Objective To investigate the effect of HSP70 gene transfection to peripheral blood cytokines in the early phase of sepsis in rats. Methods A rat model of sepsis was established by cecal ligation and puncture(CLP),and adenovirus-mediated HSP70 gene transfection was performed in the CLP rats. Serum TNF-?,IL-1?,IL-6,IL-10 were measured before and after HSP70 gene transfection. Results In CLP rats, serum TNF-? increased at 3h and peaked at 6h(t_(TNF-?)=16.506, P

Objective To make better use of the antitumor effects of TNF related apopotosis inducing ligand (TRAIL) ,we con-struct a new fusion protein including TRAIL ,for the sake of improving its purity and targeting to hepatocytes .Methods Fusion protein concluding His label protein and human recombinant TRAIL which linked by constructed specifically u-PA cleavage site . TRAIL protein was obtained by disintigrating from the fusion protein which had been purified .Results The purified His-rh-sTRAIL was demonstrated to be cleavable by u-PA and possess enhanced antitumor effect ,it′s toxicity to hepatocytes was reduced . Conclusion The fusion protein His-rhsTRAIL has antitumor effect on colon cancer cell with high efficiency ,highly targeting and low-toxicity .

Objective To evaluate the value of circular anastomotic stapler for sigmoid colostomy in laparoscopy abdominoperi-neal resection .Methods 62 patients of low colorectal cancer treated in our hospital from October 2010 to February 2013 was retro-spectively analyzed .All the patients were treated by laparoscopy abdominoperineal resection ,34 patients received sigmoid colostomy with circular anastomotic stapler(group A) ,and the other 28 patients received sigmoid colostomy with conventional suturing tech-niques(group B) .The medical records including operation time ,the time of return of bowl function ,postoperative hospital stay and postoperative complication rate were analysed statistically .Results All patients received the operations successfully .No conversion to open procedure and no operative death occured in two groups .The result of statistical analysis showed that in group A ,the opera-tion time ,the time of return of bowl function ,postoperative hospital stay time were shorter than group B ,and the rate of edema of sigmoid in group A were lower than group B(P<0 .05) .Conclusion The application of circular anastomotic stapler for sigmoid co-lostomy in LAPR is a safe ,effective and minimally invasive technique ,which can shorten operation time ,postoperative hospital stay and reduce the related complications of colostomy .

Objective To study the possibility of direct identification of pathogens from positive blood cul-tures by methods of separation gel tube -centrifugation.Methods 216 cases of positive blood culture were collected from 2015.7 to 2015.12.The bacterias were purified from blood culture bottle by separation gel tube.After washing 2 times,identified by MALDI -TOF MS.At the same time,traditional culture,smears and identification were done. Compared the results of identification by two methods.Results 216 cases of positive blood culture were single bacte-rial infection.By Gram stain,89 strains were Gram positive,119 strains were Gram negative and 8 strains were fungal spores.190 cases of positive blood culture were identified by MALDI -TOF MS,it concluded 67 Gram positive strains,111 Gram negative strains,4 anaerobe strains and 8 fungus.Compared with traditional culture,the coincidence rate reached up to 87.9%,Gram positive strains 78.8%,Gram negative strains 93.2%,anaerobe strains 100.0%and fungus 100.0%.Conclusion It takes less than 30 minutes purified from blood culture bottle by separation gel tube.And the time of identification is shorter than traditional culture.This method is good for clinical diagnosis and treatment.

Objective To observe the therapeutic efficacy of different treatment programs on the acute subjective tinnitus . Methods 327 patients with acute subjectivity tinnitus were randomly divide into physical ,medication and comprehensive treatment group .The 97 cases in physical group were treated with hyperbaric oxygen ,acupuncture and sound therapy ;the 103 cases in medica‐tion group were treated with microcirculation and oral steroids ;the 127 cases in comprehensive treatment groups were treated with combination the therapy of above what have mentioned .THI was used to evaluate three groups before the treatment ,two and four weeks after treatment .Results After two week′s treatment ,the efficiency rate for physical group is 60 .82% ,64 .78% in medica‐tion group and 70 .08% in comprehensive group .After four weeks of treatment ,the efficiency rate of each group is 70 .10% , 72 .81% and 80 .31% .At two‐week time point ,there was no significant difference between drug and physics group (P> 0 .05) , there was statistically significant difference between medication group and comprehensive group (P< 0 .05) .At four‐week time point ,the difference of physics group and drug group had no statistically significance compared with the two‐week in each group (P>0 .05) ,but there was a statistically significant difference in comprehensive group (P<0 .05) ,meanwhile ,there was statistically significant differences between comprehensive group and the physical group and drug group (P<0 .05) .Conclusion Physical and medication therapy have quite effects on acute subjective tinnitus ,but the effect was worse than the comprehensive therapy group . After four weeks of treatment ,the efficiency of purely physical and drug therapy have not been significantly improved compared with two weeks time ,but still improved of comprehensive treatment .

