helper epitope into the GRP DNA vaccine showed a markedly stronger humoral immune response against GRP and tumor rejection even than the DNA-prime protein-boost strategy. No further stronger immunogenicity of this foreign T-helper epitope modified DNA vaccine was observed even using the strategy of modified DNA vaccine-priming and HSP65-GRP6 boosting method. The data presented demonstrate that improvement of potency of anti-GRP DNA vaccine with the above two feasible approaches should offer useful methods in the development of new DNA vaccine against growth factors for cancer immunotherapy.

Strong Humoral Response Elicited by a DNA Vaccine Targeting Gastrin-releasing Peptide with Optimized Adjuvants Inhibits Murine Prostate Carcinoma Growth in Vivo

Endocrine-related Cancer. Dec, 2009  |  Pubmed ID: 19648182

Previous studies demonstrated that the elevated expression and receptor binding of gastrin-releasing peptide (GRP) in various types of cancer suggest that GRP might be a putative target for immunotherapy in neoplastic diseases. DNA vaccine for hormone/growth factor immune deprivation represents a feasible and attractive approach for cancer treatment; nevertheless, there is still a need to increase the potency of the DNA vaccine. Here, based on six copies of the B cell epitope GRP(18-27) in a linear alignment as an immunogen, we designed several anti-GRP DNA vaccines containing different combinations of immunoadjuvants, such as HSP65, tetanus toxoid(830-844) (T), pan HLA-DR-binding epitope (PADRE) (P), and mycobacterial HSP70(407-426) (M), on a backbone of pCR3.1 plasmid vector with eight 5'-GACGTT-3' CpG motifs and the VEGF183 signal peptide (VS). The effects of these immunoadjuvants in enhancing GRP-specific humoral immune response were then evaluated by comparing the respective immunogenicity and antitumor effects. Immunization of mice with pCR3.1-VS-HSP65-TP-GRP6-M2 elicited much higher levels of specific anti-GRP antibodies and more effectively inhibited the growth of a GRP-dependent tumor RM-1 in vivo. Interestingly, plasmids encoding for 2HSP70(407-426), but not the one with 1 or 3HSP70(407-426) showed stronger immune stimulatory potential as well as impressive antitumor activity, suggesting that 2HSP70(407-426) is an efficient molecular adjuvant for developing self-epitope vaccines. The highly immunogenic, potent anti-tumorigenic and antiangiogenesis activities of the anti-GRP DNA vaccine offered a novel immunotherapeutic approach in the treatment of GRP-dependent tumors and their complications.

Murine Monoclonal Antibodies Generated Against Mouse/rat Hemokinin-1

Hybridoma (2005). Aug, 2009  |  Pubmed ID: 19663698

The mouse/rat hemokinin-1 (m/rHK-1) was discovered nearly 9 years ago. This molecule is a peptide comprising 11 amino acids. The m/rHK-1 was found to be mainly expressed in central immune organs like bone marrow, and was proven to have lymphopoietic roles in B and T lymphocyte development. m/rHK-1was also reported to have analgesic roles in rat spinal cord, in addition to other functions such as relaxing activity on coronary artery. Unlike its analogues SP, NKA, and NKB, m/rHK-1 does not express in the nervous system. To further study the distribution and function of m/rHK-1, we carried out conventional immunization and cell fusion procedures to acquire the hybridomas secreting specific monoclonal antibodies to m/rHK-1. In the 17 positive clones obtained, three antibodies named 1B12, 2B4, and 4G5 were shown representative in cross-reactivity against m/rHK-1 and its analogues by indirect ELISA, competitive indirect ELISA, and immunofluorescence assays. Among the three clones, the 2B4 monoclonal antibody appeared to be the high-titered and specific clone to m/rHK-1. Monoclonal antibodies to m/rHK-1 will function as good tools in the physiological study of m/rHK-1 in the near future.

Brain Penetration of Colistin in Mice Assessed by a Novel High-performance Liquid Chromatographic Technique

Antimicrobial Agents and Chemotherapy. Oct, 2009  |  Pubmed ID: 19667287

A sensitive and reliable liquid chromatographic method was developed and validated for the determination of colistin concentrations in mouse brain homogenate. With a mobile phase consisting of acetonitrile-tetrahydrofuran-water (50:25:25 [vol/vol]) at a flow rate of 1 ml/min, a linear correlation between peak area and colistin concentration was observed over the concentration range of 93.8 to 3,000 ng/g in brain tissue (R2 > 0.994). Intra- and interday coefficients of variation were 5.1 to 8.3% and 5.8 to 8.5%, respectively, and the recovery ranged from 85% to 94%. This assay was then utilized to determine the amount of colistin that permeated the blood-brain barrier over a 2-h period following bolus intravenous administration of colistin sulfate to mice. After a single dose of 5 mg/kg of body weight to mice, brain homogenate concentrations of colistin were very low, relative to plasma colistin concentrations, suggesting that colistin permeability across the healthy blood-brain barrier is negligible during this experimental period.

Correlations Between Environmental Factors and Wild Bee Behavior on Alfalfa (Medicago Sativa) in Northwestern China

Environmental Entomology. Oct, 2009  |  Pubmed ID: 19825303

To discover the effect of environmental factors on pollinator visitation to flowering Medicago sativa, several field experiments were designed to examine the diurnal movement patterns of wild bee species in the Hexi Corridor of northwestern China. Our study results showed that Megachile abluta, M. spissula, and Xylocopa valga showed unimodal diurnal foraging behavior, whereas Andrena parvula and Anthophora melanognatha showed bimodal diurnal foraging behavior. Correlation analysis indicated that diurnal foraging activities of pollinators were significantly correlated with environmental factors. Correlations of foraging activities versus environmental factors for M. abluta, M. spissula, and X. valga best fit a linear model, whereas those of A. parvula and A. melanognatha best fit a parallel quadratic model. Results of this study indicated that solitary wild bees such as M. abluta, M. spissula, X. valga, A. parvula, and A. melanognatha are potential alfalfa pollinators in the Hexi Corridor. An understanding of the environmental factors that affect the behaviors of different wild bees foraging in alfalfa are basic to the utilization of solitary wild bees in a practical way for increased, or more consistent, pollination of alfalfa for seed production.

