Objective To observe the postnatal development and perinatal electrophysiological characteristics of oligodendrocyte precursor cell (OPC) in rats. Methods Immunohistochemistry and Western blot were applied to determine the expression of NG2 OPC in cerebral cortex and hippocampus at various developmental stages of SD rats. Electrophysiological characteristics of OPC were also recorded in slices of 7-day rats. Results The majority of hippocampal and cerebral OPC exhibited stellate shape,a small cell body with a few processus. Total population of the NG2 immunopositive OPC was numerous at P7d in cerebral cortex and hippocampus. OPC expressed in adult rats with slightly more quantity. Moreover,OPC in hippocampus of P7d rats typically exhibited small inward sodium current and weak active responses,whereas only outward potassium current and inactive responses were recorded in white matter OPC of P7d rats. Conclusion Total population of OPC and relative optical density of NG2 are the highest in P7d rats at the postnatal developmental stages. OPC in cerebrum and hippocampus of P7d rats displays electrophysiological heterogeneity.

Objective To explore the effect of hypoxia on the expression of Janus kinase (JAK) in cocultured pulmonary arterial smooth muscle cells (PASMC) Methods Rat PASMC were cocultured with lung microvascular endothelial cells (LMVEC), the expression of JAK protein in PASMC was detected with Immunocytochemical method Results Expressions of JAK1 and JAK3 Protein were seen excessive positive (++++) in the cytoplasm of simple PASMC in hypoxia, and better positive (+++) in cocultured PASMC in hypoxia Compared with those of the control group, the expressions of JAK1 and JAK3 protein obviously increased in PASMC in hypoxia for 2 h, reached highest in those with hypoxia for 6 h, and decreased in those with hypoxia for 12 hours hypoxia, but still higher than those in hypoxia for 2 h Conclusion Hypoxia modulates the signal pathways and improves cell proliferation by the enhancement of the expressions of JAK1 and JAK2 proteins in co cultured PASMC

Objective To develop new methods to cultivate, retrieve and purify oligodendrocyte progenitor cells (OPCs). Methods Primary OPCs were isolated from mixed glial cultures of newborn Sprague Dawley rat forebrains by using a selective detachment procedure, further purified by differential adhesion, and maintained in a chemical conditioned medium. The characteristic of OPCs was determined by both electrophysiological recording and immunostaining. Results Immunocytochemical analysis revealed that more than 90% of the isolated cells expressed the OPCs-specific marker NG2. OPCs exhibited a moderate input resistance, delayed rectifier K+ currents (IK) that could be antagonized partly by TEA-Cl and CsCl, small tetrodotoxin (TTX) sensitive Na+ currents that failed to generate typical action potentials. Conclusion Isolation and purification by selective detachment and differential adhesion procedure are effective to obtain highly purified OPCs. OPCs, distinct from neuron, astrocyte, oligodendrocyte or microglia, possess unique physiological properties.

Objective To investigate the change of P2X7 receptor expression in neonatal rats after hypoxic-ischemic brain damage(HIBD).Methods Twenty SD rats were randomized into 2 groups,control group and hypoxic-ischemic(HI) group.Western blotting and immunohistochemistry was used to detect the protein expression of P2X7 receptor in cerebal cortex,subcortical white matter and hippocampus before and after hypoxia-ischemia.Results In comparison to the sham operation controls,a significant down-regulation of the P2X7 receptor protein was observed in ischemic cerebral cortex,subcortical white matter(P

Objective To study the imaging findings of the chest disease due to intravenous heroin abuse. Methods Twenty five cases of clinically confirmed chest disease due to intravenous heroin abuse were retrospectively analyzed. 25 cases had conventional X ray film, 6 cases had CT scanning, and 6 cases had echocardiography scanning. Results On X ray and CT, the following signs were found: lung making manifold ( n =5), small patchy shadow ( n =15), pneumatocele ( n =16), small cavity ( n =16), small node ( n =7), pleural effusion ( n =8), pneumothorax ( n =2), hydropneumothorax ( n =6), pulmonary edema ( n =2), megacardia ( n =11), multiple shaped lesion ( n =20). On echocardiography, tricuspid vegetation ( n =4) and tricuspid insufficiency ( n =4) were found. Conclusion The X ray and CT manifestations of chest inflammation due to intravenous heroin abuse are multiple. The multiple small cavities and pneumatoceles sign are of some value in the diagnosis of lung inflammation due to intravenous heroin abuse among young patients.

