Objective:To probe essence of TCM syndrome of gastric carcinoma from gene protein expression.Methods:Based on the collected data from pre-operative patients of gastric carcinoma,their TCM syndromes were determined by means of TCM syndrome classification criteria.E-cadherin gene protein expressions in tumor tissue after operation of gastric cancer were detected with EnVision two step immunohistochemical method.Results:The expression rate of E-cadherin was 90% in 100 cases of gastric carcinoma,with significant difference among different syndrome types(P

Antiviral Agents ; Hepatitis B, Chronic ; Hepatitis C, Chronic ; Humans

Objective To establish a rapid,sensitive and specific assay based on real-time PCR combined with reverse transcription for detecting and identifying viable Listeria monocytogenes.Methods The hlyA gene of Listeria monocytogenes was chosen as target,and then the primers and TaqMan probe were designed.Both ends of probe were modified with two different fluorescence groups.The PCR reaction was optimized systematically.The mRNA of Listeria monocytogenes was extracted,and then reverse transcription was performed through random primer.The cDNA Was detected by real-time PCR.Then the specificity,sensitivity and reproducibility of real-time PCR were estimated.In final,real-time PCR was applied to detect 20 mocked double-blind samplea.Results Viable Listeria monocytogenes were detected by real-time PCR accurately and quickly,and meanwhile,none of other bacteria and non-viable Listeria monocytogenes could be identified.The sensitivity was 10 CFU/ml in pure culture and 103CFU/ml for mocked samples respectively.The coefficient of variation of intra-assay and inter-assay Was less than 5%.When this assay was applied directly to identify 20 mocked double-blind samples,10 of these were positive to viable Listeria monocytogenes,5 were negative to non-viable Listeria monocytogenes,and 5 were negative to other pathogens.Conclusion It is demonstrated that real-time PCR is a reliable,accurate and feasible assay for viable Listeria monocytogenes.The establishment of this assay provided complete data for analysis and diagnosis in the field of food safety and epidemiologic survey.

Objective To explore the relationship between disorders of lipid metabolism and avascular necrosis of femoral head after operation of femoral neck fracture .Methods 160 patients with a diagnosis of fresh femoral neck fractures treated with AO cannulated compressed screws in orthopedic department of our hospital from January 2007 to January 2012 were involved .All the patients were divided into two groups :ANFH or non ANFH group according to the features of femoral head .The levels of blood lipid were examined .Then ,Logistic regression was used to screening risk factors and evaluate the influence of blood lipid factors ac-cording to βrisk factor .Results The levels of TC ,TG ,LDL and Apo B in ANFH group were (5 .99 ± 1 .05)mmol/L ,(2 .63 ± 0 .85)mmol/L ,(4 .16 ± 1 .21)mmol/L ,(0 .99 ± 0 .28)g/L respectively ,which increased remarkably compared with the control group ;while HDL ,Apo A1 levels in the ANFH group were (0 .88 ± 0 .19)mmol/L ,(1 .20 ± 0 .17)g/L ,which was remarkably lower compared with the control group ,and there were statistically significant difference (P=0 .000) .Serum TC and LDL levels were risk factors impacted on postoperative ANFH .Conclusion Postoperative ANFH was related to disorders of lipid metabolism .Serum TC and LDL levels could be diagnostic values on postoperative ANFH .Early control of blood lipid levels may prevent the development of ANFH in patients treated with AO cannulated compressed screws .

TcdA and B toxins secreted by Clostridium difficile( CD) are two important causes of diseases in organisms. The expression of tcdA and tcdB genes is regulated by a few factors located in the pathogenicity locus ( PaLoc) .Studies have indicated that the tcdC gene is likely to act as a negative regulator of toxin gene expression.So far, it has been debatable whether tcdC gene is regarded as a negative regulator.The mechanism of tcdC gene in pathogenesis remains unclear.In this paper, the structure and function of the tcdC gene are summarized, which will help study the mechanism of tcdC gene and obtain optimal drug targets.

