Diagnostic value of FIB-4, aspartate aminotransferaseto-platelet ratio index and liver stiffness measurement in hepatitis B virus-infected patients with persistently normal alanine aminotransferase

AIM To assess the diagnostic value of FIB-4, aspartate aminotransferase-to-platelet ratio index(APRI), and liver stiffness measurement(LSM) in patients with hepatitis B virus infection who have persistently normal alanine transaminase(PNALT).METHODS We enrolled 245 patients with chronic hepatitis B: 95 in PNALT group, 86 in intermittently elevated alanine transaminase(PIALT1) group [alanine transaminase(ALT) within 1-2 × upper limit of normal value(ULN)], and 64 in PIALT2 group(ALT > 2 × ULN). All the patients received a percutaneous liver biopsy guided by ultrasonography. LSM, biochemical tests, and complete blood cell counts were performed.RESULTS The pathological examination revealed moderate inflammatory necrosis ratios of 16.81%(16/95), 32.56%(28/86), and 45.31%(28/64), and moderate liverfibrosis of 24.2%(23/95), 33.72%(29/86), and 43.75%(28/64) in the PNALT, PIALT1, and PIALT2 groups, respectively. The degrees of inflammation and liver fibrosis were significantly higher in the PIALT groups than in the PNALT group(P < 0.05). No significant difference was found in the areas under the curve(AUCs) between APRI and FIB-4 in the PNALT group; however, significant differences were found between APRI and LSM, and between FIB-4 and LSM in the PNALT group(P < 0.05 for both). In the PIALT1 and PIALT2 groups, no significant difference(P > 0.05) was found in AUCs for all comparisons(P > 0.05 for all). In the overall patients, a significant difference in the AUCs was found only between LSM and APRI(P < 0.05).CONCLUSION APRI and FIB-4 are not the ideal noninvasive hepatic fibrosis markers for PNALT patients. LSM is superior to APRI and FIB-4 in PNALT patients because of the influence of liver inflammation and necrosis.

Serum thymosin β4 levels in patients with hepatitis B virus-related liver failure

AIM:To investigate whether serum thymosinβ4 can provide diagnostic or prognostic information in liver failure patients caused by chronic hepatitis B virus(HBV) infection. METHODS:Serum thymosinβ4 levels were measured in 30 patients with acute-on-chronic liver failure(ACLF), 31 patients with chronic liver failure(CLF),30 patients with compensated liver cirrhosis(CR)and 32 patients with chronic hepatitis B and 30 healthy controls.Serum thymosinβ4 levels were measured by enzyme-linked immunosorbent assay and Child-Pugh and model for end-stage liver disease(MELD)scores were calculated for each patient on admission.RESULTS:Compared with healthy controls,serum thymosinβ4 levels in ACLF,CLF,CR and chronic hepatitis B patients were significantly lower,6.5047 (4.7879-10.5314)μg/mL vs 0.4632(0.2759-0.8768) μg/mL,0.6981(0.5209-1.2008)μg/mL,1.8053 (0.8110-2.3397)μg/mL,3.7803(1.8570-6.4722)μg/mL, respectively(P<0.001).The levels of thymosinβ4 in liver failure(ACLF or CLF)patients were markedly lower than that in CR(P<0.001),and a difference was also found between CLF and ACLF patients(P=0.038).In patients with chronic liver disease,there was a positive relationship between thymosinβ4 levels and albumin, choline esterase,and platelet(P<0.001),and negative relationship with alanine aminotransferase(P=0.020), aspartate aminotransferase,total bilirubin,international normalized ratio of prothrombin time,and Child-Pugh and MELD scores(P<0.001).Of the 61 liver failure patients,the thymosinβ4 levels of non-survivors were significantly lower than that of survivors(P=0.007). Receiver operating characteristics analysis identified a thymosinβ4 cutoff level of 0.5708μg/mL for predicting poor prognosis in all liver failure patients.The serial thymosinβ4 values were observed in 13 liver failure inpatients.Lower initial values were observed in the death.While greater improvement in thymosinβ4 value was found in those who recovered from the disease. CONCLUSION:Serum thymosinβ4 can be used as an important potential predictor for liver failure caused by chronic HBV infection.

Sex disparity in viral load, inflammation and liver damage in transgenic mice carrying full hepatitis B virus genome with the W4P mutation in the preS1 region

AIM To study sex disparity in susceptibility to hepatocellular carcinoma(HCC), we created a transgenic mouse model that expressed the full hepatitis B virus(HBV) genome with the W4P mutation.METHODS Transgenic mice were generated by transferring the p HY92-1.1 x-HBV-full genome plasmid(genotype A2) into C57 Bl/6 N mice. We compared serum levels of hepatitis B surface antigen(HBs Ag), interleukin(IL)-6, and the liver enzymes alanine aminotransferase(ALT) and aspartate transaminase(AST), as well as liver histopathological features in male and female transgenic(W4PTG) mice and in nontransgenic littermates of 10 mo of age. RESULTS W4PTG males exhibited more pronounced hepatomegaly, significantly increased granule generation in liver tissue, elevated HBs Ag expression in the liver and serum, and higher serum ALT and IL-6 levels compared to W4PTG females or littermate control groups. CONCLUSION Together, our data indicate that the W4 P mutation in pre S1 may contribute to sex disparity in susceptibility to HCC by causing increased HBV virion replication and enhanced IL-6-mediated inflammation in male individuals. Additionally, our transgenic mouse model that expresses full HBV genome with the W4 P mutation in pre S1 could be effectively used for the studies of the progression of liver diseases, including HCC.

