Objective To determine the contents of Poloxamer in contact lens care solution by spectrophotometric method,and investigate the effect of the other lubrication composition on the measurement. Methods Five brands of contact lens care solution with hydroxypropyl methyl cellulose(HPMC)were obtained.The interference of HPMC in contact lens care solution was removed by heating and rapid filtering,and the contents of Poloxamer was determined by KI-I_2 spectrophotometry. Results For the pre-treated contact lens care solution,the tested wavelength was 500 nm and the development time was 120 min.Favourable linearity was observed when Poloxamer ranged between 0.001 mg/mL and 0.020 mg/mL,with standard curve of Y=0.050 91X+0.017 15 and r~2=0.998 2.Precision(n=6)test revealed that RSD was 1.331 4%,and RSD of daytime repeatability experiment was 2.971 3%.The recovery rates of 80%,100%and 120% samples ranged between 90%to 105%. Conclusion KI-I_2 spectrophotometry is a convenient,stable and reliable method,and may sere as a mass control method for nonionic surfactant Poloxamer content in contact lens care solution.

Objective To discuss the value of hypotonic-MRCP combined with LAVA dynamic contrast-enhanced scan in diagnosis of duodenal tumor.Methods Five patients with duodenal adenocarcinomas confirmed by pathology were involved in this study.All cases underwent plain MRI and hypotonic-MRCP and LAVA dynamic contrast-enhanced scans.Results 4 of 5 cases were correctly diagnosed with MR imaging.Imaging signs included:the cavum of descending duodenum appeared as asymmetrical stenosis,masses with soft tissue signal intensity connecting the intestinal wall with wide base,the intestinal wall stiff,the intestinal mucosa destruction.The lesions would be slightly-moderately enhanced at LAVA dynamic enhancement scanning.Conclusion Hypotonic-MRCP combined with LAVA dynamic contrast-enhanced scan can display the direct and indirect sign of duodenal tumors,which was effective in identifying the circumscription of the tumor and its extension.

Objective To investigate the guidance of a new classification on the treatment methods selection for cystic dilation of bile duct (CDBD).Methods The clinical data of 213 patients with CDBD who received treatment according to the Todani 2003 classification at the Peking Union medical College Hospital from September 1968 to July 2013 were retrospectively analyzed.The CDBD was reclassified with a new classification proposed by Dong Jiahong et al,and the guidance of the new classification on the treatment methods selection for CDBD was analyzed.Patients were followed up via out-patient examination and telephone interview till August 2013.Results Of the 213 patients,139 were with Todani type Ⅰ CDBD (type C CDBD of the new classification) ; 3 were with Todani type Ⅱ CDBD (type C1 CDBD of the new classification) ; 1 was with Todani type Ⅲ CDBD (type E CDBD of the new classification); 52 were with Todani type Ⅳa CDBD (35 with type D1 and 17 with type D2 CDBD of the new classification) ; 1 was with Todani type Ⅳb CDBD (type C CDBD of the new classification) ; 8 were with Todani typeⅤ-Ⅰ CDBD (type B CDBD of the new classification) ; 9 were with Todani type Ⅴ-Ⅱ CDBD (type A CDBD of the new classification).Eighteen patients did not receive the surgical treatment.Of the 195 patients who received surgical treatment,patients with type C and D CDBD of the new classification took a large proportion.Patients with type C CDBD of the new classification received cystectomy,biliary cyst resection,Rouxen-Y cholangiojejunostomy or internal drainage.Patients with type D CDBD of the new classification received extrahepatic biliary cyst resection,Roux-en-Y cholangiojejunostomy.Patients with severe intrahepatic disease and with type D1 CDBD of the new classification received concomitant left hemihepatectomy or pancreaticoduodenectomy.Patients with type A1 CDBD of the new classification received right hemihepatectomy.Patients with type A2 CDBD of the new classification were cured by conservative treatment after split liver transplantation.Patients with type B1 CDBD of the new classification received left hemihepatectomy and Roux-en-Y cholangiojejunostomy.Patients with type B2 CDBD of the new classification received bile duct stone extraction.There was 1 patient with type E CDBD,and partial resection of the CDBD ± bile duct reconstruction was carried out.Pancreatic fistula,biliary fistula,reflux cholangitis,cholangitis and anastomotic stricture were detected on 74 patients,and they were cured by conservative treatment or lithotomy.A total of 187 patients were followed up with the median time of 85 months (range,1-432 months).One hundred of seventy-five patients recovered well,and 12 patients with canceration of the bile duct died of tumor metastasis at postoperative 1-282 months.Conclusion This new classification simplifies the typing of extrahepatic bile duct dilation,refines the typing of intrahepatic bile duct dilation,and has better guidance for surgical treatment.

