Objective To determine the best extraction process of ginkgo leaves with the total transfer rate of total flavonol glycosides and terpene lactones as the index.Methods The effects of ethanol concentration, solid-liquid ratio and extraction time on extraction process were investigated by orthogonal design method, and the contents of total flavonol glycosides and terpene lactones were detected by HPLC to calculate transfer rate.Results The optimum extraction conditions were as follows:85% ethanol refluxing and extracting for three times;the first time extracting with five-fold amount of solvent (V/W) for 3 hours;the last two times with three-fold solvent (V/W) for 2 hours.Conclusion This extraction process has the advantages of simplicity of operator, reason, energy conservation, high efficiency, and is suitable for industrial production.

Objective To assess the cesarean section scar morphology and size with transvaginal ultrasound and the healing of incision diverticulum after the repairing operation.Methods Forty cases with cesarean section scar defects needed repairing operation,40 cases of cesarean section without symptoms and 40 cases of vaginal delivery were involved.The scar condition and measured the size of cesarean section defects were observed.For the 40 cases needed repairing operation,the healing of the scar and measured the size of the defects were observed which still existed before and after the surgery.For the transvaginal delivery cases the thickness of uterine isthmus were measured.Results After the scar defects repairing operation,there were 9 cases who still had diverticulum,but the defects were smaller than that before operation (P <0.05).The symptoms were relieved.Among the 40 asymptomatic cases,there were 1 1 cases had defects,but the diverticulum were smaller than that of needed operation patients (P < 0.05 ). Conclusions The transvaginal ultrasound is a noninvasive and convenient method to observe the cesarean section scar.

【Objective】To construct the c-myb antisense RNA recombinant retroviral vector and its packaging cell line.【Methods】The segment of c-myb gene was cloned into pUC19 with TA cloning method after amplication by RT-PCR,and then was subcloned into retroviral vector pDOR.The recombinant retroviral vector named pDOR-myb was transfected into retroviral package cell line PA317 after selection with G418.【Results】Sequencing data indicated that the c-myb gene was exactly identical to the sequence in the GenBank.The segment of c-myb gene was inserted directionally into pDOR.Resistant colonies were obtained and the titers of pDOR-myb were 5.2×104～9.5×104 CFU/mL.【Conclusion】The recombinant retroviral vector containing c-myb gene is successfully constructed and its packaging cell line PA317/pDOR-myb was established.

98%. In the pDOR-myb infected HSCs, c-myb expression levels, the cell proliferation, and ? 1-Ⅰ collagen mRNA expression were repressed significantly. CONCLUSIONS: c-myb plays a key role in the activation and proliferation of HSC. c-myb antisense RNA can inhibit cell proliferation and ? 1-Ⅰ collagen mRNA expression in the infected HSC. These data suggest that inhibition of c-myb gene expression would be a potential way for the treatment of liver fibrosis.

Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; mortality ; Humans ; Infant ; Male

AIM: To Screen and identify human single-chain variable fragment (ScFv) specific to hepatitis B virus core protein and determine its gene sequence. METHODS: The recombinant phages were panned by HBcAg coated in a 96-pore plate and 48 clones were identified specific to HBc after three rounds of panning. The specificity of ScFv from the positive clone was determined by ELISA. Then, the soluble ScFv was expressed in E.coli. HB2151 and secreted in the supernatant. Subsequently, SDS-PAGE and dot blot were performed to identify the ScFv in the supernatant and cell lysate. The gene of ScFv specific to hepatitis B virus core protein was sequenced. RESULTS: The ScFv screened from phage antibodies has a specific combination character with hepatitis B virus core antigen. Soluble ScFv was confirmed to express in E.coli. HB2151 and secrete in the supernatant. The sequence of ScFv gene conformed to that of heavy chain and kappa chain of human immunoglubulin. CONCLUSION: Human ScFv specific to hepatitis B virus core protein has been identified by means of the phage display technology, and its gene sequence has been determined.

AIM: To investigate the function and morphological changes of long-term cultured primary rat hepatocytes. METHODS: Rat primary hepatocytes were isolated by two-step in situ collagenase perfusion method, and then were purified by density and grade centrifugal method with Percoll. Cell viability was observed by 0.4% trypan blue. The hepatocytes were seeded into 6 wells plate with HepatoZYME-SFM medium. ALT, AST, albumin and urea levels in the supernatant were measured, CYPⅠA1 was detected with EROD method. RESULTS: (2-3)?108 cells per whole liver were obtained with viability and purity above 90% after purified with Percoll. Hepatocytes cultured in HepatoZYME-SFM grew well with normal hepatocyte morphometrics. ALT, AST levels in the supernatant decreased after 3-day culture, and kept at a stable level after 6-9 days. Albumin secretion and urea synthesis were maintained at high levels in 18 days, while CYPⅠA 1 enzyme activity was only detected in 3-6 days. CONCLUSIONS: Percoll was used to increase the viability and purity of freshly isolated rat hepatocytes. Hepatocyte morphometrics and their biological synthesized function are effectively maintained in HepatoZYME-SFM medium.

