Diabetes mellitus affects 347 million people worldwide, and over 80 % of diabetes deaths occur in low- and middle-income countries. Type 1 diabetes (T1D) is characterized by the attacks of the body's own immune system on the pancreatic ?-cells. In this work, we present a new CTLA-4 Ig targeting at the surface of ?-cell and prepare it from Escherichia coli aiming at clearing activated T cells around ?-cells and avoiding all-round decline in systematic immunity. This fusion protein is composed of CTLA-4-Ig part and ?-cell-targeting part, with properties of the therapeutic effect of CTLA-4-Ig and selective binding to ?-cells. In preliminary biological activity assay, our results verified the feasibility of ?-cell-targeting strategy and its activity of CTLA-4-Ig part. The fusion protein recognizes and binds specifically to CD80(+) and CD86(+) cells as well as ?-cell, but not to control cells, displaying the potential to be used as a feasible and effective treatment of T1D with lessened side effect.
Liver fibrosis is a wound-healing response to chronic liver injury that could lead to liver failure, but treatment remains ineffective. In this study, we investigated anti-hepatic fibrosis activity of n-hexane, chloroform, ethyl acetate, and methanol extracts of mycelia from six commercially available medicinal mushrooms in submerged culture, namely Antrodia camphorata, Cephalosporium sinensis, Cordyceps mortierella, Hericium erinaceus, Ganoderma lucidum, and Armillaria mellea. Their anti-fibrotic activities were evaluated via inhibition against accumulation of TGF-?1-induced collagen deposition in CFSC-8B cells. Hex, Chl, and MeOH extracts of A. camphorata and Hex extract of A. mellea significantly decreased collagen production. Bioactivity-guided fractionation led to the identification of seven compounds using UPLC-Q-TOF-MS from the Hex Fr.2 of A. camphorata. At the molecular level, Hex Fr.2 of A. camphorata suppressed ?-SMA, Collagen I, Collagen III, and Fibronectin expression induced by TGF-?1 in CFSC-8B cells as indicated by qRT-PCR analysis. They also inhibited ?-SMA and Collagen I protein expression according to western blot analyses. Mechanistically, Hex Fr.2 of A. camphorata negatively regulates TGF-?1/Smad2/3 signaling. Our studies demonstrate that A. camphorata has in vitro anti-hepatofibrotic activity and that there is great potential for the discovery of new drugs for the treatment of liver fibrosis by screening more medicinal mushrooms.
A novel silicon-on-insulator (SOI) polarization splitter-rotator (PSR) with a large fabrication tolerance is proposed based on cascaded multimode interference (MMI) couplers and an assisted mode-evolution taper. The tapers are designed to adiabatically convert the input TM0 mode into the TE1 mode, which will output as the TE0 mode after processed by the subsequent MMI mode converter, 90-degree phase shifter (PS) and MMI 3 dB coupler. The numerical simulation results show that the proposed device has a < 0.5 dB insertion loss with < -17 dB crosstalk in C optical communication band. Fabrication tolerance analysis is also performed with respect to the deviations of MMI coupler width, PS width, slab height and upper-cladding refractive index, showing that this device could work well even when affected by considerable fabrication errors. With such a robust performance with a large bandwidth, this device offers potential applications for CMOS-compatible polarization diversity, especially in the booming 100 Gb/s coherent optical communications based on silicon photonics technology.
To observe the myocardial protective effect of Danhong Injection evaluated by velocity vector imaging (VVI) in patients with unstable angina pectoris after percutaneous coronary intervention (PCI) and elucidate its possible mechanism.
T-wave alternans (TWA) represents myocardial instability. The present study was to determine the impact of cardiac resynchronization therapy (CRT) on TWA and left ventricular ejection fraction (LVEF) in heart failure patients.
To investigate the effects of boron on growth performance and meat quality, 10-day-old Africa ostrich chicks were randomly divided into 6 groups with 6 replicates in each group. For 80 days, birds in the treatments were fed the same basal diet but given different concentrations of boron-supplemented water. The highest final BW (33.4 ± 0.30 kg), ADFI (376 ± 1.83 g), and ADG (224 ± 1.01 g) appeared in the group receiving 160 mg/L boron (group 4). 160 mg/L boron also decreased drip loss (2.20 ± 0.59), cooking loss (35.3 ± 1.14), and elevated pH value (6.13 ± 0.28) of meat (P < 0.05). Ostrich chicks in the 640 mg/L treatment group (group 6) had the lowest final BW (30.8 ± 1.05 kg) and ADG (208 ± 0.74 g) (P < 0.05). The highest ash (1.35 ± 0.01%) and pH (6.18 ± 0.03) and the lowest protein (20.4 ± 1.74%), drip loss (2.10 ± 0.76%), cooking loss (35.0 ± 0.41%), C18:1 (28.2 ± 0.65%), and C18:3?3 (2.60 ± 0.51%) appeared in group 6 (P < 0.05) as well. Overall, the optimum concentration of 160 mg/L supplemental boron improved ostrich growth performance and meat quality; however, high concentrations of boron decreased both performance and meat quality.
Abstract The treatment outcome and development of new mutations in imatinib- and/or nilotinib-failure Ph+ leukemia patients with highly nilotinib-resistant mutations (Y253H, E255K/V and F359V/C) were assessed on dasatinib. A total of 111 Ph+ leukemia patients were grouped into 3 cohorts by baseline BCR-ABL kinase domain mutation status: no mutation (n=44), non-nilotinib-resistant mutations (n=26), or nilotinib-resistant mutations (n=41). The frequencies of hematological, cytogenetic and molecular responses and clinical resistance to dasatinib were similar among the 3 cohorts during dasatinib therapy. In dasatinib-resistant patients, new mutations were most frequently observed in the cohort with nilotinib-resistant mutations (P=0.001). Multivariate analyses revealed that the advanced disease and harboring the Y253H or F359V mutation before dasatinib were independent predictors of developing new mutations. We concluded that Ph+ leukemia patients with the Y253H or F359V mutation have a high likelihood of developing new mutations in the setting of dasatinib resistance.
The GDF3 gene plays a fundamental role in embryonic morphogenesis. Recent studies have indicated that GDF3 plays a previously unrecognised role in cardiovascular system development. Non-syndromic CHDs might be a clinically isolated manifestation of GDF3 mutations. The purpose of the present study was to identify potential pathological mutations in the GDF3 gene in Chinese children with non-syndromic CHDs, and to gain insight into the aetiology of non-syndromic CHDs.
We propose in this Letter a single-mode graphene-coated nanowire surface plasmon waveguide. The single-mode condition and modal cutoff wavelength of high order modes are derived from an analytic model and confirmed by numerical simulation. The mode number diagram of the proposed waveguide in the wavelength-radius space is also demonstrated. By changing the Fermi level of graphene, the performance of the proposed waveguide could be tuned flexibly, offering potential application in tunable nanophotonic devices.
The maldigestion and malabsorption of fat in infants fed milk formula results due to the minimal production of pancreatic lipase. Thus, to investigate lipid digestion and absorption and mimic the situation in newborns, a young porcine exocrine pancreatic insufficient (EPI) model was adapted and validated in the present study. A total of thirteen EPI pigs, aged 8 weeks old, were randomised into three groups and fed either a milk-based formula or a milk-based formula supplemented with either bacterial or fungal lipase. Digestion and absorption of fat was directly correlated with the addition of lipases as demonstrated by a 30 % increase in the coefficient of fat absorption. In comparison to the control group, a 40 and 25 % reduction in total fat content and 26 and 45 % reduction in n-3 and n-6 fatty acid (FA) content in the stool was observed for lipases 1 and 2, respectively. Improved fat absorption was reflected in the blood levels of lipid parameters. During the experiment, only a very slight gain in body weight was observed in EPI piglets, which can be explained by the absence of pancreatic protease and amylase in the gastrointestinal tract. This is similar to newborn babies that have reduced physiological function of exocrine pancreas. In conclusion, we postulate that the EPI pig model fed with infant formula mimics the growth and lipid digestion and absorption in human neonates and can be used to elucidate further importance of fat and FA in the development and growth of newborns, as well as for testing novel formula compositions.