Objective To probe the indication of biliary tract reconstruction without T-tube in orthotopic liver transplantation.Methods We put forward indications of biliary tract reconstruction without T-tube in orthotopic liver transplantation since January 2004 and there were 102 patients who underwent liver transplantation in our hospital without a T-tube in place after biliary tract reconstruction.The incidence of biliary tract complication was observed in these patients.Results All patients were followed up for more than 6 months.The incidence of biliary tract complication was 4.9 percent(5/102)in this group with 3 patients of intrahepatic difluse bile duct stenosis necessitating liver re-transplantation.The other 2 patients with common hepatic duct nonanastomotic stenosis were healed by ERCP plus stent placement.Conclusions Biliary tract reconstruction without T-tube placement helps to decrease the incidence of biliary tract complications resulting from the T-tube removal.

Objective To develop a rapid, sensitive and specific assay based on multiplex real-time PCR for detecting and identifying Escherichia coli O157: H7. Methods The lipopolysaccharide gene (rJbE) and H7 flagellar antigen gene(fliC) of Escherichia coli O157:H7 was chosen as targets, and then the primers and TaqMan-MGB probe were designed. The 5'end of probes was labeled with FAM and HEX fluo-resceins respectively; the 3'end of probes was labeled with MGB. The PCR reaction was optimized systemati-cally. Then the specificity, sensitivity and reproducibility of multiplex real-time PCR were estimated. Final-ly, multiplex real-time PCR was applied to detected clinical specimens. Results Escherichia coil O157:H7 were detected by multiplex real-time PCR accurately and quickly, which could distinguish Escherichia coli O157:H7 from O157: non-H7. Meanwhile, none of other bacteria could be identified. The sensitivity was 10 CFU/ml in pure culture. The coefficient of variation of intra-assay and inter-assay was less than 5%. When this assay was applied directly to identify 66 clinical specimens, the results showed that t5 were positive to Escherichia coil O157:H7 and 2 were positive to Escherichia coil O157: non-H7, in which 16 was the same to the results obtained from the conventional assays. The coincidence was 98.49%. Conclusion It is showed that multiplex real-time PCR is a reliable, accurate and feasible assay for detecting and identifying Escherich-ia coli Oi57: H7, The assay reported here provided a tool for analysis and diagnosis in the field of detecting clinical pathogens, epidemiologic survey and food safety monitoring.

Objective In comparison of the performances for the detection of Clostridium difficile toxin B genes from stool between BD MAX Cdiff assay and a laboratory-developed (LD) assay.The LD assay was evaluated in clinical application.Methods This study was a clinical application research.A total of 147 stool specimens from patients with diarrhea in Hangzhou First Hospital affiliated with Zhejiang Chinese Medical University were detected by the two assays from 1 July to 30 September 2014.DNA extraction and amplification of the tcdB gene were performed automatically on the BD MAX platform.Meanwhile, the tcdA and tcdB gene were detected by the LD real-time PCR assay after DNA extraction.Then, the results were analyzed by use of SPSS 10.0.Results A total of 147 stool samples were collected.There were 33 C.difficile positive cases and 114 negative cases detected by both of two assays.However, there were four stool samples had incongruent results.In comparison with BD MAX, the LD assay had a sensitivity of 93.94% (31/33), a specificity of 98.25% (112/114), a positive predictive value of 93.94% (31/33), and negative predictive value 98.25% (112/114).Furthermore, the results of the LD assay were statistically coherent with that of the BD assay (Kappa=0.922, P<0.01).Conclusions The LD assay was highly sensitive and accurate as BD MAX Cdiff assay in the detection of toxigenic Clostridium difficile.Furthermore, this LD assay could be also applied to detection of clinical stool samples directly with low cost.The assay will be more promising in diagnosis of toxigenic C.difficile in clinical application in China due to no additional instrument needed.

