目的调查重庆地区男男性行为者(men who have sex with men,MSM)EB病毒(Epstein-Barr virus)血清流行病学现状,推测MSM人群EB病毒感染和再活化情况。方法采用非概率抽样法,对1 082例MSM(来源为2012-2015年重庆地区进行临床药物试验预防...目的调查重庆地区男男性行为者(men who have sex with men,MSM)EB病毒(Epstein-Barr virus)血清流行病学现状,推测MSM人群EB病毒感染和再活化情况。方法采用非概率抽样法,对1 082例MSM(来源为2012-2015年重庆地区进行临床药物试验预防艾滋病感染的MSM人群)和1 059名健康成人血清,采用酶联免疫吸附测定(ELISA)方法检测EB-VCA-IgG、EB-NA-IgG和EB-VCA-IgM,使用χ~2检验对结果进行分析,以P<0.05为差异有统计学意义。结果1 082例MSM中HIV阳性130例,HIV阴性952例:(1)EB病毒既往感染率在MSM、HIV阳性、HIV阴性者中分别为1 002例(92.6%)、115例(88.5%)、887例(93.2%),其中MSM组、HIV阴性MSM组与对照组952例(89.9%)差异有统计学意义(P<0.05)。(2)EB病毒未感染率在MSM、HIV阳性MSM、HIV阴性MSM组分别为5例(0.5%)、1例(0.8%)、4例(0.4%),3组均显著低于对照组53例(5.0%),差异有统计学意义(P<0.05)。HIV阳性与HIV阴性间差异无统计学意义(P>0.05)。(3)EB病毒血清学再活化率在MSM、HIV阳性MSM、HIV阴性MSM组中分别为52例(4.8%)、13例(10.0%)、39例(4.1%),其中HIV阳性MSM同对照组40例(3.8%)、HIV阴性MSM组39例(4.1%)差异有统计学意义(P<0.005)。结论(1)MSM人群EB病毒感染率高于普通人群,其发生EB病毒感染相关疾病的威胁更大。(2)非HIV感染的MSM人群EB病毒再活化率可能与正常人无差异。(3)HIV感染的MSM人群EB病毒血清学再活化率明显增高,可能同HIV感染后机体免疫力下降有关,发生EB病毒感染相关疾病的风险更大。展开更多
[Objective]The research aimed to test and identify the physicochemical characters and antiviral activity in vitro of semi-finished product of the recombinant porcine rPoIFN-α. [Method]HEp-2/VSV system was used to tes...[Objective]The research aimed to test and identify the physicochemical characters and antiviral activity in vitro of semi-finished product of the recombinant porcine rPoIFN-α. [Method]HEp-2/VSV system was used to test the antiviral activity of three batches of rPoIFN-α. Using recombinant human IFN-α as reference,the titer of interferon was measured. The semi-finished product of rPoIFN-α with the known titer were treated with 0.25% trypsin,HCl and mouse anti-porcine IFN-α monoclonal antibody. And the anti-viral activity of each batch of rPoIFN-α was detected. The physicochemical characters of rPoIFN-α were evaluated. The inhibition of induced cytopathic effect of rPoIFN-α on PPV （Porcine parvovirus） and PRV （Porcine pseudorabies） on swine kidney cell （PK-15） was determined. And the antiviral activity of rPoIFN-α in vitro was observed. [Result]The titers of semi-finished products of rPoIFN-α titrated by HEp-2/VSV system could reach 1.5×105 IU/ml,with the specific activity of 1.1×106 IU/mg. The residual rate of the tier of rPoIFN-α treated by 0.25% trypsin at 37 ℃ for 1 h was less than 1%. And that treated with HCl （pH of 2.0） for 72 h was up to 95%. And that treated at 56 ℃ for 30 min and that treated by mouse anti-porcine IFN-α monoclonal antibody at 37 ℃ for 1 h were higher than 47% and about 1% respectively. The antiviral test in vitro showed that 50 and 500 IU/ml rPoIFN-α could inhibit the induced cytopathic effect of PRV and PPV on PK-15 cell lines. [Conclusion]rPoIFN-α had the basic physicochemical characters of IFN-α. And it could inhibit the induced cytopathic effect of PRV and PPV on PK-15 cell lines,but there was dosage difference.展开更多
AIM： To detect the plateletoactivating factor （PAF） and the plasma or serum levels of tumor necrosis factoroa （TNF-α） malondialdehyde （MDA）, endotoxin （ET） and to discuss their significance in various types ...AIM： To detect the plateletoactivating factor （PAF） and the plasma or serum levels of tumor necrosis factoroa （TNF-α） malondialdehyde （MDA）, endotoxin （ET） and to discuss their significance in various types of viral hepatitis. METHODS： PAF, TNF-α, MDA, and ET levels in 60 controls, 16 cases of acute viral hepatitis, 71 cases of chronic viral hepatitis, 19 cases of severe viral hepatitis were detected by reverse phase high-performance liquid chromatography （rHPLC）, bio-assay, ELISA, thiobarbituric acid （TBA）, and limulus lysate test （LLT）, respectively. RESULTS： The rHPLC was more sensitive and specific than bio-assay （r = 0.912, P〈0.01）. The plasma levels of PAF, TNF-α, MDA, and ET in patients with viral hepatitis were higher than those in controls （P〈0.01）. CONCLUSION： rHPLC is more reliable and accurate for the detection of PAF.展开更多
