Objective To isolate and identify embryonic stem (ES) cells of rhesus monkey from mature blastocysts cultured in vitro . Methods Rhesus monkey blastocysts were obtained after in vitro maturation of oocytes, fertilization and maturation of early embryos. After the blastocysts were hatched naturally from the zone pellucida, the inner cell mass (ICM) was dislodged from blastocysts using the sealed end of a finely drawn Pasteur pipette and co cultured with the feeder cells. The ICM, resembling embryonic stem cell colony, was isolated, cultured, and identified. Results A total of 92 normal GV oocytes were obtained from 4 FSH prime rhesus monkeys. Six blastocysts with high quality were selected from 22 oocytes after culture in HECM 10 medium. One of the rhesus monkey ES cell lines, i.e. RS5 cell line, was finally isolated from 3 of the inner cell mass isolated from the 6 blastocysts. The RS5 cells presented a high nucleus/cytoplasm ratio, prominent nucleoli, and flatter colonies with individual and distinct cells. After successive passages for 5 months passage, the RS5 cells remained a normal karyotype, i.e. 42 of chromosomes and alkaline phosphatase (AP) positive, which meant the RS5 ES cell colony remained undifferentiated. After high density culture for a longer time, the ES cell could differentiate into multi types of cells. Conclusion The RS5 cell line has the ability to self renew and potential to differentiate. Thus, RS5 cell line belongs to embryonic stem cells.

OBJECTIVE:To optimize the formulation of Dimemorfan phosphate tablets. METHODS:Using 60 min dissolution rate of dimemorfan phosphate as index,L9(34) orthogonal test was used to optimize the amount of starch,microcrystalline cellu-lose,croscarmellose sodium and concentration of HPMC E5 solution. The friability,hardness,60 min dissolution rate and main component were detected. The similarity of dissolution curves of Dimemorfan phosphate tablets was compared with that of imported tablets in 0.1 mol/L hydrochloric acid,pH 6.8 phosphate buffer,water and pH 4.0 acetate buffer. RESULTS:The optimized formu-lation of Dimemorfan phosphate tablet(1 000 tablets)was composed of dimemorfan phosphate 10 g,starch 60 g,microcrystalline cellulose 40 g,10% HPMC E5 solution and croscarmellose sodium 25 g. The friability,hardness,60 min dissolution rate and main component of 3 batches of Dimemorfan phosphate tablets prepared by optimized prescription were 0.42%-0.58%,9.8-10.5 kg,94.89%-96.21% and 99.21%-99.52%,respectively. In 4 dissolution mediums,similar factors f2 of dissolution curves between prepared tablets and imported tablets were above 50. CONCLUSIONS:Dimemorfan phosphate tablets were prepared successfully. The optimized formulation is rational. The dissolution behavior of prepared tablets is similar to that of imported tablets in vitro.

OBJECTIVE:To prepare borneol-puerarin liposomes,and to investigate its brain-targeting. METHODS:Film dis-persion ultrasonic method was used to prepare borneol-puerarin liposomes. The morphology of liposomes was observed by TEM;the particle size and Zeta potential were measured by laser particle size analyzer;the entrapment efficiency were measured by sepha-dex gel filtration method. Compared with Puerarin injection,brain-targeting of borneol-puerarin liposomes and puerarin liposomes via intravenous injection of mice tail was studied with relative intake rate and peak concentration ratio. RESULTS:Borneol-puerarin liposomes were spherical or quasi-circular;its mean particle size,polydispersity index and Zeta potential were 226 nm,0.263 and-21.3 mV respectively. The entrapment efficiency were(65.32±2.13)%. Compared with Puerarin injection,relative intake rate of puerarin liposome and borneol-puerarin liposome were 1.68 and 2.58,and peak concentration ratio were 1.15 and 1.42. CONCLU-SIONS:Brain-targeted borneol-puerarin liposomes are prepared successfully.

