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Objective@#To study the functional effects of killer cell lectin-like receptor subfamily G member 1 (KLRG1) expression on natural killer cells (NK cell) in chronic hepatitis B virus infection (HBV).@*Methods@#Peripheral blood mononuclear cells (PBMC) were extracted from 120 patients with chronic hepatitis B virus infection and 19 healthy persons. The frequency of NK cells and KLRG1+ NK cells in peripheral blood was detected by flow cytometry. Interferon-γ levels secreted by NK cells were detected in peripheral blood. Statistical analysis of experimental data was performed using GraphPad Prism 6.03 software.@*Results@#The frequency of NK cells in HBV-infected group (16.92% ± 7.9%) was not significantly different from that in healthy controls (10.57% ± 6.5%). The frequency of KLRG1+NK cells in HBV-infected group was significantly higher (49.43% ± 21.2%) than that to healthy control group (31.60% ± 17.9%), (t = 7.347 6, P < 0.001). IFN-γ secretion of KLRG1 + NK cells in HBV-infected patients (2.59% ± 1.0%) were significantly lower than healthy controls (5.96% ± 2.4%), (P = 0.009).@*Conclusion@#HBV infection can increase the expression of KLRG1 in NK cells and further reduce the secretion of IFN-γ in NK cells, which may be an important cause for chronic HBV infection.
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Objective@#To explore the relationship between sialic-acid-binding immunoglobulin-like lectin 7 (Siglec-7) expressed on NK cells and hepatitis B virus-related cirrhosis.@*Methods@#Peripheral venous blood samples were collected from 23 healthy controls and 31 patients with hepatitis B virus-related cirrhosis (Child-Pugh A, n = 7; Child-Pugh B, n = 12; Child-Pugh C, n = 12). Peripheral blood mononuclear cells (PBMCs) were obtained by using Ficoll-Hypaque density gradient centrifugation and the expression of Siglec-7 and NK cells phenotype and their subpopulations were detected by flow cytometry. Comparisons between various groups were performed using t -test, one-way analysis of variance (ANOVA), and correlations between variables were analyzed using Pearson’s-correlation coefficient.@*Results@#(1) There was no significant difference in the percentage of NK cells and in their subpopulations with HBV-related cirrhosis and healthy controls. (2) Siglec-7 expression on NK cells in patients with HBV-related cirrhosis(62.44±13.45%)was significantly down-regulated than that to healthy controls(75.39±12.19%)while the frequency of Siglec-7+ NK cells were negatively correlated with Child-Pugh score. (3) Subpopulation analysis showed that Siglec-7 expression on CD56brightCD16-NK cells(66.99±15.93%)was significantly lower than CD56dimCD16+NK cells(76.54±13.9%) in HBV-related cirrhosis. However, the expression of Siglec-7 in healthy controls showed no difference in these two NK cell subsets. (4) Phenotypic analysis showed that Siglec-7+ NK cells express higher levels of activating receptor CD16, CD38, NKp46 and lower levels of inhibitory receptor CD158b. Indeed, the frequency of CD16 and CD38 on Siglec-7+ NK cells in HBV-related cirrhosis was lower than that in healthy controls.@*Conclusion@#The disease progression in patients with hepatitis B virus-related cirrhosis is associated to decreased frequencies of Siglec-7+NK cells.
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OBJECTIVE:To observe clinical efficacy and safety of Mailuoning injection in adjunctive treatment of acute cere-bral infarction. METHODS:A total of 65 patients with acute cerebral infarction selected from neurology department of our hospital were divided into control group(32 cases)and observation group(33 cases)according to random number table. Control group was given conventional treatment. Observation group was additionally given Mailuoning injection 10 mL added into 0.9% sodium chlo-ride injection 250 mL intrnrenously,ivgtt,qd,on the basis of control group. Both group were treated for 15 d. Clinical efficacy as well as serum levels of ox-LDL,BNP and MMP-9,NIHSS score before and after treatment,the occurrence of ADR were com-pared between 2 groups. RESULTS:The response rate of observation group was 90.91%,which was significantly higher than 65.63%,with statistical significance(P0.05). After treatment,serum levels of ox-LDL,BNP and MMP-9,NIHSS score in 2 groups were all decreased significantly,and the observation group was significantly lower than the control group,with statis-tical significance (P0.05). CONCLU-SIONS:Mailuoning injection has significant therapeutic efficacy for acute cerebral infarction,can significantly reduce serum levels of ox-LDL,BNP and MMP-9,promotes neurological function and the recovery of patients with cerebral infarction with good safety.