Intracellular RNA viruses are sensed by receptors retinoic acid-inducible gene 1 （RIG-I）/melanoma differentiation-associated gene 5 （MDA5） that trigger the formation of MAVS signal complex on mitochondria. Consequ...Intracellular RNA viruses are sensed by receptors retinoic acid-inducible gene 1 （RIG-I）/melanoma differentiation-associated gene 5 （MDA5） that trigger the formation of MAVS signal complex on mitochondria. Consequently, this leads to the activation of TANK-binding kinase 1 （TBK1） and phosphorylation of interferon regulatory factor 3 （IRF3）, both of which constitutively associate with cytosolic chaperone Hsp90. It remains largely unknown how MAVS activates TBK1/IRF3. In this study, we identified translocases of outer membrane 70 （Tom70）, a mitochondrial import receptor, to interact with MAVS upon RNA virus infection. Ectopic expression or knockdown of Tom70 could enhance or impair IRF3-mediated gene expression, respectively. Mechanistically, the clamp domain （R192） of Tom70 interacts with the C-terminal motif （EEVD） of Hsp90, thus recruiting TBK1/IRF3 to mitochondria. Disruption of this interaction or mislocation of Tom70 sharply impairs activation of TBK1 and IRF3. Furthermore, host antiviral responses are significantly boosted or crippled in the presence or absence of Tom70. Collectively, our study characterizes Tom70 as a critical adaptor linking MAVS to TBK1/IRF3, revealing that mitochondrion is evolutionarily integrated with innate immunity.展开更多
We present a case of fetal liver failure caused by the activation of larnivudine-resistant hepatitis B virus （HBV） nine months after lamivudine treatment. A 57-year old man visited our hospital for the treatment of ...We present a case of fetal liver failure caused by the activation of larnivudine-resistant hepatitis B virus （HBV） nine months after lamivudine treatment. A 57-year old man visited our hospital for the treatment of decornpensated chronic hepatitis B. Lamivudine was started in December 2001. Subsequently, serum HBV was negative for HBV DNA with seroconversion from HBeAg to anti-HBe and improvement of liver function. However, HBV DNA and HBeAg were again detected in September 2002. He was complicated by breakthrough hepatitis and admitted to our hospital in November for severely impaired liver function. Vidarabine treatment was started and serum HBV DNA and alanine aminotransferase （ALT） decreased transiently. However, after the start of m-interferon treatment, HBV DNA level increased and liver function deteriorated. He died 1 mo after admission. An analysis of amino acid sequences in the polymerase region revealed that rtM204I/V with rtLSOI/V occurred at the time of viral breakthrough. After the start of antiviral treatment, rtL180M was detected in addition to rtM204I/V and rtLSOI/V, and became predominant in the terminal stage of the disease. HBV clone with a high replication capacity may be produced by antiviral treatment leading to the worsening of liver function. Antiviral therapy for patients with breakthrough hepatitis in advanced liver disease should be carefully performed.展开更多
