Effects of antibiotic treatment and antibiotics combined with surgery treatment on the prognosis of patients with infective endocarditis / 中华传染病杂志

Objective:To investigate the effects of antibiotic treatment and antibiotics combined with surgery treatment on the prognosis of patients with infective endocarditis (IE).Methods:The clinical data and prognosis of all patients diagnosed as IE discharged from Shanghai Jiao Tong University Affiliated Sixth People′s Hospital from June 2011 to May 2021 were collected. There were 240 IE patients, divided into antibiotic treatment group and the antibiotics combined with surgery group according to the treatment methods. The clinical characteristics and prognosis of the IE patients were compared between the two groups, so as to investigate the timing of surgery for IE patients and to analyze the effects of the two treatment methods on the prognosis of IE patients.Statistical analysis methods including Wilcoxon rank sum test, chi-square test, Kaplan-Meier survival analysis and Cox regression analysis were used when appropriate.Results:Of the 240 patients with IE, 63 cases were only treated with antibiotics and 177 cases were treated with antibiotics combined with surgery. After propensity score matching (PSM), one-year mortality rate of the IE patients in the antibiotics combined with surgery group was 11.1%(4/36), which was significantly lower than that in the antibiotic treatment group (33.3%(12/36), χ2=5.14, P=0.023). The median values of left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDD) and left ventricular fractional shortening (LVFS) in the antibiotics combined with surgery group were 59%, 47 mm and 31%, respectively, which were significantly lower than those before surgery (63%, 54 mm and 34%, respectively, Z=6.19, 9.36 and 6.11, respectively, all P<0.001). The most common surgical indication was moderate to severe heart failure, and there was no significant difference between the early operation group and the late operation group (both P>0.050). The one-year cumulative survival rate of antibiotics combined with surgery group was 94.9%, which was significantly higher than that in the antibiotic treatment group (83.2%, χ2=7.38, P=0.007). Heart failure and Pitt bacteremia scores≥4 were the independent risk factors for one-year all-cause death of the IE patients (hazard ratio ( HR)=5.668 and 19.392, respectively, both P<0.050). Hospital days and antibiotics combined with surgery were independent related factors for reducing the risks of one-year all-cause death ( HR=0.931 and 0.299, respectively, both P<0.050). Pitt bacteremia scores≥4 had the greatest impact on one-year prognosis of the IE patients. Conclusions:Surgery could significantly improve cardiac function and one-year prognosis of the IE patients. IE patients with heart failure and Pitt bacteremia score≥4 should be actively treated.

In vitro study of dendritic cell derived exosomes transporting hepatitis D antigen induced specific cytotoxic T lymphocyte response / 中华传染病杂志