Immunotherapy of Autoimmune Diabetes by Nasal Administration of Tandem Glutamic Acid Decarboxylase 65 Peptides

Immunological Investigations. 2009  |  Pubmed ID: 19860582

Mucosally induced tolerance is an attractive strategy for immunotherapy of autoimmune diseases. Treatment of nonobese diabetic (NOD) mice with a mixture of autoantigen peptides is better geared toward slowing the progression of late stage type 1 diabetes (T1D) than treatment with any of the peptides alone. In this study, we constructed a fusion protein CTB-GADIII. It contains cholera toxin B subunit (CTB) and three tandem peptides derived from glutamic acid decarboxylase 65 (GAD65), p217-236, p524-538 and p290-306. The purified renatured pentamer fusion protein was effective in inhibiting the development of diabetes in NOD mice when the mice were nasally immunized three times (8w, 10w and 12w). Prevention of diabetes was associated with special humoral immune tolerance against tandem peptides GADIII. These data indicate that using CTB as a vaccine carrier, tandem GAD65 peptides can prevent T1D in NOD mice at the late stage of disease.

The Fusion Protein of HSP65 with Tandem Repeats of Beta-hCG Acting As a Potent Tumor Vaccine in Suppressing Hepatocarcinoma

International Immunopharmacology. Feb, 2010  |  Pubmed ID: 19913113

It has been demonstrated that the beta-subunit of human chorionic gonadotropin (beta-hCG) is ectopically expressed on a variety of human cancers of different histological types and has been used as an antigenic target in anti-cancer vaccines. We engineered a fusion protein by fusing 10 tandemly repeated copies of the 10-residue sequence of beta-hCG (109-118) (in CTP37) combined with beta-hCG C-terminal 37 peptides to mycobacterial heat-shock protein 65 and immunized mice via subcutaneous injection. Humoral immune and cellular immune responses were effectively elicited. High titer of anti-beta-hCG antibody was detected in immunized mice sera by ELISA and verified by Western blot analyses. The fusion protein of HSP65-X10-beta-hCGCTP37 effectively inhibited the growth of tumor both protective and therapeutic anti-tumor immunity in hepatocellular carcinoma tumor models in mice. Meanwhile, it also attenuated tumor-induced angiogenesis in intradermal tumor model in mice. Taken together, these results demonstrate that immune responses are effectively induced by a novel fusion protein vaccine targeting beta-hCG, suppressing the growth of hepatocellular carcinoma in mice. The beta-hCG-targeted vaccine holds promise for the treatment of a number of cancers and merits further study.

HSP65 Serves As an Immunogenic Carrier for a Diabetogenic Peptide P277 Inducing Anti-inflammatory Immune Response in NOD Mice by Nasal Administration

Vaccine. Apr, 2010  |  Pubmed ID: 20226247

Mucosal administration of autoantigen HSP65 can induce anti-inflammatory immune response and decrease organ-specific inflammation and disease in several models of autoimmunity, such as arthritis and atherosclerosis. We have been interested in whether the HSP65 serves as an immunogenic carrier for a diabetogenic peptide P277 can also induce anti-inflammatory immune response in NOD mice by mucosal administration. Thus, the dual functions of anti-type 1 diabetes of HSP65 and P277 will be obtained. To test this hypothesis, we examined the effect of intranasal vaccination with P277 tandem repeat sequences carried by HSP65 in the absence of adjuvants on autoimmune diabetes in NOD mice. We found a significant decrease in the incidence of diabetes, inhibition of insulitis, reduction in IgG2a isotype antibodies to P277 and proinflammatory cytokines IFN-gamma and IL-2 secretion, increased IgG1, IgG2b subclass antibodies to P277 and anti-inflammatory cytokines IL-10 and IL-4 secretion, and reduced proliferation in nasal administration of the fusion protein HSP65-6xP277. Our results demonstrate that HSP65 may serve as a particularly advantageous carrier for P277-based vaccines and mucosal administration may be a therapeutic approach for treatment of type 1 diabetes.

Genetic Diversity and Population Structure of a Diverse Set of Rice Germplasm for Association Mapping

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik. Aug, 2010  |  Pubmed ID: 20364375

Germplasm diversity is the mainstay for crop improvement and genetic dissection of complex traits. Understanding genetic diversity, population structure, and the level and distribution of linkage disequilibrium (LD) in target populations is of great importance and a prerequisite for association mapping. In this study, 100 genome-wide simple sequence repeat (SSR) markers were used to assess genetic diversity, population structure, and LD of 416 rice accessions including landraces, cultivars and breeding lines collected mostly in China. A model-based population structure analysis divided the rice materials into seven subpopulations. 63% of the SSR pairs in these accessions were in LD, which was mostly due to an overall population structure, since the number of locus pairs in LD was reduced sharply within each subpopulation, with the SSR pairs in LD ranging from 5.9 to 22.9%. Among those SSR pairs showing significant LD, the intrachromosomal LD had an average of 25-50 cM in different subpopulations. Analysis of the phenotypic diversity of 25 traits showed that the population structure accounted for an average of 22.4% of phenotypic variation. An example association mapping for starch quality traits using both the candidate gene mapping and genome-wide mapping strategies based on the estimated population structure was conducted. Candidate gene mapping confirmed that the Wx and starch synthase IIa (SSIIa) genes could be identified as strongly associated with apparent amylose content (AAC) and pasting temperature (PT), respectively. More importantly, we revealed that the Wx gene was also strongly associated with PT. In addition to the major genes, we found five and seven SSRs were associated with AAC and PT, respectively, some of which have not been detected in previous linkage mapping studies. The results suggested that the population may be useful for the genome-wide marker-trait association mapping. This new association population has the potential to identify quantitative trait loci (QTL) with small effects, which will aid in dissecting complex traits and in exploiting the rich diversity present in rice germplasm.

Characterization of a Direct Detection Device Imaging Camera for Transmission Electron Microscopy

Ultramicroscopy. Jun, 2010  |  Pubmed ID: 20382479

The complete characterization of a novel direct detection device (DDD) camera for transmission electron microscopy is reported, for the first time at primary electron energies of 120 and 200 keV. Unlike a standard charge coupled device (CCD) camera, this device does not require a scintillator. The DDD transfers signal up to 65 lines/mm providing the basis for a high-performance platform for a new generation of wide field-of-view high-resolution cameras. An image of a thin section of virus particles is presented to illustrate the substantially improved performance of this sensor over current indirectly coupled CCD cameras.

