中华实验和临床病毒学杂志最新文献

[Establishment of localization ultrathin section for cytopathic cells]. 【细胞病变细胞定位超薄切片的建立】。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Jing-Dong Song, Jian-Guo Qu, Tao Hong

引用次数: 0

[Expression and purification of four single-stranded DNA-binding proteins and their binding on HCV RNA]. [四种单链dna结合蛋白的表达纯化及其与HCV RNA的结合]。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Hai-Yan Shi, Yong-Jun Li, Ji-Min Gao

{"title":"[Expression and purification of four single-stranded DNA-binding proteins and their binding on HCV RNA].","authors":"Hai-Yan Shi, Yong-Jun Li, Ji-Min Gao","doi":"","DOIUrl":"","url":null,"abstract":"Objective: Express and purify four single-stranded DNA-binding (SSB) proteins, and evaluate the binding of SSB proteins on HCV RNA.Methods: The expression plasmids of four SSB proteins were conducted, termed TTH, SSOB, KOD and BL21, respectively. The BL21 (DE3) was transformed by the expression plasmid of TTH, Transetta (DE3) were transformed by the expression plasmid of SSOB, KOD and BL21, then protein expression was induced with IPTG, the expression products were analysised by SDS-PAGE. To evaluate the binding of SSB on HCV RNA, RNA-SSB protein complexes were applied to a 1.2% TAE agarose gel.Results: Suitable competent cells were transformed with the expression plasmids, induced by IPTG. SSB proteins were purified by affinity chromatography, to visualize their purity all SSB proteins were applied to SDS-PAGE analysis. All four proteins showed single clear bands. We have successfully obtained the SSB protein expression plasmid, expressed and purified SSB protein. TAE agarose gel electrophoresis was used to confirm SSB protein-RNA binding activity. The each of SSB-RNA complex migrated more slowly than the sole RNA, which suggested SSB protein could specifically bind to RNA.Conclusions: We have expressed and purified four SSB proteins, and for the first time found that SSB protein can bind HCV RNA. Our results may provide a basis for future studies of the novel functions of SSB proteins on RNA.","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"354-6"},"PeriodicalIF":0.0,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32187770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[The comparative study on the regulation of apoptosis by hepatitis B virus X protein between B and C genotype]. [乙型肝炎病毒X蛋白在B、C基因型中调控细胞凋亡的比较研究]。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Yue Ming, Qi-Feng Xie, Lin Yang

{"title":"[The comparative study on the regulation of apoptosis by hepatitis B virus X protein between B and C genotype].","authors":"Yue Ming, Qi-Feng Xie, Lin Yang","doi":"","DOIUrl":"","url":null,"abstract":"Objective: To investigate the apoptosis regulation on hepatoma cells by HBx between genotype B and C.Methods: Genotype B and C HBx gene fragments were amplified and inserted into green fluorescent protein (GFP) eukaryotic expression vector pEGFP-C1 to construct recombinant pGFP-XB and pGFP-XC. The pEGFP-C1, pGFP-XB and pGFP-XC were introduced into Bel-7402 cells by Fugene HD to obtain Bel-7402 cells expressing GFP. The transcription and expression of HBx gene were demonstrated by RT-PCR and Western Blot analysis. Bel-7402, Bel-7402/GFP, Bel-7402/GFP-XB, Bel-7402/GFP-XC cells were treated with adriamycin (2.5 microg/ml), and the apoptosis of the cells was determined by trypan blue exclusion, and flow cytometry analysis.Results: RT-PCR and Western Blot analysis showed that HBx genes of genotypes B and C were transcribed and expressed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Trypan blue exclusion showed adriamycin induced time-dependent cell death in Bel-7402, Bel-7402/GFP cells while no significant cell death was observed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Flow cytometry analysis indicated that no significant differences of apoptosis rates of Bel-7402/GFP-XB (3.87%) and of Bel7402/GFP-XC (4.01%) were observed (P > 0.05), moreover, no significant differences of Bel-7402/ GFP-XB (3.87%), Be17402/GFP-XC (4.01%) and of the untreated cells. Apoptosis rates in Bel-7402/GFP-XB (3.87%), Bel-7402/GFP-XC (4.01%) cells were significantly lower than those in Bel-7402 (27.05%) and Bel-7402/GFP (29.14%) cells at 48 hours after the adriamycin treatment (P < 0.01).Conclusions: Bel-7402 cell lines expressing GFP, GFP-XB and GFP-XC fusion proteins were successfully established. HBV X protein blocks adriamycin-induced apoptosis of Bel-7402 cells. There is no difference between HBx of genotype B and C in inhibiting apoptosis induced by adriamycin.","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"344-7"},"PeriodicalIF":0.0,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32188343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Expression of TGF-beta1/Smad protein in rat liver fibrosis model and the role of IFN-gamma]. [tgf - β 1/Smad蛋白在大鼠肝纤维化模型中的表达及ifn - γ的作用]。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Xiao-Qing Fu, Shou-Rong Liu, Jian-Chun Guo, Jian-Feng Bao