Objective:To explore the effect and mechanism of dendritic cell derived exosomes (Dexs) loading ubiquitinated (Ub) hepatitis D antigen (HDAg) on activating specific cytotoxic T lymphocytes (CTL).Methods:Ub-S-HDAg-Dexs were co-cultured with dendritic cells (DC) which were from the femora of C57BL/6 mice for 48 h, then flow cytometry was used to detect the maturity of DC (CD86, CD80 and major histocompatibility complex (MHC) Ⅱ). The spleen-derived T lymphocytes from C57BL/6 mice were added in vitro to activate DC and co-cultivated for 72 h. The T cells were divided into Ub-S-HDAg-Dexs group (add 50 μg/mL Ub-S-HDAg-Dexs), Blank-Dexs group (add 50 μg/mL DC derived exosomes without plasmid transfection), Con-Dexs group (add 50 μg/mL DC derived exosomes transfected by cantrol plasmid), PBS group (add 50 μL/mL phosphate buffered saline), and Ub-S-HDAg-Dexs+ AG490 group (add 50 μg/mL Ub-S-HDAg-Dexs, DC and T lymphocytes stimulated by exosomes, and 50 μmol/L AG490 was also added to the cell mix). Flow cytometry was used to detect CD8 + T cells secreting interferon-gamma, non-radioactive lactate dehydrogenase release test to detect the killing activity of specific CTL. Real-time quantitative polymerase chain reaction (PCR) and Western blotting were used to detect the mRNA and protein expressions of JAK kinase (JAK) 2, GATA-binding protein 3 (GATA3), T-bet, signal transduction and activator of transcription (STAT) 1 and STAT4. Independent sample t test were used for statistical analysis. Results:The positive rates of the surface molecules CD80, CD86, MHCⅡof DC stimulated by Ub-S-HDAg-Dexs were 83.850%±0.219%、68.910%±0.134%、84.320%±0.445%, respectively.In the Ub-S-HDAg-Dexs group, the rate of CD8 + T cells secreting interferon-gamma was 6.420%±0.028%, which was higher than those of other groups, including PBS group, Blank-Dexs group, Con-Dexs group and Ub-S-HDAg-Dexs+ AG490 group ( t=90.78, 30.32, 63.06 and 85.42, respectively, all P<0.001). The cytotoxicity of T cells in the Ub-S-HDAg-Dexs group was 82.4%±3.9%, which was higher than those of other groups ( t=17.28, 9.74, 3.95 and 15.89, respectively, all P<0.050). The relative mRNA expressions of JAK2, T-bet, STAT1, STAT4 in Ub-S-HDAg-Dexs group were higher than those in other groups, including Con-Dexs group ( t=10.74, 32.34, 13.00 and 16.28, respectively, all P<0.001), Blank-Dexs group ( t=15.05, 21.51, 6.46 and 13.12, respectively, all P<0.050), PBS group ( t=21.83, 41.42, 7.30 and 17.50, respectively, all P<0.050), Ub-S-HDAg-Dexs+ AG490 group ( t=35.75, 20.69, 17.02 and 17.07, respectively, all P<0.001), and the differences were all statistically significant. The protein expressions of T-bet, STAT1, STAT4 in Ub-S-HDAg-Dexs group increased compared with those in PBS group ( t=346.70, 57.54 and 55.81, respectively, all P<0.001), with statistical significance. In the presence of AG490, the protein expressions of T-bet, STAT1 and STAT4 decreased compared with those in Ub-S-HDAg-Dexs group, and the differences were statistically significant ( t=355.40, 52.79 and 126.10, respectively, all P<0.001). Conclusions:Ubiquitinated HDAg transported by exosomes could effectively promote DC maturation, induce T lymphocyte differentiation, and generate specific CTL responses, which provides a new idea for the treatment of hepatitis D.

Effect of ubiquitination hepatitis B virus core antigen on inducing dendritic cells autophagy to enhance specific cytotoxic T lymphocyte responses / 中华传染病杂志

