In order to identify whether functional L-type calcium channels are expressed in neural stem cells(NSCs) from rat embryonic hippocampus, and whether L-type calcium channels participate in the modulation of proliferation and differentiation of NSCs, the rat embryonic hippocampal tissue was dispersed into a single cell suspension, and the dissociated cells were cultured in serum-free DMEM/F12 medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), N-2 and B27 supplement. Immunofluorescent labeling showed an expression of nestin-positive cells. Following five-days culture in differentiation medium, neuron-like and astrocyte-like cells were observed, which expressed ?-tubulin Ⅲ(Tuj1) and Glial fibrillary acidic protein (GFAP), respectively. Western blot analysis showed an expression of Cav1.2?1C subunits in NSCs, but no Cav1.3?1D subunits. Moreover, L-type calcium channel currents were recorded in those cells by using whole-cell patch clamp techniques. It was found that activation of L-type calcium channels promotes proliferation and differentiation to neuronal type of NSCs. The results indicated that rat embryonic hippocampal NSCs express functional L-type calcium channels, L-type calcium channels modulate proliferation and differentiation of NSCs.

Objective To kown X-ray and clinic feature of thoracic metastasis of primary malignant tumor of female genital organs,so that to improve diagnostic accuracy.Methods All 204 cases of female genital tumor were confirmed by operation and pathology.The thoracic films of postero-anterior and lateral projections were performed before and after operation and chemotherapy in all patients, CT scan was performed in 30 cases CA-125 in 48 cases and HCG in 23 cases were done.All data of patients were reviewed and analysed by 3 doctors.Results 25 cases presented pleural fluid,CA-125 rised to 125-3765 u/ml;12 cases presented pulmonal shadow,hCG rised to 92-200000 mIU/ml.One of Choriocarcinoma had appeared pulmonal metastasis in normal pregnancy.Conclusion The metastasis of pleural fluid is most seen in ovary carcinoma,pulmonary metastatic tubercle is most manifestion in choriocarcinoma.Pulmonary metastasis of choriocarcinoma can't be ignored in normal pregnancy.

Objective In this study,we elucidated the role of high mobility group box chromosomal protein 1 (HMGB1) in collagen-induced arthritis (CIA) rat and the antagonist role of ethyl pyruvate by using a rat model of CIA as the research object by comparing the expression of HMGB1 in normal control group,CIA model group and ethyl pyruvate group.Methods Thirty-six rats were randomly divided into 3 groups (n=12):normal control group,CIA group and ethyl pyruvate group.Then the 6 rats were dissected at the 6th,9th week respectively.Thc expression of HMGB1 was analyzed by immunohistochemistry and Pathology-image analysis software in the cytoplasma.The expression of HMGB1 mRNA with real time-polymerse chain reaction (PCR) was evaluate,and the HMGB1 expression of each group were compared with t-test.Results The immunohistochemical results of HMGB1 showed that the expression intensity in the normal control group,CIA model group and ethyl pyruvate group was 2.1±0.6,7.3±1.2,6.0±1.2 respectively at the 6th week; and 2.2±0.7,12.4±4.5,5.5±1.0 at the 9th week respectively.The HMGB1 mRNA real time-PCR results had shown that the relative quantification of the normal control group and CIA model group were 1,2.865,2.602respectively at the 6th week and 1.005,4.694,1.729 at the 9th week.At those two points, the HMGB1 expressions of HMGB1 antagonist group were significantly higher than those of the normal controls (P＜0.05).In addition,there was statistical significant difference(P＜0.05) in the HMGB1 expression when compared with the placebo group.Furthermore, when the degree of HMGB1 expression among the three groups was compared,the HMGB1 antagonist group was decreased significantly (P＜0.05).Conclusion The results has demonstrated that HMGB1 could induce inflammation in the synovial tissue of CIA rats,and has provided the rationale that HMBG 1 could be the target of treating rheumatoid arthritis (RA).The results of this study have shown that ethyl pyruvate could antagonize the effect of HMGB1.This finding may provide a new therapeutic target for the treatment of RA.