Objective： This study was designed to investigate the relationship between p16 protein expression and gastric carcinogenesis,depth of invasion, lymph node metastasis, and evaluate the role of deletion and mutation of p16 gene in exon 2 in gastric carcinoma. Methods： p16 protein expression in gastric carcinoma and precancerous lesion was examined by streptavidin-peroxidase conjugated(S-P) method; The deletion and mutation of p16 gene were examined respectively by polymerase chain reaction(PCR) and polymerase chain reaction single-strand conformation polymorphism analysis(PCR-SSCP) in gastric carcinoma. Results： ① The positive rates of p16 protein expression were 96.25％ (77/80) in normal gastric mucosa, 92.00％ (45/50) in dysplastic gastric mucosa, and 47.54％ (58/122) in gastric carcinoma. The positive rate of p16 protein expression in gastric carcinoma was significantly lower than that in normal gastric mucosa and in dysplastic gastric mucosa (P<0.05). ② The positive rate of p16 protein expression in mucoid carcinoma (10.00％ ,1/10) was significantly lower than that of poorly differentiated carcinoma (51.22％ ,21/41), undifferentiated carcinoma (57.69％ ,15/26), and signet ring cell carcinoma (62.50％ ,10/16) (P< 0.05). ③ The positive rates of p16 protein in 30 cases paired primary and lymph node metastatic gastric carcinoma were 46.67 ％ (14/30) in primary gastric carcinoma,16.67％ (5/30) in lymph node metastatic gastric carcinoma. The positive rate of lymph node metastatic carcinoma was significantly lower than that of primary carcinoma(P<0.05). ④ Evaluation of mutation and deletion of p16 gene： There was no mutation of p16 gene in exon 2, but there were 5 cases displayed deletion of p16 gene in exon 2 in the 25 primary gastric carcinoma. Conclusions： ① The expression loss of p16 protein is related to carcinogenesis, histopathological subtypes,and lymph metastasis of gastric carcinoma. ② The mutation of p16 gene in exon 2 may not be involved in gastric carcinogenesis. But the deletion of p16 gene in exon 2 might be involved in gastric carcinogenesis.

Objective To explore associations between SG13S114A/T and SG13S32A/C polymorphisms of ALOX5AP gene and the genetic susceptibility of ischemic cerebrovascular diseases (ICVD) in Henan Han population.Methods Two hundred and forty-six ICVD patients and 245 healthy controls were recruited from Han population in Henan province. Polymorphisms of SG13S114A/T and SG13S32A/C in ALOX5AP gene were genotyped in these samples by SnaPshot minisequencing method.Each genotype frequency and allele frequency were statistically analyzed and compared between ICVD group and control group using SPSS16.0 software.Haplotype and linkage disequilibrium were analyzed by SHEsis software.Results The SG13S114 AA genotype frequency ( 18.7％ ) and A allele frequency (41.3％) in ICVD group were significantly higher than those in control group (9.0％ and 32.7％,respectively; P =0.002 and P =0.005 ).It was also found that in male ICVD group and in younger ICVD group ( ＜50 years old),the SG13S114 AA genotype frequencies (22.1％ and 22.0％,respectively) and A allele frequencies (42.1％ and 42.7％,respectively) were significantly higher than those in male control group and younger control group (SG13S114 AA genotype:9.0％ and 8.9％ ; P =0.010 and P =0.006,respectively) ;A allele frequencies,34.0％ and 32.0％ ; P =0.048 and P =0.020,respectively.Finally,the prevalence of A-A haplotype in ICVD group was significantly higher than that in control group(30.4％ vs 23.5％,OR =1.419,95％ CI 1.068-1.885,P =0.015).T-C haplotype frequency of ICVD group was significantly lower than that in control group (22.0％ vs 28.8％,OR =0.698,95％ CI 0.523-0.932,P =0.014 ).Conclusions The A allele in SG13S114 loci of ALOX5AP may be a genetic risk factor for ICVD in Han population in Henan province.The association is predominant in ICVD patients of male and younger than 50 years old.Maybe A-A haplotype increases the risk of ICVD and T-C haplotype and has a protective effect against ICVD in Henan Han population.