{"title":"[Expression of TGF-beta1/Smad protein in rat liver fibrosis model and the role of IFN-gamma].","authors":"Xiao-Qing Fu, Shou-Rong Liu, Jian-Chun Guo, Jian-Feng Bao","doi":"","DOIUrl":"","url":null,"abstract":"Objective: To study the impact of IFN-gamma on liver fibrosis and its possible mechanism. Thirty healthy male SD rats were randomly divided into two groups: fibrosis model group, IFN-gamma treatment group. Experimental liver fibrosis was induced by subcutaneous injection of CCl4. After 12-week-treatment, serum hyalurnic acid and TGF-beta1 was examined, histopathological changes and degrees of fibrosis were observed by optical microscopy. Meanwhile, the expression of TGF-beta1, TbetaR- I and Smad2/3 proteins was detected by immunohistochemistry and quantified by using computerized image analysis.Results: (1) Pathological observation of hepatic specimens: histological examination showed that there were significant difference between normal group and fibrosis model group by comparing with the degrees of inflammation and fibrosis (P < 0.05). And the difference between fibrosis model group and IFN-gamma treatment group was significant (P < 0.05). (2) Changes of the hepatic fibrosis index (serum HA and TGF-beta1): the levels of serum HA, TGF-beta1 in fibrosis model group were higher than IFN-gamma treatment groups (P < 0.05). (3) Changes of gene protein levels about TGF-beta1/Smad: the expressions of TGF-beta1, TbetaR- I and Smad2/3 in rat hepatic tissue were detected with immunohistochemistry techniques. The expressions of the three items in model group were higher than normal group (P < 0.01). The difference between model group and IFN-gamma treatment group was significant (P < 0.05);Conclusion: IFN-gamma treatment group had significant results on treating experimental hepatic fibrosis. By the way of inhibiting expressions of TGF-beta1, TbetaR- I, Smad2/3, IFN-gamma treatment group exerted its anti-fibrosis effect.","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"340-3"},"PeriodicalIF":0.0,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32188342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Formation and identification of virus-like particles of poliovirus type I]. [I型脊髓灰质炎病毒样颗粒的形成和鉴定]。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Xiao-Wen Wang, Wang Sheng, Yi Zeng

引用次数: 0

[Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification]. A/H1N1流感病毒HA在果蝇S2细胞株的可溶性表达及其生物活性鉴定

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Si-Min Yao, Qiao Lin, Guo-Liang Zhang, Hui Yang, Xiao-Feng Deng, Guang Nie, Xue-Bao Zheng, Ying-Xia Liu

{"title":"[Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification].","authors":"Si-Min Yao, Qiao Lin, Guo-Liang Zhang, Hui Yang, Xiao-Feng Deng, Guang Nie, Xue-Bao Zheng, Ying-Xia Liu","doi":"","DOIUrl":"","url":null,"abstract":"Objective: To express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.Methods: HA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.Results: We successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.Conclusion: We expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"360-2"},"PeriodicalIF":0.0,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32187772","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Clinical research of early, enough methylprednisone combined with immunoglobulin in treatment of severe hand-foot-mouth disease]. 【早期、足量甲基强的松联合免疫球蛋白治疗重症手足口病的临床研究】。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Ming Jiang, Xing-Chuan Wang