Preparation, Characterization and in Vivo Pharmacodynamic Evaluation of Thymopentin Loaded Poly(lactide Acid)/poly(lactide-co-glycolide Acid) Implants

International Journal of Pharmaceutics. Oct, 2010  |  Pubmed ID: 20674730

To avoid the clinical inconvenience of repeated injection of the immune modulator thymopentin (TP5), biodegradable implants comprising a mixed polymer matrix of poly(lactide acid) (PLA) and poly(lactide-co-glycolide acid) (PLGA) were produced using a simple extrusion method. Drug release from these TP5-loaded implants was characterized both in vitro and in vivo. Pharmacodynamic studies were carried out in immunosuppressed rats using the ratio of CD4(+)/CD8(+) cells, determined by flow cytometry, as an index of immunity. The results indicated that the entrapment efficiency of the implants was greater than 98%, but the release rate of TP5 depended on the drug loading. Implants containing less than 10% TP5 showed consistent release over 30 days, with low burst-release both in vitro and in vivo. Improved immunity and survival rates were observed in rats treated by TP5 injection and in rats given middle-to-high dose implants. When the release of TP5 exceeded 0.1 mg/kg body weight/day the CD4(+)/CD8(+) ratios increased in the 3 weeks after implantation, reaching a maximum (91.6% of the normal level) by the end of the third week. The TP5-loaded implants presented here provide a promising alternative to injections and the results support the further development of controlled-release TP5 formulations.

Molecular Dissection of the Migrating Posterior Lateral Line Primordium During Early Development in Zebrafish

BMC Developmental Biology. 2010  |  Pubmed ID: 21144052

Development of the posterior lateral line (PLL) system in zebrafish involves cell migration, proliferation and differentiation of mechanosensory cells. The PLL forms when cranial placodal cells delaminate and become a coherent, migratory primordium that traverses the length of the fish to form this sensory system. As it migrates, the primordium deposits groups of cells called neuromasts, the specialized organs that contain the mechanosensory hair cells. Therefore the primordium provides both a model for studying collective directional cell migration and the differentiation of sensory cells from multipotent progenitor cells.

[Clinical Analysis on Causes of Dental Implant Failure: Report of 32 Cases]

Zhonghua Kou Qiang Yi Xue Za Zhi = Zhonghua Kouqiang Yixue Zazhi = Chinese Journal of Stomatology. Dec, 2010  |  Pubmed ID: 21211235

To analyze the causes associated with the failure of dental implant restoration.

[Clinical Retrospective Study on Membrane Guided Bone Tissue Regeneration Technique in Dental Implants in the Anterior Maxilla]

Shanghai Kou Qiang Yi Xue = Shanghai Journal of Stomatology. Dec, 2010  |  Pubmed ID: 21431255

The purpose of this study was to investigate the effect of Bio-Gide in MGBR (membrane guided bone regeneration ) in the anterior maxilla dental implant therapy.

RNA-binding Protein Quaking, a Critical Regulator of Colon Epithelial Differentiation and a Suppressor of Colon Cancer

Gastroenterology. Jan, 2010  |  Pubmed ID: 19686745

Colon cancer is one of the best understood neoplasms from a genetic perspective, yet it remains the second most common cause of cancer-related death. Post-transcriptional regulation mediated by RNA-binding proteins or microRNAs coordinately targets multiple genes, holding promise involved in colon cancer initiation and development. Here we studied the role of RNA-binding protein quaking (QKI) in colon cancer.

Complex Mutation and Weak Selection Together Determined the Codon Usage Bias in Bryophyte Mitochondrial Genomes

Journal of Integrative Plant Biology. Dec, 2010  |  Pubmed ID: 21106008

Mutation and selection are two major forces causing codon usage biases. How these two forces influence the codon usages in green plant mitochondrial genomes has not been well investigated. In the present study, we surveyed five bryophyte mitochondrial genomes to reveal their codon usage patterns as well as the determining forces. Three interesting findings were made. First, comparing to Chara vulgaris, an algal species sister to all extant land plants, bryophytes have more G, C-ending codon usages in their mitochondrial genes. This is consistent with the generally higher genomic GC content in bryophyte mitochondria, suggesting an increased mutational pressure toward GC. Second, as indicated by Wright's Nc-GC3s plot, mutation, not selection, is the major force affecting codon usages of bryophyte mitochondrial genes. However, the real mutational dynamics seem very complex. Context-dependent analysis indicated that nucleotide at the 2nd codon position would slightly affect synonymous codon choices. Finally, in bryophyte mitochondria, tRNA genes would apply a weak selection force to fine-tune the synonymous codon frequencies, as revealed by data of Ser4-Pro-Thr-Val families. In summary, complex mutation and weak selection together determined the codon usages in bryophyte mitochondrial genomes.

Impact of P-glycoprotein Inhibition and Lipopolysaccharide Administration on Blood-brain Barrier Transport of Colistin in Mice

Antimicrobial Agents and Chemotherapy. Feb, 2011  |  Pubmed ID: 21115788

The aim of this study was to investigate the factors limiting the blood-brain barrier (BBB) transport of colistin in healthy mice and to assess the impact of systemic inflammation on the transport of this antibiotic across the BBB. Colistin sulfate (40 mg/kg) was administered subcutaneously to Swiss outbred mice as single and multiple doses to determine any relationship between brain uptake and plasma concentrations of colistin. To assess the effect of P-glycoprotein (P-gp) on BBB transport, colistin sulfate (5 mg/kg) was concomitantly administered intravenously with PSC833 or GF120918 (10 mg/kg). Systemic inflammation was induced by three intraperitoneal injections of lipopolysaccharide (LPS; 3 mg/kg), and BBB transport of colistin was subsequently measured following subcutaneous administration and by an in situ brain perfusion. The brain uptake of colistin was low following single and multiple subcutaneous doses, with brain-to-plasma concentration ratios ranging between 0.021 and 0.037, and this was not significantly enhanced by coadministration of GF120918 or PSC833 (P > 0.05). LPS significantly increased the brain uptake of subcutaneously administered colistin with area under the brain concentration time curve (AUC(brain)) values of 11.7 ± 2.7 μg·h/g and 4.0 ± 0.3 μg·h/g for LPS- and saline-treated mice, respectively (mean ± standard deviation). Similarly, in situ perfusion of colistin led to higher antibiotic brain concentrations in LPS-treated animals than in saline-treated animals, with colistin brain-to-perfusate concentration ratios of 0.019 ± 0.001 and 0.014 ± 0.001, respectively. This study demonstrates that the BBB transport of colistin is negligible in healthy mice; however, brain concentrations of colistin can be significantly enhanced during systemic inflammation, as might be observed in infected patients.