Background:The effect of adiposity on response to cardiac resynchronization therapy (CRT) and long-term outcome in patients undergoing CRT has not been previously reported. This study assessed the impact of baseline body mass index (BMI) on cardiac reverse remodeling and prognosis following CRT.Methods?and?Results:A total of 247 CRT patients were included and divided into 4 groups according to baseline BMI. During 6-month follow-up, overweight and obese patients (BMI, 24-28 kg/m(2), ?28 kg/m(2), respectively) were inclined to have better clinical and echocardiographic improvements (P<0.05) as well as higher response rate (P<0.001) than underweight and normal weight patients (BMI, <18.5 kg/m(2), 18.5-24 kg/m(2), respectively). During long-term follow-up, overweight and obese patients had lower all-cause mortality (P=0.015) and combined endpoint of death or HF hospitalizations (P=0.001) than underweight and normal weight patients. Compared with normal weight patients, underweight patients had a 2.29-fold increase in risk of combined endpoint events whereas overweight and obese patients had a reduction in the risk of death (66% and 58%, respectively) and combined endpoint events (52% and 38%, respectively).Conclusions:Patients with obesity and overweight derived more benefit from CRT. Higher BMI was independently associated with better clinical outcome in CRT patients.
Follicular helper T (TFH) cells and B cells are linked to the pathogenesis of ankylosing spondylitis (AS). Follicular regulatory T (TFR) cells suppress TFH cell and germinal center B cell numbers in vivo. The role of TFR cells in AS is unknown. The frequency of peripheral blood inducible FOXP3+CXCR5+CD4+TFR cells and CXCR5+CD4+TFH cells were taken from 20 onset AS patients and 10 healthy controls, and were examined by flow cytometry, their disease activity were measured by the bath ankylosing spondylitis disease activity index(BASDAI). The concentrations of serum IL-21, IgG, IgA, IgM, CRP were examined, and the values of eESR were measured. The frequency of peripheral blood FOXP3+CXCR5+CD4+TFR cells, CXCR5+CD4+TFH cells, the ratio of FOXP3+CXCR5+CD4+TFR/ CXCR5+CD4+TFH cells, and the concentration of serum IL-21 in the AS patients were significantly higher than those in the HC (p<0.0001, p=0.0027, p<0.0001, p=0.0039, respectively). The frequency of FOXP3+CXCR5+CD4+TFR cells and the ratio of FOXP3+CXCR5+CD4+TFR/ CXCR5+CD4+TFH cells still significantly rose in those patients after standard treatment (p=0.0006, p<0.0001), the concentration of serum IL-21 decreased after treatment (p=0.0049), accompanied by significantly minimized disease activities. Furthermore, the TFR cells were negatively correlated with serum IgA in those patients before treatment (r=-0.582, p=0.0071) and the frequency of TFR cells was negatively correlated with that of TFH cells and the concentration of serum IL-21 after treatment (r=-0.550, p=0.046; r=-0.581, p=0.0371). TFR cells may participate in the pathogenesis of AS and may be responsible for controlling the autoantibodies, the frequency and function of TFH cells to inhibit the development of AS. This article is protected by copyright. All rights reserved.
A viable pathway is proposed for the formation of the triplet state of the GC Watson-Crick base pair. It includes the following steps: (a) a low-energy electron is captured by cytosine in the GC pair, forming the cytosine base-centered radical anion GC(-•); and (b) photoradiation with energy around 5 eV initiates the electron detachment from either cytosine (in the gas phase) or guanine (in aqueous solutions). This triggers interbase proton transfer from G to C, creating the triplet state of the GC pair. Double proton transfer involving the triplet state of GC pair leads to the formation of less stable tautomer G(N2-H)(•)C(O2H)(•). Tautomerization is accomplished through a double proton transfer process in which one proton at the N3 of C(H)(•) migrates to the N1 of G(-H)(•); meanwhile, the proton at the N2 of G transfers to the O2 of C. This process is energetically viable; the corresponding activation energy is around 12-13 kcal/mol. The base-pairing energy of the triplet is found to be ?3-5 kcal/mol smaller than that of the singlet state. Thus, the formation of the triplet state GC pair in DNA double strand only slightly weakens its stability. The obtained highly reactive radicals are expected to cause serious damage in the DNA involved in biochemical processes, such as DNA replication where radicals are exposed in the single strands.
The etiology of an outbreak of gastroenteritis in humans cannot always be determined and ?25% of outbreaks remain unsolved in New Zealand. It is hypothesised that novel viruses may account for a proportion of unsolved cases, and new unbiased high-throughput sequencing methods hold promise for their detection. Analysis of the faecal metagenome can reveal the presence of viruses, bacteria and parasites which may have evaded routine diagnostic testing. Thirty one faecal samples from 26 gastroenteritis outbreaks of unknown etiology occurring in New Zealand between 2011 and 2012 were selected for de novo metagenomic analysis. A total dataset of 193 million sequence reads of 150 bp in length were produced on an Illumina MiSeq. The metagenomic dataset was searched for virus and parasite sequences with no evidence of novel pathogens found. Eight viruses and one parasite were detected, each already known to be associated with gastroenteritis including adenovirus, rotavirus, sapovirus and Dientamoeba fragilis. In addition, we also describe the first detection of human parechovirus 3 (HPeV3) in Australasia. Metagenomics may thus provide a useful audit tool when applied retrospectively to determine where routine diagnostic processes may have failed to detect a pathogen.
To evaluate the cleaning effect of the C-shaped canal treated by manual K file and ProTaper rotary endodontic file combined with ultrasonic cleaning, and find a better cleaning program for the C-shaped root canal.
Based on the representative articles in recent years, the different mechanisms of decitabine on immune regulation in patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (HSCT) are summarized. Decitabine improves the expression of WT1 gene to stimulate specific cytotoxic T cells which can enhance graft versus leukemia effect (GVL) and improve the expression of FOXP3 gene to stimulate regulatory T cells so as to inhibit the acute graft versus host disease (GVHD). Through the above-mentimed mechanisms, decitabine can improve both therapeutic effect and quality of life in the patients with AML after allogeneic HSCT.
Polychlorinated biphenyls (PCBs) were measured in house dust from an e-waste site and urban site in the Pearl River Delta, southern China. The PCB concentrations in house dust at the e-waste site ranged from 12.4 to 87 765 ng x g(-1), with an average of 10 167 ng x g(-1). There was no significant difference in the PCB concentrations between indoor and outdoor dust. The PCB homologue pattern was dominated by tri-, penta-, hexa-, and tetra-CBs, which was not similar to that in Chinese technical PCB product. There was also no significant difference in the PCB compositions between indoor and outdoor dust. PCB sources in house dust at the e-waste site were apportioned by chemical mass balance (CMB) model. The results showed that the PCBs were derived primarily from Aroclor 1262 (36.7% ), Aroclor 1254 (26.7%), Aroclor 1242 (21.4%), and Aroclor 1248 (18.5%). The daily exposure doses were 42, 17, and 2.9 ng x (kg x d)(-1) for toddlers, children/adolescents, and adults in the e-waste area, respectively. Risk assessment indicated that the hazard quotients were higher than 1 for toddlers and children/adolescents indicating adverse effects for them. The lifetime average excess carcinogenic risk for population in the e-waste area was 4.5 x 10(-5), within the acceptable range of U. S. Environmental Protection Agency. The mean concentrations of PCBs in house dust in Guangzhou was 48.7 ng x g(-1). The low PCB level is consistent with the fact that technical PCBs were not widely used in China in the past. The risks of exposure to PCBs via house dust in Guangzhou are very low.