BACKGROUND: NOV protein encoded by nephroblastoma overexpression gene(NOV) is IGF(insulin-like growth factor) -binding protein. What is its impact on human neural stem cell(hNSC) proliferation and differentiation?OBJECTIVE: To investigate the impacts of NOV protein on hNSCs proliferation and differentiation.DESIGN: A single factor analysis of variance experimental study using cells as subjectsSETTING: Department of histology and embryology, and department of neurobiology in a military medical university.MATERIALS: Study was conducted in the Department of Histology and Embryology of the Third Military Medical University of Chinese PLA. Subjects were hNSCs cultured from 10 to 14 weeks human embryo cerebral cortex.INTERVENTIONS: COS-7 cells were transfected by NOV gene recombined plasmid. COS-7 cell and COS-7 cell modified by NOV gene conditioned culture media(COS-CM and NOV-CM) were collected and reacted with the cultured HNSCs.MAIN OUTCOME MEASURES: hNSCs proliferation was detected by 3H-TdR scintillation analysis, and hNSCs differentiation was detected by immunocytochemistry and flow cytometer(FCM).RESULTS: Both COS-CM and NOV-CM could significant promote the intake of 3H-TdR by HNSCs, of which the 1/minute of NOV-CM group was significantly higher than that of COS-CM group(P ＜ 0.05), which indicated that NOV-CM contained component that could facilitate hNSCs proliferation, and moreover, there was certain dose-effect relationship in NOV-CM' s facilitation of cellular proliferation. The results of immunocytochemistry and FCM revealed that there were more NF-200 positive cells in NOV-CM group, while many glial fibrillary acidic protein positive cells could be seen in COS-CM group.CONCLUSION: NOV protein might have facilitative effects on hNSCs proliferation and differentiation into neurons.

Objective To eonstruct three eukaryotic expression vectors containing wilde-type hGHR gene and its mutants(hGHR-E42K and hGHR-H56R) related to congenital growth hormone insensitivity, then check their expression in CHO cells. Methods With the available PUC-hGHR vector, two mutate hGHR genes (hGHR-E42K and hGHR-H56R) were obtained through mutagenesis. Then three recombinants were cloned into eukaryotic expression vectors pcDNA3.1/zeo(+) with restriction enzymatic reactions.Then with Lipofectamine2000, we trans-fected expression vectors to CHO cells and screened the stably expressed CHO cells by Zeocin. RT-PCR and/or Western blotting were used to examine hGHR and STAT5-P. Results After sequencing, two mutations were introduced to hGHR, three eukaryotic expression vectors were identified. The transfected CHO cells expressed vd-hGHR or its mutants. Compared with hGH-wt, two mutate cells (E42K and H56R) had decreased phosphorylated STAT5 levels. Conclusion Three CHO cells which stably expresses wide type hGHR and its mutants were successfully established, E42K and H56R partly interfered the phosphorylation of STAT5.

Objective To construct three eukaryotic expression vectors containing wilde-type hGHR gene and its mutants(hGHR-E42K and hGHR-H56R) related to congenital growth hormone insensitivity,then check their expression in CHO cells. Methods With the available PUC-hGHR vector,two mutate hGHR genes (hGHR-E42K and hGHR-H56R) were obtained through mutagenesis. Then three recombinants were cloned into eukaryotic expression vectors pcDNA3.1/zeo(+) with restriction enzymatic reactions. Then with Lipofectamine2000,we transfected expression vectors to CHO cells and screened the stably expressed CHO cells by Zeocin. RT-PCR and/or Western blotting were used to examine hGHR and STAT5-P.Results After sequencing,two mutations were introduced to hGHR,three eukaryotic expression vectors were identified. The transfected CHO cells expressed wt-hGHR or its mutants. Compared with hGH-wt,two mutate cells (E42K and H56R) had decreased phosphorylated STAT5levels. Conclusion Three CHO cells which stably expresses wide type hGHR and its mutants were successfully established,E42K and H56R partly interfered the phosphorylation of STAT5.