AIM To research the antiasthmatic effects of Zuogui Pills,Yougui Pills and Yupingfeng Powder and their mechanism of action.METHODS Male Brown Norway rats were randomly divided into five groups.OVA was used to induce the model of chronic asthma.Five groups of asthmatic rats were given Zuogui Pills,Yougui Pills,Yupingfeng Powder by gavage once daily for 14 d,respectively.The normal and the model group were given normal saline.Penh value,serum Thl,Th2 inflammatory cytokines,and CROT and ACTH protein levels were determined.Pathological changes of airway remodeling were measured.RT-PCR method was used to measure the expression of CRH mRNA in hypothalamus,and TGF-β1 and Smad3 mRNA in lung tissues.RESULTS Penh values of Yupingfeng Powder group and Yougui Pills group were significantly lower than that of Zuogui Pills group.The Thl inflammatory cytokine of Yupingfeng Powder group was significantly higher than that of Zuogui Pills group,while the Th2 cytokine was significantly lower than that of Zuogui Pills group.In Yougui Pills group,only IL-5 was significantly lower than that of Zuogui Pills group.Serum CROT,ACTH protein and CRH mRNA of Yupingfeng Powder group were significantly higher than those in Zuogui Pills group.No significant difference was found between Yupingfeng Powder group and Zuogui Pills group with regard to TGF-β1 and Smad3 mRNA.Smad3 mRNA of Yougui Pills group was significantly higher than that of the model group.Airway remodeling was significantly reduced by Yupingfeng Powder and Zuogui Pills.The number of goblet cells in Yougui Pills group was significantly lower than that in Zuogui Pills group.CONCLUSION All the three formulaes possess significant antiasthma effects.Regulation effects of Yupingfeng Powder on Thl/Th2 balance,HPA and TGF-β1/Smad3 pathway are strong.Compared with Zuogui Pills,Yougui Pills possesses weaker TGF-β1/Smad3 regulative effects and stronger HPA regulative effects.

A glassy carbon electrode (GCE) modified with gold nanoparticles loading on the reduced graphene oxide (rGO)-multi-walled carbon nanotubes (MWCNTs) composite film was fabricated by a two-step procedure.Firstly, rGO-MWCNTs composite were prepared by in-situ chemical reduction method with hydrazine as a reducing agent.Then, AuNPs were deposited on the surface of rGO-MWCNTs using simple cyclic voltammetry.This modified electrode was characterized using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS) and electrochemical methods.Furthermore, the electrochemical behavior of bisphenol A (BPA) was also investigated using this modified electrode.The results showed that the modified electrode had high electrochemical activity for the oxidation of BPA.In 0.10 mol/L phosphate buffer solution (PBS, pH 7.0), the linear range for the determination of BPA with differential pulse voltammetry (DPV) was in the range of 5.0 × 10-9 -1.0 × 10-7 mol/L and 1.0 × 10-7-2.0 × 10-5 mol/L.The detection limit was 1.0 × 10-9 mol/L (S/N=3).The as-prepared modified electrode was successfully used to determine BPA in river water and the shopping receipt samples with recovery ranges of 97%-110% and 98%-104%, respectively.

Objective To study the recombination type of B/C genotype in hepatitis B virus.Methods The PCR was applied to amplifed the whole genes of HBV through the serums of four chronic HBV carriers who come from Jinghong distict,Yunnan province.The whole HBV genomes were ligated with pMD18-T vector and trasformed to E.coli JM109.After the positive colones were picked up,the HBV genotypes and recombinated sites were discoved through sequenced the acquired positive colones.Results All the acquired sixteen HBV sequences from the four HBV carriers were genotype B which were combinated with genotype C in some region.There are two ways of the combinations.For the first one,a 496 bp fragment from genotype C taked place the genotype B at the place of nt1825 to nt2320 of precore C/C region.For the second way,a 695 bp fragment of genotype B taked place at the both sites of nt822 to nt1020 of P gene region and nt1825 to nt2320 of precore C/C region.Conclusion A new recombination type of B/C genotype in hepatitis B virus was reported for the first time.The new Bj subgenotype was combinated with genotype C not only at the region of precore C/C but also at the place of P gene region.