Novel analgesics that do not suppress the respiratory drive are urgently needed. Glutamate signaling through ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors plays important roles in central pain circuits. AMPAkines augment AMPA receptor function and have been shown to stimulate the respiratory drive to oppose opioid-induced hypoventilation. However, their role in chronic pain states remains unknown.
Cr(VI) biotreatment has attracted a substantial amount of interest due to its cost effectiveness and environmental friendliness. However, the slow Cr(VI) bioreduction rate and the formed organo-Cr(III) in solution are bottlenecks for biotechnology application. In this study, a novel strain, Acinetobacter sp. HK-1, capable of reducing Cr(VI) and immobilizing Cr(III) was isolated. Under optimal conditions, the Cr(VI) reduction rate could reach 3.82 mg h(-1) g cell(-1). To improve the Cr(VI) reduction rate, two quinone/graphene oxide composites (Q-GOs) were first prepared via a one-step covalent chemical reaction. The results showed that 2-amino-3-chloro-1,4-naphthoquinone-GO (NQ-GO) exhibited a better catalytic performance in Cr(VI) reduction compared to 2-aminoanthraquinone-GO. Specifically, in the presence of 50 mg L(-1) NQ-GO, a Cr(VI) removal rate of 190 mg h(-1) g cell(-1), which was the highest rate obtained, was achieved. The increased Cr(VI) reduction rate is mainly the result of NQ-GO significantly increasing the Cr(VI) reduction activity of cell membrane proteins containing dominant Cr(VI) reductases. X-ray photoelectron spectroscopy analysis found that Cr(VI) was reduced to insoluble Cr(III), which was immobilized by glycolipids secreted by strain HK-1. These findings indicate that the application of strain HK-1 and NQ-GO is a promising strategy for enhancing the treatment of Cr(VI)-containing wastewater.
Stem cell therapy is an emerging therapeutic modality in the treatment of stroke. We assessed the safety and feasibility of the co-transplantation of neural stem/precursor cells (NSPCs) and mesenchymal stromal cells (MSCs) in patients with ischemic stroke. Eight patients were enrolled in this study. All patients had a hemisphere with infarct lesions located on one side of the territories of the cerebral middle or anterior arteries as revealed with cranial magnetic resonance imaging (MRI). The patients received one of the following two types of treatment: the first treatment involved 4 intravenous injections of MSCs at 0.5x10(6)/kg body weight; the second treatment involved one intravenous injection of MSCs at 0.5x10(6)/kg weight followed by three injections of MSCs at 5x10(6)/patient and NSPCs at 6 x 10(6)/patient through the cerebellomedullary cistern. The patients' clinical statuses were evaluated with the National Institutes of Health Stroke Scale (NIHSS), the modified Rankin Scale (mRS), and the Barthel Index (BI). Six patients were given 4 cell transplantations. The most common side effect of stem cell transplantation in these 6 cases was low fever that usually lasted 2-4 days after each therapy. One patient exhibited minor dizziness. All side effects appeared within the first 2-24 hours of cell transplantation and they resolved without special treatment. There was no evidence of neurological deterioration or neurological infection. Most importantly, no tumorigenesis was found at a 2-year follow-up. The neurological functions, disability levels, and daily living abilities of the patients in this study were improved. While these observations support the use of the combination transplantation of NSPCs and MSCs as safe and feasible method of improving neurological function, further studies that include larger samples, longer follow-ups, and control groups are still needed. This manuscript is published as part of the International Association of Neurorestoratology (IANR) special issue of Cell Transplantation.
Fas signaling promotes metastasis of gastrointestinal (GI) cancer cells by inducing epithelial-mesenchymal transition (EMT), and EMT acquisition has been found to cause cancer chemoresistance. Here, we demonstrated that the response to chemotherapy of GI cancer patients with higher expression of FasL was significantly worse than patients with lower expression. Fas-induced activation of the ERK1/2-MAPK pathway decreased the sensitivity of GI cancer cells to chemotherapeutic agents and promoted the expression of P-glycoprotein (P-gp). FasL promoted chemoresistance of GI cancer cell via upregulation of P-gp by increasing ?-catenin and decreasing miR-145. ?-catenin promoted P-gp gene transcription by binding with P-gp promoter while miR-145 suppressed P-gp expression by interacting with the mRNA 3'UTR of P-gp. Immunostaining and qRT-PCR analysis of human GI cancer samples revealed a positive association among FasL, ?-catenin, and P-gp, but a negative correlation between miR-145 and FasL or P-gp. Altogether, our results showed Fas signaling could promote chemoresistance in GI cancer through modulation of P-gp expression by ?-catenin and miR-145. Our findings suggest that Fas signaling-based cancer therapies should be administered cautiously, as activation of this pathway may not only lead to apoptosis but also induce chemoresistance.
Abstract In a classical drop-loser (or drop-arm) design, patients are randomized into to all arms (doses) and at the interim analysis, inferior arms are dropped. Therefore, compared to the traditional dose-finding design, this adaptive design can reduce the sample size by not carrying over all doses to the end of the trial or dropping the losers earlier. However, all the doses have to be explored. For unimodal (including linear or umbrella) response curves, we proposed an effective dose-finding design that allows adding arms at the interim analysis. The trial design starts with two arms, depending on the response of the two arms and the unimodality assumption; we can decide which new arms to be added. This design does not require exploring all arms (doses) to find the best responsive dose; therefore it can further reduce the sample size from the drop-loser design by as much as 10-20%.
To study the roles of stromal interaction molecule 2 (STIM2) and transient receptor potential canonical 3 (TRPC3) in extracellular Ca(2+)-sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO) in human umbilical vein endothelial cells (HUVEC).
miRNA-16 (miR16) plays an important role in modulating the drug resistance of SGC7901 cell lines to adriamycin (ADR). A variety of viral carriers have been designed for miRNA delivery. However, the safety concerns are currently perceived as hampering the clinical application of viral vector-based therapy. Herein a type of magnetic nanoparticles (MNPs) was designed and synthesized using poly(ethylene glycol) (PEG)-coated Fe3O4 nanoparticles as a miRNA delivery system for the purpose of reducing drug resistance of gastric cancer cells by enforcing miR16 expression in SGC7901/ADR cells. The MNPs with good biocompatibility were synthesized by thermal decomposition, and then conjugated with miRNA via electrostatic interaction producing miR16/MNPs. After co-culture with miR16/MNPs, ADR-induced apoptosis of SGC7901/ADR was examined by MTT and TUNEL. miR16/MNPs treatment significantly increased cell apoptosis in vitro. SGC7901/ADR(fluc) tumor-bearing nude mice under ADR therapy were treated with miR16/MNPs by tail vein injection for in vivo study. After intraperitoneal injection of ADR, tumor volume measurement and fluorescence imaging were performed to for the death of SGC7901/ADR cells in vivo. Results showed that miR16/MNPs were able to significantly suppress SGC7901/ADR tumor growth, probably through increasing SGC7901/ADR cells' sensitivity to ADR. Our results suggest the efficient delivery of miR16 by MNPs as a novel therapeutic strategy for drug resistant tumor treatment.