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OBJECTIVE:To observe the effects of Shenqi fuzheng injection combined with Nimodipine tablet on blood indica-tors of cerebral infarction patients in the recovery period. METHODS:58 patients diagnosed as cerebral infarction in the recovery period were collected and randomly divided into control group and trial group,with 29 cases in each group. Control group was giv-en Nimodipine tablet 30 mg,tid;trial group was additionally given Shenqi fuzheng injection 250 ml,qd,ivgtt. Both group re-ceived 14 d of treatment. After treatment,the levels of serum hs-CRP,Fractalkine,tPA,PAI-1,blood rheology index and plasma fibrinogen(FIB)were observed in 2 groups. RESULTS:After treatment,the levels of serum hs-CRP,Fractalkine,blood rheology index PAI-1 and FIB decreased in trial group,while tPA activity increased;there was statistical significance between trial group and control group (P<0.05). CONCLUSIONS:Shenqi fuzheng injection combined with Nimodipine tablet can significantly im-prove the serum hs-CRP and Fractalkine levels,blood rheology indicators,PAI-1 and FIB,and improve tPA activity.
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Objective:To study the role of γδ T cells and Vδ1 and Vδ2 T subsets played in patients with chronic hepatitis C.Methods:The percentage of peripheral bloodγδT cells and Vδ1 and Vδ2 T subsets cells was assessed by flow cytometry ( FACS) . Results:There were no significant differences in the percentage of circulating γδ T cells and Vδ1 and Vδ2 T subsets between HCV-infected patients and healthy controls ( HCs).However,The number of peripheral blood Vδ2 T cells from HCV-infected patients was positively correlated with serum alanine aminotransferase ( ALT) levels,but showed no correlation with serum HCV RNA.Peripheral Vδ2 T cells from HCV-infected patients were in activated status.The expression of CD107a was enhanced in Vδ2 T cells from HCV-infected patients compared to HCs.Conclusion:Vδ2 T cells were involved in liver injury in chronic HCV-infected patients.
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Objective To investigate the effects of interleukin -15 ( IL-15 ) on the cytotoxicity of Vδ2 γδ(Vδ2) T cells against K562 cells.Methods PBMCs were separated and cultured with zoledronate and interleukin-2 ( IL-2) to induce the proliferation of Vδ2 T cells.The obtained Vδ2 T cells were in vitro stimulated with IL-15.Flow cytometry analysis was performed to evaluate the changes of phenotypes of Vδ2 T cells and their cytotoxicity against K562 cells.Results Zoledronate effectively induced the massive prolifer -ation of V2δT cells.The Vδ2 T cells were highly activated and the expression of CD 107a and IFN-γby Vδ2 T cells were upregulated upon IL-15 stimulation.The cytotoxicity of Vδ2 T cells against K562 cells was greatly enhanced by the treatment with IL-15.Conclusion IL-15 could activate the Vδ2 T cells and en-hance their cytotoxicity against K562 cells.
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<p><b>OBJECTIVE</b>To generate a mouse model of chronic hepatitis B (CHB) infection by performing in vivo transduction of hepatitis B virus (HBV) covalently closed circular (ccc)DNA.</p><p><b>METHODS</b>Nude mice were injected with HBV cccDNA at doses of 1.5, 1.0 or 0.5 mug/ml. A control group was generated by giving equal injection volumes of physiological saline. The serum levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) on post-injection days 1 and 3, weeks 1-6, 8 and 10 were assayed by reflection immunoassay. At post-injection week 10, all animals were sacrificed and liver tissues were collected. Copies of HBV DNA in serum and liver tissue were detected by real-time PCR. HBV antigens in liver tissue were detected of by immunohistochemistry. Pathological analysis of liver tissue carried out with hematoxylin-eosin staining. Linear correlation of data was determined by statistical analysis.</p><p><b>RESULTS</b>HBsAg and HBeAg were detected in sera from all three groups of cccDNA-injected mice staring at post-injection day 1 and lasting through week 10. The levels of HBsAg over the 10-week period showed two patterns of increase-decrease;the lowest level was detected at week 4 and the highest level was detected at week 8. In contrast, the levels of HBeAg over the 10-week period showed three patterns of increase-decrease; the lower levels were detected at weeks 2 and 4 and the higher levels at weeks 3 and 6. HBV DNA copies in liver tissues showed a cccDNA dose-dependent descending trend over the 10-week study period (1.5 mug/ml:1.14E+07 ± 6.51E+06 copies/g, 1.0 mug/ml:9.81E+06 ± 9.32E+06 copies/g, and 0.5 mug/ml:3.72E+06 ± 2.35E+06 copies/g; Pearson's r =0.979). HBV DNA copies in sera showed the pattern of 1.0 mug/ml cccDNA more than 1.5 mug/ml cccDNA more than 0.5 mug/ml cccDNA, and in general were higher than those detected in the liver tissues. Liver tissues from all cccDNA-injected mice showed positive immunohistochemistry staining for both HBsAg and HBeAg. HE staining showed that the liver tissues of all cccDNA-injected mice had severe fatty and vacuolar degeneration and less obvious structure of liver lobules (compared to the liver tissues from control mice).</p><p><b>CONCLUSION</b>The CHB mouse model successfully established in this study by in vivo transduction of HBV cccDNA may represent a useful tool to study the pathogenic mechanisms and potential antiviral treatments of human CHB.</p>