Objective To investigate the biological functions of IncRNA-BG on the radiosensitivity of normal human bronchial epithelial cell line Beas-2B.Methods Three IncRNA-BG siRNAs were designed,synthesized and traasfected into Beas-2B cells via lipofectamine.The RNA transcription level of BG was detected by quantitative real time-PCR to confirm the siRNA transfection efficiency.The experiment was divided into control group,control siRNA transfected group,and BG transfected group.Cell survival was detected by clonogenic assay,and the cell cycle distribution was determined by flow cytometry assay.The γ-H2AX foci formation after irradiation was visualized via immunofluorescence.Western blot assay was performed to detect the protein expressions of RAD50,p-P53,KU70,KU80,MDM2,CDK2 and RB.Results BG-siRNA transfection significantly reduced the BG transcription level (t =8.32-15.29,P ＜0.05) and increased cell survival after irradiation at 0.5,1,2,4 and 6 Gy.Analyzed with the multi-target model,the SERD0 of Beas-2B cells and control siRNA transfected cells were calculated to be 0.80 and 0.82,respectively.In addition,BG-siRNA transfection enhanced radiation-induced cell cycle arrest at G2 phase so that,after 4 Gy irradiation,the cells in G2 phase was increased from (37.37 ±0.63) ％ of control siRNA cells to (64.19 ± 1.01) ％ (t =30.65,P ＜ 0.05).Meanwhile,the γ-H2AX foci of BG-siRNA transfected cells was decreased from 76 ± 1.78 per 100 cells to 59-± 3.49 per 100 cells (t =13.72,P ＜0.05).The expressions of DNA damage related proteins including KU70,KUS0,CDK2 and RB were increased,but the expressions of p-P53 and RAD50 were decreased.Conclusions LncRNA-BG could regulate the radiosensitivity of the normal human bronchial epithelial cells,probably through inducing cell cycle G2 phase arrest and promoting DNA damage repair after irradiation.

Objective To clone and construct the recombinant plasmid containing Jo-1 of HepG2 cells,then purify the protein and identify the immunoreactivity of the recombinant protein.and establish the enzyme linked immunosorbent assay(ELSA)to detect Jo-1 autoantigen correlative antibodies in diagnosis of polymyositis/dermatomyositis.Methods The constructed plasmid was transformed into E.coli.DH5α and BL21(DE3).This fusion protein was purified by Ni-NTA chromatography and its immunnoreactivity was identified by SDS-PAGE and Western blot.ELISA with the fusion protein was established to detect the Jo-1 autoantigen correlative antibodies in sernm samples of 75 patient with PM/DM,30 patients with SLE.30 patients with RA,10 patients with SS and 30 normal controls.Results The sequence of Jo-1 autoantigen gene Was the same as the sequence reported on the literatures.SDS-PAGE gel analysis showed the molecular weisat of fusion protein was approximately 55 000 Da. Western blotting analysis showed that the fusion protein had the same immunoreactivity as human Jo-1 autoantigen.The results of ELISA indicated that the positive rate of anti-Jo-1 antibody was 28%.but the antibody was negative in other controls.There was significant difierence of positivity of the autoantibody between PM/DM and disease controls or normal controls (x2=31.84,P＜0.01).Conclusions The plasmid containing Jo-1 is successfully cloned into E.coli.DH5α and BL21 (DE3).EUSA analysis shows its good antigenicity and specificity.