Neuropathic pain is a challenge for physicians and basic science researchers because it often does not respond to routine treatment. The administration of morphine has been considered one of the effective recommended treatments, but its wide application is limited because of the development of antinociceptive tolerance. In general, basic science studies focus on neuropathic pain and morphine tolerance separately. However, we tried to investigate the effect of microglial activation on morphine tolerance in spinal nerve ligation (SNL) rats during the maintenance period of neuropathic pain. This study produced the following results. First, the repeated administration of morphine induces the development of antinociceptive tolerance during the maintenance period of neuropathic pain. Second, during the development of morphine tolerance, microglial activation, which is related to the analgesic effect of morphine, decreases in the first few days, but this pattern is reversed in the following days with the development of morphine tolerance. Third, the repeated administration of minocycline, a microglial activation inhibitor, does not influence the antinociceptive effect of a single dose of morphine. Fourth, the pre-administration of minocycline can delay the development of morphine tolerance, but repeated minocycline administration cannot reverse existing morphine tolerance. We concluded that microglial activation contributes to the morphine tolerance of SNL rats in the maintenance period of neuropathic pain and that minocycline delays the development of morphine tolerance but does not reverse existing morphine tolerance during the maintenance period of neuropathic pain in rats. These findings might be useful for clinical pain management. This article is protected by copyright. All rights reserved.
The regulation of light-dependent anthocyanin biosynthesis in Brassica rapa subsp. rapa cv. Tsuda turnip was investigated using an EMS-induced mutant R30 with light-independent pigmentation. TILLING and subsequent analysis showed that a stop codon was inserted in the R2R3-MYB transcription factor BrMYB4 and that the encoded protein (BrMYB4mu) had lost its C-terminal region. In R30, anthocyanin accumulated in the underground portion of the storage root of 2-mo-old plants. In 4-d-old seedlings and 2-mo-old plants, expression of BrMYB4 was similar between R30 and wild type (WT), but the expression of the cinnamate 4-hydroxylase gene (BrC4H) was enhanced markedly in R30 in the dark. In turnip seedlings, BrMYB4 expression was suppressed by UV-B irradiation in the WT, but this negative regulation was absent in R30. Concomitantly, BrC4H was repressed by UV-B irradiation in the WT, but stayed at high levels in R30. A gel-shift assay revealed that BrMYB4 could directly bind to the promoter region of BrC4H, but BrMYB4mu could not. The BrMYB4-eGFP protein could enter the nucleus in the presence of BrSAD2 (an importin ?-like protein) nuclear transporter, but BrMYB4mu-eGFP could not. These results showed that BrMYB4 functions as a negative transcriptional regulator of BrC4H and mediates UV-B-dependent phenylpropanoid biosynthesis, while BrMYB4mu has lost this function. In the storage roots, the expression of anthocyanin biosynthesis genes was enhanced in R30 in the dark and in sunlight in both the WT and R30. However, in the WT, anthocyanin-inducing sunlight did not suppress BrMYB4 expression. Therefore, sunlight-induced anthocyanin biosynthesis does not seem to be regulated by BrMYB4.
We report a patient harboring a de novo m.5540G>A mutation affecting the MT-TW gene coding for the mitochondrial tryptophan-transfer RNA. This patient presented with atonic-myoclonic epilepsy, bilateral sensorineural hearing loss, ataxia, motor regression, ptosis, and pigmentary retinopathy. Our proband had an earlier onset and more severe phenotype than the first reported patient harboring the same mutation. We discuss her clinical presentation and compare it with the only previously published case.
The maintenance of a neural stem cell (NSC) population in mammalian postnatal and adult life is crucial for continuous neurogenesis and neural repair. However, the molecular mechanism of how NSC populations are maintained remains unclear. Gangliosides are important cellular membrane components in the nervous system. We previously showed that ganglioside GD3 plays a crucial role in the maintenance of the self-renewal capacity of NSCs in vitro. Here, we investigated its role in postnatal and adult neurogenesis in GD3-synthase knock-out (GD3S-KO) and wild-type mice. GD3S-KO mice with deficiency in GD3 and the downstream b-series gangliosides showed a progressive loss of NSCs both at the SVZ and the DG of the hippocampus. The decrease of NSC populations in the GD3S-KO mice resulted in impaired neurogenesis at the granular cell layer of the olfactory bulb and the DG in the adult. In addition, defects of the self-renewal capacity and radial glia-like stem cell outgrowth of postnatal GD3S-KO NSCs could be rescued by restoration of GD3 expression in these cells. Our study demonstrates that the b-series gangliosides, especially GD3, play a crucial role in the long-term maintenance NSC populations in postnatal mouse brain. Moreover, the impaired neurogenesis in the adult GD3S-KO mice led to depression-like behaviors. Thus, our results provide convincing evidence linking b-series gangliosides deficiency and neurogenesis defects to behavioral deficits, and support a crucial role of gangliosides in the long-term maintenance of NSCs in adult mice.
Cerenkov luminescence endoscopy (CLE) is an optical technique that captures the Cerenkov photons emitted from highly energetic moving charged particles (?(+) or ?(-)) and can be used to monitor the distribution of many clinically available radioactive probes. A main limitation of CLE is its limited sensitivity to small concentrations of radiotracer, especially when used with a light guide. We investigated the improvement in the sensitivity of CLE brought about by using a ?(-) radiotracer that improved Cerenkov signal due to both higher ?-particle energy and lower ? noise in the imaging optics because of the lack of positron annihilation.
Spermatogenesis is a complex process closely associated with the phosphorylation-orchestrated cell cycle. Elucidating the phosphorylation-based regulations should advance our understanding of the underlying molecular mechanisms. Here we present an integrative study of phosphorylation events in the testis. Large-scale phosphoproteome profiling in the adult mouse testis identified 17,829 phosphorylation sites in 3,955 phosphoproteins. Although only approximately half of the phosphorylation sites enriched by IMAC were also captured by TiO2, both the phosphoprotein datasets identified by the two methods significantly enriched the functional annotation of spermatogenesis. Thus, the phosphoproteome profiled in this study is a highly useful snapshot of the phosphorylation events in spermatogenesis. To further understand phosphoregulation in the testis, the site-specific kinase-substrate relations (ssKSRs) were computationally predicted for re-constructing kinase-substrate phosphorylation networks (KSPNs). A core sub-KSPN among the spermatogenesis-related proteins was retrieved and analyzed to explore the phosphoregulation during spermatogenesis. Moreover, network-based analyses demonstrated that a number of protein kinases such as MAPKs, CDK2 and CDC2 with statistically more ssKSRs might have significantly higher activities and play an essential role in spermatogenesis, and the predictions were consistent with previous studies on the regulatory roles of these kinases. In particular, the analyses proposed that the activities of POLO-like kinases (PLKs) might be dramatically higher, while the prediction was experimentally validated by detecting and comparing the phosphorylation levels of pT210, an indicator of PLK1 activation, in testis and other tissues. Further experiments showed that the inhibition of PLKs decreases cell proliferation by inducing G2/M cell cycle arrest. Taken together, this systematic study provides a global landscape of phosphoregulation in the testis, and should prove to be of value in future studies of spermatogenesis.
The transition/transversion (Ti/Tv) ratio and heterozygous/nonreference-homozygous (het/nonref-hom) ratio have been commonly computed in genetic studies as a quality control (QC) measurement. Additionally, these two ratios are helpful in our understanding of the patterns of DNA sequence evolution.
A traditional neuro-fuzzy system is transformed into an equivalent fully connected three layer neural network (NN), namely, the fully connected neuro-fuzzy inference systems (F-CONFIS). The F-CONFIS differs from traditional NNs by its dependent and repeated weights between input and hidden layers and can be considered as the variation of a kind of multilayer NN. Therefore, an efficient learning algorithm for the F-CONFIS to cope these repeated weights is derived. Furthermore, a dynamic learning rate is proposed for neuro-fuzzy systems via F-CONFIS where both premise (hidden) and consequent portions are considered. Several simulation results indicate that the proposed approach achieves much better accuracy and fast convergence.