Exacerbated Brain Damage, Edema and Inflammation in Type-2 Diabetic Mice Subjected to Focal Ischemia

Journal of Neurochemistry. Feb, 2011  |  Pubmed ID: 21133923

One of the limiting factors in stroke therapeutic development is the use of animal models that do not well represent the underlying medical conditions of patients. In humans, diabetes increases the risk of stroke incidence as well as post-stroke mortality. To understand the mechanisms that render diabetics to increased brain damage, we evaluated the effect of transient middle cerebral artery occlusion in adult db/db mice. The db/db mouse is a model of type-2 diabetes with four times higher blood sugar than its normoglycemic genetic control(db/+ mouse). Following transient middle cerebral artery occlusion, the db/db mice showed significantly higher mortality, bigger infarcts, increased cerebral edema, worsened neurological status compared to db/+ mice. The db/db mice also showed significantly higher post-ischemic inflammatory markers (ICAM1(+) capillaries, extravasated macrophages/neutrophils and exacerbated proinflammatory gene expression) compared to db/+ mice. In addition, the post-ischemic neuroprotective heat-shock chaperone gene expression was curtailed in the db/db compared to db/+ mice.

Cavity Margin Status is an Independent Risk Factor for Local-regional Recurrence in Breast Cancer Patients Treated with Neoadjuvant Chemotherapy Before Breast-conserving Surgery

The American Surgeon. Dec, 2011  |  Pubmed ID: 22273234

The objection of this study is to investigate whether the cavity margin (CM) status has different predictive efficacy for local-regional recurrence (LRR) in patients who have received or have not received neoadjuvant chemotherapy (NAC) before breast-conserving surgery. We identified 61 patients who received NAC before breast-conserving surgery. A nonrandomized unmatched cohort of 295 patients without history of receiving NAC were also included in this study. Clinicopathological features and follow-up data were abstracted and analyzed. Patients in the NAC-treated group had more advanced diseases when compared with patients in the nonNAC-treated group. With a median follow-up of 42 months, the LRR-free survival rate of patients with positive CMs was significantly lower than that of patients with negative CMs in the NAC-treated group. This distinction was not observed in the nonNAC-treated group. Univariate and multivariate analysis revealed that positive CM was the only independent predictive factor for LRR in the NAC-treated group but not in nonNAC-treated patients. CM status had different predictive efficacy for LRR in different settings. Association between CM status and LRR was observed in NAC-treated patients rather than nonNAC-treated patients. More extensive surgical treatment might be needed in NAC-treated patients when their CMs are positive.

A Single-center, Prospective and Randomized Controlled Study: Can the Prophylactic Use of Lamivudine Prevent Hepatitis B Virus Reactivation in Hepatitis B S-antigen Seropositive Breast Cancer Patients During Chemotherapy?

Breast Cancer Research and Treatment. Jun, 2011  |  Pubmed ID: 21445574

Over the past four decades, chemotherapy has played an important role in prolonging survival in breast cancer patients. However, it may also result in undesirable side effects such as hepatitis B virus (HBV) reactivation seen in this study. With the increasing use of chemotherapy paralleling the rise in breast cancer incidence, the occurrence of HBV reactivation is likely to further increase. Several strategies use lamivudine to deal with this problem. Initially, lamivudine had been used to treat patients who developed alanine transaminase elevation attributable to HBV reactivation during chemotherapy. However, using this strategy, fatal reactivation has also been reported. Later studies have suggested that prophylactic lamivudine significantly reduces HBV reactivation and its associated morbidity. However, these studies were based mainly on patients with lymphoma, whereas studies on breast cancer patients were few. Moreover, these studies were retrospective. Recently, a prospective study has recommended that deferred preemptive lamivudine could be a comparable alternative to the prophylactic strategy. However, it was not a randomized controlled study. In this study, it was examined the efficacy of the prophylactic strategy in hepatitis B s-antigen seropositive breast cancer patients during chemotherapy using a prospective, randomized controlled study. Two groups were studied. One group consisted of 21 patients who were treated with prophylactic lamivudine, the other group consisted of 21 patients who were not treated with prophylactic lamivudine. The results showed that the prophylactic lamivudine strategy significantly decreased the incidence of HBV reactivation (0 vs. 28.6%, P = 0.021). It was conclude that the prophylactic lamivudine strategy significantly reduces the incidence of HBV reactivation for hepatitis B s-antigen seropositive breast cancer undergoing chemotherapy.

Oral Administration of Lactococcus Lactis Delivered Heat Shock Protein 65 Attenuates Atherosclerosis in Low-density Lipoprotein Receptor-deficient Mice

Vaccine. May, 2011  |  Pubmed ID: 21497632

Lactococcus is a genus of Gram positive food-grade bacteria that has been widely used as a vaccine platform for the safe delivery of heterologous antigens. Many reports support the involvement of inflammation and immunity in atherosclerosis as well as the role of autoimmunity to heat shock proteins (HSPs) in the progression of atherogenesis. In this study, experiments were specifically designed to investigate the effect of oral administration of mycobacterial heat shock protein 65 (HSP65) delivered by Lactococcus lactis (L. lactis) on atherogenesis. Two types of HSP65-encoding plasmids for intracellular expression or secretion were constructed, and then transformed into L. lactis NZ9000. Oral administration of two recombinant L. lactis strains both induced suppression of HSP65-specific proliferation, accompanied by elevation of Interleukin-10 (IL-10) production and reduction of interferon-gamma (IFN-γ) level. Inducible HSP65-specific tolerance exerted a protective effect on atherosclerotic lesion formation and endothelial damage in low-density lipoprotein receptor-deficient (LDL-RD) mice model, while no obvious pathological abnormalities were observed. In conclusion, delivery of HSP65 at the intestinal mucosa by recombinant L. lactis provides a novel approach for the prevention of atherosclerosis. The results further illustrate the potential of using genetically modified L. lactis as a safe and effective vaccine delivery to elicit antigen-specific tolerance for treatment of autoimmune diseases.

