Objective To investigate the effect of tea polyphenols on global cerebral ischemia reperfusion injury in rats.Method Forty-five pathogen-free male SD rats weighing 180-220 g were randomly divided into 3 groups( n =15 each):sham operation group (group S),cerebral ischemia reperfusion group (group IR) and tea polyphenols group (group TP).Global cerebral ischemia reperfusion injury was establish by four-vessel occlusion method.At 24 h of reperfusion,five rats were chosen and Evan's blue(EB) was injected iv,and then sacrificed and brain was removed for determination of EB content; another five rats were sacrificed and brain was removed for determination of water content; five rats were chosen for Morris water maze test.Result Compared with group S,EB content and water content in brain tissue were increased in groups IR and 'rP,and escape latency was prolonged,frequency of crossing the original platform was reduced in group IR ( P ＜ 0.05 ).Compared with group IR,EB content and water content in brain tissue were decreased,and escape latency was shortened,frequency of crossing the original platform was increased in group Tp ( P ＜ 0.05).Conclusion Tea polyphenols can attenuate global cerebral ischemia reperfusion injury in rats.

C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.

Objective To investigate the effect of ketamine on depression-like behaviors at different developmental stages of offspring rat exposed to prenatal restraint stress (PRS).MethodsPregnant SD rats were randomly divided into control group (n=6) and PRS group (n=8).The dams of PRS group received three times(45 minutes/time)restraint stress every day.The anxiety-like and depression-like behaviors of the offsprings of the two groups were tested in the stage of juvenile,adolescence and early adulthood.Then the antidepressant effect of ketamine on prenatal stress rats at different developmental stages was observed.ResultsIn the open-field test,the time in the central area of the offspring rats in PRS group at different developmental stages (juvenile(2.50±0.43)s,adolescence(9.17±1.05)s,early adulthood(8.33±0.92)s) were significantly lower than those of the control group((8.33±1.05)s,(19.17±1.06)s,(18.83±1.30)s,all P<0.05).In the forced swimming test,the immobility time in the offspring rats of PRS group at the different developmental stages (juvenile(192.50±10.82)s,adolescence(182.75±10.12)s,early adulthood(199.88±9.20) s)were significantly higher than those of control group((76.00±19.00)s,(96.30±12.91)s,(108.30±10.98)s,all P<0.05).Ketamine could quickly and strongly reduce the immobility time of the offsprings exposed to PRS in the stage of adolescence and early adulthood (P<0.01),but the effect was weaker in the juvenile offsprings (P<0.05).ConclusionPRS leads to persistent anxiety-like and depression-like behavior in offsprings and ketamine exerts a good antidepressant effect on the offspring rats in the stage of adolescence and early adulthood.

BACKGROUNDThe feasibility of the mapping of quantitative cerebral blood flow (CBF) named flow-sensitive alternating inversion recovery exempting separate T1 measurement (FAIREST) is still controversial. This study aimed to evaluate the reliability of FAIREST in the measurement of regional CBF (rCBF) in healthy volunteers.

METHODSEighteen healthy volunteers underwent magnetic resonance (MR) scanning with the sequence of FAIREST. While they were at rest, rCBF values were obtained in various brain regions of interest (ROIs). The same scheme was repeated on every subject after two weeks. Statistical analysis was made to determine the effect of location, scan and side on the measurement of rCBF.

RESULTSThe mean CBF values were (122+/-28) ml x (100 g)(-1) x min(-1) and (43+/-10) ml x (100 g)(-1) x min(-1) in the gray and white matter respectively. There was significant main effect of location (t=-12.5, P<0.01), but no significant effect of side. Paired t-test of ROIs in the same slice showed no significant difference in most sites between two scans, except in the gray matter of the bilateral frontal lobes (t=2.18-2.34, P <0.05). However, the rCBF values of the same structure obtained from different slices showed a significant difference (t=-3.49, P<0.01).

CONCLUSIONFAIREST is a reliable technique in the measurement of rCBF, but different imaging slice may affect the agreement of rCBF across the scans.