Murine norovirus (MNV) was first discovered in mice in 2003. MNV is a member of the genus Norovirus in the family Caliciviridae. It is one of the most important and prevalent pathogens of laboratory mice, and almost all mouse strains are susceptible to MNV infection. In this study, a MNV strain was isolated from the cecal contents of infected mice and identified by the cytopathic effect (CPE) assay, virus plaque assay, 50% tissue culture infectious dose (TCID50) assay, electron microscopy, indirect immunofluorescence assay (IFA) and nucleotide sequencing. On infection, the RAW264.7 cell line showed obvious cytopathic effects within 24 to 48 hours post-inoculation, as infected cells became rounded, bright and shrunken, with ultimate disintegration of the cell sheet. After the isolation of the MNV virus, the virus was plaque-purified in RAW264.7 cells. The TCID50 of the virus was 10(5.25/0.1 mL. Electron microscopic observations of the purified virus showed the presence of spherical and non-enveloped viral particles that were 30 to 35 nm in diameter. According to the identification results, the isolate was named as MNV Guangzhou/K162/09/CHN. Thereafter, five overlapping gene fragments that covered the entire open reading frame (ORF) were amplified by RT-PCR, and the 3'-untranslated region (UTR) and 5'-UTR were amplified using the 3'-rapid amplification of cDNA ends (RACE) and the 5'-RACE method, respectively. Each of the gene fragments were cloned and sequenced, and whole genome sequences of the strain were obtained by assembling the cDNA fragment sequences. The results showed that the length of the complete genome was 7 380 nucleotides (GenBank accession number: HQ317203). The comparison of nucleotide and deduced amino acid sequences of the isolate was performed against other MNV strains in the GenBank database. A phylogenetic tree based on VP1 nucleotide sequences was constructed using MEGA5.0 software. The homology of nucleotides between the MNV Guangzhou/K162/09/CHN strain and other MNV isolates ranged from 87.4% to 89.7%. Phylogenetic analysis showed that there was a close genetic relationship between the Guangzhou/K162/09/CHN strain and MNV strains isolated from Japan (S7-P2 and S7-PP3 isolates), Korea (K4 isolate), and Germany (Berlin/04/06/DE and Berlin/05/06/DE isolates). This is the first report of the isolation and identification of MNV in China, and the first report of the genetic analysis of its complete genome.
By integrating the clinically used endoscope with the emerging Cerenkov luminescence imaging (CLI) technology, a new endoscopic Cerenkov luminescence imaging (ECLI) system was developed. The aim is to demonstrate the potential of translating CLI to clinical studies of gastrointestinal (GI) tract diseases. We systematically evaluated the feasibility and performance of the developed ECLI system with a series of in vitro and pseudotumor experiments. The ECLI system is comprised of an electron multiplying charge coupled device (EMCCD) camera coupled with a clinically used endoscope via an optical adapter. A 1951-USAF test board was used to measure the white-light lateral resolution, while a homemade test chart filled with (68)Ga was employed to measure the CL lateral resolution. Both in vitro and pseudotumor experiments were conducted to obtain the sensitivity of the ECLI system. The results were validated with that of CLI using EMCCD only, and the relative attenuation ratio of the ECLI system was calculated. Results showed that The white-light lateral resolution of the ECLI system was 198 µm, and the luminescent lateral resolution was better than 1 mm. Sensitivity experiments showed a theoretical sensitivity of [Formula: see text] ([Formula: see text]) and [Formula: see text] ([Formula: see text]) for the in vitro and pseudotumor studies, respectively. The relative attenuation ratio of ECLI to CLI was about 96%. The luminescent lateral resolution of the ECLI system was comparable with that of positron emission tomography (PET). The pseudotumor study illustrated the feasibility and applicability of the ECLI system in living organisms, indicating the potential for translating the CLI technology to the clinic.
Hydrothermal carbonization (HTC) of carbohydrate is an interesting candidate for the preparation of carbon materials, as it provides an easy, inexpensive and environmental friendly route. However, it is difficult to prepare porous carbon materials by a straight HTC process. Herein, the solubilising technology of micelles was introduced to direct the HTC of fructose by using an amphiphilic block copolymer, poly-(4-vinylpyridine)-block-poly-(ethylene glycol) (P4VP-PEG), as a structure-directing agent. By this strategy, hierarchical porous carbon materials with tunable properties were prepared. It was found that P4VP-PEG micelles could solubilize fructose and confine the formation of primary carbon domains during a sol-gel process. And the micelle size could be adjusted easily by changing the preparation conditions. Accordingly, the particle size of the obtained carbon materials was effectively tuned from 20 to 100 nm by the direction of the primary micelle size. After calcination, the hierarchical porous carbon materials were evidenced as effective electrode materials for supercapacitor with a capacitance of ?197 F at 1 A g(-1), which was almost four times higher than the carbon materials prepared by a straight HTC process.
The kinetic adsorption profile at the DNA-gold nanoparticle (AuNP) interface is probed by following the binding and organization of thiolated linear DNA and aptamers of varying chain lengths (15, 30, 44, and 51 mer) to the surface of AuNPs (13.0 ± 1.0 nm diameter). A systematic investigation utilizing dynamic light scattering has been performed to directly measure the changes in particle size during the course of a typical aging-salting thiolated DNA/AuNP preparation procedure. We discuss the effect of DNA chain length, composition, salt concentration, and secondary structure on the kinetics and conformation at the DNA-AuNP interface. The adsorption kinetics are chain-length dependent, composition independent, and not diffusion rate limited for the conditions we report here. The kinetic data support a mechanism of stepwise adsorption of thiols to the surface of AuNPs and reorganization of the thiols at the interface. Very interestingly, the kinetic increases of the particle sizes are modeled accurately by the pseudo-second-order rate model, suggesting that DNA could possess the statistically well-defined conformational evolution. Together with other experimental evidence, we propose a dynamic inner-layer and outer-tail (DILOT) model to describe the evolution of the DNA conformation after the initial adsorption of a single oligonucleotide layer. According to this model, the length of the tails that extend from the surface of AuNPs, capable for hybridization or molecular recognition, can be conveniently calculated. Considering the wide applications of DNA/AuNPs, the results should have important implications in sensing and DNA-directed nanoparticle assembly.
To systematically collect differentially expressed genes (DEGs) from rodent models of chronic stress (CS) and apply them to research of stress-related disease. CS is an important environmental factor that may affect numerous complex diseases. Its relevant DEGs identified from rodent models provide valuable information for understanding the mechanisms underlying stress-related diseases. Currently, no suitable data tool have been developed to use such data.
Blood vessel degeneration is critically involved in nearly all types of degenerative diseases. Therefore strategies to enhance blood vessel protection and survival are highly needed. In this study, using different animal models and cultured cells, we show that PDGF-CC is a potent vascular protective and survival factor. PDGF-CC deficiency by genetic deletion exacerbated blood vessel regression/degeneration in various animal models. Importantly, treatment with PDGF-CC protein not only increased the survival of retinal blood vessels in a model of oxygen-induced blood vessel regression but also markedly rescued retinal and blood vessel degeneration in a disease model of retinitis pigmentosa. Mechanistically, we revealed that heme oxygenase-1 (HMOX1) activity is critically required for the vascular protective/survival effect of PDGF-CC, because blockade of HMOX1 completely abolished the protective effect of PDGF-CC in vitro and in vivo. We further found that both PDGF receptors, PDGFR-? and PDGFR-?, are required for the vasoprotective effect of PDGF-CC. Thus our data show that PDGF-CC plays a pivotal role in maintaining blood vessel survival and may be of therapeutic value in treating various types of degenerative diseases.