The Physiological Mechanism of a Drooping Leaf2 Mutation in Rice

Plant Science : an International Journal of Experimental Plant Biology. Jun, 2011  |  Pubmed ID: 21497711

Here we characterized a classic rice (Oryza sativa) drooping leaf2 mutant (named dl2). The dl2 allele affects both the midrib development and the total leaf venation pattern. Leaf anatomy results revealed the central vein lacks both clear cells and the adaxial small vascular bundle in dl2 mutant, which seemed to cause the drooping leaf phenotype. The dl2 leaves contain more small veins, and the size of the vascular cylinder in dl2 leaf is also altered. Furthermore, similar anatomy alteration was found in the dl2 roots. A reduction in the number of xylem and phloem poles in the central vascular cylinder in dl2 roots was observed and the diameter of cortical cell is also reduced. In addition, the alterations of the vegetative development such as the longer leaf blade and fewer adventitious and lateral roots were also observed in dl2. The physiological mechanism underlying the morphological and vascular alterations of dl2 was further studied. The result demonstrated that the dl2 vascular patterning distortions are strictly associated with a defective PAT (polar auxin transport) activity and sensitivity to different classes of polar auxin transport inhibitors. Finally, the drooping leaf phenotype of dl2 is coupled to a defective response to auxin.

Vaccination of Non-obese Diabetic Mice with a Fragment of Peptide P277 Attenuates Insulin-dependent Diabetes Mellitus

International Immunopharmacology. Sep, 2011  |  Pubmed ID: 21530685

P277 is a peptide derived from the HSP60 regions, have potent immunological effect on insulin-dependent diabetes mellitus (IDDM) and its phase III clinical trials are currently under investigation. However, we recently discovered a positive correlation between anti-P277 autoantibodies and the presence of endothelial cells damage in inducing vascular leak syndrome. Therefore, the aim of our study was to demonstrate the critical peptide epitope of P277 to IDDM and to highlight the effects of this peptide therapy on inflammation of the islets. Groups of 4-week old female non-obese diabetic (NOD) mice were immunized one time every three weeks for three times with a residue of P277, showing a significant effect of down-regulating immunity to P277 protein and preventing the development of IDDM. Immunologic results including the suppression of T-cell proliferation, the increase of interleukin-10 (IL-10) production and reduction of interferon-γ (IFN-γ) production caused immune tolerance to P277. Hence, a functional role of the key epitope in P277 peptide capable of preventing IDDM is suggested, which could be modified to develop a novel safe and effective peptide vaccine against IDDM by reconstructing P277 in the further studies.

E2F1 and RNA Binding Protein QKI Comprise a Negative Feedback in the Cell Cycle Regulation

Cell Cycle (Georgetown, Tex.). Aug, 2011  |  Pubmed ID: 21768773

pRb/E2F1 activity is coordinately regulated during the cell cycle progression, while the molecular strategies safeguarding this pathway are not fully understood. We have previously shown that RNA binding protein QKI inhibits the cell proliferation and promotes the differentiation of gastrointestinal epithelium, suggesting a role of QKI in cell cycle regulation. Here we found that with the cell entry into S phase, QKI expression increased both at the mRNA and protein levels, which was reminiscent of cyclin E expression. Forced expression of E2F1 increased the endogenous level of QKI. Promoter luciferase assay and ChIP analysis identified that the -542~-538 E2F1 binding site was responsible for the upregulation. Increased QKI expression by E2F1, in turn, reduced the E2F1 activity and delayed S-phase entry, forming a negative feedback. As a gene expression regulator, QKI overexpression increased p27, while it decreased cyclin D1 and c-fos expression. Molecularly, p27 and c-fos were direct targets of QKI, while cyclin D1 reduction might be an indirect effect. Taken together, our results reveal that E2F1 directly transcribes QKI, which, in turn, negatively regulates the cell cycle by targeting multiple cell cycle regulators, forming an E2F1-QKI-pRb/E2F1 negative feedback loop.

IL-17A Upregulates Keratin 17 Expression in Keratinocytes Through STAT1- and STAT3-dependent Mechanisms

The Journal of Investigative Dermatology. Dec, 2011  |  Pubmed ID: 21796151

Psoriasis, an immunological skin disease, is characterized by epidermal hyperproliferation, chronic inflammation, and an accumulation of infiltrating T cells. IL-17A is a key cytokine that has a critical role in the pathogenesis of psoriasis. Keratin 17 (K17) is strongly expressed in psoriatic lesions but not in normal skin. Thus, K17 expression is regarded as a hallmark of psoriasis. We previously reported that the K17/T cells/cytokine autoimmune loop was involved in psoriasis. However, the relationship between IL-17A and K17 has yet to be determined. In the present study, IL-17A-induced K17 expression was confirmed in cultured keratinocytes in both mRNA and protein levels. In addition, increased K17 expression was found in the epidermis of IL-17A-injected mouse skin. The regulatory mechanism of K17 expression was further investigated. We found that both the signal transducer and activator of transcription (STAT) 1 and STAT3 pathways were involved in the upregulation of K17 expression induced by IL-17A, and that such regulation could be partially suppressed by STAT1 or STAT3 small interfering RNA and inhibitor. Our data suggest that IL-17A can upregulate K17 expression in keratinocytes in a dose-dependent manner through STAT1- and STAT3-dependent mechanisms. The results indicate that IL-17A might be an important cytokine in the K17/T cells/cytokine autoimmune loop associated with psoriasis.

Screening of Cd Tolerant Genotypes and Isolation of Metallothionein Genes in Alfalfa (Medicago Sativa L.)