Adolescent ; Adult ; Cerebrovascular Circulation ; Female ; Humans ; Magnetic Resonance Imaging ; Male

OBJECTIVETo construct a eukaryotic expression vector for expressing hepatitis C virus (HCV) recombinant NS5B-EGFP fusion protein and obtain a stable transfected HepG2 cell line.

METHODSThe coding region of NS5B gene of HCV was amplified by PCR and was digested by Xho I/Kpn I. This fragment was inserted into pEGFPN3 with T4 ligase and transformed E. coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into HepG2 cell by Lipofectin AMINE 2000. Cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level NS5B-EGFP fusion protein was obtained.

RESULTSThe eukaryotic expression vector named pEGFPN3-ns5b was successfully constructed and the stable transfected HepG2 cell line expressing NS5B-EGFP fusion protein was obtained.

CONCLUSIONThe stable transfected HepG2 cell line could express NS5B-EGFP fusion protein, could be used for anti-HCV infection with ns5b gene as the target.

Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Viral Nonstructural Proteins ; genetics ; metabolism

This prospective study was conducted to assess the rate of resolution of second trimester placenta previa in women with anterior placenta and posterior placenta,and that in women with and without previous cesarean section.In this study,placenta previa was defined as a placenta lying within 20 mm of the internal cervical os or overlapping it.We recruited 183 women diagnosed with previa between 20+0 weeks and 25+6 weeks.They were grouped according to their placenta location (anterior or posterior) and history of cesarean section.Comparative analysis was performed on demographic data,resolution rate of previa and pregnancy outcomes between anterior group and posterior group,and on those between cesarean section group and non-cesarean section group.Women with an anterior placenta tended to be advanced in parity (P=0.040) and have increased number of dilatation and curettage (P=0.044).The women in cesarean section group were significantly older (P=0.000) and had more parity (P=0.000),gravidity (P=0.000),and dilatation and curettage (P=0.048) than in non-cesarean section group.Resolution ofprevia at delivery occurred in 87.43％ women in this study.Women with a posterior placenta had a higher rate of resolution (P=0.030),while history of cesarean section made no difference.Gestational age at resolution was earlier in posterior group (P=0.002) and non-cesarean section group (P=0.008) than in anterior group and cesarean section group correspondingly.Placenta location and prior cesarean section did not influence obstetric outcomes and neonatal outcomes.This study indicates that it is more likely to have subsequent resolution of the previa when the placenta is posteriorly located for women who are diagnosed with placenta previa in the second trimester.

Objective To observe the outcomes of living donor liver transplantation (LDLT) for children with biliary atresia (BA) and to summarize the clinical experiences. Methods Forty-four BA patients (26 boys and 18 girls) underwent LDLT between October 2006 and December 2010. Mean (SD) and median (range) age at operation was (12.1 ± 9.0) months and 9 (6-60) months,respectively. The 44 donors were lineal relatives to the consorted recipients. Their mean (SD) and median (range) age at operation was (32. 7 ± 8. 0) months and 31 (20～54) years, respectively. All donor graft types were the left lateral segments with compatible ABO blood groups. Clinical data,including pre-operative evaluations, surgical technique, postoperative management and outcomes in all donors and recipients were retrospectively analyzed. Results All donors were followed up for (17. 5 ± 13. 3) months. No donor mortality was encountered, with a minimal morbidity and no long-term sequelae. Nine out of 44 recipients died. Three patients died of portal vein thrombosis (PVT), one of hepatic artery thrombosis (HAT), two of biliary complications, one of surgical site infections, one of abdominal bleeding and one of pulmonary infection. The overall 1-year and 2-year cumulative survival rate in recipients was 81. 2% and 76. 1 %, respectively. No re-transplantation was done. Postoperative complications included PVT, HAT, biliary leakage and refluxing cholangitis, pulmonary infections,surgical site infections and acute rejection. Conclusion LDLT has been the effective treatment for pediatric recipients with BA and provides favorable prognosis. To improve prognosis of recipients, the key points are pre-operative evaluations, surgical technique, and postoperative management