Objective:To clarify the effect of ubiquitination hepatitis B virus core antigen (Ub-HBcAg) on dendritic cells (DC) autophagy, and to explore the mechanism of autophagy in enhancing DC antigen presentation and inducing hepatitis B virus-specific cytotoxic T lymphocyte (CTL) responses.Methods:Ub-HBcAg lentiviral vector (LV-Ub-HBcAg), lentiviral vector-hepatitis B virus core antigen (LV-HBcAg) and no-load plasmid LV (LV) were constructed and packaged. DC2.4 cells were divided into LV-Ub-HBcAg group, LV-HBcAg group and LV group. The blank control group (NC group) was also set. The protein expression of autophagy-related protein P62, microtubule associated protein 1 light chain 3 beta (LC3B), autophagy related 5(ATG5) and Beclin-1 were detected by Western blotting. The expressions of co-stimulatory molecules such as CD86, CD80 and major histocompatibility complex (MHC)-Ⅱ were detected by flow cytometry. Cell counting kit-8 (CCK-8) method was used to detect T lymphocytes proliferation. The non-radioactive lactic acid dehydrogenase (LDH) release method was applied to detect the killing ability of CTL. Statistical analysis was conducted by independent sample t test. Results:The relative protein expressions of LC3B-Ⅱ/LC3B-Ⅰ, Beclin-1 and ATG5 in NC group were 0.445±0.076, 0.522±0.026 and 0.761±0.038, respectively, which were all lower than those in LV-Ub-HBcAg group (0.926±0.021, 0.919±0.016 and 1.451±0.028, respectively). The relative protein expression of P62 in the NC group was higher than that in LV-Ub-HBcAg group ((1.875±0.016) vs (0.647±0.121)). The differences were all statistically significant ( t=6.102, 9.842, 17.490 and 10.590, respectively, all P<0.01). The expressions of CD86 (75.51%), CD80 (83.35%), MHC-Ⅱ (66.66%) in the LV-Ub-HBcAg group were high, and those in the NC group were 8.03%, 7.49%, 0.04%, respectively. The specific CTL killing rate ((65.310±2.091)%) of the LV-Ub-HBcAg group was significantly higher than both NC group ((14.400±0.497)%) and LV-HBcAg group ((54.870±1.443)%), and the differences were both statistically significant ( t=23.690 and 4.111, respectively, both P<0.05). Conclusion:Ub-HBcAg promotes the DC autophagy, up-regulates the expressions of costimulatory molecules on cell surface of DC to induce the maturation and activation, and then stimulates T lymphocyte to induce a stronger specific CTL response under the effort of ubiquitination.

Activation of Rip1 promotes necroptosis in LNCaP-AI cells via inhibiting SHARPIN / 中国病理生理杂志

[ ABSTRACT] AIM:To explore the role of SHARPIN in regulation of Rip1 in castration-resistant prostate cancer LNCaP-AI cells.METHODS:The LNCaP-AI cells were treated with TNF-α+Z-VAD ( an inhibitor of pan-caspase) to activate necroptosis, which were compared to the cells treated with TNF-α+Z-VAD+Nec-1 ( an inhibitor of Rip1 ) .A blank group and a TNF-α-treated group were set up as controls.The cell viability in each group was measured by MTS as-say.In addition, SHARPIN was knocked down by siRNA, and the inhibitory efficiency was evaluated by RT-qPCR.The expression of Rip1 at mRNA and protein levels after knocking down SHARPIN was determined by RT-qPCR and Western blot to explore the underlying mechanism of regulatory network of necroptosis in prostate cancer.RESULTS: Compared with blank control group and TNF-α-treated group, the viability of LNCaP-AI cells treated with TNF-α+Z-VAD decreased by 28%(P LNCaP-AI cells.CONCLUSION:Necroptosis is an important way of cell death .Inhibition of oncogenic factor SHARPIN enhances necroptosis via activating Rip1 in LNCaP-AI cells.

Predictors for outcome of exchange arthroplasty for periprosthetic joint infections after primary total knee arthroplasty / 中华传染病杂志