Environmental Pollution (Barking, Essex : 1987). Dec, 2011  |  Pubmed ID: 21868142

In order to evaluate Cd tolerance in wide-ranging sources of alfalfa (Medicago sativa) and to identify Cd tolerant genotypes which may potentially be useful for restoring Cd-contaminated environments, thirty-six accessions of alfalfa were screened under hydroponic culture. Our results showed that the relative root growth rate varied from 0.48 to 1.0, which indicated that different alfalfa accessions had various responses to Cd stress. The candidate fragments derived from differentially expressed metallothionein (MT) genes were cloned from leaves of two Cd tolerant genotypes, YE and LZ. DNA sequence and the deduced protein sequence showed that MsMT2a and MsMT2b had high similarity to those in leguminous plants. DDRT-PCR analysis showed that MsMT2a expressed in both YE and LZ plants under control and Cd stress treatment, but MsMT2b only expressed under Cd stress treatment. This suggested that MsMT2a was universally expressed in leaves of alfalfa but expression of MsMT2b was Cadmium (Cd) inducible.

Quality of Life and Related Factors in Adult Patients with Epilepsy in China

Epilepsy & Behavior : E&B. Oct, 2011  |  Pubmed ID: 21871838

The objective of this work was to study the quality of life (QOL) of adult patients with epilepsy in northern China.

Assessing Second Echelon Lymph Nodes During Sentinel Lymph Node Biopsy: Can We Have More Accurate Axillary Treatment for Breast Cancer Patients?

Medical Hypotheses. Dec, 2011  |  Pubmed ID: 21908107

Sentinel lymph node biopsy (SLNB) is the standard treatment for breast cancer patients with clinically negative axilla. For patients with positive sentinel lymph nodes, axillary lymph node dissection (ALND) was required. However, approximately a half of the SLNs-positive patients were found to have clear axillary lymph nodes after ALND, indicating that they had received unnecessary ALND without therapeutic benefit. Therefore, we propose a hypothesis for solution of this clinical problem. We defined the second echelon lymph nodes (SELNs) as those nodes receiving lymphatic drainage directly from the SLNs. For patients with positive-SLNs, SELNs can be biopsy and assessed. If SELNs are negative, no more ALND was needed in these patients even if their SLNs are positive. If our hypothesis were confirmed to be true, we can tailored our axillary treatment to more breast cancer patients, avoiding unnecessary ALND and its complications.

Sequencing-based Expression Profiling in Zebrafish

Methods in Cell Biology. 2011  |  Pubmed ID: 21924174

Gene expression profiling is a powerful technique for studying biological processes, especially tissue/organ-specific ones, at the molecular level. With the rapid development of the next-generation sequencing techniques, high throughput sequencing-based expression profiling techniques have been more and more widely adopted in molecular biology studies. In this chapter, we described a protocol for applying one of the sequencing-based expression profiling techniques, Digital Gene Expression (DGE), for zebrafish research. The protocol provides guidelines for wet-bench experimental procedures as well as for bioinformatics data analyses. We also discuss potential issues/challenges with the use of DGE.

Safety Study of Axillary Reverse Mapping in the Surgical Treatment for Breast Cancer Patients

Journal of Cancer Research and Clinical Oncology. Dec, 2011  |  Pubmed ID: 21935615

With the purpose of minimizing arm lymphedema after axillary staging surgeries in breast cancer patients, the axillary reverse mapping (ARM) technique has been developed to identify and preserve arm drainage system during axillary surgery. This study aimed to clarify risk factors for metastasis in arm lymphatic drainage system in breast cancer patients with clinically negative axillary nodes.

[Intranasal Vaccination with Mycobacterial 65-kD Heat-shock Protein Can Prevent Insulitis and Diabetes in Non-obese Diabetic Mice]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi = Chinese Journal of Cellular and Molecular Immunology. Nov, 2011  |  Pubmed ID: 22078438

To study the efficacy of heat shock protein 65 kDa (HSP65) of Mybobacterium tuberculosis var. bovis in prevention of autoimmune diabetes by intranasal.

[Case of Phantom Limb Pain]

Zhongguo Zhen Jiu = Chinese Acupuncture & Moxibustion. Nov, 2011  |  Pubmed ID: 22136033

[Three-dimension Finite Element Analyses of Interior Stress of Two Kinds of Replace Implant]

Hua Xi Kou Qiang Yi Xue Za Zhi = Huaxi Kouqiang Yixue Zazhi = West China Journal of Stomatology. Oct, 2011  |  Pubmed ID: 22165110

To establish three-dimension finite element model of mandible with two kinds of Replace implant, and to study stress of implant and abutment.

Antifeedant and Insecticidal Effects of Mandelic Acid on the Brown Planthopper Nilaparvata Lugens Stål

Zeitschrift Für Naturforschung. C, Journal of Biosciences. Sep-Oct, 2011  |  Pubmed ID: 22191216

To study the effects of mandelic acid (MA) on the brown planthopper (BPH), Nilaparvata lugens, the survival rate and behaviour of BPH fed on an artificial diet with different dosages of MA was observed. The survival rate of BPH decreased with the increase of the MA concentration and feeding time. In contrast to the control, the survival rate of BPH 72 h after feeding decreased significantly. Electrical penetration graph (EPG) data indicated that MA absorbed by the rice plant from Kimura B solution significantly affected the feeding behaviour of BPH. At the concentrations of 0.1, 0.5, and 1.0 mg/ml, duration of the phloem ingestion of BPH decreased from 115.34 min (control) to 30.41, 7.63, and 0.36 min, respectively. Periods of xylem ingestion of MA-treated BPH were significantly shorter than those of the control (50.44 min). Moreover, BPH spent more time walking around or being at rest on MA-treated rice plants, as well as in stylet activities. The GST (glutathione S-transferase) activity of BPH increased with the increasing MA concentration, while the GPX (glutathione peroxidases) activity did not change significantly. The results indicate that MA has an antifeedant and insecticidal effect on BPH.