Cytomegalovirus enteritis is most usually associated with patients positive for human immunodeficiency virus or immunosuppressed transplant patients. The gastrointestinal tract may be affected anywhere from the esopha...Cytomegalovirus enteritis is most usually associated with patients positive for human immunodeficiency virus or immunosuppressed transplant patients. The gastrointestinal tract may be affected anywhere from the esophagus to the colon, but the small bowel involvement is rare. We report a case of cytomegalovirus ileitis in an immunocompetent adult, which was confirmed by histopathologic findings through colonoscopic biopsy.展开更多
AIM： To study the effects of hepatitis C virus （HCV） core and non-structural 5A （NS5A） proteins on nuclear factor- k B （NF- k B） activity for understanding their biological function on chronic hepatitis caused ...AIM： To study the effects of hepatitis C virus （HCV） core and non-structural 5A （NS5A） proteins on nuclear factor- k B （NF- k B） activity for understanding their biological function on chronic hepatitis caused by HCV infection. METHODS： Luciferase assay was used to measure the activity of NF-kB in three different cell lines cotransfected with a series of deletion mutants of core protein alone or together with NS5A protein using pNF- k B-Luc as a reporter plasmid. Western blot and indirect immunofluorescence assays were used to confirm the expression of proteins and to detect their subcellular localization, respectively. Furthermore, Western blot was also used to detect the expression levels of NF- k B/p65, NF- k B/p50, and inhibitor k B-a（k B-a）. RESULTS： The wild-type core protein （C191） and its mutant segments （C173 and C158） could activate NF- k B in Huh7 cells only and activation caused by （C191） could be enhanced by NS5A protein. Moreover, the full-length core protein and its different deletion mutants alone or together with NS5A protein did not enhance the expression level of NF- k B. The NF- k B activity was augmented due to the dissociation of NF-k： B-I k： B complex and the degradation of Ik B-a. CONCLUSION：NF- k B is the key transcription factor that can activate many genes that are involved in the cellular immune response and inflammation. Coexpression of the full-length core protein along with NS5A can enhance the NF- k B activation, and this activation may play a significant role in chronic liver diseases including hepatocellular carcinoma associated with HCV infection.展开更多
AIM: To investigate the mechanism of interleukin (IL)-6 secretion through blocking the IL-17A/IL-17A recepto (IL-17RA) signaling pathway with a short hairpin RNA (shRNA) in hepatic stellate cells (HSCs) in vitro . MET...AIM: To investigate the mechanism of interleukin (IL)-6 secretion through blocking the IL-17A/IL-17A recepto (IL-17RA) signaling pathway with a short hairpin RNA (shRNA) in hepatic stellate cells (HSCs) in vitro . METHODS: HSCs were derived from the livers of adul male Sprague-Dawley rats. IL-6 expression was evalu ated using real-time quantitative polymerase chain reaction and enzyme linked immunosorbent assay. The phosphorylation activity of p38 mitogen activated pro tein kinases (MAPK) and extracellular regulated pro tein kinases (ERK) 1/2 upon induction by IL-17A and suppression by IL-17RA shRNA were examined using Western blotting.RESULTS: IL-6 expression induced by IL-17A was significantly increased compared to control in HSCs (P < 0.01 in a dose-dependent manner). Suppression of IL17RA using lentiviral-mediated shRNA inhibited IL-6 expression induced by IL-17A compared to group with only IL-17A treatment (1.44 ± 0.17 vs 4.07 ± 0.43, P < 0.01). IL-17A induced rapid phosphorylation of p38 MAPK and ERK1/2 after 5 min exposure, and showed the strongest