Objective:To prepare monoclonal antibody(McAb) against human augmenter of liver regeneration(hALR) by hybridoma technique with unpurified recombinant protein expressed by E.coli as screening antigen.Methods:BALB/c mice were immunized with hALR thioredoxin(hALR Thi) fusion protein twice.The splenocytes were fused with myeloma cells by polyethylene glycol and the hybridomas were selected in HAT medium.The positive hybridomas secreting anti hALR McAb were screened by means of ELISA,with the lysates of E.coli containing the recombinant plasmid pQE30 hALR induced by IPTG as screening antigen and containing plasmid pQE30 as controlling antigen.The reactivity of McAb secreted by the positive hybridoma to hALR expressed in eukaryocyte and native hALR in human serum was confirmed by means of ELISA and Western blot methods.Results:A positive hybridoma was screened successfully;the specific antigen antibody reaction was confirmed between the McAb prepared and hALR expressed in eukaryocyte and native hALR in human serum.Conclusion:Without any purification procedure,recombinant protein expressed by E.coli can be used for screening bybridomas in preparing of McAb with the expressed product of empty plasmid as control;anti hALR McAb can be used to the further study of hALR.

Acidic environment of organelles of eukaryotic cells plays an important role in endocytosis, the secretion of lysoso-mal enzymes and other physiological activities.V-ATPase (vacuolar ATPases) and CLC (chloride channel) family are widely distribu-ted and present in virtually all eukaryotic cells in intracellular membranes and in the plasma membrane.They are responsible for the a-cidification of the vesicular interior.In this paper, the distribution, structure, mechanism of action and physiological and pathological significance of V-ATPase and CLC protein are discussed.

10 000 u,5 000~10 000u and 10 000 u group,and the gene expression was slightly increased in molecular weight 5 000～10 000 u group,but no significant difference of gene expression and protein secretion in 10 000 u uremic toxin,through promoting the gene expression and protein excretion of TGF-?_1 of renal tubular epithelial cells in patients with chronic renal failure.

A total of 210 female patients with overactive bladder (OAB) were divided into M receptor blockage (M,n =71),pelvic floor muscle stimulation physiotherapy (S,n =70) and combination (C,n =69) groups.After one course of treatment,the OAB symptom scores were compared to pretreatment values (P ＜ 0.05).After a second course,the overall response rate was 69％ in group S.Pelvic floor muscle stimulation physiotherapy provided excellent sustained outcomes.

Objective Molecular targeted drugs have anti-angiogenesis and anti-tumor effects.The study was to investigate the role of sunitinib on treating metastatic renal cell carcinoma ( mRCC) and its long-term efficacy and adverse reactions. Methods From November 2007 to March 2013, 281 mRCC patients were divided into two groups according to their own willingness.102 patients in the experimental group received sunitinib 50 mg per day.Pain score, metastatic focuses before and after targeted therapies were compared.179 patients in control group was treated by immunotherapy.Kaplan-Meier survival analysis was used to draw the survival curves and Cox regression model was applied in multivariate analysis. Results There was no difference between the two groups on baseline (P>0.05).In the experimental group, 78 cases were clear cell carcinoma, with remission rate 29.4%, stablity rate 53.8%and total effective rate 83.2%after treatment;15 cases were papillary renal cell carcinoma, with remission rate 33.3%, stablity ratio 46.7%and total effective rate 80.0% after treatment;9 cases were collecting duct carcinoma, with the remission rate 22.2%, the stablity ratio 55.6% and the total effective rate 77.8% after treatment.The bone scan after 1 cycle of treatment showed 2 cases progressed(10.5%), 5 cases relieved(26.3%) and 12 cases sta-blized(63.2%), the total effective rate of bone metastasis amounting to 89.5%, the effective rate of lung metastasis 81.7%, the effective rate of liver metastasis 60.0%and the effective rate of lymph node metastasis 72.8%.The median overall survival in the experimental group was 28.9 months, 20.7 months in control group(P<0.05).Cox regression analysis showed age, tumor classification, liver, lung, bone and lymph node metastasis were prognostic factors influencing mRCC.The common averse events were white blood cells, thrombocytopenia, neutropenia and hypoalbuminemia. Conclusion Sunitinib for the treatment of mRCC can diminish the metastatic focuses , ease the pain , improve the quality of life, delay the progression of the disease, and prolong the overall survival.