We isolated a new strain of endophytic Pseudomonas G5 from the stems of Chinese parsley (Coriandrum sativum L.), and it is tentatively identified as Pseudomonas aurantiaca according to analysis of the entire substrate utilization profiles using BIOLOG Microstation system (BIOLOG, Inc, Hayward CA). An array of evidence established that many Gram-negative bacteria employ Quorum sensing (QS) system to regulate gene expression in response to cell density using small diffusible signal molecules, N-acyl homoserine lactones (AHLs), and control diverse phenotypic traits in plant-associated bacteria. In this study, we showed that Pseudomonas sp. strain G5 can produce several types of AHLs at a detectable level using Thin Layer Chromatography (TLC) analysis combined with bioreporter Chromobacterium violaceum CV026 bioassay, and N-hexanoyl-homoserine lactone (HHL, C6-HSL) with Rf value 0.4 is the major signal molecule. Furthermore, we have identified its quorum sensing system composed of PhzI and PhzR by cloning and sequencing of phzI-phzR. PhzI is responsible for synthesis of AHLs signal molecules, and PhzR is a transcriptional regulator. Finally, we heterologously expressed the recombinant plasmid pMD-phzIR in Escherichia coli JM109 and verified it using C. violaceum CV026 bioassay. The phylogenetic analysis using MEGA4 revealed highly similarities exist among the phzIR homologs, suggesting it is evolutionary well conserved in the genus Pseudomonas.
4-Butyrolactone ; analogs & derivatives ; metabolism ; Acyl-Butyrolactones ; metabolism ; Amino Acid Sequence ; Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Phylogeny ; Pseudomonas ; classification ; genetics ; isolation & purification ; Trans-Activators ; biosynthesis ; genetics

Objective To investigate the prevalence of fatigue in rheumatoid arthritis (RA) and its relationship with other clinical and functional parameters used for the evaluation of disease activity and health related quality of life. Methods The fatigue was assessed in 230 patients with RA using visual analogue scale (VAS). The correlation between fatigue and the clinical disease activity including morning stiffness, pain, PGA, physician's global assessment, TJC, TJI, SJC, SJI, DAS28, HAQ and health-related quality of life were assessed. Results The prevalence of fatigue was 85.7% and the fatigue score of 51.7% patients was higher than 50 mm. After controlled for the possible confounding factors such as age, gender and disease duration, it was found that fatigue was highly correlated with pain, disease activity, functional disability, physical health and mental health. Conclusion Fatigue is an important symptom of RA and is correlated with pain, disease activity, functional disability and health-related quality of life.

Objective To evaluate the clinical efficacy and safety of methotrexate(MTX),cyclophosphamide(CTX)and MTX plus CTX in patients with active rheumatoid arthritis(RA).Methods In a randomized,single-blinded,controlled study,90 patients were randomly assigned to receive MTX(10～15 mg/w)or CTX(400 mg/2～3 w)or MTX plus CTX(MTX 10～15 mg/w+CTX 400 mg/2～3 w).The primary end point was the proportion of patients meeting the American College of Rheumatology 20% improvement criteria(achieving an ACR20 response,)at week 24.The secondary end points were responses of the ACR50 and ACR70 improvement criteria,and the European League Against Rheumatism(EULAR)response criteria.The change from baseline in duration of pain,patient's global assessment,physician's global assessment,tender joint count/index,swollen joint count/index,health assessment questionnaire(HAQ),erythrocyte sedimentation rate(ESR)were also evaluated.The clinical efficacy and safety were analyzed at baseline,6,12 and 24 weeks respectively.Results The ACR response rate was significantly higher in the MTX plus CTX treatment group compared with MTX or CTX group at week 24.The MTX plus CTX group,MTX group and CTX group showed 81%,56% and 35% in ACR20,58%,41% and 12% in ACR50 and 19%,11% and 0 in ACR70,.respectively.At week 24,the proportion of patients achieving the EULAR moderate response in those who received combination treatment were significantly higher than those who received either MTX or CTX.The incidence of adverse events(AEs)was not significantly higher in MTX plus CTX group than MTX or CTX group.Conclusion MTX plus CTX effectively reduces the signs and symptoms of RA and is generally well tolerated by patients without significant increase in the rate of adverse events compared with monotherapy.

Objective To analyze the quality of life of patients with active rheumatoid arthritis (RA) and its relationship with other clinical and functional parameters used for the evaluation of disease activity. Methods The quality of life was assessed in 127 patients with active RA using SF-36 and was compared with non-active RA and the general population. The correlation between the quality of life and the clinical measures of disease, including morning stiffness, pain, fatigue, patient's global assessment (PGA) physician's global assessment , SJC/SJ1, TJC/TJI, DAS28, HAQ were measured. Results The patients with active RA reported significantly decreased scores in all dimensions of SF-36. Fatigue, PGA, physician's global assess-ment, DAS28 and HAQ significantly correlated with the scores in all dimensions of SF-36. Pain was correlated with the scores in all dimensions of SF-36 except RE. TJI was correlated with six dimensions. TJC was correlated with five dimensions. ESR was correlated with three dimensions. Conclusion The quality of life in patients with active RA is significantly declined compared with non-active RA and the general population. Disease activity and functional status of patients with active RA is closely correlated with the quality of life.