levels of phosphorylation of p38 MAPK and ERK1/2 at 15 min in IL-17A-treated HSCs. IL-6 mRNA expression induced by IL-17A (100 ng/mL) for 3 h exposure was inhibited by preincubation with specific inhibitors of p38 MAPK (SB-203580) and ERK1/2 (PD-98059) compared to groups without inhibitors preincubation (1.67 ± 0.24, 2.01 ± 0.10 vs 4.08 ± 0.59, P < 0.01). Moreover, lentiviral-mediated IL-17RA shRNA 1 inhibited IL-17A-induced IL-6 mRNA expression compared to random shRNA in HSCs (1.44 ± 0.17 vs 3.98 ± 0.68, P < 0.01). Lentiviral-mediated IL17RA shRNA 1 inhibited phosphorylation of p38 MAPK and ERK1/2 induced by 15 min IL-17A (100 ng/mL) exposure. CONCLUSION: Down-regulation of the IL-17RA receptor by shRNA decreased IL-6 expression induced by IL-17A via p38 MAPK and ERK1/2 phosphorylation in HSCs. Suppression of IL-17RA expression may be a strategy to reduce the inflammatory response induced by IL-17A in the liver.展开更多
文摘目的调查重庆地区男男性行为者(men who have sex with men,MSM)EB病毒(Epstein-Barr virus)血清流行病学现状,推测MSM人群EB病毒感染和再活化情况。方法采用非概率抽样法,对1 082例MSM(来源为2012-2015年重庆地区进行临床药物试验预防艾滋病感染的MSM人群)和1 059名健康成人血清,采用酶联免疫吸附测定(ELISA)方法检测EB-VCA-IgG、EB-NA-IgG和EB-VCA-IgM,使用χ~2检验对结果进行分析,以P<0.05为差异有统计学意义。结果1 082例MSM中HIV阳性130例,HIV阴性952例:(1)EB病毒既往感染率在MSM、HIV阳性、HIV阴性者中分别为1 002例(92.6%)、115例(88.5%)、887例(93.2%),其中MSM组、HIV阴性MSM组与对照组952例(89.9%)差异有统计学意义(P<0.05)。(2)EB病毒未感染率在MSM、HIV阳性MSM、HIV阴性MSM组分别为5例(0.5%)、1例(0.8%)、4例(0.4%),3组均显著低于对照组53例(5.0%),差异有统计学意义(P<0.05)。HIV阳性与HIV阴性间差异无统计学意义(P>0.05)。(3)EB病毒血清学再活化率在MSM、HIV阳性MSM、HIV阴性MSM组中分别为52例(4.8%)、13例(10.0%)、39例(4.1%),其中HIV阳性MSM同对照组40例(3.8%)、HIV阴性MSM组39例(4.1%)差异有统计学意义(P<0.005)。结论(1)MSM人群EB病毒感染率高于普通人群,其发生EB病毒感染相关疾病的威胁更大。(2)非HIV感染的MSM人群EB病毒再活化率可能与正常人无差异。(3)HIV感染的MSM人群EB病毒血清学再活化率明显增高,可能同HIV感染后机体免疫力下降有关,发生EB病毒感染相关疾病的风险更大。
基金Supported by Natural Science Research Project of Anhui Educational Committee (2004kj218 )Major Special Program of Science and Technology Grand Plan of Anhui Province (08010302179)~~
文摘[Objective]The research aimed to test and identify the physicochemical characters and antiviral activity in vitro of semi-finished product of the recombinant porcine rPoIFN-α. [Method]HEp-2/VSV system was used to test the antiviral activity of three batches of rPoIFN-α. Using recombinant human IFN-α as reference,the titer of interferon was measured. The semi-finished product of rPoIFN-α with the known titer were treated with 0.25% trypsin,HCl and mouse anti-porcine IFN-α monoclonal antibody. And the anti-viral activity of each batch of rPoIFN-α was detected. The physicochemical characters of rPoIFN-α were evaluated. The inhibition of induced cytopathic effect of rPoIFN-α on PPV （Porcine parvovirus） and PRV （Porcine pseudorabies） on swine kidney cell （PK-15） was determined. And the antiviral activity of rPoIFN-α in vitro was observed. [Result]The titers of semi-finished products of rPoIFN-α titrated by HEp-2/VSV system could reach 1.5×105 IU/ml,with the specific activity of 1.1×106 IU/mg. The residual rate of the tier of rPoIFN-α treated by 0.25% trypsin at 37 ℃ for 1 h was less than 1%. And that treated with HCl （pH of 2.0） for 72 h was up to 95%. And that treated at 56 ℃ for 30 min and that treated by mouse anti-porcine IFN-α monoclonal antibody at 37 ℃ for 1 h were higher than 47% and about 1% respectively. The antiviral test in vitro showed that 50 and 500 IU/ml rPoIFN-α could inhibit the induced cytopathic effect of PRV and PPV on PK-15 cell lines. [Conclusion]rPoIFN-α had the basic physicochemical characters of IFN-α. And it could inhibit the induced cytopathic effect of PRV and PPV on PK-15 cell lines,but there was dosage difference.