Objective To explore the clinical significance of tumor pathological length in evaluation of prognosis of esophageal squamous cell carcinoma.Methods 421 patients with esophageal squamous cell carcinoma were selected as study subjects.Each patient's data of gender,age,pathological examination(tumor pathological length),survival time,cancer stage,operation mode were collected.The patients were divided into two groups:≤4 cm group(189 cases) and >4 cm group(232 cases), and the constituent ratios of gender,age,or operation mode and the survival rate between the two groups were analyzed.Results There were no significant differences in gender composition,age structure and operation mode between the two groups(x2=2.03,2.41,0.06,all P>0.05);There was positive correlation between tumor pathological length and TNM stage(CMH=68.07,P<0.01).The 25%,50% and 75% quantiles of survival period in the group whose tumor length was less than or equal to 4 cm were 28,40 and 58 months,respectively,which in the group whose tumor length was more than 4 cm were 19,31 and 61 months,and there was significantly difference between the two groups(x2=44.88,P<0.01), the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm.In the further stratification analysis by TNM stage,the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm in theⅠstage patients(x2=5.61,P<0.05);the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm in theⅡstage patients(x2=4.62,P<0.05);the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm in the Ⅲ stage patients(x2=4.88,P<0.05).Conclusion Tumor pathological length is an effective prognostic factor for esophageal squamous cell carcinoma,and the critical value can be determined as 4 cm.

Objective To explore the effect of puncture the left or right branch of portal vein on the incidence of postoperative hepatic encephalopathy in transjugular intrahepatic portosystemic shunt (TIPS). Methods A literature search was conducted through investigating PubMed, Cochrane Library, CNKI digital library and Wanfang databases. Randomized controlled trials and cohort studies related to the effect of puncture the left or right branch of portal vein on the incidence of postoperative hepatic encephalopathy in TIPS were included in this review. According to the methods for Cochrane systematic review, the data were extracted, the quality of the literatures was evaluated, statistical analysis was conducted and the data were analyzed with RevMan 5.0. Results One randomized controlled trial and four cohort studies, including 552 patients with liver cirrhosis received TIPS, were finally included in this review. Random-effects model meta-analysis showed that puncture of the left branch of portal vein in TIPS caused lower incidence of postoperative hepatic encephalopathy than puncture of the right branch of portal vein (RR=0.51, 95%CI was 0.39 to 0.66, P<0.01). Conclusion Puncture of the left branch of portal vein in TIPS can significantly reduce the incidence of postoperative hepatic encephalopathy in comparison with puncture of the right branch of portal vein.

Objective:To determine the content of chloroform, ethyl acetate and DMF in dimemorfan phosphate by gas chromatog-raphy (GC). Methods:The capillary gas chromatography was used with a PEG-20M column, programmed temperature, water as the solvent and FID as the detector. Results:The three organic solvents were separated and showed good linear relationship (r>0. 999 0). The detection limit of chloroform, ethyl acetate and DMF was 0. 63,0. 60 and 8. 92μg·ml-1 , respectively. The residues of the organ-ic solvents in three batches of the samples all met with the requirements of ICH. Conclusion: The method is sensitive, accurate and reliable, and can be used in the quality control of dimemorfan phosphate.

Objective:To establish a method for the determination of dichloromethane, methanol and ethanol in fenticonazole ni-trate. Methods:The samples were detected by headspace GC. The column was OV-1301(30 m × 0. 53 mm,3. 0 μm), the detector was FID with nitrogen as the carrier gas, the detector temperature was 250 ℃ and the injector temperature was 200 ℃. Results:The linear range of dichloromethane, methanol and ethanol was 2. 436-21. 924(r=0. 998 8), 12. 268-110. 412(r=0. 999 5) and 20. 052-180. 468 μg·ml-1(r=0. 996 9) with the average recovery of 99. 30% (RSD=2. 36%), 100. 21%(RSD=1. 07%) and 100. 15%(RSD=1. 21%)(n=9), respectively. Conclusion:The detection method is sensitive, accurate and reliable, and can be used in the quality control of fenticonazole nitrate.

Objective To analyze the genomic evolution characteristics of pathogenicity islands (PAIs)in Deng strain of enteropathogenic Escherichia coli (E.coli,EPEC Deng).Methods EPEC Deng was isolated from infant stool specimen,serotypes were identified and antimicrobial susceptibility testing was performed;whole-genome se-quencing was performed by Illumina 2000 system,the locations of prophages(PPs)in the chromosome were detected using PHAST software,collinearity analysis was performed by MUMmer software,phylogenetic trees of homolo-gous gene were constructed in order to understand the evolutional rule of homology gene.PAIs prediction was per-formed using PAI finder software,the homologous evolutionary rule of PAIs core region(LEE)and core genes were clarified,genetic polymorphism was analyzed.Results The serotype of EPEC Deng strain was O119:H6,the strain was resistant to ciprofloxacin,levofloxacin,and ampicillin,but sensitive to other antimicrobial agents.The complete circular chromosome contained 5 025 482 bp with a GC content of 50.52 %,and the plasmid contained 207 564 bp with a GC content of 49.50%.A total of 17 PPs in the chromosomal genome were discovered,phyloge-netic trees analysis suggested that EPEC Deng strain was highly homologous with O26:H11 and O111 :H strains;PAIs and core genes were highly homologous with RDEC-1 and O26:H413/89-1 strains;genetic diversity analysis showed that the intimin (eae)and its receptor tir had high polymorphism,with the pi (π)value>0.10,the genes in type III secretion system was relatively stable.Conclusion The study clarified the genomic evolution characteris-tics of EPEC Deng genome and it’s PAIs,and is helpful for understanding genetic characteristics of native EPEC.

