1.Effect of HSP70 gene transfection to rat peripheral blood cytokines in the early phase of sepsis
Yingdong CHENG ; Ping LIANG ; Chaojun ZHANG ; Zhenxiang YAO ; Congbing YANG
Chinese Journal of General Surgery 1993;0(03):-
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
2.Application of circular anastomotic stapler for sigmoid colostomy in laparoscopy abdominoperineal resection
Dan MA ; Chaojun ZHANG ; Zulin CHEN ; Yingdong CHENG ; Hua YANG
Chongqing Medicine 2014;(5):556-557,560
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 .
3.The experimental study of the inhibitory effect of human recombinant sTRAIL targeting activated by u-PA on the colon cancer
Dan MA ; Chaojun ZHANG ; Zulin CHEN ; Hua YANG
Chongqing Medicine 2013;(33):3992-3994
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 .
4.Expression of TβRⅡRH mRNA in PBMCs of patients with lupus nephritis
Chen ZHAO ; Yongzhe LI ; Dinghua LIU ; Chaojun HU ; Dawei TONG ; Shulan ZHANG ; Yang ZHAO
Chinese Journal of Rheumatology 2008;12(6):395-397
Objective To investigate the association of TGF-β receptor typeⅡ(TβRⅡ)mRNA with lupus nephritis (LN) and disease activity by testing its expression levelin peripheral blood mononuclear cells (PBMCs).Methotis Forty-four patients with LN were included in this study.They were all had active LN.Twepty-eight LN patients were taking glueocorticoids and/or immunosuppressive agents and sixteen had never taken steroids or immunosuppressive agents.The expression levels of T13R H mRNA were semi-quantitativelydetermined by reverse transcription-polymerase chain reaction(RT-PCR).Resuits The expression levels of TβRⅡ mRNA in PBMCs from LN patients(1.7±1.0)were lower than those of non-lupus nephritis(4.0±3.1) and healthy subiects(4.1±2.5),(P<0.01).The difference of the expression levels between patients who took and had never taken glucocorticoids and/or immunosuppressive drugs was significantly statistically(P<0.05).The expression levels of TβRⅡ mRNA in PBMCs of patients with LN were correlated significantly with the systemic lupus erythematosus disease activity index (SLEDAI)scores(r-0.309.P<0.05),titers of anti-dsDNA antibody(r=-0.401,P<0.01)and serum complement C3 level(r=0.621,P<0.01).Conclusion This study suggests that TβRⅡ may be involved in the development of LN,and the TβRⅡ mRNA expression levels in PBMCs from patients with SLE are significantly correlated with LN activity.Glucocortieoids or immunosuppressive drugs can increase the expression levels of TβRⅡ mRNA and ameliorate renal damage.
5.Quantitative study of cochlear hair cells injuries in guinea pigs exposed to blast underpressure
Chaojun LI ; Zhaohua LIU ; Peifang ZHU ; Zhengguo WANG ; Cheng YANG ; Haibin CHEN ; Jihong ZHOU ; Xin NING
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(03):-
OBJECTIVE To explore the characteristics of cochlear hair cell injuries in guinea pigs exposed to blast underpressure(BUP). METHODS The guinea pigs were killed 14 days after exposure to experimental BUP. Their basilar membranes were stained by silver nitrate and the hair cell injuries were quantitatively assessed using the light microscope. RESULTS The outer hair cells(OHCs) of the guinea pigs clearly appeared to be injured following exposure to the experimental BUP at a peak underpressure varying between -22.4kPa and -63.3kPa. The most obvious injury was in the second turn and OHC loss was smallest in the first row and more severe in the second and third rows. OHC loss was seen in all of three experimental groups of animals following exposure to BUP. Furthermore, the higher the peak of underpressure, the more obvious the injury of OHCs. Quantitative morphological analysis of cochlear hair cells showed that the total OHC loss rates in all the experimental groups of guinea pigs exposed to BUP were significantly higher than that of control group animals(P
6.Expression of costimulating factor B7-H4 in patients with primary billary cirrhosis
Chen ZHAO ; Yongzhe LI ; Yang ZHAO ; Chaojun HU ; Dinghua LIU ; Dawei TONG ; Shulan ZHANG
Chinese Journal of Microbiology and Immunology 2008;28(9):851-855
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.
7.Cloning and expression of histidyl-tRNA synthetase autoanfigen gene and its clinical application
Shanshan LI ; Yongzhe LI ; Zhixian ZHAO ; Dawei TONG ; Shulan ZHANG ; Chaojun HU ; Weiping YANG
Chinese Journal of Laboratory Medicine 2008;31(2):138-142
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.
8.The discussion on therapeutic method and curative effect of acute subjectivity tinnitus
Guijun YANG ; Minghua GAO ; Chaojun LI ; Zhongwan LI ; Shaojing KUANG ; Fan ZOU ; Shuping SU
Chongqing Medicine 2015;(31):4352-4354
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 .
9.Study of direct identification of pathogens from positive blood cultures by methods of separation gel tube -centrifugation
Chenrui HOU ; Yun YANG ; Ziyang LI ; Chunyan GAO ; Miao SU ; Chaojun LANG
Chinese Journal of Primary Medicine and Pharmacy 2016;23(14):2119-2122
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.
10.Effect of suppression of long non-coding RNA-BG on radiosensitivity of normal human bronchial epithelial cell line Beas-2B
Chang LIU ; Chaojun PENG ; Lili WANG ; Wei ZHU ; Jiaying XU ; Yang JIAO
Chinese Journal of Radiological Medicine and Protection 2014;34(5):323-328
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.