基金Supported by a Health Foundation of Zhejiang Province, No.2004C083, 2004B068
文摘AIM： To detect the plateletoactivating factor （PAF） and the plasma or serum levels of tumor necrosis factoroa （TNF-α） malondialdehyde （MDA）, endotoxin （ET） and to discuss their significance in various types of viral hepatitis. METHODS： PAF, TNF-α, MDA, and ET levels in 60 controls, 16 cases of acute viral hepatitis, 71 cases of chronic viral hepatitis, 19 cases of severe viral hepatitis were detected by reverse phase high-performance liquid chromatography （rHPLC）, bio-assay, ELISA, thiobarbituric acid （TBA）, and limulus lysate test （LLT）, respectively. RESULTS： The rHPLC was more sensitive and specific than bio-assay （r = 0.912, P〈0.01）. The plasma levels of PAF, TNF-α, MDA, and ET in patients with viral hepatitis were higher than those in controls （P〈0.01）. CONCLUSION： rHPLC is more reliable and accurate for the detection of PAF.
文摘Intracellular RNA viruses are sensed by receptors retinoic acid-inducible gene 1 （RIG-I）/melanoma differentiation-associated gene 5 （MDA5） that trigger the formation of MAVS signal complex on mitochondria. Consequently, this leads to the activation of TANK-binding kinase 1 （TBK1） and phosphorylation of interferon regulatory factor 3 （IRF3）, both of which constitutively associate with cytosolic chaperone Hsp90. It remains largely unknown how MAVS activates TBK1/IRF3. In this study, we identified translocases of outer membrane 70 （Tom70）, a mitochondrial import receptor, to interact with MAVS upon RNA virus infection. Ectopic expression or knockdown of Tom70 could enhance or impair IRF3-mediated gene expression, respectively. Mechanistically, the clamp domain （R192） of Tom70 interacts with the C-terminal motif （EEVD） of Hsp90, thus recruiting TBK1/IRF3 to mitochondria. Disruption of this interaction or mislocation of Tom70 sharply impairs activation of TBK1 and IRF3. Furthermore, host antiviral responses are significantly boosted or crippled in the presence or absence of Tom70. Collectively, our study characterizes Tom70 as a critical adaptor linking MAVS to TBK1/IRF3, revealing that mitochondrion is evolutionarily integrated with innate immunity.
文摘We present a case of fetal liver failure caused by the activation of larnivudine-resistant hepatitis B virus （HBV） nine months after lamivudine treatment. A 57-year old man visited our hospital for the treatment of decornpensated chronic hepatitis B. Lamivudine was started in December 2001. Subsequently, serum HBV was negative for HBV DNA with seroconversion from HBeAg to anti-HBe and improvement of liver function. However, HBV DNA and HBeAg were again detected in September 2002. He was complicated by breakthrough hepatitis and admitted to our hospital in November for severely impaired liver function. Vidarabine treatment was started and serum HBV DNA and alanine aminotransferase （ALT） decreased transiently. However, after the start of m-interferon treatment, HBV DNA level increased and liver function deteriorated. He died 1 mo after admission. An analysis of amino acid sequences in the polymerase region revealed that rtM204I/V with rtLSOI/V occurred at the time of viral breakthrough. After the start of antiviral treatment, rtL180M was detected in addition to rtM204I/V and rtLSOI/V, and became predominant in the terminal stage of the disease. HBV clone with a high replication capacity may be produced by antiviral treatment leading to the worsening of liver function. Antiviral therapy for patients with breakthrough hepatitis in advanced liver disease should be carefully performed.
文摘Cytomegalovirus enteritis is most usually associated with patients positive for human immunodeficiency virus or immunosuppressed transplant patients. The gastrointestinal tract may be affected anywhere from the esophagus to the colon, but the small bowel involvement is rare. We report a case of cytomegalovirus ileitis in an immunocompetent adult, which was confirmed by histopathologic findings through colonoscopic biopsy.