Prion diseases are composed of a group of fatal neurodegenerative disorders resulting from misfolding of cellular prion (PrP(C)) into scrapie prion (PrP(Sc)). Sirt1, a class III histone deacetylase, has been reported to protect neuronal cells against PrP (106-126)-induced cell death. To address the potential role of Sirt1 during prion infection, the levels and enzyme activities of Sirt1 in the brains of scrapie-infected rodents, including hamsters infected with strain 263K, mice infected with strains 139A and ME7, and in prion infected SMB-S15 cells, were analyzed. Western blots revealed that endogenous Sirt1 levels were significantly decreased in all tested scrapie-infected models. Dynamic assays of brain Sirt1 levels in 263K-infected hamsters during incubation period showed a time-dependent decrease. The acetylating forms of Sirt1 target proteins, P53, PGC-1, and STAT3, markedly increased both in the brains of scrapie-infected rodents and in SMB-S15 cells, representing decreased Sirt1 activity. Immunofluorescent assays illustrated that Sirt1 predominately localized in cytosol of SMB-S15 cells but clearly distributed in nucleus of its normal partner cell line, SMB-PS. Moreover, accompanying with increase of Sirt1 level and decrease of acetyl-P53 level, treatments with Sirt1 activators SRT1720 and resveratrol in SMB-S15 cells significantly reduced PrP(Sc); at the same time, the cellular distribution of PrP proteins became normal, and the cell proliferating state was slightly improved. These data indicate that prion infection notably attenuates the Sirt1 activity in host cells. Sensitivity of the PrP(Sc) to Sirt1 activators highlights a potential role of Sirt1 in prion therapeutics.
Macrophage polarization is emerging as an important area of research for the development of novel therapeutics to treat inflammatory diseases. Within the current study, the role of Notch1R in macrophage differentiation was investigated as well as downstream effects in THP-1 monocytes cultured in "inflammation mimicry" media. Interference of Notch signaling was achieved using either the pharmaceutical inhibitor DAPT or Notch1R SiRNA and Notch1R expression, macrophage phenotypes, and anti- and pro-inflammatory cytokine expression were evaluated. Data presented shows that Notch1R expression on M1 macrophages as well as M1 macrophage differentiation is significantly elevated during cellular stress (p<0.05). However, under identical culture conditions, interference to Notch signaling via Notch1R inhibition mitigated these results as well as promoted M2 macrophage differentiation. Moreover, when subjected to cellular stress, macrophage secretion of pro-inflammatory cytokines was significantly heightened (p<0.05). Importantly, Notch1R inhibition not only diminished pro-inflammatory cytokine secretion but also enhanced anti-inflammatory protein release (p<0.05). Our data suggest that Notch1R plays a pivotal role in M1 macrophage differentiation and heightened inflammatory responses. Therefore, we conclude that inhibition of Notch1R and subsequent downstream signaling enhances monocyte to M2 polarized macrophages outcomes and promotes anti-inflammatory mediation during cellular stress.
Naringin (COG) is a flavanone with various bioactivities including an expectorant effect, antitussive effect, and inhibitory effect on asthma and acute lung injury. The aims of the present study were to investigate the antiarthritis activities of COG and elucidate the underlying mechanisms with regard to its molecular basis of action for the best combination. Arthritis was induced by intradermal injection of complete Freund's adjuvant. Sprague-Dawley rats were randomly divided into five groups with 10 rats in each group: (1) control group, (2) AA group, (3) AA?+?dexamethasone (AA?+?Dex, 2 mg/kg), (4) AA?+?COG (20 mg/kg), (5) AA?+?COG (40 mg/kg). Paw swelling was measured, and the production of tumor necrosis factor-? (TNF-?), interleukin (IL)-1?, and IL-6 was detected by enzyme-linked immunosorbent assay (ELISA) in serum. Pathological changes of joints tissues were observed by hematoxylin and eosin (HE) staining. Apoptosis of synovial tissues was measured by terminal dUTP nick-end labeling (TUNEL) assay. The expressions of apoptosis-related molecules, including Bcl-2 and Bax, were determined by Western blotting. In both COG and Dex treatments, COG and Dex significantly suppressed paw swelling in AA rats. Moreover, COG and Dex significantly suppressed the production of TNF-?, IL-1?, and IL-6 in serum. HE staining study demonstrated that COG and Dex significantly suppressed pathological changes of joints tissues; TUNEL assay demonstrated that COG and Dex induced apoptosis of AA via regulation of the protein expression of Bcl-2 and Bax. These data suggest that COG may have therapeutic potential for RA.
With the aim of maximally reducing imaging dose while meeting requirements for adaptive radiation therapy (ART), we propose in this paper a new cone beam CT (CBCT) acquisition and reconstruction method that delivers images with a low noise level inside a region of interest (ROI) and a relatively high noise level outside the ROI. The acquired projection images include two groups: densely sampled projections at a low exposure with a large field of view (FOV) and sparsely sampled projections at a high exposure with a small FOV corresponding to the ROI. A new algorithm combining the conventional filtered back-projection algorithm and the tight-frame iterative reconstruction algorithm is also designed to reconstruct the CBCT based on these projection data. We have validated our method on a simulated head-and-neck (HN) patient case, a semi-real experiment conducted on a HN cancer patient under a full-fan scan mode, as well as a Catphan phantom under a half-fan scan mode. Relative root-mean-square errors (RRMSEs) of less than 3% for the entire image and ~1% within the ROI compared to the ground truth have been observed. These numbers demonstrate the ability of our proposed method to reconstruct high-quality images inside the ROI. As for the part outside ROI, although the images are relatively noisy, it can still provide sufficient information for radiation dose calculations in ART. Dose distributions calculated on our CBCT image and on a standard CBCT image are in agreement, with a mean relative difference of 0.082% inside the ROI and 0.038% outside the ROI. Compared with the standard clinical CBCT scheme, an imaging dose reduction of approximately 3-6 times inside the ROI was achieved, as well as an 8 times outside the ROI. Regarding computational efficiency, it takes 1-3?min to reconstruct a CBCT image depending on the number of projections used. These results indicate that the proposed method has the potential for application in ART.
Systemic sclerosis (SSc) is a kind of autoimmune disease characterized by inflammatory and endothelial dysfunction. Asymmetric dimethylarginine (ADMA), as an endogenous nitric oxide synthase inhibitor, can cause or contribute to the inflammatory syndrome and endothelial dysfunction. Recently, increased ADMA levels have been demonstrated in SSc, revealing that ADMA might play an important role for the associated manifestations of SSc. Besides, ADMA may play a significant role in the level of NO, which is produced by arginine. In the review, we discuss the role of arginine and ADMA in patients with SSc.
Although the prevalence and incidence of diabetes have increased in the United States in recent decades, no studies have systematically examined long-term, national trends in the prevalence and incidence of diagnosed diabetes.
Anxiety is usually generated because of the threatened feeling. The data of electrocardio, respiration, blood volume pulse and skin conductance signals were collected. The arithmetic of Relief were used for the feature selection and combined with k-Nearest Neighbor (kNN) arithmetic and Support Vector Machine (SVM) arithmetic for classification. The results show that the combination of Relief-SVM is better than combination of Relief-kNN on the recognition of anxiety state. The emotion recognition based on multi-physiological signals is superior to that based on one single signal.