Objective To characterize the clinical course of biliary complications after right lobe living donor liver transplantation (RL-LDLT) and to identify the independent risk factors for biliary strictures.Methods 105 consecutive RL-LDLT recipients operated from April 2007 to April 2010 were followed up. The clinical and operative data were reviewed. The biliary complications and independent risk factors of biliary stricture were studied.Results The median follow-up duration was 49.5 months ranging from 562 to 1675 days.A total of 40 patients (38.1 ％) experienced 11 bile leak episodes (10.4％ ) and 37 (35.2％) biliary stricture episodes after transplantation.Bile leaks occurred at a median time of 9 days ranging from 4 to 54 days after transplantation.For biliary strictures,the occurring time was delayed and scattered wide with a median of 7.6 months ranging from 12 to 790 days after transplantation. Moreover, the biliary stricture incidence in the first year after transplantation was significantly higher than later.The independent risk factors for biliary strictures were CMV infection,bile leaks and bile duct size (≤3 mm).Conclusion The independent risk factors for biliary strictures after RL-LDLT were CMV infection,bile leaks and bile duct size (≤3mm).In order to avoid biliary complications,careful preoperative evaluations are necessary. The dissection of bile ducts should be meticulous to protect its blood supply.CMV infection should be prevented after transplantation.Close surveillance of biliary complications should be given to RL-LDLT recipients during the first year after transplantation.

Objective To explore the protective effects of diazoxide on injury of human renal tubular cell(HK-2)induced by serum obtained from neonates with asphyxia.Methods HK-2 cells was used as the target cel1.The attacking concentration of serum obtained from neonates with asphyxia was 200 mL/L.The experiment was designed as 3 groups.HK-2 cells were divided into control group,asphyxia group,and diazoxide group.Control group:joined nutrient fluid including 100 mL/L embryo cow blood serum.Asphyxia group:joined nutrient fluid including the isometric 200 mL/L serum obtained from neonates with asphyxia.Diazoxide group:the diazoxide was joined nutrient including the isometric 200 mL/L serum obtained from neonates with asphyxia fluid.The diazoxide density finally was 100 ?mol/L.Then the change of morphology was observed and photographed under inverted microscope,and the cell viability was measured by methyl thiazolyl tetrazolium method,and the leakage rate oflactate dehydrogenase(LDH)was determined by biochemical methods.Results Under inverted microscopy,HK-2 cells in control group pastes the wall to be good,assumes the paving stone type,into flat polygon,fission many,the cell arrangement was close,connection large expanse,quantity were many.Compared with control group,the HK-2 cell to suffer injury obviously,the shape changed,become the anomalous circular or the ellipse by the model flat polygonal cell,the intercellular space crevice enlarged,the connection was loose,intercellular space obviously many cell fragmented.Living cell quantity reduced obviously,the cell vigor dropped,and the leakage rate of LDH increased significantly in asphyxia group(P

OBJECTIVETo identify and characterize the epitope associated with the virus attachment protein (VAP) of hantaan virus.

METHODSThe monoclonal antibody 3G1 was used as the ligand to biospan from a phage-displayed 12-amino acid peptide library, then the positive phage clones were chosen and sequenced. The amino acid sequences of them were compared with that of hantaan virus G2 in homology. The characteristics of positive phage were studied by IFA and ELISA. A decapeptide combining to cell membrane was observed under laser scanning confocal microscope (LSCM).

RESULTSThe conservative motif PX(1-2) HX(0-2) H displaying on positive clones shared homologous amino acid sequence with G2 96YPWHTAKCHY105.

CONCLUSIONG2 96YPWHTAKCHY105 might play some roles in virus binding to host cell, and might be a possible key epitope of hantaan virus VAP.

Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; immunology ; metabolism ; Hantaan virus ; immunology ; Microscopy, Confocal ; Oligopeptides ; immunology ; metabolism ; Peptide Library ; Vero Cells ; Viral Proteins ; immunology ; metabolism