基金Supported by the Ph D Program Foundation of Ministry of Education of China, No. 20010486015
文摘AIM： To study the effects of hepatitis C virus （HCV） core and non-structural 5A （NS5A） proteins on nuclear factor- k B （NF- k B） activity for understanding their biological function on chronic hepatitis caused by HCV infection. METHODS： Luciferase assay was used to measure the activity of NF-kB in three different cell lines cotransfected with a series of deletion mutants of core protein alone or together with NS5A protein using pNF- k B-Luc as a reporter plasmid. Western blot and indirect immunofluorescence assays were used to confirm the expression of proteins and to detect their subcellular localization, respectively. Furthermore, Western blot was also used to detect the expression levels of NF- k B/p65, NF- k B/p50, and inhibitor k B-a（k B-a）. RESULTS： The wild-type core protein （C191） and its mutant segments （C173 and C158） could activate NF- k B in Huh7 cells only and activation caused by （C191） could be enhanced by NS5A protein. Moreover, the full-length core protein and its different deletion mutants alone or together with NS5A protein did not enhance the expression level of NF- k B. The NF- k B activity was augmented due to the dissociation of NF-k： B-I k： B complex and the degradation of Ik B-a. CONCLUSION：NF- k B is the key transcription factor that can activate many genes that are involved in the cellular immune response and inflammation. Coexpression of the full-length core protein along with NS5A can enhance the NF- k B activation, and this activation may play a significant role in chronic liver diseases including hepatocellular carcinoma associated with HCV infection.
基金Supported by The Zhejiang Extremely Key Subject of SurgeryThe Wenzhou Key Laboratory Project in Surgery
文摘AIM: To investigate the mechanism of interleukin (IL)-6 secretion through blocking the IL-17A/IL-17A recepto (IL-17RA) signaling pathway with a short hairpin RNA (shRNA) in hepatic stellate cells (HSCs) in vitro . METHODS: HSCs were derived from the livers of adul male Sprague-Dawley rats. IL-6 expression was evalu ated using real-time quantitative polymerase chain reaction and enzyme linked immunosorbent assay. The phosphorylation activity of p38 mitogen activated pro tein kinases (MAPK) and extracellular regulated pro tein kinases (ERK) 1/2 upon induction by IL-17A and suppression by IL-17RA shRNA were examined using Western blotting.RESULTS: IL-6 expression induced by IL-17A was significantly increased compared to control in HSCs (P < 0.01 in a dose-dependent manner). Suppression of IL17RA using lentiviral-mediated shRNA inhibited IL-6 expression induced by IL-17A compared to group with only IL-17A treatment (1.44 ± 0.17 vs 4.07 ± 0.43, P < 0.01). IL-17A induced rapid phosphorylation of p38 MAPK and ERK1/2 after 5 min exposure, and showed the strongest levels of phosphorylation of p38 MAPK and ERK1/2 at 15 min in IL-17A-treated HSCs. IL-6 mRNA expression induced by IL-17A (100 ng/mL) for 3 h exposure was inhibited by preincubation with specific inhibitors of p38 MAPK (SB-203580) and ERK1/2 (PD-98059) compared to groups without inhibitors preincubation (1.67 ± 0.24, 2.01 ± 0.10 vs 4.08 ± 0.59, P < 0.01). Moreover, lentiviral-mediated IL-17RA shRNA 1 inhibited IL-17A-induced IL-6 mRNA expression compared to random shRNA in HSCs (1.44 ± 0.17 vs 3.98 ± 0.68, P < 0.01). Lentiviral-mediated IL17RA shRNA 1 inhibited phosphorylation of p38 MAPK and ERK1/2 induced by 15 min IL-17A (100 ng/mL) exposure. CONCLUSION: Down-regulation of the IL-17RA receptor by shRNA decreased IL-6 expression induced by IL-17A via p38 MAPK and ERK1/2 phosphorylation in HSCs. Suppression of IL-17RA expression may be a strategy to reduce the inflammatory response induced by IL-17A in the liver.