引用次数: 0

[Activation of Kupffer cell and related signal pathway proteins in the liver of high fat and high fructose diet induced NAFLD mice]. [高脂高果糖饮食诱导NAFLD小鼠肝脏Kupffer细胞及相关信号通路蛋白的激活]。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Ming-Li Zhu, Jing Liu, Yin-Lan Liu, Wen-Jun Yang, Yan Luo, Zhen-Jie Zhuang, Qi-Bin Jiao, Jian-Yu Chen, Jian Yan, Dong-Xue Bian, Xiao-Jie Ma, Yun-Hao Xun, Jun-Ping Shi

{"title":"[Activation of Kupffer cell and related signal pathway proteins in the liver of high fat and high fructose diet induced NAFLD mice].","authors":"Ming-Li Zhu, Jing Liu, Yin-Lan Liu, Wen-Jun Yang, Yan Luo, Zhen-Jie Zhuang, Qi-Bin Jiao, Jian-Yu Chen, Jian Yan, Dong-Xue Bian, Xiao-Jie Ma, Yun-Hao Xun, Jun-Ping Shi","doi":"","DOIUrl":"","url":null,"abstract":"Objective: To investigate the expression of F4/80, NF-kappaB, p-AKT, AKT in the liver of nonalcoholic fatty liver disease (NAFLD) mice. To determine the role of Kupffer cells (KCs) in the development of NASH (non-alcoholic steatohepatitis), and understand the pathogenic mechanism of NASH.Methods: Five C3H/HeN mice fed with normal diet were served as controls, while fifteen fed with high fat, high fructose, high fat combined fructose diet respectively for 16 weeks were as NAFLD mice models. The liver inflammation and hepatic damage were examined, and the expression of F4/80, NF-Kb, p-AKT, AKT and the content of lipid in the liver were also detected.Results: Chronic intake of high fat and 30% fructose solution caused a significant increase in hepatic steatosis in animals in comparison to water controls. Liver F4/80 and NF-kappaB were significantly higher in high fat and high fat combined fructose diet fed mice than that in controls (P < 0.01, P < 0.01), F4/80 protein were higher in high fat diet treated mice than those in fructose and high fat combined fructose groups (P < 0.01, P < 0.01). Markers of insulin resistance (e. g, hepatic phospho-AKT, AKT) were only altered in fructose-fed or high fat combined fructose animals (P < 0.01, P < 0.01).Conclusion: High fat and fructose diet may induce NAFLD in C3H/HeN mice. Kupffer cells and signal pathway proteins were activated, and they may play key roles in the initiation and progression of NASH.","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"325-7"},"PeriodicalIF":0.0,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32188338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Rapidly detect and distinguish between norovirus G I and G II type with a pair of primers]. [用一对引物快速检测和区分诺如病毒G型和G型]。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Jian-Kang Han, Xiao-Fang Wu, De-Shun Xu, Li-Ping Chen, Lei Ju

引用次数: 0

[Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection]. [多重PCR检测非o157 H7 STEC毒力基因的建立及应用]。

中华实验和临床病毒学杂志 Pub Date : 2013-10-01

Xiao-Guang Wang, Ying-Hua Zhang, Ping Wang, Xiu-Hua Chen, Ling-Fei Luo, Yun Liu, Ji-Qian Liu, Chi-Ping Song, Yang Lin Ou, Guo-Qiang Chen

{"title":"[Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection].","authors":"Xiao-Guang Wang, Ying-Hua Zhang, Ping Wang, Xiu-Hua Chen, Ling-Fei Luo, Yun Liu, Ji-Qian Liu, Chi-Ping Song, Yang Lin Ou, Guo-Qiang Chen","doi":"","DOIUrl":"","url":null,"abstract":"Objective: Traditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance. Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.Methods: Six virulence genes of non-O157:H7 (stx1, stx2, eae, hly, etpD, katP6) were detected by two groups of trebling PCRs. The multiplex PCRs were optimized by melting curve analysis in SYBR Green I real-time PCR. Testing result of multiplex PCR was consistent with serological testing.Results: The sensitivity limits of the multiplex PCR for stx1, stx2, eaeP, etpD, katP, and hly were 10 ng/ml, 120 ng/ml, 110 ng/ml,165 ng/ml, 85 ng/ml, and 15 ng/ml, respectively, which is similar with that of single PCR. When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection, 13 cases were detected for non-O157 positive.Conclusion: The method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"388-91"},"PeriodicalIF":0.0,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32190334","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0