Hispolon is isolated from Phellinus igniarius and exhibits antitumor activity. Here, we explored the effects of hispolon on the lung cancer A549 and H661 cells. Cells were incubated with various concentrations of hispolon (0, 5, 10, 20, 40, 80 or 160?M) for 12, 24, 48 or 72h. Cell viability was examined by MTT assay. Cell cycle and apoptosis assay were assessed by flow cytometry. Hispolon decreased cell viability in a dose- and time-dependent manner. The cell cycle distribution showed that hispolon enhanced the accumulations of the cells in G0/G1 phase. Mechanically, hispolon decreased the expression of G1-S transition-related proteins: Cyclin D1, cyclin E, CDK2, CDK4 and CDK6, but increased the expression of CDK inhibitor p21(CIP1) and p27(KIP1). Moreover, hispolon induced cell apoptosis through activation of the mitochondrial pathway, evidenced by the loss of mitochondrial membrane potential, the release of cytochrome c into cytosol, and the cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP) in hispolon-treated cells. Additionally, hispolon enhanced the expression of p53, specific silencing of which almost completely reversed hispolon-mediated antitumor activity. Moreover, hispolon treatment was more effective on H661 cells than on A549 cells in inhibiting cell viability and inducing cell apoptosis. Our results indicate that hispolon inhibits the cell viability, induces G0/G1 cell cycle arrest and apoptosis in lung cancer cells and p53 plays a critical role in hispolon-mediated antitumor activity.
The aim of this study is to determine if contrast enhanced (CE) sampling perfection with application-optimized contrasts using different flip-angle evolutions (SPACE) images can provide clearer pituitary adenoma margin than conventional CE T1-weighted spin echo (T1-SE) sequence for cavernous sinus (CS) invasion evaluation.
The binding of antigen to the B cell receptor (BCR) stimulates the assembly of a signaling complex (signalosome) composed initially of the kinases Lyn, spleen tyrosine kinase (Syk), and Bruton's tyrosine kinase (Btk), as well as the adaptor protein B cell linker (BLNK). Together, these proteins recruit and activate phospholipase C-?2 (PLC-?2), a critical effector that stimulates increases in intracellular Ca(2+) and activates various signaling pathways downstream of the BCR. Individuals with one copy of a mutant PLCG2 gene, which encodes a variant PLC-?2 that lacks the autoinhibitory C-terminal Src homology 2 (cSH2) domain, exhibit PLC-?2-associated antibody deficiency and immune dysregulation (PLAID). Paradoxically, although COS-7 cells expressing the variant PLC-?2 show enhanced basal and stimulated PLC-?2 activity, B cells from PLAID patients show defective intracellular Ca(2+) responses upon cross-linking of the BCR. We found that the cSH2 domain of PLC-?2 played a critical role in stabilizing the early signaling complex that is stimulated by BCR cross-linking. In the presence of the variant PLC-?2, Syk, Btk, and BLNK were only weakly phosphorylated and failed to stably associate with the BCR. Thus, BCRs could not form stable clusters, resulting in dysregulation of downstream signaling and trafficking of the BCR. Thus, the cSH2 domain functions not only to inhibit the active site of PLC-?2 but also to directly or indirectly stabilize the early BCR signaling complex.
RunX2 has been identified to crucially regulate the osteolysis in giant cell tumor of bone. MiR-30a is an intronic miRNA identified as tumor suppressor, but little is known about its role in giant tumor cell of bone. In our research, we reported miR-30a was down-regulated in GCT whereas RunX2 was highly expressed. Further research proved that miR-30a can regulate the expression of RunX2 by binding to its 3'-UTR, which influence the osteoclast differentiation and osteolysis formation. Thus, these results suggest that miR-30a could directly target RunX2 and participate in osteolysis in GCT.
Alcohol consumption is accounted for a large proportion in patients with multiple sclerosis (MS) and may be a modifiable lifestyle factor that affects the risk of developing the disease. The epidemiological studies about the association between MS and alcohol consumption have got corresponding studies during the last decade. It has been suggested that alcohol consumption was associated with mood disorders, disability and even onset of MS, but a common theme is lacking. To make an understanding of the effect of alcohol consumption on MS, the related epidemiological evidence and potential mechanisms are reviewed.
Baicalin extremely fine powder was made by using ball-mill and the effect of micronization on the micromeritics properties of baicalin was studied and analyzed. The microstructures of baicalin ordinary and extremely fine powder were compared by scanning electron microscope, differential scanning calorimeter and X-ray diffraction and the powder characteristic of them was investigated. The hygroscopicity was studied. The effect of micronization on the dissolution of baicalin was investigated. The results showed that the chemical constituents of baicalin were not changed after micronization with better compressibility. It was confirmed that micronization technology had a certain application value in promoting the insoluble component of baicalin absorption with higher dissolution.
The synaptic vesicle protein synaptotagmin-1 (SYT) is required to couple calcium influx to the membrane fusion machinery. However, the structural mechanism underlying this process is unclear. Here we report an unexpected circular arrangement (ring) of SYT's cytosolic domain (C2AB) formed on lipid monolayers in the absence of free calcium ions as revealed by electron microscopy. Rings vary in diameter from 18-43 nm, corresponding to 11-26 molecules of SYT. Continuous stacking of the SYT rings occasionally converts both lipid monolayers and bilayers into protein-coated tubes. Helical reconstruction of the SYT tubes shows that one of the C2 domains (most likely C2B, based on its biochemical properties) interacts with the membrane and is involved in ring formation, and the other C2 domain points radially outward. SYT rings are disrupted rapidly by physiological concentrations of free calcium but not by magnesium. Assuming that calcium-free SYT rings are physiologically relevant, these results suggest a simple and novel mechanism by which SYT regulates neurotransmitter release: The ring acts as a spacer to prevent the completion of the soluble N-ethylmaleimide-sensitive factor activating protein receptor (SNARE) complex assembly, thereby clamping fusion in the absence of calcium. When the ring disassembles in the presence of calcium, fusion proceeds unimpeded.
Congenital heart disease (CHD) is one of the most common birth defects in newborns. The etiology of CHD has remained largely unknown, but it is assumed to result from the combined effects of genetic and environmental factors. Recent investigations have detected potentially pathogenic copy number variations (CNVs) in a proportion of patients with CHD. The present case-control study aimed at evaluating whether CNVs in the GATA4 and NKX2-5 genes contribute to the pathogenesis of CHD in Chinese fetuses (n = 117), by comparing the results to non-CHD control subjects (n = 100).
This study showcases the qualitative and quantitative source apportionments of size-dependent polycyclic aromatic hydrocarbons (PAHs) in road deposited sediment by means of molecular diagnostic ratio (MDR) and positive matrix factorisation (PMF) approaches. The MDR was initially used to narrow the PAH source candidates. PMF modelling was subsequently used to provide more precise source apportionment with the assistance of a multiple linear regression analysis. Through a combined qualitative and quantitative source apportionment, different potential source contributors were identified at different size fractions. Explicitly, three major contributors to sorption at the size fraction of 1000-400?m were tentatively identified as incineration (26%), coal combustion (53%) and gasoline-powered vehicle (20%). Four major contributors to the size fraction of 400-100?m were identified as gasoline-powered vehicle (25%), surface pavement (15%), diesel-powered vehicle (37%) and industrial boiler (24%). Four major contributors to the size fraction of 100-63?m were identified as cogeneration emission (13%), diesel-powered vehicle (28%), tire debris (45%) and wood combustion (14%). The potential contributors in the size fraction 63-0.45?m were identified as diesel-powered vehicle (21%), heterogeneous sources (41%) and biomass burning (38%). In addition, the highest ?16PAH concentration was found in the smallest size fraction of 63-0.45?m, which is also where the highest BaPE and TEF values for potential risk assessment occurred.
Acute liver failure (ALF) is a condition with high mortality and morbidity. Fibrosis in chronic liver disease were extensively researched, whereas fibrosis and underlying mechanism in acute liver failure remains unclear.