Objective To evaluate the overall failure rate of one or two-stage exchange arthroplasty for infections in total knee arthroplasty (TKA) and the predictors affecting the outcome of exchange.Methods Thirty-nine cases received one or two-stage exchange arthroplasty for periprosthetic joint infections after primary TKA in Shanghai Sixth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine and Southeast Hospital affiliated to Xiamen University from January 2012 to November 2014 were reviewed.Periprosthetic tissue and articular fluid of all patients were analyzed by bacterial culture.All patients were followed up for more than one year.C-reactive protein (CRP),erythrocyte sedimentation rate (ESR),procalcitonin (PCT) and blood routine were tested every four weeks,and the evaluation on pain,total periprosthetic function,range of motion and deformation of arthroplasty were conducted.Differences between groups were analyzed using chi-square test or Student's t test when appropriate.A stepwise selection approach in logistic regression analysis was used to screen key predictors for outcome of one or two-stage exchange for infections in TKA.Results There were 39 patients who had undergone one or two-stage exchange for infections in TKA,including 20 males (51.3％) and 19 females (48.7％) with an average age of (62.4±11.7) years.Among the 39 patients,18 gram-positive strains were isolated from specimens,and 6 gram negative strains,2 Mycobacterium tuberculosis and 1 candida albicans.Ten of 39 reimplantations developed reinfection.Between the success and failure groups,there were significant differences in the time from primary TKA to revision (P =0.023),operative time (P =0.029),multidrug resistant organisms (P =0.045),the preoperative and post-operative ESR (P=0.002 and P＜0.001,respectively) and post-operative CRP (P=0.018).Multivariable logistic regression analysis demonstrated that time from primary TKA to revision (OR =0.96,95％CI:0.92-1.00,P=0.025),preoperative ESR (OR=0.97,95％CI:0.95-1.00,P=0.045) and post-operative ESR (OR =0.94,95％ CI:0.91-0.98,P =0.002) were independent indicators associated with the outcome of one or two-stage revision.Conclusions The failure rate after revision for infected TKA is relatively high.The time from primary TKA to revision,preoperative and post-operative ESR could predict the outcome of one or two-stage revision effectively.

Knockdown of SALL4 expression regulates cell proliferation and apoptosis in prostate cancer LNCaP cells / 中国病理生理杂志

[ ABSTRACT] AIM: To investigate the SALL4 expression, proliferation and apoptosis in the LNCaP cells after transfection of SALL4 siRNA.METHODS: The expression of SALL4 at mRNA and protein levels was detected by real-time PCR and Western blotting.MTS assay, colony formation assay and flow cytometry were used to determine the prolifer-ation, colony formation ability and apoptosis of the LNCaP cells.The effect of SALL4 on the expression of Bax and Bcl-2 was analyzed by Western blotting.RESULTS:Compared with negative control group, the expression of SALL4 at mRNA and protein levels in LNCaP cells was down-regulated by transfection of SALL4 siRNA ( P<0.05 ) .The proliferation rate and colony formation ability were decreased, while apoptosis rate increased in si-SALL4 group (P<0.05).Higher expres-sion of Bax and lower expression of Bcl-2 in si-SALL4 group were observed ( P<0.05 ) .CONCLUSION:Down-regula-tion of SALL4 by siRNA not only suppresses LNCaP cell proliferation and colony formation, but also inhibits Bcl-2 expres-sion and activates Bax expression to induce apoptosis.

The fusion protein of cytoplasmic transduction peptide (CTP)-HBcAg18-27-Tapasin enhances specific immune response to hepatitis B virus and inhibits viral replication in transgenic mice / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of protein transduction domain-hepatitis B virus core antigen (CTP-HBcAg18-27)-Tapasin fusion protein-induced specific cytotoxic T lymphocyte (CTL) response on hepatitis B virus (HBV) replication in HBV transgenic mice.</p><p><b>METHODS</b>Twenty HBV-transgenic mice were randomly divided into two groups for a 3-week course of once weekly subcutaneous immunizations with either CTP-HBcAg18-27-Tapasin fusion protein or CTP-HBcAg18-27. Mice administered isotonic saline served as blank controls. Expressions of cytokines in splenocytes were analyzed by flow cytometry. Serum levels of hepatitis B surface antigen (HBsAg) and HBV DNA were determined by microparticle enzyme immunoassay and real-time fluorescent PCR assay, respectively. Expression of HBsAg in hepatic tissues was detected by immunohistochemistry.</p><p><b>RESULTS</b>Immunization with 100 mug of CTP-HBcAg18-27-Tapasin fusion protein led to a significant increase in proportions of CTLs in spleen (2.70%+/-0.20% vs. 50 mug of CTP-HBcAg18-27-Tapasin: 1.66%+/-0.53%, 50 mug of CTP-HBcAg18-27: 1.26%+/-0.56%, and blank controls: 0.75%+/-0.71%; F = 741.45, P = 0.000) and up-regulation of inflammatory cells in hepatic tissue. In addition, both immunizations of CTP-HBcAg18-27-Tapasin led to significant decreases in serum HBsAg and HBV DNA levels compared to those in the CTP-HBcAg18-27 group.</p><p><b>CONCLUSION</b>HBV-related modification of the expression of the molecular chaperone Tapasin may affect its interaction with intracellular antigen peptides, thereby leading to increases the number of specific CTLs in the spleen, decreases in serum HBsAg and HBV DNA levels, and down-regulation of HBsAg expression in hepatic tissue. These results obtained in HBV-transgenic mice suggest that the CTP-HBcAg18-27-Tapasin fusion protein has anti-HBV activity.</p>