Objective To study the clinical characteristics which are related to malignancy in pancreatic intraductal papillary mucinous neoplasm (IPMN) with an aim to provide evidence for clinical practice.Methods Using PubMed,all pancreatic IPMN related articles with positive pathologic results before July 30th,2011 were studied.A pooled analysis was carried out on the morphological features of the disease.The analysis included gender,diameter of main pancreatic duct,diameter of cystic lesion,mural nodules and histological types (benign/malignant) of the neoplasm.Results 98 articles (including 1902 cases) were collected and analyzed.1025 cases were benign (53.89％) and 877 cases (46.11 ％) were malignant.Morphologically,there were a correlation between main pancreatic duct dilatation (≥5 mm),cystic lesion of large size (≥30 mm),presence of mural nodules and malignancy.The OR (95％ CI) were 5.591 (3.657-8.548),3.633 (2.626-5.027) and 4.983 (3.872-6.412) respectively.Conclusions A main pancreatic duct dilatation (≥5 mm),cystic lesions of large size (≥30 mm) and presence of mural nodules prompt the tumor to be malignant.In clinical work,the management of pancreatic IPMN should be made prudently based on comprehensive analysis of clinical features and the patient's status and intent.

Objective To investigate the relationship between the immunohistochemical types and the pathological types of pancreatic intraductal papillary mucinous neoplasms (IPMN).Methods Literatures on the diagnosis and treatment of pancreatic IPMN published before June 30,2011 in the PubMed database were retrieved.The literatures adopted were scored by the case reports quality assessment list.The correlation analysis between the immunohistochemical types and the gender,pathological types,characteristics of benign or malignant tumor and morphological types were analyzed.All data were analyzed using the Pearson chi-square test or multiple regression analysis.Results Thirteen literatures were adopted,the scores were 31-45 (full mark:50),and the mean score was 37.The clinical data of 826 pancreatic IPMN patients who were comfirmed by pathological examination were collected,and there were 4 immunohistochemical types:(1) The gastric type (363 patients).Of the 271 patients who underwent gender analysis,there were 195 males and 76 females,with the median age of 65.6 years.Of the 225 patients who underwent pathological types analysis,there were 146 cases of adenoma,34 cases of borderline tumor,24 cases of carcinoma in situ,21 cases of invasive carcinoma,the benign tumor was accounted for 86.68％ (293/338).Of the 215 patients who underwent morphological type analysis,there were 34 cases of main pancreatic duct type,151 cases of branch duct type and 30 cases of mixed type.(2) The intestinal type (327 patients).Of the 269 patients who underwent gender analysis,there were 184 males and 85 females,with the median age of 64.5 years.Of the 262 patients who underwent pathological types analysis,there were 28 cases of adenoma,43 cases of borderline tumor,91 cases of carcinoma in situ,100 cases of invasive carcinoma,the benign tumor was accounted for 29.21％ (85/291).Of the 151 patients who underwent morphological type analysis,there were 63 cases of main pancreatic duct type,54 cases of branch duct type and 34 cases of mixed type.(3) The pancreatobiliary type (92 patients).Of the 78 patients who underwent gender analysis,there were 41 males and 37 females,with the median age of 69.2 years.Of the 81 patients who underwent pathological types analysis,there were 4 cases of adenoma,1 case of borderline tumor,21 cases of carcinoma in situ,55 cases of invasive carcinoma,the benign tumor was accounted for 5.75％ (5/87).Of the 34 patients who underwent morphological type analysis,there were 7 cases of main pancreatic duct type,18 cases of branch duct type and 9 cases of mixed type.(4) The oncocytic type (44 patients).Of the 37 patients who underwent gender analysis,there were 24 males and 13 females,with the median age of 60.3 years.Of the 33 patients who underwent pathological types analysis,there were 18 cases of carcinoma in situ,15 cases of invasive carcinoma,and all of them were with malignant tumors.Of the 36 patients who underwent morphological type analysis,there were 9 cases of main pancreatic duct type,16 cases of branch duct type and 11 cases of mixed type.The immunohistochemical type of pancreatic IPMN was correlated with gender,pathological type and characteristics of maglignant or benign tumors (x2=10.626,281.839,333.212,r =0.097,0.569,0.625,P ＜ 0.05).The result of the chi-square test between the immunohistochemical types and the morphological types was statistically significant (x2 =50.732,P ＜ 0.05),but there was no correlation between them (r =0.010,P ＞ 0.05).Conclusion The immunohistochemical type of pancreatic IPMN is correlated with gender,pathological type and the characteristics of maglignant or benign tumors,which provide references for the diagnosis and prognosis evaluation of pancreatic IPMN.