Somatic embryogenesis receptor kinase (SERK) proteins play pivotal roles in regulation of plant development and immunity. The rice genome contains two SERK genes, OsSerk1 and OsSerk2. We previously demonstrated that OsSerk2 is required for rice Xa21-mediated resistance to Xanthomonas oryzae pv. oryzae (Xoo) and for normal development. Here we report the molecular characterization of OsSerk1. Overexpression of OsSerk1 results in a semi-dwarf phenotype whereas silencing of OsSerk1 results in a reduced angle of the lamina joint. OsSerk1 is not required for rice resistance to Xoo or Magnaporthe oryzae. Overexpression of OsSerk1 in OsSerk2-silenced lines complements phenotypes associated with brassinosteroid (BR) signaling defects, but not the disease resistance phenotype mediated by Xa21. In yeast, OsSERK1 interacts with itself forming homodimers, and also interacts with the kinase domains of OsSERK2 and BRI1, respectively. OsSERK1 is a functional protein kinase capable of auto-phosphorylation in vitro. We conclude that, whereas OsSERK2 regulates both rice development and immunity, OsSERK1 functions in rice development but not immunity to Xoo and M. oryzae.
Suppressive oligodeoxynucleotides (Sup ODN) express repetitive TTAGGG motifs that have proven useful in the treatment/prevention of numerous inflammatory and autoimmune diseases. The mechanism underlying the immunosuppressive activity of Sup ODN is incompletely understood. Regulatory T cells (Treg) play a key role in controlling a variety of pathologic autoimmune responses. Treg are generated from activated CD4(+) T cells in a process that involves the phosphorylation of STAT family members. Current studies demonstrate that Sup ODN promote the differentiation of CD4(+)CD25(-) T cells into functionally active iTreg in vitro. When administered in vivo, Sup ODN promote the generation of iTreg in response to peptide challenge. Central to this effect is the ability of Sup ODN to block the phosphorylation of STAT1. These findings clarify the mechanism underlying the therapeutic activity of Sup ODN and support their use in Treg-based immunotherapy.
Intraductal administration of cytotoxic agents has been shown to inhibit the development of breast cancer in animal models. The object of this study was to demonstrate the safety of intraductal delivery cytotoxic agents in patients prior to mastectomy. This method is hopeful to be developed as a chemoprevention approach in patients with pre-malignant or non-invasive ductal lesions to prevent breast cancer which will be further developed.
As an integral part of the innate immunity, the complement system has been reported to involve in the pathogenesis of prion diseases (PrD). However, the states of expression and activity of complement proteins in experimental models of scrapie infection are still not fully understood. Herein, the state of complement activation, the presence, and distribution as well as localization of C3 and membrane attack complex (MAC) in the brains of several scrapie-infected rodents were comparatively assessed through various methodologies. Our data illustrated a significant increase in the total complement activity (CH50, U/ml) in several scrapie-infected rodent brains at the terminal stage and a time-dependent upregulation of C1q in 263K-infected hamsters during the incubation period, intimating the sustained and progressive activation of the classical pathway during PrD progression. Confocal microscopy revealed robust activation of C3 and its localization to various central nervous system (CNS) cells with differential morphology in the brain tissues of both 263K-infected hamsters and 139A-infected C57BL/6 mice at disease end stages. Dynamic analyses of MAC in the brains of 263K-infected hamsters and 139A-infected C57BL/6 mice demonstrated remarkably time-dependent deposition during the incubation period, which may highlight a persistently activated terminal complement components. Moreover, immunofluorescent assays (IFAs) showed that MAC-specific signals appeared to overlap with morphologically abnormal neurons rather than proliferative astrocytes or activated microglia throughout the CNS of both 263K-infected hamsters and 139A-infected C57BL/6 mice. Overall, these results indicate that the activation of the complement system and the subsequent localization of the complement components to neurons may be a hallmark during prion infection, which ultimately contribute to the neurodegeneration in PrD.
Hepatocellular carcinoma (HCC) is one of the most common cancer types, and chemotherapy plays an important role in treatment of HCC. However, long?term treatment with chemotherapeutic drugs such as 5?fluorouracil (5?FU) often results in chemoresistance, and the underlying mechanisms remain unclear. In this study, we showed that the annexin A2 (ANXA2) protein is highly expressed in hepatoma cells compared to healthy cells. Knockdown of the ANXA2 gene inhibited hepatoma cell growth, and the underlying mechanism may involve cell cycle inhibition through downregulation of ??catenin and cyclin D1. We also investigated the role of ANXA2 in chemotherapeutic treatment with 5?FU. 5?FU inhibited hepatoma cell growth, while ANXA2 overexpression reduced, and knockdown enhanced, the effects of 5?FU on hepatoma cell growth. Furthermore, ??catenin and cyclin D1 were asscociated with the ANXA2?induced resistance. Taken together, our data suggest that the ANXA2 protein is a critical factor in HCC and that its downregulation can enhance chemotherapeutic treatment with 5?FU. ANXA2 may thus constitute a new therapeutic target for HCC.
Hyaluronic acid binding protein 1 (HABP1/gC1qR/p32), a ubiquitous multifunctional protein belonging to the hyaladherin family, has been implicated in the tumorigenesis, progression, invasion, and metastasis of several malignant tumors. However, the role of HABP1 in endometrial cancer has not yet been studied. This study aimed to detect the expression of HABP1 in endometrial cancer and explore its role in the clinicopathological features and prognosis of endometrial cancer. We analyzed HABP1 expression by immunohistochemistry in 188 endometrial cancer specimens, 43 benign endometrial lesion specimens, and 41 normal endometrium specimens and assessed using Western blot analysis. Statistical analysis showed that HABP1 was overexpressed in endometrial cancer and benign endometrial lesion compared with normal endometrium (P?0.001 and P?=?0.012, respectively). In addition, HABP1 expression was significantly higher in endometrial cancer than in benign endometrial lesion (P?0.001). High HABP1 expression was significantly associated with advanced International Federation of Gynecology and Obstetrics stage (P?=?0.019), higher histologic grade (P?0.001), deep myometrial invasion (P?=?0.013), lymphovascular space invasion (P?=?0.010), lymph node metastasis (P?=?0.015), and recurrence (P?=?0.009). Patients with high HABP1 expression had a poorer overall survival (OS) and disease-free survival (DFS) than patients with low HABP1 expression (P?=?0.015 and P?=?0.012, respectively). Multivariate Cox regression analysis showed that the HABP1 expression status was an independent prognostic factor of OS and DFS (P?=?0.025 and P?=?0.022, respectively) in patients with endometrial cancer. Our results indicated that overexpression of HABP1 may serve as a new biomarker to predict the progression and prognosis of endometrial cancer.
Tissue factor (TF)/VIIa/protease?activated receptor 2 (PAR2) has been shown to trigger the ERK1/2 signaling pathway. This was shown to be closely associated with the proliferation and migration of SW620 colon cancer cells; however, the detailed mechanisms remain unclear. The aim of the present study was to elucidate the effects of calcium signaling on the proliferation and migration of SW620 cells induced by coagulation factor VIIa. The results demonstrated that VIIa and PAR2 agonist PAR2?AP increased [Ca2+]i in SW620 cells. In addition, VIIa?and PAR2?AP?induced ERK1/2 activation was inhibited by thapsigargin (TG)?induced depletion of intracellular Ca2+ stores and EGTA?mediated removal of extracellular Ca2+. It was also identified that VIIa and PAR2?AP?induced proliferation and migration of SW620 cells was modulated by EGTA and TG. Taken together, the present results indicate that VIIa triggers calcium signaling in SW620 cells, in a TF?dependent manner, which is critical for VIIa?induced ERK1/2 activation in SW620 cells. These results suggested that calcium signaling had a vital role in the proliferation and migration of SW620 cells.
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