Objective To evaluate the decreased level of serum inhibin B(INHB)treated by gunadotropin releasing hormone agonist(GNRH-a)in predicting ovarian response and pregnancy in in vitro fertilization-embryo transfer(IVF-ET).Methods The prospective study enrolled 124 women given by GnRH-a+recombine follicle stimulating hormone(rFSH)+human chorionic gonadotrophin(hCG)long term stimulation protocol undergone their first cycle of IVF-ET treatment.The following predictive factors were collected and analyzed,such as age,basal level of follicle stimulating hormone(FSH),the ratio of FSH/luteinizing hormone(LH),the concentration of INHB after down-regulation,total number of antral follicle count(AFC)and mean ovarian volume. Ovarian response was evaluated by the number of oecytes obtained.A multiple regression analysis and logistic regression model were used for all possible prognostic variables to evaluate the value of difierent hormones in predicting ovariall response and pregnancy after IVF-ET.Receiver operating characteristic(ROC) analysis was used to evaluate the level of INHB in predicting the number of oocytes obtained.The sensitivity and specificity were calculated at the discriminating cut-off point Results The concentration of INHB after down-regulation showed a highly significant positive correlations with the number of oocytes obtained(r=0.435,P<0.01).The multiple regression analyses showed INHB was the most significant predictor of the number of retrieved oocytes,but INHB was not associated with IVF-ET outcome significantly(P>0.05).ROC analyses showed INHB after down-regulation had the largest area under curve(AUC)0.933(95%CI:0.878-0.988).When a threshold of 15 ng/L of INHB was established,95.5%sensitivity and 50.0% specificity in ovarian response were observed.Conclusions The level of INHB Was the best factor in predicting ovarian response in IVF-ET.Decreased level of INHB Was the early sign of ovarian reserve function failure,however,useless in predicting IVF-ET outcome.

OJECTIVE To investigate the preventive effect of Fengliao extract on ulcerative colitis of mice. METHODS Using the intestinal propulsion rate experiment and senna induced diarrhea model , the intestinal propulsion rate, diarrhea rate and index of diarrhea were observed. Mice were randomly divided into normal group,model group,mesalazine hydrochloride group and Fengliao extract 11.7, 23.4 and 46.8 g · kg-1 group. The mouse colitis model was induced by 4% dextran sulfate sodium. The mice were administraed once daily for 7 d while the disease activity index(DAI)score was calculated and the activity of tumor necrosis factor α(TNF-α),interleukin-1β(IL-1β), myeloperoxidase(MPO) and content of malondialdehyde(MDA) and nitric monoxide (NO) in colon tissue were determined. RESULTS Fengliao extract 46.8 g · kg-1 inhibited the intestinal propulsion rate(P<0.05),reduced the frequency of diarrhea and the diarrhea index(P<0.05). Results of colitis showed that the body mass of mice in the model group was significantly decreased but the DAI score increased compared with normal group(P<0.05). The activity of MPO and the contents of IL-1β,TNF-α,MDA and NO in colon mucosa were increased(P<0.01). Compared with the model group,Fengliao extract 46.8 g·kg-1 decreased the DAI score(P<0.05)while Fengliao extract 46.8 and 23.4 g · kg-1 reduced MPO activity in colonic mucosa and content of IL-1β,TNF-α,MDA and NO in colonic homogenate(P<0.05). CONCLUSION Fengliao extract can significantly improve the DSS induced colitis in mice,which is probably associated with its antispasmodic and anti-diarrheal effect as well as the reduced release of inflammatory mediators and antioxidants.