Objective To detect the value of autoantibedy profile in primary biliary cirrhosis (PBC). Methods 96 serum samples of patients with primiary biliary cirrhosis, 100 serum samples of other antoimmune disease and 49 serum samples of healthy were tested for anti- M2, anti-3E(BPO), anti-Sp100,anti-PML,anti-gp210,anti-LKM-1 ,anti-LC-1 ,anti-SLA/LP by EUROLine. Results The positive of the anti-M2,anti-BPO, anti-Sp100, anti-PML and anti-gp210 for PBC was 76. 0%, 84.4%, 32. 3%, 28. 1% and 35.4% ,respectively. The positive of the anti-M2, anti-BPO, anti-Sp100, anti-PML and anti-gp210 for other autoimmune disease was 13.0% ,9. 0% ,3.0% ,2.0% and 1. 0%, respectively. The sensitivity of the anti-M2 for PBC was 76. 0% ,with specificity of 87. 0%. The sensitivity of the anti-BPO for PBC was 84. 4%, with specificity of 91.0% ;the sensitivity of the anti-Sp100 for PBC was 32. 3%, with specificity of 97.0%. The sensitivity of the anti-PML for PBC was 28. 1% ,with specificity of 98.0%. The sensitivity of the anti-gp210 for PBC was 35.4%, with specificity of 99. 0% . Anti-LKM-1, anti-LC-1, anti-SLA/LP positive patients with PBC were not detected;the incidence rate of liver function failure in anti-gp210 positive serum higher than anti-gp210 negative serum (χ2 = 11.17, P < 0. 01). Conclusions Multiple autoantibedies can be detected in the sera of PBC patients. The detection of autoantibody profile is useful for the diagnosis and differential diagnosis of PBC, and may he helpful for therapy and prognosis of PBC.

Objective To study the relationship between the expression of eostimulating factor B7-H4 in patients with primary biliary cirrhosis (PBC) and the pathogenesis of PBC. Methods The expression of B7-H4 mRNA on peripheral blood mononuclear cell (PBMC) of 65 patients with PBC was tested by real-time PCR. Serum levels of IL-2 were assayed by ELISA. CD4+, CD8+ T lymphocytes expression level and B7-H4 expression rate before and after activation were measured by three-color flow cytometry (FCM). Re-sults (1) Expression of B7-H4 mRNA and BT-H4 percentage in PBC group were significantly lower than that in none-PBC group and healthy controls(P<0.01);(2)After 72 h activation, the percentage of CD4+, CD8+, and CD4+ CD8+T lymphoeytes and serum levels of IL-2 decreased (P<0.05), and the percentage of CD4+, CD4+ CD8+T lymphocytes and serum levels of IL-2 were significantly higher than that of none-PBC group and healthy controls(P<0.01);(3)Levels of alanine aminotransferase(ALT), aspartate amin-otransferase(AST), alkaline phosphatase(ALP) and γ-glutamyl transpeptidase(GGT) of patients with posi-tive anti-mitochondrial antibody(AMA)-M2 rose. There was no significant difference of B7-H4 expressions on T cells between patients either with or without AMA-M2 antibody. Conclusion The costimulating factor B7-H4 can express on T lymphocytes which is activated by phytohemagglatinin(PHA) aad plays a negative role on T cells responses.

Objective To investigate the effect of endovascular treatment in iliac femoral artery occlusive disease of TransAtlantic Inter-Society Consensus (TASC) C,D lesions in elderly patients.Methods Totally 104 elderly TASC C,D patients with iliac femoral artery occlusion were treated,including TASC C (n =73),TASC D (n =31).Occlusive length range were 4.25-17.61 cm,with an average of (13.46±1.24)cm.The curative effect,complications and following up after surgery were analyzed.Results Ninty-four patients were treated successfully,and the technical success rate was 90.38％ (94/104);12 patients underwent balloon dilation,92 patients underwent balloon dilation combined stent.There were 8 cases (8/104,7.69 ％) with periprocedural complications,included arterial thrombosis (n =2),pseudoaneurysm (n=1),heart failure (n=4),and radiographic contrast nephropathy (n=1).Doppler ankle-brachial index increased from 0.32±0.12 to 0.81±0.11 (P＜0.01) at 1 week after treatment.The patency rates were 88.46 ％ (69/78) and 81.13％ (43/53) at 6 and 12 months followed-up respectively.Conclusion Minimally invasive endovascular treatment has high technical success rate,low complication rate,and satisfactory clinical curative effect for iliac femoral artery occlusive disease of TASC C,D lesions in elderly patients.