AIM: To construct an eukaryotic expression vector of human single-chain variable fragment ~against hepatitis B virus core protein (anti-HBc ScFv) and detect its expression in HepG2 cells. METHODS: Anti-HBc ScFv genes were amplified from the plasmids abstracted from positive clone and inserted into pEGFP-c1 vector that contained green fluorescent protein gene. The recombinant plasmids were transfected into HepG2 cells, and resistant clones were obtained by G418 selection. The expression of the gene of fusion protein was determined by fluorescent invert microscope and ELISA. RESULTS: Recombinant plasmids were successfully constructed. The plasmid transfected HepG2 cells were obtained by G418 selection. Specific fluorescence was observed in HepG2 cells 48 hours after transfection. ELISA analysis confirmed the expression of anti-HBc ScFv in the cells. CONCLUSION: The construction of human anti-HBc ScFv eukaryotic expression vector and its expression in HepG2 cells lay the foundation for advanced research of intracellular anti-HBc ScFv.

Objective To explore the clinical effect of transvaginal repair of cesarean scar diverticulum (CSD). Methods Totally 64 patients of CSD in the First Maternity and Infant Hospital, Tongji University between Mar. 2013 and Sept. 2014 underwent transvaginal repair of CSD were reviewed retrospectively and followed. Result All the patients had a prolonged period, and the duration was (14.8± 3.5) days; all the patients were received the transvaginal repair of CSD, there was no intra-operative complications, the procedures were successfully performed in all patients. The mean operation time was (67± 12) minutes, the mean blood loss was (53±32) ml, and the mean length of hospital stay was (4.0±1.1) days. All patients were followed after the operation, the duration of menstruation was (8.1 ± 3.5) days shorter in average, which was statistically significant (P<0.01);the operation effective rate was 94%(60/64) to assess the clinic syptoms, the operation effective rate was 95%(61/64) for anatomic assessment. The distance of the CSD from the serosa became thicker after surgery significantly, the distance was thicker (3.4 ± 0.4) mm compared with preoperation (P<0.01). Conclusions Transvaginal repair of CSD offers minimal invasiveness, good exposure and accurate resection. It is worth to be popularized in the treatment of patients with CSD.

Objective Toexploretheriskfactorsofchronicinjuryofmedialcollateralligament(MCL)inpatientswithosteoarthritis(OA). Methods Thestudywasconductedretrospectivelyandatotalof191patientsinourhospitalfromFebruary2017toApril2018were collected,amongthem,86casesofOApatientswithchronicinjuryofMCLastheobservationgroup,42casesofpatientswithacute MCLinjurycausedbytraumaasthecontrolgroup1,63casesofOApatientswithoutMCLinjuryascontrolgroup2.TheMRImanifestationsof kneejointsintheobservationgroupandthecontrolgroup1werecompared,thedifferencesofthedamagemechanismsbetweenthe twogroupswereanalyzed.TheMRImanifestationsandrelatedclinicaldataoftheobservationgroupandthecontrolgroup2wereanalyzed,and thefactorsthatmightcausethechronicinjuryofMCL wereidentifiedbyunivariateregressionanalysis,thenwereincludedinthe non-conditional L o g istic regression m odel for m ultivariate analysis and the risk factors and protective factors of chronic injury of MCLwerefinallyscreenedout.Results Thedifferencesinthe MRImanifestationsbetweentheobservationgroupandthecontrol group1,includingthegradingoftheMCLinjury,thegradingandthedislocationofmedialmeniscus,thedistributionofbonemarrow edema,theanteriorandposteriorcruciateligamentinjuriesandthestenosistypesofthejointspacewerestatisticallysignificant(P<0.05).The resultsrevealedthatthechronicinjuryofMCLwasrelatedtoage,sex,medialmeniscusdislocation,thegradingofmedialmeniscus, osteophyte,anteriorcruciateligamentinjury,posteriorcruciateligamentinjury,andthestenosistypeofthejointspaceusingtheunivariateanalysis (P<0.05).Theresultsrevealedthatosteophyte(OR=38.231,95%CI:6.573-222.370),medialmeniscusdislocation (OR=6.504, 95%CI:1.508-28.046),anteriorcruciateligamentinjury(OR=7.236,95%CI:1.188-44.090)wereriskfactorsforchronicinjury ofMCLinOApatientsandlateraljointspacestenosis(OR=0.014, 95%CI:0.002-0.092)andpatella-femoraljointspacestenosis (OR=0.006,95%CI:0.000-0.071)wereprotectivefactorsusing multiple L o g istic regression model.Conclusion Thepathogenicfactorsaredifferentbetweenchronicinjuryof MCLin OA patients andacuteinjuryofMCL.Osteophyte,medialmeniscusdislocation,anteriorcruciateligamentinjuryareriskfactorsforchronicinjury ofMCLinOApatients,andlateraljointspacestenosisandpatella-femoralspacestenosisareprotectivefactors.