To construct a cDNA subtractive library of genes transactivated by hepatitis C virus core protein with suppression subtractive hybridization technique. mRNA was isolated from HepG2 cells transfected with pcDNA3 1(-)-core and pcDNA3 1(-) empty vector,respectively, then cDNA was synthesized. After restriction enzyme RsaI digestion, small sized cDNA were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, and then it was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain JM109. The cDNA were sequenced and analyzed in GenBank with Blast search after PCR. The subtractive library of genes transactivated by HCV core protein was constructed successfully. The amplified library contained 233 positive clones. Colony PCR product showed that 213 clones contained 100~ 1 000 bp inserts. Sequence analysis was performed in 63 clones. Six of the sequences were unknown genes before. The full length sequences were obtained with bioinformatics method,which had been accepted by GenBank. It suggested that six novel cDNA sequences might be target genes transactivated by HCV core protein.

Crohn disease (CD) is a relatively infrequent disease in China. CD and intestinal tuberculosis (IT) are lack of specific clinical manifestations and are generally identified by endoscopic and histological examination. However, it is sometimes difficulty to distinguish them by endoscopy and histology. Their characteristics were investigated to offer more evidences and data for their differential diagnosis. The records of inpatients were reviewed from 1980～2001 in our hospital, all enrolled subjects were verified by endoscopy(or operation) and histology. The characteristics and key points of differential diagnosis of 28 CD cases and 14 IT cases were comparatively analyzed. The results showed that abdominal pain, mass, and hematochezia occurred more significantly in CD than in IT ﹙ P

To screen anti idiotypic single chain variable fragments(anti Id scFv)against hepatitis C virus NS4A(HCV NS4A)so as to lay a foundation for developing anti Id scFv vaccine againist the hepatitis C virus. The recombinant phage antibody library was panned by hepatitis C virus NS4A monoclonal antibody which was coated in a microwell plate. After five rounds of biopanning, 82 clones specific to HCV NS4A antibody were determined with the enzyme linked immunoadsorbent assay(ELISA). The specificity of anti idiotypic scFv was identified by ELISA and competitive inhibition assay. The DNA sequence of the positive clone was determined. The result showed that HCV NS4A anti Id scFv had a specific combination character with hepatitis C virus NS4A monoclonal antibody. The DNA sequence data showed that the anti Id scFv coding gene included 789 bp. The results suggested that the anti Id scFv fragments to HCV NS4A monoclonal antibody could be successfully selected by recombinant phage antibody library screening technique, which might pave a way for the study of new preventive and therapeutic strategy of hepatitis C using anti Id scFv.

To screen HCV core mimotopes,HCV core monoclonal antibody was used as immobilized molecule,and a 12 mer phage peptide library was biopanned and positive clones were selected by enzyme linked immunoadsorbent assay(ELISA), competitive inhibition assay, and DNA sequencing. 10 positive clones were chosen for DNA sequencing. From the experiment and sequencing comparison results,one epitope was confirmed as a mimotope of HCV core protein. The study might provide a new approach for HCV therapy and vaccine development.

For investigating the effectiveness and safety of oral sodium phosphate in preparation for colonoscopy,108 cases from both outpatient and inpatient clinics from June,1995 to March,1996 were randomized and blinded to receive either oral sodium phosphate or standardized electrolyte lavage solu- tion as control.The questionaire was designed for scoring by patients and doctors regarding to its toler- ance,taste,side effects and cleaning degree etc.Results showed that the tolerance rates in oral sodium phosphate and electrolyle lavage were 87.5% and 69.2% and the rates of preference in using them were 32.2% and 15.4% respectively(P

Objective To investigate the safety and feasibility of electrocoagulation of gallbladder artery in laparoscopic cholecystectomy ( LC) . Methods A total of 376 patients with gallbladder benign diseases underwent LC in our hospital from May 2004 to September 2013.The gallbladder artery was treated by electrocoagulation . Results Because of unclear gallbladder triangle due to abdominal adhesion , conversion to laparotomy was performed in 9 patients.In the remaining 367 patients, three-port LC with electrocoagulation of the gallbladder artery was conducted successfully .Laparoscopic appendectomy was conducted simultaneously in 12 patients.An additional fenestration and drainage of the left renal cyst was performed in 1 patient.Postoperatively, a secondary bile duct exploration was conducted in 1 patient because of bile duct obstruction caused by common bile duct stones . Conclusion The electrocoagulation of gallbladder artery is safe and feasible in LC for the treatment of gallbladder benign diseases .

Objective:In order to investigate the clinical efficacy of acupuncture plus cupping method for treating chronic pharyngitis. Methods: Sixty patients with chronic simple pharyngitis were randomly divided into two groups. Thirty cases in the treatment group were treated by acupuncture plus cupping method and 30 cases in the control group by routine anti-inflammatory and antiviral therapy. The clinical effects were observed in the two groups. Results: The total effective rate was 93.3% in the treatment group and 80.0% in the control group. The therapeutic effect was significantly better in the treatment group than in the control group (P<0.05). Conclusion: Acupuncture plus cupping method has a good effect for chronic simple pharyngitis.

Objective To investigate the expression of epidermal growth factor receptor (EGFR) and multidrug resistance protein, and it's relationship with efficacy of chemotherapy in NSCLC.MethodsThe immunohistochemical (EnVision method) was used to detect the expression of EGFR and LRP, P-gp, MRP, GST-π in 95 cases of NSCLC tissues and 15 cases of normal lung samples.ResultsEGFR in NSCLC tissues was significantly correlated with the history of smoking, pathological types of cancer(squamous cell carcinoma vs adenocarcinoma), presence of lymph node metastasis, and TNM stages (P< 0.05).The positive expression rate of LRP and MRP in adenocarcinoma were higher than squamous cell Carcinoma (P< 0.05), and MRP in high differentiate cancer was higher than low differentiate cancer (P< 0.05).The expression of MRP and LRP, PGP and GST-π in NSCLC were positive correlation(r=0.341, P=0.001;r=0.213, P=0.038), there is no correlation between EGFR with LRP, P-gp, MRP, GST-π.EGFR-negative chemotherapy efficiency (62.5%) was significantly higher than EGFR-positive(37.5%, P< 0.05).ConclusionEGFR and multi drug resistance proteins are overexressed in NSCLC tissues.multi drug resistance proteins have a synergistic to drug resistance.EGFR could be used to objectively predict chemotherapy effect and prognosis of NSCLC.

Apoptosis ; Down-Regulation ; Electron Transport Complex IV ; chemistry ; genetics ; metabolism ; Humans ; Mitochondria ; metabolism ; Mutation ; Neoplasms ; genetics ; metabolism ; pathology

Objective To investigate the effect of metformin ( MET) on learning and memory behavior in HFD-induced insulin-resistant rats.Methods Thirty male Sprague-Dawley (SD) rats were divided into three groups to receive either a normal diet (Control group) or a high-fat diet (two HFD groups) for four weeks(HFD+MET).From two HFD groups, one received vehicle ( HFD group ) alone and other MET administration ( HFD+MET group ) .MET was dissolved in drinking water at a concentration of 2 mg/ml.All rats were subjected to the glucose tolerance test ( GTT) and behavioral tests using the elevated plus maze ( EPM ) , open field test ( OFT ) , Morris water maze ( MWM ) test and the step-through passive avoidance test ( PA) after four-week consecutive MET treatment .Blood samples were collected for determination of glucose. Results MET attenuated the glucose resistant condition and improved cognitive behavior in MWM and PA, vs the HFD group. Conclusion MET can improve the impaired learning and memory behavior in HFD-induced insulin-resistant rats.