OBJECTIVE:To establish a method for the bacterial endotoxin test of vinpocetine raw material. METHODS:Ac-cording to the bacterial endotoxin test in the Chinese Pharmacopoeia(2010 edition,Ⅱ)Appendix Ⅺ E,the samples with different batches were used for interference test and bacterial endotoxin test by tachypleus amebocyte lysate from 2 manufacturers. RE-SULTS:The vinpocetine solutions with high concentration have interference effect on the agglutination reaction of bacterial endotox-in and the interference can be eliminated by diluting. CONCLUSIONS:The non-interference concentration of vinpocetine solutions is 0.25 mg/ml. Bacterial endotoxin test can be used to control the quality.

Objective To analyze the curative effect of blood transfusion on 100 patients undergoing liver transplantation,to efficiently enhance hemostasis and coagulation function of the patients,to lower the incidence of the operative complications,and to put forward the guideline of blood transfusion to the patients undergoing liver transplantation.Methods The volumes of red blood cells concentrates(RCCs),fresh frozen plasma(FFP),cryopreserved platelets,and cryoprecipitate transfused to the patients undergoing liver transplantation were measured before,during and after the operation respectively.The volumes of blood components transfused to the 50 patients in the former stage and 50 patients in the later stage of liver transplantation in our hospital were also measured respectively.Results The mean volume of the blood components transfused to each patient was 3 334ml.The ratios of the transfused blood volumes to 100 patients were 4.3%,53.4% and 42.3% before,during and after operation respectively.The rate of blood components transfusion was 100% and the ratio of transfused blood volumes to the patients between RCCs and FFP was 1.45:1.The mean volumes of the transfused blood and the operative complications of 50 patients in the former stage of liver transplantation of our hospital were significantly higher than those of the 50 patients in the later stage.Conclusion It is very vital in the scientific,safe and reasonable blood transfusion for the success of liver transplantation,which significantly lowers the incidence of operative complications,such as massive haemorrhage during and after operation,and thrombosis at inosculation of liver artery and portal vein.

Objective to construct the high titers rat Hes1 adenovirus expression vector (Ad-Hes1).Methods With the rat cDNA as a template,the Hes1 fragment was amplified by PCR,which constructed pShuttle-CMV-Hes1 shuttle plasmid by directly clone.Based on pShuttle-CMV-Hes1,pAdeno-Hes1 virus plasmid was constructed,pAdeno-Hes1 was transfected into 293 cells to package Ad-Hes1,virus titers were determined by modified TCID50.Hes1 was detected by Western blot after Ad-Hes1 infected with H9c2 myocardial cells.Results pShuttle-CMV-Hes1 shuttle plasmid and pAdeno-Hes1 plasmid were constructed successfully,with a general titer of 1.6 × 1011 PFU,Ad-Hes1 can be expressed in H9c2 myocardial cells,and its MOI value was 30.Conclusion Ad-Hes1 is successfully constructedand packaged,thus provide basis for further research on the protection effect of Hes1 on myocardium.

0.05).Conclusion The 15 STR loci of Miao ethnic group show the characteristics of high polymorphism and hereditary stability.Therefore,the obtained data can be of applicable value in human population of Guangxi Miao genetics investigation,and forensic application,as well as individual identification and parent checkup.

Objective To evaluate the safety and efficacy of self-expanding metallic stents (SEMS) in one-stage surgery for patients with left-sided malignant colorectal obstruction. Methods The clinical data of 97 patients with left-sided colorectal cancer who had been admitted to the First Affiliated Hospital of Zhejiang Chinese Medical University from May 2005 to September 2008 were retrospectively analyzed. Forty-six patients with left-sided malignant colorectal obstruction were in the test group, after the placement of SEMS, they received one-stage resection and anastomosis; 51 patients with left-sided colorectal cancer (without obstruction) were in the control group, and they received one-stage surgery. The defecation and anal exhaust time, hospital stay and incidence of postoperative complications between the 2 groups were analyzed. All data were analyzed via t test or chi-square test. Results One patient in test group had acute diffuse peritonitis which was induced by intestinal perforation at postoperative day 4, and was treated with radical resection for sigmoid cancer. One-stage surgery was successfully performed on 45 patients, and they were discharged at postoperative day 11-16. One patient died of respiratory infection and heart failure at postoperative day 10. All patients were followed up for 4-48 months. There was no significant difference in defecation and anal exhaust time, hospital stay, incidence of perioperative complications, recurrence, metastasis and survival within follow-up interval between the 2 groups (t=0.164, 1.358, χ~2 = 0.252, 1.200, 0.580, P >0.05). Conclusion SEMS is effective and safe in the treatment of left-sided malignant colorectal obstruction.

Objective To establish the A549 cell line with stable expression of HIF1α by using lentiviral vector system.Methods Primers were designed and synthesized with human HIF1α gene coding sequence by the National Center of Biotechnogical Information(NCBI) as the template.HIF1α was amplified by PCR.The HIF1α fragment recycled by enzyme digestion was recombined with prepared lentiviral vector HBLV-RFP-Puro.The recombinant plasmid was identified by PCR and gene sequencing.The recombinant plasmid and the auxiliary plasmid were co-transfect into 293T cell.After filtration and concentration of packaged virus,the viral titer was detected by using the dilution counting method.The prepared lentivirus was infected A549 cells.The drug screening was adopted to stabilize the transfected cell line.The transfection effect was detected and observed by fluorescence microscope and Western blotting.Results The HIF1α fragment amplified by PCR was successfully verified and the recombinant plasmid was successfully constructed by PCR and gene sequencing identification.High-titer LV-HIF1α was obtained by successful package.After LV-HIF1α infecting A549 cells,the cells showed the red fluorescence by fluorescence microscope.The expression level of HIF1α in the LV-HIF1α group was significant higher than that in the control group by Western blot.Conclusion The 549 cell line with HIF1α stable expression mediated by lentivirus is constructed successfully.