Concerns About Targeting Cancer Stem Cell for Cancer Therapy

Medical Hypotheses. Mar, 2011  |  Pubmed ID: 21288650

Codon Usage Bias and Determining Forces in Green Plant Mitochondrial Genomes

Journal of Integrative Plant Biology. Apr, 2011  |  Pubmed ID: 21332641

The phenomenon of codon usage bias has been observed in a wide range of organisms. As organisms evolve, how their codon usage pattern change is still an intriguing question. In this article, we focused on the green plant mitochondrial genomes to analyze the codon usage patterns in different lineages, and more importantly, to investigate the possible change of determining forces during the plant evolution. Two patterns were observed between the separate lineages of green plants: Chlorophyta and Streptophyta. In Chlorophyta lineages, their codon usages showed substantial variation (from strongly A, T-biased to strongly G, C-biased); while in Streptophyta lineages, especially in the land plants, the overall codon usages are interestingly stable. Further, based on the Nc-GC3s plots and Akashi's scaled χ(2) -tests, we found that lineages within Chlorophyta exhibit much stronger evidence of deviating from neutrality; while lineages within Streptophyta rarely do so. Such differences, together with previous reports based on the chloroplast data, suggests that after plants colonized the land, their codon usages in organellar genomes are more reluctant to be shaped by selection force.

Characterization of an in Vitro Differentiation Assay for Pancreatic-like Cell Development from Murine Embryonic Stem Cells: Detailed Gene Expression Analysis

Assay and Drug Development Technologies. Aug, 2011  |  Pubmed ID: 21395400

Embryonic stem (ES) cell technology may serve as a platform for the discovery of drugs to treat diseases such as diabetes. However, because of difficulties in establishing reliable ES cell differentiation methods and in creating cost-effective plating conditions for the high-throughput format, screening for molecules that regulate pancreatic beta cells and their immediate progenitors has been limited. A relatively simple and inexpensive differentiation protocol that allows efficient generation of insulin-expressing cells from murine ES cells was previously established in our laboratories. In this report, this system is characterized in greater detail to map developmental cell stages for future screening experiments. Our results show that sequential activation of multiple gene markers for undifferentiated ES cells, epiblast, definitive endoderm, foregut, and pancreatic lineages was found to follow the sequence of events that mimics pancreatic ontogeny. Cells that expressed enhanced green fluorescent protein, driven by pancreatic and duodenal homeobox 1 or insulin 1 promoter, correctly expressed known beta cell lineage markers. Overexpression of Sox17, an endoderm fate-determining transcription factor, at a very early stage of differentiation (days 2-3) enhanced pancreatic gene expression. Overexpression of neurogenin3, an endocrine progenitor cell marker, induced glucagon expression at stages when pancreatic and duodenal homeobox 1 message was present (days 10-16). Forced expression (between days 16 and 25) of MafA, a pancreatic maturation factor, resulted in enhanced expression of insulin genes, glucose transporter 2 and glucokinase, and glucose-responsive insulin secretion. Day 20 cells implanted in vivo resulted in pancreatic-like cells. Together, our differentiation assay recapitulates the proceedings and behaviors of pancreatic development and will be valuable for future screening of beta cell effectors.

Association Mapping of Grain Color, Phenolic Content, Flavonoid Content and Antioxidant Capacity in Dehulled Rice

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik. Mar, 2011  |  Pubmed ID: 21161500

Phytochemicals such as phenolics and flavonoids in rice grain are antioxidants that are associated with reduced risk of developing chronic diseases including cardiovascular disease, type-2 diabetes and some cancers. Understanding the genetic basis of these traits is necessary for the improvement of nutritional quality by breeding. Association mapping based on linkage disequilibrium has emerged as a powerful strategy for identifying genes or quantitative trait loci (QTL) underlying complex traits in plants. In this study, genome-wide association mapping using models controlling both population structure (Q) and relative kinship (K) were performed to identify the marker loci/QTLs underlying the naturally occurring variations of grain color and nutritional quality traits in 416 rice germplasm accessions including red and black rice. A total of 41 marker loci were identified for all the traits, and it was confirmed that Ra (i.e., Prp-b for purple pericarp) and Rc (brown pericarp and seed coat) genes were main-effect loci for rice grain color and nutritional quality traits. RM228, RM339, fgr (fragrance gene) and RM316 were important markers associated with most of the traits. Association mapping for the traits of the 361 white or non-pigmented rice accessions (i.e., excluding the red and black rice) revealed a total of 11 markers for four color parameters, and one marker (RM346) for phenolic content. Among them, Wx gene locus was identified for the color parameters of lightness (L*), redness (a*) and hue angle (H (o)). Our study suggested that the markers identified in this study can feasibly be used to improve nutritional quality or health benefit properties of rice by marker-assisted selection if the co-segregations of the marker-trait associations are validated in segregating populations.

Production and Characterization of Polyclonal and Monoclonal Abs Against the RNA-binding Protein QKI

Applied Biochemistry and Biotechnology. Jun, 2011  |  Pubmed ID: 21165711

RNA-binding protein QKI, a member of the Signal Transduction and Activation of RNA family, is found to be essential in the blood vessel development and postnatal myelination in central nervous system (Woo et al., Oncogene 28:1176-1186, 2009; Lu et al., Nucleic Acids Res 31(15):4616-4624, 2003; Bohnsack et al., Genesis 44(2):93-104, 2006). However, its wide expression pattern suggests other fundamental roles in vivo (Kondo et al., Mamm Genome 10(7):662-669, 1999). To facilitate the understanding of QKI function in various systems, we prepared the polyclonal and monoclonal antibodies against QKI. To obtain the antigen, recombinant His-tagged QKI was expressed in Escherichia coli and highly purified by Ni(2+)-chelated column combined with hydrophobic and ion exchange methods. Following three types of immunizations with different adjuvants, including Freund's, PAGE gel, and nitrocellulose membrane, only the antiserum produced with Freund's adjuvant is effective for Western blot detection. Several McAb clones are able to recognize both endogenous and over-expressed QKI with high affinity in Western blot and immunofluorescence. The specificity of Ab was validated as weakening, and no specific signals were observed in cells with QKI knocking down. Immunohistochemistry analysis further showed positive staining of QKI in kidney where QKI mRNA was abundantly expressed, ensuring the wide applications of the QKI Abs in the ongoing mechanistic studies.

