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.

Objective To analyze the correlation between the epithelial‐mesenchymal transition and the clinicopathologic features of IgA nephropathy .Methods A total of 168 patients diagnosed as IgA nephropathy by renal biopsy in Xinqiao hospital from Janu‐ary 2011 to December 2013 were divided into high expression of dual‐positive Snail and a‐SMA group (Group A ,117 cases) and low expression of dual‐positive Snail and a‐SMA group (Group B ,51 cases) according to results of immunohistochemical method .The clinical parameters (age ,gender ,course of disease ,BMI and chemical indicators) and renal pathology grade were compared by statis‐tical analysis .Results There were difference between group A and group B in the course of disease and BMI(P<0 .05) .There were differences between group A and group B in the incidences of creatinine ,blood urea nitrogen ,serum triglycerides and 24‐hour urine protein amount (P<0 .05) .The percentage of Lee′s grade Ⅰ ,Ⅱ ,Ⅲ in group B was significant ,while percentage of Lee′s gradeⅣ + Ⅴin group A was significant .The expression of Snail and a‐SMA in grade Ⅳ + Ⅴ was more than that in grade Ⅰ + Ⅱ (40 .2%vs .9 .4% ) ,and the difference between two groups was statistically significant (P< 0 .05) .Conclusion The expression of Snail and‐SMA were related with 24‐hour urine protein amount and kidney function in IgA nephropathy ;and Lee′s grade was severe in patients with high expression of Snail and‐SMA .

OBJECTIVE:To comparison the clinical efficacy and safety of sitagliptin and Repaglinide in the treatment of new-ly diagnosed patients with type 2 diabetes mellitus. METHODS:214 newly diagnosed patients with type 2 diabetes mellitus were randomly divided into study group and control group with 107 cases in each group according to random number table method. Both group received routine diabetes diet,health education and suitable exercise. Control group was treated with Repaglinide tablet 1.0 mg,tid;while study group was treat with Sitagliptin tablet 100 mg,qd. Lab indexes,FBG,PBG,HbA1c and BMI were observed in 2 groups before and after treatment. ADR were compared between 2 groups. RESULTS:There was no statistical significance in lab indexes between 2 groups before and after treatment (P>0.05). After treatment,FBG,PBG,HbA1c were significantly de-creased significantly in 2 groups,with statistical significance (P<0.05),there was no statistical significance between 2 groups (P>0.05). BMI of 2 groups were increased significantly,and the study group was higher than the contrd group,with statistical sighificance (P<0.05). Patients in the study group had no hypoglycemia,while 3 patients in the control group suffered from it, there was statistical significance (P>0.05). CONCLUSIONS:Sitagliptin is similar with repaglinide in therapeutic efficacy;both can effectively reduce blood glucose of elderly patients with newly diagnosed type 2 diabetes mellitus,and the incidence of hypo-glycemia was low,the saterty was good,but sitagliptin has a better control of BMI in patients.

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.

Objective To observe the effect of electroacupuncture (EA) on dopaminergic neurons in the mesencephalic substantia nigra(MSN) of rats with Parkinson′s disease (PD). Methods A model of PD was induced in rats by injecting 6-hydroxydopamine (6-OHDA) into the MSN. Acupuncture was then used to treat the model rats. To examine the apoptosis of dopaminergic neurons, the number of tyrosine hydroxylase (TH) positive neurons and the concentration of dopamine (DA) were measured in a normal control group、a sham-operation group、the EA group and the PD group which received no EA. Results In the EA group, markedly less apoptosis of dopaminergic neurons was observed, and the number of TH positive neurons and the concentration of DA were significantly higher. Conclusion Electroacupuncture may mitigate injury to the substantial nigra in Parkinson′s disease.