Effects of cytoplasmic transduction peptide-HBcAg18-27-Tapasin induced murine bone marrow derived dendritic cell maturation on T lymphocyte proliferation in vitro / 中华传染病杂志

Objective To observe the effects of cytoplasmic transduction peptide (CTP)-HBcAg18-27-Tapasin induced murine bone marrow-derived dendritic cell (DC) maturation on T lymphocyte proliferation in vitro,Methods Bone marrow derived DC isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleutin (IL)-4 for 5 days followed by lipopolysaccharide added to induce DC maturation.10 μg/L CTP-HBcAg18-27-Tapasin,50 μg/L CTP-HBcAg18-27-Tapasin,10 μg/L CTP-HBcAg18-27 or RPMI-1640 were added into culture medium to induce DC maturation.DC phenotypes were analyzed by flow cytometry.The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay.The proliferation of.T lymphocytes was performed by using cell counting kit-8 and intracellular cytokine of proliferative T cells were analyzed by flow cytometry.The means among groups were compared using one-way ANOVA and those between two groups were compared by least significant difference test.Results DC were cultured and induced successfully.The molecules on DC surface,such as CD80,CD86 and major histocompatibility antigen-Ⅰ were upregulated by CTP-HBcAg18-27-Tapasin.IL-12p70 level induced by 50 μg/L CTP-HBcAg18-27-Tapasin was (61.12±10.25) pg/mL,which was higher than those induced by 10 μg/L CTP-HBcAg18-27-Tapasin (50.43±10.42) pg/mL,10μg/L CTP-HBcAg18-27 (40.17±8.54) pg/mL and medium control (30.51±8.03) pg/mL (F=15.85,P=0.030 and 0.037).The proliferation of T lymphocytes induced by CTP- HBcAg18-27 -Tapasin was higher than control groups.The amounts of cytotoxic T lymphocyte (CTL) induced by 50 μg/L CTP-HBcAg18-27-Tapasin [(2.05±0.41) ％] and 10 μg/L CTP-HBcAg18-27-Tapasin [(1.06 ±0.10 )％] were both significantly higher than the 10 μg/L CTP-HBcAg18-27 group [(0.45±0.11)％] and medium group [(0.09±0.02)％,F=60.22,P=0.003].Conclusions CTP HBcAg18- 27 Tapasin could promote the differentiation and maturation of DC,and enhance the ability of DC stimulating T lymphocytes proliferation and increase CTL expression effectively.

The observation of clinical effectiveness of adefovir combined with Anfate in treatment of patients with decompensated cirrhosis / 中国基层医药

Objective To observe the effects and safety of adefovir combined with anfate(compound fetal cow liver extract tablets)in treament of patients with decompensated cirrhosis caused by chronic hepatitis B.Methods 45 decompensated cirrhosis patients with HBsAg, HBeAb-Positive HBcAb-Positive( 103Copies/ml ＜ HBV-DNA Level ＜10

Effects of PTD-HBcAg induced murine bone marrow-derived dendritic cells maturation on T lymphocyte proliferation in vitro / 中华传染病杂志

