Objective:To study the effectiveness of Loop electrosurgical excision procedure (LEEP) in the diagnosis and management of cervical intraepithelial neoplasia(CIN). Methods: 155 patients with CIN were diagnosed by cervical biopsy under colposcopy. All cases were treated with LEEP and the operative time, bleeding volume, cervical biopsy results before and after LEEP were evaluated, and the effects and cervical regeneration were followed up. Results: The mean operative time of LEEP was 6. 8 minutes. The mean bleeding volume was 9.6m1. Pathological diagnosis of CINI CIN Ⅱ CIN Ⅲ after LEEP were positively correlated to directed biopsy by colposcopy. The correlation coefficient was 0.785 ( P < 0.01). The cure rate of LEEP was 98.6%. Among the 151 cases, 134 (88. 7%) were satisfactory cervical regeneration. Conclusions: LEEP is one of the effective and safe methods for the diagnosis and treatment of CIN.

Objective To evaluate the effect of probiotics on inducing and maintaining remission of ulcerative colitis (UC). Methods PubMed, EMBase, Web of Science and The Cochrane Central Register of Controlled Trials from 1966 to 2009 (up to August), and China Journal Full-text Database, Chinese Technologic Journal Database (Weipu), Wan Fang Digital Journal Full-text Database from 1978 to 2009 (up to August) were retrieved in order to collect clinical randomized controlled trials regarding the effect of probiotics in remission induction and maintenance in UC patients. Statistical analysis was performed by meta-analysis using Review Manager 4.2.10. Results Eleven randomized controlled trials met the inclusion criteria, of which five studies were included in the control trial of probiotics and placebo to evaluate the clinical remission rate, seven studies were included to compare probiotics with placebo (3 trials) or with mesalazine (4 trials) to evaluate the clinical relapse rate (one of the trials was included in the induction treatment group and the maintenance treatment group). Meta-analysis showed that: a) On the basis of combining with conventional therapy, probiotics were superior to placebo in clinical remission rate (OR=0.28, 95%CI: 0.16-0.49, P

Objective To estimate the sensitivity of malignant cells from ascites of gastric cancer to anticancer agents by in vitro sensitivity test, and to study its value in individualized intraperitoneal chemotherapy. Methods Forty-seven cases of gastric cancers with malignant ascites were selected. The gastric cancer cells were isolated from the ascites of 19 patients with gastric cancers, and in vitro sensitivity tests to carboplatin, taxol, fluorouracil, cisplatin, adriamycin, hydroxycamptothecine, methotrexate, mitomycin and dacarbazine were performed by ATP bioluminescence assay. One of the most sensitive drugs was applied to intraperitoneal chemotherapy. The rates of the patients with complete remission of ascites and with disappearance of intraperitoneal cancer cells in the sensitivity test group were compared to those of 28 patients in control group who all were treated with intraperitoneal cisplatin. Results In sensitivity test group, the sensitive cases to carboplatin, taxol, fluorouracil, cisplatin, adriamycin, hydroxycamptothecine, and mitomycin were 7,6,6,6,5,5 and 5 respectively. No case was sensitive to methotrexate or dacarbazine. The rates of patients with complete remmission of ascites and the rates of patients with disappearance of intraperitoneal cancer cells in the sensitivity test group were significantly higher than those in cisplatin control group(68.4% vs. 32.1%, P

Objective:To investigate the change of apoptosis-associated genes in human lung adenocarcinoma cell lines A549/DDP cells,which were induced by the recombinant human interleukin-24(rhIL-24) combined with Cisplatin (DDP). Methods: Six genes expression level by GeXP genetic analysis system at the same time,after rhIL-24,DDP and rhIL-24+DDP were used to intervene in A549/DDP cells. Results:rhIL-24 could induce Bax gene,Caspase3 gene and Rb gene transcription up regulation;Bcl-2 gene and survivin gene transcription down regulation. But no regular change in the genes expression level of P53. Bax,Survivin and Rb were more obviously changed after rhIL-24 combined with DDP. Conclusion: RhIL-24 can induce apoptosis of A549/DDP cells through upregulated the genes expression level of Bax,Caspase3,Rb and down regulated of Bcl-2,Survivin.