The Sense Strand Pre-cleaved RNA Duplex Mediates an Efficient RNA Interference with Less Off-target and Immune Response Effects

Applied Microbiology and Biotechnology. Apr, 2011  |  Pubmed ID: 21191787

RNA interference is an appealing and promising therapeutic approach in cancer and other diseases. Designing novel strategies aiming to increase the efficiency, duration, and reduce the off-target silencing by sense strand is of great significance for its future application clinically. Here, we report that RNA duplex with the sense strand pre-cleaved at the base between base 10 and 11 relative to the 5' end of the antisense strand induced a target-specific RNA silencing effectively. Furthermore, different from the canonical RNA duplex, this novel RNA duplex rarely inhibits the luciferase activity in the reporter, bearing the target sequence corresponding to the sense strand, suggesting a less off-target effects of this novel strategy. Furthermore, the immune response of the novel RNA duplex induced a much milder immune response as seen from the NFkappaB activity. In addition, our newly designed RNA duplex should be easier for preservation than the asymmetric RNA duplex. Our results establish a novel method to design a new class of RNA duplex for improved RNA interference.

Synthesis, Characterization and Cytotoxicity Studies of Chitosan-coated Tea Polyphenols Nanoparticles

Colloids and Surfaces. B, Biointerfaces. Feb, 2011  |  Pubmed ID: 20888740

Chitosan nanoparticles (CS-NPs) were prepared by ionic gelation method using carboxymethyl chitosan and chitosan hydrochloride as carriers of tea polyphenols. The characteristics of chitosan-coated tea polyphenols nanoparticles (CS-TP NPs) were determined by using transmission electron microscopy (TEM) and FT-IR spectroscopy. It was found that the synthesized CS-TP NPs were non-spherical in shape with an average size of 407±50nm. Meanwhile, the drug content and encapsulation rate of the nanoparticles was 8-16% and 44-83%, respectively. These CS-TP NPs also demonstrated sustained release of tea polyphenols in PBS. The antitumor of CS-TP NPs towards HepG2 cancer cells was investigated. The result showed that CS-TP NPs retained significant antitumor activities.

Inhibition of Tumor-induced Angiogenesis and Its Mechanism by Ardipusilloside I Purified from Ardisia Pusilla

Journal of Asian Natural Products Research. Jan, 2012  |  Pubmed ID: 22263594

The aim of this study was to evaluate the effects of ardipusilloside I isolated from Ardisia pusilla on tumor angiogenesis and its mechanism of action. The anti-angiogenic effect in vivo was evaluated on xenograft in the athymic mice model and the chicken chorioallantoic membrane (CAM) neovascularization model, the inhibition of growth in vitro was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and the mechanism was demonstrated through detecting microvessel density (MVD), vascular endothelial growth factor (VEGF), VEGF receptor 2 (VEGFR2) and P-VEGFR2 protein expressions, as well as mRNA expressions of VEGF and VEGFR2. The results showed that ardipusilloside I had a good inhibitory effect on A549 xenografted tumor growth, angiogenesis of CAM, and A549 cell growth. Compared to the negative control, MVD protein and mRNA expressions of VEGF and VEGFR were significantly inhibited by ardipusilloside I in a dose-dependent manner. These findings suggested that ardipusilloside I might be a promising candidate as angiogenesis inhibitors.

Comparison of ER/PR and HER2 Statuses in Primary and Paired Liver Metastatic Sites of Breast Carcinoma in Patients with or Without Treatment

Journal of Cancer Research and Clinical Oncology. Jan, 2012  |  Pubmed ID: 22290394

PURPOSE: The aim of this study was to determine whether estrogen receptor (ER)/progesterone receptor (PR) and human epidermal growth factor receptor type 2 (HER2) statuses between primary tumors and paired liver metastatic localizations of breast carcinoma were modified by treatment or during the natural metastatic process. METHODS: ER, PR, and HER2 expressions were analyzed on paired tissue specimens taken from the primary and the liver metastatic tumors in breast cancer patients. The first group included 46 women who presented with T1-T4, N0-N3, M0 breast carcinoma when first diagnosed and were treated by neoadjuvant therapy or directly underwent surgery, then received postoperative treatment and developed liver metastasis several months/years later. The second group included 12 patients with liver metastatic breast carcinoma when first diagnosed for breast cancer. HER2 status was determined by immunohistochemistry as well as fluorescence in situ hybridization. RESULTS: Among the 46 patients in the first group, the ER/PR and HER2 statuses (when considered as a whole histological subtype) were changed between primary tumor and liver metastatic lesions in 12 patients (26.1%). While ER and PR status were modified in 14 (30.4%) and 25 (54.3%) patients, respectively, there were only 5 (10.9%) cases showed a discrepancy in the HER2 status. In the second group, the ER/PR and HER2 statuses (when considered as a whole subtype) were consistent between primary and liver metastatic tumor in 10 of 12 (83.3%) patients. ER, PR, and HER2 statuses were modified in 0 of 12 (0%), 4 of 12 (33.3%), and 1 of 12 (8.3%) cases, respectively. CONCLUSIONS: ER/PR and HER2 statuses between primary and liver metastatic lesions of breast carcinoma can be modified after treatment but are stable in most cases during the natural metastatic process.

MiR-30c-1* Promotes Natural Killer Cell Cytotoxicity Against Human Hepatoma Cells by Targeting the Transcription Factor HMBOX1

Cancer Science. Feb, 2012  |  Pubmed ID: 22320217

Natural killer (NK) cells play a critical role in antitumor immunity, and the activation of NK cells is regulated by a series of NK cell receptors. Here, we show that crosslinking CD226, an important NK cell receptor, with the anti-CD226 mAb LeoA1 on NKL cells, regulated the expression of several microRNA and transmembrane tumor necrosis factor-α. Among them, miR-30c-1* was noticed because overexpression of miR-30c-1(*) triggered upregulation of transmembrane tumor necrosis factor-α expression and enhanced NK cell cytotoxicity against hepatoma cell lines SMMC-7721 and HepG2. Furthermore, we proved that the inhibitory transcription factor HMBOX1, which depressed the activation of NK cells, was the direct target gene of miR-30c-1(*) . In conclusion, our results revealed a novel regulatory mechanism: miR-30c-1(*) promoted NK cell cytotoxicity against hepatoma cells by targeting HMBOX1. (Cancer Sci, doi: 10.1111/j.1349-7006.2012.02207.x, 2012).