Objective To observe the effects of PTD-hepatitis B core antigen (HBcAg) induced murine bone marrow-derived dendritic cells (DCs) maturation on T lymphocyte proliferation in vitro.Methods Bone marrow derived DCs isolated from BALB/c mice were cultured with recombinant granu|ocyte-macrophage colony-stimulating factor (rGM-CSF) and recombinant interleutin-4 (rIL-4)for 5 days.Tumor necrosis factor (TNF)-a,HBcAg and PTD-HBcAg were added to induce DCs maturation.The distribution and localization of intracellular immunofluorescence were observed by confocal microscopy,and DCs phenotypes were analyzed by flow cytometry.The level of IL-12 p70 in the supernatant was detected by enzyme linked immunosorhent assay (ELISA).The proliferation of T lymphocytes was performed by using cell counting kit-8 (CCK-8).All data were analyzed using t test.Results DCs were cultured and identified successfully.Recombinant PTD-HBcAg could penetrate into DCs cytoplasm while recombinant HBcAg was detected on the surface of cells.DCs surface molecules,such as CD80,CD86 and major histocompability complex (MHC) II were upregulated by PTDHBcAg;IL-12 p70 levels induced by 50 mg/L and 100 mg/L recombinant PTD-HBcAg were (142.50±18.31) ng/L and (124.30±15.12) ng/L,respectively,which were significantly higher than those induced by recombinant HBcAg [(42.31±4.21 ) ng/L,t = 9.234 and 9.045,respectively,P＜0.05].The proliferation of T lymphocytes induced by PTD-HBcAg was much higher than that in HBcAg group or positive control TNF-a group.Conclusions PTD-HBcAg could penetrate membrane of DCs and promote the differentiation and maturation of DCs.PTD-HBcAg could up-regulate the expressions of costimulatory molecules on cell surface of DCs,and enhance the ability of DCs on stimulating T lymphocytes proliferation and IL-12 p70 production.

Trans-activator of transcription protein transduction domain mediated cell membrane penetration of HBcAg / 中华传染病杂志

0bjective To observe the cell membrane penetration of protein transduction domain (PTD)-HBeAg fusion protein in vitro.Methods The sequence of trans-activator of transcription (Tat)-PTD was synthesized and the whole HBcAg gene was amplified by polymerase chain reaction (PCR).Overlap extension PCR was employed to fuse Tat-PTD and whole HBcAg gene.Then the fusion gene was cloned into prokaryotic expression vector pMAL-c2X.The correct vector was transformed into E.coli Rosetta-gamiTM 2(DE3),and the protein was induced by isopropyl β-D-1-thiogalactopyranoside(IPTG).Western blot was used tO identify the protein. Furthermore,the fusion protein PTD-HBcAg was purified by affinity chromatography.HBcAg protein expressed using the same methods was employed as eontr0l.The purified protein was added tO HuH-7 cell culture,then the transduction of PTD-HBcAg and HBcAg in cells were detected by indirect immunofluorescence assay (IFA).Results The fusion protein was effectively expressed in E. Coli and purified by affinity chromatography.Both purified PTD-HBcAg and HBcAg could be recognized by HBeAg monoclonal antibody in Western blot analysis.IFA visualization showed that PTD-HBeAg could be introduced into HUH-7 ceils while HBcAg only could not be detected in cells.Conclusions PTD-HBcAg fusion protein can be expressed effectively and purified in prokaryotic expression system.PTD could mediate HBcAg penetrating eell membrane into the cells.