AIM:To investigate the effect of RNA interference (RNAi)-mediated insulin-like growth factor 1 receptor ( IGF1R) gene silencing on the growth , migration, and invasion of hepatocellular carcinoma cells .METHODS:The most effective siRNA targeting IGF1R gene was designed and screened .After lentiviral expression vector pLVX-shR-NA2-IGF1R carrying the most effective siRNA sequence was constructed , it was transfected into 293T cells and packed into pLVX-shRNA2-IGF1R lentivirus.Huh7 and Hep3B cells were infected with the pLVX-shRNA2-IGF1R lentivirus to screen the positive clone Huh7 cells and Hep3B cells with the lentivirus .These Huh7 cells and Hep3B cells were cultured to ana-lyze the mRNA level of IGF1R, cell proliferation, cell cycle, cell apoptosis, cell migration/invasion, and the protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL.RESULTS:The mRNA expression of IGF1R in Huh7 cells and Hep3B cells with pLVX-shRNA2-IGF1R lentivirus was significantly reduced .The proliferation of these cells was remarkably inhibited , and the number in G 1 phase was increased significantly .The percentages of apop-totic cells were increased markedly , and the number of cell migration/invasion was decreased markedly .The protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL were decreased significantly compared with the blank control group and negative control group .CONCLUSION:The RNAi-mediated IGF1R gene silencing sig-nificantly suppresses the growth and the malignant biological characteristics of Huh 7 cells and Hep3B cells, which may be involved in the reduced protein levels of the above genes induced by down -regulation of IGF1R expression.

AIM: To construct HBx eukaryotic expression vector pEYFP-C1-X and eukaryotic expression vector pGL3-P4 driven by P4 promoter of human IGF-II gene and to investigate the effect of HBx on the transcription activity of IGF-II gene P4 promoter. METHODS: HBx gene and P4 promoters were cloned into pEYFP-C1 and pGL3-basic vectors respectively by gene recombination techniques to construct recombinant plasmids pEYFP-C1-X and pGL3-P4. HepG2 cells were transfected with pEYFP-C1-X and the resistant cell clones were selected by G418. Then methylated pGL3-P4 was transiently transfected into the above cell clones, and the transcription activity of P4 promoter was determined by dual-luciferase reporter assay system. RESULTS: (1) Aim fragments HBx gene and P4 promoter that were cloned were 465 bp and 1 246 bp, respectively and the DNA sequences were accordant with GenBank data confirmed by restricted enzyme digestion and sequencing. (2) HepG2-EYFP-X cells that expressed HBx protein were obtained. (3) Luciferase activity of methylated P4 promoter in the HepG2-EYFP-X was more than that of control cell HepG2-EYFP (P<0.01). CONCLUSION: HBx may enhance the transcription activity of the P4 promoter.

AIM To evaluate the beneficial effects of isoliensinine on paraquat(PQ)-induced acute lung injury and pulmonary fibrosis and explore the mechanism of its action. METHODS PQ (45 mg·kg-1, ip)-induced acute lung injury and PQ (100 mg·kg-1, ig)-induced pulmonary fibrosis were prepared. At 8, 24 and 48 h after PQ administration, the effects of isoliensinine (20 mg·kg-1, ig, 3 times a day, from 24 h before PQ administration to the end of experiment) on activity of superoxide dismutase (SOD), alkaline phosphatase (ALP) and the content of malondialdehyde (MDA) in plasma and bronchoalveolar lavage fluid (BALF) of acute lung injury groups were evaluated respectively. On the 14 d following PQ ingestion, the effects of isoliensinine (10, 20 and 40 mg·kg-1, ig, twice a day, from 24 h before PQ administration to the end of experiment) on hydroxyproline content, transforming growth factor β1 (TGF-β1) and matrix metalloproteinase-2 (MMP-2) expressions and the histopathological changes in lung tissues of pulmonary fibrosis groups were observed. RESULTS In the acute lung injury model, isoliensinine (20 mg·kg-1) significantly increased SOD activity, and decreased MDA content and ALP activity, as well as ameliorated the histopathological damage of lung tissue compared with PQ group. However, the indexes mentioned above in isoliensinine alone group did not change obviously compared with normal saline group. In the pulmonary fibrosis model, isoliensinine (10, 20 and 40 mg·kg-1) resulted in a dose-dependent decrease of hydroxyproline content compared with PQ group [(2.11±0.21), (1.94±0.24) and (1.89±0.26), respectively, vs (2.44±0.33) mg·g-1 wet tissue]. The expressions of TGF-β1 and MMP-2 in the lung tissue of the isoliensinine 40 mg·kg-1+PQ group were significantly less than those of the PQ group. Furthermore, isoliensinine could improve the histopathological changes of fibrosis as comparison with PQ group. CONCLUSION Isoliensinine has protective effects on PQ-induced acute lung injury and pulmonary fibrosis.