Objective To investigate the clinical efficacy and safety of intravenous immunoglobulin and interferon in the treatment of hand foot and mouth disease complicated with viral encephalitis .Methods 80 cases of hand foot and mouth disease complicated with viral encephalitis were randomly divided into the three groups according to the random number table .27 cases in the control group were given comprehensive symptomatic treatment ,27 cases in the study group 1 were given gamma globulin ,and 26 cases in the study group 2 were given interferon .The clinical efficacy,improvement of disease ,incidence of adverse reactions of the three groups were compared .Results Defer-vescence time ,seizure control time ,time of skin rash subsided ,time of psychiatric symptoms relieved and the average hospitalization time in the study 1 group were (3.65 ±0.28)d,(4.04 ±0.33)d,(3.86 ±0.27)d,(5.83 ±0.36)d and (7.53 ±0.83)d,which were significantly less than those in the control group (t=8.43,8.58,9.15,9.80,8.96, all P<0.05).Those in the study 2 group were (3.92 ±0.29)d,(4.21 ±0.32)d,(4.27 ±0.30)d,(6.32 ±0.43)d and (8.10 ±0.72)d,which were significantly less than the control group (t=7.99,8.17,8.54,9.18,8.55,all P<0.05);however,there were no significant differences between the study 1 group and the study 2 group(t=1.12, 2.04,1.67,1.38,2.21,all P>0.05).The incidence rate of adverse reaction among the three groups showed no sta-tistically significant difference (χ2 =2.17,P>0.05).Conclusion Intravenous immunoglobulin and interferon have significant effect in the treatment of hand foot mouth disease complicated with viral encephalitis , which can quickly improve symptoms ,shorten treatment time and have high safety and good clinical application value .

Aim To investigate the effects of Panax Notoginsenosides(PNS) on the gene expression and protein excretion of TGF-?_1 and CTGF of human renal tubular epithelial cell induced by uremic serum in vitro.Methods Forty sera from CRF patients and twenty sera from healthy volunteers were collected,gently mixed,inactived and separated respectively in steriled condition.HK-2 Cells were cultured in RPMI-1640 medium with 10% newborn calves serum and subcultured routinely.They were differentiated by phase contrast microscope and scanning electron microscope detection and cytokeratin18(CK-18) immunohistochemistry method.The protein levels of TGF-?_1 were examined by enzyme-linked immunoadsordent assay(ELISA).The protein levels of CTGF were examined by Western Bloting assay.The gene expression of TGF-?_1 and CTGF was detected by Semi-quantitative reverse trainscriptase polymerase chain reaction(RT-PCR).Results TGF-?_1 and CTGF gene expression and protein level were increased in 10% uremic serum groups compared with that of normal control group(P

10 000 D uremic toxin through promoting the gene and protein expression of CTGF in renal tubular epithelial cells in patients with chronic renal failure.

Objective To investigate the relationship between vascular cognitive impairment(VCI) and the angiotensinogen(AGT) gene ( G-6A and M235T) polymorphism. Methods Randomnized controled study was ap- plied in the study. AGT gene G-6A and M235T genotypes of 67 cases with VCI and 71 normal controls were deter- mined by polymerase chain reaction (PCR). Results For the location of M235T, the frequencies of T allele(0.73 I and TT genotype ( 0.52 ) were observed in VCI compared with control group ( 0.68,0.45, P > 0.05 ). The odds ratio associated with TT/MM genotype was 0.544 ( 95% CI 0.208～1. 424 ,P > 0.05 ). For the location of G-6A ,the fre- quencies of A allele(0.69) and AA genotype (0.48) were observed in VCI compared with control subjects (0.63, 0.39,P > 0.05). The odds ratio associated with AA/GG genotype was 0.602 ( 95% CI 0.252～1. 738, P > 0.05 ). There was no difference in allele distribution between 67 VCI patients and the controls. Conclusion There is no correlation between vascular cognitive impairment and AGT gene polymorphisms of M235T and G-6A. AGT gene pol- ymorphism is not included in the risk factors for development vascular cognitive impairment.