Objective To investigate the expression of phosphorylation-mammalian target of rapamycin (p-mTOR) in endometrial carcinoma. Methods The expressions of p-mTOR protein in tissues from 45 cases of endometrial adenocarcinoma, 7 endometrial atypical hyperplasia, and 6 normal endometrium were detected by immunohistochemistry SP method. Results The expression of p-mTOR protein maily restricted to cytoplasm. Compared with normal endometrium, the expression of p-mTOR protein in the cases of endometrial atypical hyperplasia and endometrial adenocarcinoma was significantly up-regulation(2.36 ± 0.76vs 6.21 ± 1.19, 15.82 ± 2.64)( F = 11.37, P ＜ 0.05 ). There was significant difference of p-mTOR protein in different histology class in endometrial adenocarcinoma (F = 8.27, P ＜ 0.05), but there was no significant difference of p-mTOR protein in different pathological grade in endometrial adenocarcinoma (P ＞0.05).Conclusion The expression of p-mTOR protein may participate in the occurrence and development of endometrial carcinoma.

Antigens, CD20 ; metabolism ; CD79 Antigens ; metabolism ; Heart Atria ; Heart Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Lymphoma, Large B-Cell, Diffuse ; diagnostic imaging ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed

AIM:ToinvestigatetheeffectofIkarosisoformsontheproliferationofhumanovariancancerSK-OV3 cells.METHODS:Three isoforms of Ikaros, IK1, IK2 and IK6, were transfected into ovarian cancer SKOV3 cells. CCK-8 assay and cell counting were used to detect the effects of Ikaros isoforms on the proliferation of SKOV 3 cells.The cell cycle was analyzed by flow cytometry .The cell cycle-related proteins were detected by Western blot .RESULTS:IK1 and IK2 expression inhibited SKOV 3 cells proliferation .Flow cytometry analysis indicated that IK 1 and IK2 induced SK-OV3 cell cycle arrest at the G 1 phase.IK6 isoform exerted no obvious effect on the proliferation or cell cycle of SKOV 3 cells.Compared with control EV group , IK1 group and IK2 group showed a dramatic elevation in the expression of the cell cycle inhibitor p21, along with a substantial decrease in the expression of the cell cycle inducers cyclin D 1 and cyclin D2, which did not change in IK 6 group.CONCLUSION:IK1 and IK2 significantly inhibit the proliferation of ovarian cancer SKOV3 cells and induce cell cycle arrest at G 1 phase by regulation of cell cycle-related proteins cyclin D1, cyclin D2 and p21, while IK6 isoform exerts no obvious effect on the proliferation and cell cycle of SKOV 3 cells.

Objective To analyze the molecular characteristics and prognosis in acute myeloid leukemia patients with AML1/ETO.Methods The clinical data of 63 cases of acute myeloid leukemia (AML) patients with AML1/ETO positive were analyzed retrospectively.56 cases of AML patients with AML1/ETO negative in the same period were analyzed as control.Characteristics in morphology,immunology,cytogenetics,molecular biology and the clinical effects of treatment were studied and analyzed.Results M2a was 57.12 ％ (36/63),M2b was 33.33 ％ (21/63) in AML with AML1/ETO.The percent of initial marrow blasts was 0.46±0.16.The positive rate of CD34,CD13,CD33,CD19,CD7 and CD56 was 67.21％,52.46 ％,40.98 ％,63.93 ％,4.92 ％ and 50.82 ％,respectively.The rate of t(8;21) translocation was 82.54 ％.There was 4.76 ％ with additional chromosome abnormality,three cases with EV1 1and one case with MLL/AT9.The overall CR rate,the relapse rate,the 3-year and the 5-year overall survival rate was 71.43 ％,51.11％,(43.01±5.31) ％ and (32.79±3.81) ％,respectively.There was no significant difference compared with the control group (P ＞ 0.05).But extramedullary infiltration,the expression of CD56 and additional chromosome abnormality had statistical effects on overall survival (P ＜ 0.05).Conclusions There has unique characteristics in AML with AML1/ETO.The effects of treatment and the prognosis are affected by many factors,so the efficacy and prognosis of AML with AML1/ETO couldn' t just depend on AML1/ETO.