AIM: To construct the shuttle plasmid vector for thymidine kinase (tk) and EGFP fusion protein gene driven by IGF - Ⅱ P3 promoter, and investigate the specific killing effect of the HSV - tk/GCV system on hepatocellular carcinoma(HCC) cells in vitro. METHODS: Recombinant shuttle plasmid vector was constructed by techniques of genetic recombination and screening, and identified by restriction digestion and sequencing analysis. Then the recombinant shuttle plasmid was transfected into HepG2 and HeLa cells by techniques of lipofectamine transfection and its expression was detected by fluorescence microscope and RT -PCR. Cell killing after ganciclovir(GCV) application was determined by MTT. RESULTS: Identification of pDC316 -tkEGFP- P3 by enzyme digestion and sequencing analysis showed that the length, inserted location and direction of the target genes which were inserted into the recombinant were correct. It was found that enhanced green fluorescence protein could only be seen in HepG2 cells, but not in HeLa cells. The results of RT -PCR showed that only two bands could be seen in the samples of pDC316 -tkEGFP- P3 transfected HepG2 cells. The MTT test showed the selective cytotoxicity of GCV to the transfected HepG2 cells. CONCLUSION: The shuttle plasmid vector carrying the tkEGFP fusion protein gene driven by IGF - Ⅱ P3 promoter has been constructed successfully and its specific expression in HepG2 cells provided a sound basis for targeted gene therapy for HCC.

ObjectiveTo evaluate the efficacy and safety of the angiotensin Ⅱ receptor blockers (ARB) in reducing portal hypertension ( PHT) in patients with cirrhosis.Methods PubMed,EMBASE,Web of Science,The Cochrane Central Register of Controlled Trials,Chinese BiomedicalDatabase,ChineseJournals Full-text Database and WanFang Digital Journal Full-text database were searched.Statistical analysis was performed by meta-analysis using RevMan4.2 software.ResultsAmong 8 randomized controlled trials ( RCT) including 282 patients met the inclusion criteria,4 trials were analyzed to compare the ARB with the placebo or no treatment and the other 4 trials were analyzed to compare the ARB with propranolol.Meta-analysis results were as follows.(1) The ARB resulted in more significant hepatic venous pressure gradient ( HVPG) reduction as compared with the placebo or no treatment [ WMD =1.87 mm Hg (1 mm Hg =0.133 kPa),95％CI ( 0.86-2.87 )mmHg,P =0.00003 ].Andthe ARB were similar to propranolol in reducing HVPG [ WMD =0.92 mm Hg,95％ CI ( - 0.41-2.26)mm Hg,P =0.17 ].(2)The ARB led to more significant reduction in mean arterial pressure than the placebo or no treatment [ WMD =8.89 mm Hg,95％ CI( 7.16-10.62)mm Hg,P ＜ 0.00001 ],but they were similar to propranolol had no significant difference.And the ARB had no significant effect on the heart rate of the patients,which was similar to no treatment group ( P ＞ 0.05 ).Whereas,propranolol could greatly decrease heart rate of the patients ( WMD =- 21.25,95％ CI - 25.83-16.68,P ＜ 0.000 01 ).( 3 ) No significant differences were found in serum bilirubin and creatinine levels between the ARB and the placebo or no treatment groups ( P ＞0.05).The rate of nonspecific adverse events was higher in the ARB groups than in the placebo or no treatment groups ( P =0.03 ),but it showed there was no difference between the ARB and propranolol groups (P =0.72).ConclusionThe ARB is effective in reducing portal hypertension in patients with cirrhosis,which is similar to propranolol.Their effects on mean arterial pressure is similar to propranolol without significant effects on hear rate,liver functionand kidney function,and with less nonspecific adverse events.The ARB could become a new choice for the treatment of portal hypertension.

AIM: To obtain a single-chain antibody with high affinity for hepatocellular carcinoma (HCC). METHODS: A second single-chain antibody mutant library was established by using error-prone PCR and DNA shuffling. Single-chain antibodies with high affinity for hepatocellular carcinoma were selected from phage antibody library by using ELISA. RESULTS: The content of the second single-chain antibody mutant library was about 4.5?10~7. Two selected mutants M25 and M36 were obtained after 3 rounds of panning and ELISA. Immunoassay showed that M25 and M36 bound to human HCC cells specifically. The relative affinity of M25 was 2.0 folds higher than that of the original antibody, and M36 was 2.4 folds higher than the original antibody. CONCLUSION: Error-prone PCR combined with DNA shuffling is an effective method to improve affinity of antibodies isolated from phage antibody library.