Effects of Astragalus Polysaccharide on Differentiation and Maturation of Dendritic Cells in Vivo / 中国中医药信息杂志

Objective To investigate the effect of astragalus polysaccharide,a component of an aqueous extract of Astragalus Membranaceus roots,on differentiation and maturation of dendritic cells in vitro. Methods 30 BALB/c mice were randomly divided into three groups,normal control group,100,200 mg/kg APS intraperitoneal injection groups. After one week,weight the mouse spleen,account the splenetic index. Collect the mouse bone marrow cell,induced and cultured with rmGM-CSF and rmIL-4. With inverted microscope to investigate the morphous of DC cell. The phenotypes of DC were detected by flow cytometry and the expression of the GM-CSF protein in serum was tested by ELISA. Results Astragalus polysaccharide injection had obvious effects on the spleen weight of mice. The degree of CD11c and MHC-Ⅱ expression in 100 mg/kg and 200 mg/kg groups on flow cytometry were advanced significantly compared with that in normal control group,but the degree of CD80 and CD86 was not increased. And the expression of the GM-CSF protein in serum in 100 mg/kg group and 200 mg/kg group were both not increased significantly compared with nomal control group. Conclusion The intraperitoneal injection of astragalus polysaccharide could stimulate the proliferation of the pre DC in bone marrow. The angtigen presentation of DC might be enhanced,but this effects was not positive correlation with concentration of GM-CSF.

CD_(25) Expression in Peripheral Blood T Lymphocytes from Patients with Chronic Hepatitis B and Its Significance / 中华医院感染学杂志

OBJECTIVE To investigate CD25 expression in peripheral blood T lymphocytes from patients with chronic hepatitis B,and explore its significance in the pathogenesis of chronic hepatitis B.METHODS To detect CD25 expression in peripheral blood T lymphocytes of patients with chronic hepatitis B by means of flow cytometry.CD25 expression was observed in chronic hepatitis B patients.In the meantime,CD25 expression in T cells from severe chronic hepatitis or acute hepatitis B patients and asymptomatic carriers of HBV was also observed.RESULTS Varying degrees of CD25 were expressed in T cells from hepatitis B patients.The expression in CD3+ T and CD8+T cells was higher than that in CD4+T cells.CD25 expression in CD4+T was lower.The average of CD25 expression in CD3+T cells from patients with chronic hepatitis B,acute hepatitis B,and chronic severe hepatitis B and asymptomatic carriers of HBV was(2.92?0.13)%,(0.51?0.36)%,(1.60?0.07)%,and(0.95?0.23)%,respectively.The average of CD25 expression in CD4+T cells from patients with chronic hepatitis B,acute hepatitis B and chronic severe hepatitis B and asymptomatic carriers of HBV was(2.58?0.50)%,(0.34?0.07)%,(1.45?0.02)%,and(0.83?0.13)%,respectively.CD3+T and CD4+T CD25 expression in patients with chronic hepatitis and,severe chronic hepatitis B was increased compared with that of acute hepatitis B patients and asymptomatic carriers of HBV.Compared with chronic severe hepatitis B,the expression of chronic hepatitis B was higher.CONCLUSIONS CD4+ CD25+T cells in chronic hepatitis B virus infection are increased compared with acute hepatitis,CD4+ CD25+T cells may be related to immune tolerance.

Construction of human anti-HBc ScFv eukaryotic expression vector and expression of anti-HBc ScFv in HepG2 cells / 中国病理生理杂志

AIM: To construct an eukaryotic expression vector of human single-chain variable fragment ~against hepatitis B virus core protein (anti-HBc ScFv) and detect its expression in HepG2 cells. METHODS: Anti-HBc ScFv genes were amplified from the plasmids abstracted from positive clone and inserted into pEGFP-c1 vector that contained green fluorescent protein gene. The recombinant plasmids were transfected into HepG2 cells, and resistant clones were obtained by G418 selection. The expression of the gene of fusion protein was determined by fluorescent invert microscope and ELISA. RESULTS: Recombinant plasmids were successfully constructed. The plasmid transfected HepG2 cells were obtained by G418 selection. Specific fluorescence was observed in HepG2 cells 48 hours after transfection. ELISA analysis confirmed the expression of anti-HBc ScFv in the cells. CONCLUSION: The construction of human anti-HBc ScFv eukaryotic expression vector and its expression in HepG2 cells lay the foundation for advanced research of intracellular anti-HBc ScFv.