Objective To establish reference methods for the measurement of catalytic activity concentrations of enzymes at 37℃ which have been published by IFCC and evaluate accuracy of reference methods.Methods Six reference methods for the measurement of catalytic activity of enzymes were established with two sets of apparatus systems of PE and Agilent according to International Federation of Clinical Chemistry(IFCC)37℃ reference procedures in two reference labs respectively.The commercial Roche calibrator c.f.a.s was used to monitor the precision of two reference labs as quality control material.Certified Reference Materials(CRMs)represented an efficient tool to assess the analytic performance for the verification of trueness and in two labs.The measurement accuracy of the assays for catalytic activity concentrations of 6 enzymes [alanine aminotransferase(ALT),aspartate aminotransferase(AST),lactate dehydrogenase(LDH),creatine kinase(CK),gamma-glutamyhransferase(GGT),amylase(AMY)]was further verified and validated by international ring trial program.Results The within-laboratory variations of 6 enzymes in both of the reference lab were ranged from 0.5%-1.9%.Their results showed fully agreement with deviation less than 2.1%.The value of CRM was in the tolerant limit and analytic accuracy was verified.The results of four enzymes(ALT,AST,GGT,AMY)lay within (x)±s.However,the result of CK and LDH lay within (x)±2s.Except sample A for the LDH testing,we did not find any deviation variable in the detection of other enzymes.Conclusions The reference methods for the measurement of catalytic activity of enzymes(ALT,AST,LDH,CK,GGT,AMY)at 37℃ in the two labs by use of two sets of apparatus systems of PE and Agilent have been established and these methods showed excellent stability and accuracy.

OBJECTIVEA method for determined oxygen permeability (Dk) of finished soft contact lens.

METHODSOxygen permeability(Dk) of 10 group finished soft contact lens was determined by polarographic method, and the values were linearly calibrated using the measured and established Dk values of the specified reference materials.

RESULTSOnly one Dk value fell outside of the requirement of GB 11417.3-2012 tolerance for Dk (± 20%).

CONCLUSIONIt is great practicability when determined the oxygen permeability (Dk) of finished soft contact lens by polarographic method which were linearly calibrated using the measured and established Dk values of the specified equal thickness reference materials without the measured lens customized 4 different thickness.

Objective To investigate the protective effect of n-butanol extract from the roots of Potentilla anserina (NP) on hypoxic hippocampal neurons in neonatal rats.Methods Primary cultured hippocampal neurons were pretreated with different concentration of NP (0.25,0.0625,and 0.0156 mg/mL) before incubation in a low oxygen (0.1％) environment for 4 h.Cell viability was evaluated by Trypan blue staining assay.Lactate dehydrogenase (LDH) released by neurons into the medium was measured.The activity of superoxide dismutase (SOD) in cell cytosol was determined using nitroblue tetrazolium.Morphological changes and mitochondrial function were observed by transmission electron microscopy.Results Hypoxic injury could decrease the cells viability of neuron,enhance LDH release (P ＜ 0.05),decrease SOD activity,and increase mitochondrial injury.Pretreatment with NP significantly increased cell viability,decreased LDH release (P ＜ 0.05),promoted SOD activity (P ＜ 0.05),and remarkably improved cellular ultra-microstructure compared with the model group.Conclusion NP could protect the primary hippocampal neurons from hypoxic injury by attenuating mitochondrial cell death.

Objective To construct an eukaryotic expression vector of human B-cell translocation gene 2 (BTG2), to express the FLAG-tagged BTG2 protein in HeLa cells,and to supply an experimental tool for investigating the function of BTG2 gene.Methods The full-length BTG2 fragment was obtained by PCR and inserted into the multiple cloning site of pcDNA3.1 (+)vector. Oligo DNA encoding FLAG tag was designed and inserted into pcDNA3.1(+)-BTG2 to construct another vector pcDNA3.1(+)-FLAG-BTG2.The HeLa cells were divided into pcDNA3.1(+)empty vector group,pcDNA3.1(+)-BTG2 group and pcDNA3.1(+)-FLAG-BTG2 group.The HeLa cells were transfected with recombinant plasmids.Western blotting using anti-FLAG antibody was performed to detect the expression of FLAG-BTG2 protein in HeLa cells.Results The sequence of the vector was verified by both BamH Ⅰ endonuclese digestion and DNA sequencing. The Western blotting analysis confirmed that FLAG-fused BTG2 was detected in pcDNA3.1(+)-FLAG-BTG2 group but not in empty vector or pcDNA3.1(+)-BTG2 groups. Conclusion The eukaryotic expression vector pcDNA3.1(+)-FLAG-BTG2 is successfully constructed and FLAG-tagged BTG2 protein is expressed in HeLa cells.

The COVID-19 outbreak has drawn attention to viral infectious diseases once again, and the development of antiviral drugs for both known and potentially emerging viruses is of great significance. In recent years, peptides and protein drugs are becoming a hot spot in the field of antiviral drug research and development. Phage display technology, as a powerful tool for screening peptides and protein drugs, has been increasingly concerned in the academic and industrial fields. The present review introduced the basic principle of phage display technology, summarized phage display libraries often used in antiviral drug discovery and their applications, discussed the challenges and future direction of antiviral drug research and development based on phage display technology.