Objective To establish a gas chromatograph method for determing Chloroform, ethyl acetate and N, N-dimethyl formamide in butoconazole nitrate. Methods The samples was detected by Headspace Gas Chromatography. Temperature programming method was adpoted and FID was used as detector. The injector temperature was 200 ℃, and the detector temperature was reach 250 ℃. Nitrogen was used as carrier gas in the experiment. Results In the mentioned chromatographic conditions, Chloroform, ethyl acetate and N, N-dimethyl formamide had good linear relationships in the ranges of 0. 066-0. 588,0. 062-0. 556 and 0. 896-8. 061 μg·mL-1 respectively. The average recoveries were 99. 18%,102. 84% and 98. 71%. RSD were 3. 87%,4. 33% and 3. 71%. Conclusion The detection method is sensitive, accurate, reliable, and can be used as a quality control for butoconazole nitrate.

Objective:To investigate the clinical efficacy and prognosis of transjugular intrahepatic portosystemic shunt (TIPS) and drug combined with endoscopic treatment in patients with liver cirrhosis and esophagogastric variceal bleeding (EGVB).Methods:From January 2012 to December 2013, at the First Affiliated Hospital of Xi′an Jiaotong University, the data of 147 patients with liver cirrhosis and EGVB undergoing TIPS or drug combined with endoscopic treatment were retrospectively collected, with 87 cases in TIPS treatment group and 60 in drug combined with endoscopic treatment group.The 5 years follow-up data were analyzed, and the overall survival rates, rebleeding-free survival rates and hepatic encephalopathy-free survival rates at 6 weeks, 1 year, 2 years and 5 years after treatment of two groups were compared. Independent sample t test, Mann-Whitney U test, chi-square test, Fisher exact test, Z test, log-rank test and trend test were used for statistical analysis. Results:There were no significant differences in age, gender, etiology, Child-Pugh classification, initial liver function, coagulation function, liver ascites, previous history of hepatic encephalopathy, blood pressure and preoperative blood transfusion history between the TIPS treatment group and combination of drugs and endoscopy treatment group (all P>0.05). Forty-one patients died within 5 years, of which 20 (48.8%) died of rebleeding and 6 (14.6%) died of hepatic encephalopathy. There were no significant differences in 6-week, 1-year and 2-year overall survival rates between the TIPS group and drug combined with endoscopic treatment group (all P>0.05), however the 5-year overall survival rate of the TIPS treatment group was higher than that of the drug combined with endoscopic treatment group (78.4% vs. 63.2%), and the difference was statistically significant ( Z=2.06, P=0.048). The 6-week, 1-year, 2-year, 5-year rebleeding-free survival rates of the TIPS group were 97.7%, 96.5%, 88.9% and 70.9%, respectively, which were all higher than those of the drug combined with endoscopic treatment group (86.7%, 53.3%, 43.3% and 27.1%), and the differences were statistically significant ( Z=2.35, 6.39, 6.26 and 4.80, all P<0.05). There were no significant differences in hepatic encephalopathy-free survival rates at 6 weeks, 1 year and 2 years after treatment between the TIPS group and drug combined with endoscopic treatment group (all P>0.05), however the 5-year hepatic encephalopathy-free survival rate of the TIPS treatment group was lower than that of the drug combined with endoscopic treatment group (67.7% vs. 86.7%), and the difference was statistically significant ( Z=2.28, P=0.030). The lower the Child-Pugh classification, the higher the cumulative 5-year survival rate ( χ2=6.75, P<0.01). There was no statistically significant difference in the 5-year overall survival rate in patients with the same Child-Pugh classification between the TIPS group and the drug combined with endoscopic treatment group (all P>0.05). Conclusions:The efficacy of TIPS is better than that of the drug combined with endoscopic treatment in treating EGVB. Even the long-term risk of hepatic encephalopathy of TIPS is higher, the short-term, middle-term and long-term rebleeding rate are decreased. Patients with Child-Pugh grade C do not need to avoid TIPS when choosing the treatment, the earlier the TIPS used, the better survival benefit will be obtained.