Objective To study the differences in body composition and blood lipids between the pre-and post-menopausal women of Maonan ethnicity , and to explore the correlations between body fat content , fat distribution and blood lipids.Methods Totally 200 Maonan pre-and post-menopausal women were randomly selected from Huanjiang county in Guangxi.Body composition were measured by bioelectrical impedance analysis (BIA), and blood lipids were tested from blood samples .Results Compared with the pre-menopausal women , the visceral fat level (/area ) , waist-hip ratio (WHR), left (/right) lower limbs fat, total cholesterol (TC), triglyceride (TG) and low-density lipoprotein( LDL-C) in post-menopausal women were significantly higher ( P <0.01 ) , and the detection rate of hypercholesterolemia , mixed hyperlipidemia and dyslipidemia in postmenopausal group was also significantly higher ( P<0.01 ) .All the blood lipids were closely related to WHR and visceral fat content (P<0.05 or P<0.01).In addition, TG, high-density lipoprotein ( HDL-C) and LDL-C except TC were significantly correlated to %BF, BMI and subcutaneous fat content ( P<0.05 or P<0.01).Conclusion The accumulation of visceral and abdominal fat in Maonan postmenopausal women is significantly correlated to dyslipidemia .The results may provide references for making preventive program for the Maonan women .

Objective To assess the methodology of papers on systematic reviews and meta-analysis of interventions published in Journal of Evidence-Based Medicine .Methods Basic data were extracted from 70 papers on systematic reviews and meta-analysis of interventions published in Journal of Evidence-Based Medicinefrom 2001 to 2011 .Methodology used in these papers was assessed according to the AMSTAR Scale.The data were input into the Excelland analyzed using the SPSS7.0 and Meta-Analyst software.Results The methodology used in 34 papers (48.6%) was assessed using the Cochrane bias risk assessment tools.Fund support, number of authors and their affiliated institutions did not sig-nificantly affect the total score of methodology used in the 70 papers .Conclusion The methodology used in papers on systematic reviews and meta-analysis of interventions published in Journal of Evidence-Based Medicine is not quite valid as its early design program and retrieval strategies are imperfect , and it does not provide the excluded literature list and the interest conflict.

Objective To investigate the distribution of six short tandem repeats(STR) loci in Maonan in Guangxi province and to study genetic relationship among 6 minorities.Methods Two hundreds unrealated individulas were analyed by STR genescanning.Genetic distance was computed and phylogenetic tree was construted for studying genetic relationship among 6 Ethnic groups.Results The average heterozygosity(H) in the six STR loci was 0.7825;the average polymorphism information content(PIC) was 0.7324;the accumulative discrimination power(DP) was 0.99999951,and the accumulative probability of paternity exclusion(EP) was 0.994575.All 6 populations were clustered into 2 group.Conclusion Allele 16 in D3S1358,allele 11 in D5S818,allele 11 in D7S820,8 in D13S317,14 in vWA,22 in FGA are probably the most primitive alleles in repective locus.The data obtained indicate highly genetic polymorphisms.Genetic relation between Maonan and Mulao were close.

OBJECTIVETo construct rat N1ICD lentiviral over-expression vector (LV-N1ICD) and N1ICD lentivirus interference vector (LV-N1ICD-shRNA.

METHODSWith the rat cDNA as a template, the N1ICD fragment was amplified by PCR to construct pGC-FU-N1ICD-3Flag shuttle plasmid by directly clone. Four pairs of N1ICD-shRNA oligonucleotide sequences were syn- thesized to construct the GVC112-N1ICD-shRNA interference plasmid. pGC-FU-N1ICD-3Flag and GVC112 -N1ICD-shRNA plasmids were co-transfected into 293T cells to screen for the best interference plasmid in the 4 GVC112-N1ICD-shRNA plasmids by detecting Flag expression. pGC-FU-N1ICD-3Flag or GVC112-N1ICD-shRNA plasmid along with with pHelper 1.0 and pHelper 2.0 plasmids were co-transfect into 293T cells to package LV-N1ICD and LV-N1ICD-shRNA, and the virus titer was determined by real-time PCR and drug screening method, respectively. H9c2 cells infected with LV-N1ICD and LV- N1ICD-shRNA respectively were assessed for cell viability using CCK-8 assay.

RESULTSpGC-FU-N1ICD-3Flag and GVC112- N1ICD-shRNA plasmid were verified by PCR, gene sequencing and Western blotting. Co-transfection of the plasmids with pHelper 1.0, and pHelper 2.0 plasmids into 293T cells obtained high-titer LV-N1ICD and LV-N1ICD-shRNA. LV-N1ICD was capable of promoting the cell viability and LV-N1ICD-shRNA produced an opposite effect.

CONCLUSIONThe vectors LV-N1ICD and LV-N1ICD-shRNA have been successfully constructed and packaged, which have the biological functions of Notch1 signaling.