Expression of B7-H4 and hepatitis B virus X in hepatitis B virus-related hepatocellular carcinoma

AIM: To investigate the expression and clinical significance of B7-H4 and hepatitis B virus X(HBx) protein in hepatitis B virus-related hepatocellular carcinoma(HBV-HCC).METHODS: The expression of B7-H4 in the human HCC cell lines Hep G2 and Hep G2.2.15 were detected by western blot, flow cytometry, and immunofluorescence. The expression of B7-H4 and HBx in 83 HBV-HCC was detected by immunohistochemistry, and the relationship with clinicopathological features was analyzed. Paraffin sections were generated from 83 HBV-HCC patients(22 females and 61 males) enrolled in this study. The age of these patients ranged from 35 to 77 years, with an average of 52.5 ± 11.3 years. All experiments were approved by the Ethics Committees of the Second Affiliated Hospital, Zhejiang University School of Medicine.RESULTS: B7-H4 was significantly upregulated in Hep G2.2.15 cells compared to Hep G2 cells. Specifically, the protein expression of B7-H4 in the lysates of Hep G2 cells was more than that in Hep G2.2.15 cells. In addition, HBx was expressed only in Hep G2.2.15 cells. Similar data were obtained by flow cytometry. The positive rates of B7-H4 and HBx in the tissues of 83 HBV-HCC patients were 68.67%(57/83) and 59.04%(49/83), respectively. The expression of HBx was correlated with tumor node metastases(TNM) stage, and the expression of B7-H4 was positively correlated with HBx(rs = 0.388; p < 0.01). The expression level of B7-H4 in HBx-positive HBV-HCC tissues was substantially higher than that in HBx-negative HBV-HCC tissues. The expression level of B7H4 was negatively related to tumor TNM stage.CONCLUSION: Higher expression of HBx and B7-H4 was correlated with tumor progression of HBV-HCC, suggesting that B7-H4 may be involved in facilitating HBV-related hepatocarcinogenesis.

Anti-viral role of toll like receptor 4 in hepatitis B virus infection: An in vitro study

AIM Toll like receptors plays a significant anti-viral role in different infections. The aim of this study was to look into the role of toll like receptor 4(TLR4) in hepatitis B virus(HBV) infection.METHODS Real time PCR was used to analyze the transcription of TLR4 signaling molecules, cell cycle regulators and HBV DNA viral load after triggering the Hep G2.2.15 cells with TLR4 specific ligand. Nuclear factor(NF)-κB translocation on TLR4 activation was analyzed using microscopic techniques. Protein and cell cycle analysis was done using Western Blot and FACS respectively.RESULTS The present study shows that TLR4 activation represses HBV infection. As a result of HBV suppression, there are several changes in host factors which include partial release in G1/S cell cycle arrest and changes in host epigenetic marks. Finally, it was observed that anti-viral action of TLR4 takes place through the NF-κB pathway.CONCLUSION The study shows that TLR4 activation in HBV infection brings about changes in hepatocyte microenvironment and can be used for developing a promising therapeutic target in future.

Interleukin-28 and hepatitis C virus genotype-4:Treatment-induced clearance and liver fibrosis

AIM:To investigate the association between interleukin-28B(IL28B) genotype and response to treatment and hepatic fibrosis in patients with hepatitis C virus(HCV) genotype 4.METHODS:Two hundred and one HCV-genotype 4 patients were included.All patients were treated with Peginterferon alph2a/Ribavirin for 48 wk.End of treatment response(ETR) was defined as loss of detectable serum HCV RNA at the end of treatment.Sustained viral response(SVR) was defined as loss of detectable serum HCV RNA at the end of 24 wk follow up.Genotyping of IL28B rs12979860 was performed using the TaqMan assay.We used logistic regression to estimate the adjusted odds ratio(aOR) and 95%CI.RESULTS:The study included 201 HCV-genotype 4 patients.The majority of patients were men(89.6%),with a median age of 47 years,inter-quartile range(40-51).Approximately 62.5% of patients had ETR,and 49.6% had SVR.Individuals who achieved SVR were more likely to be younger(χ 2 = 4.91,P = 0.027),and less likely to have fibrosis(χ 2 = 15.54,P < 0.0001),or inflammation(χ 2 = 7.58,P = 0.006).The genotype distribution of rs12979860 was 36.2%,49.0% and 14.8% for genotypes CC,CT,and TT,respectively.In these participants,rs12979860 genotype distribution did not differ by gender(P = 0.466),pretreatment viral load(P = 0.600),inflammation(P = 0.435),or fibrosis(P = 0.291).The frequencies of IL28B rs12979860 genotypes were TT(14.8%),CT(49.0%),and CC(36.2%).Compared to rs12979860 genotype TT,aORs(95%CI) for ETR and SVR were:CC genotype,[17.55(5.34-57.69) and 5.92(2.09-16.76),respectively];CT genotype,[5.15(1.80-14.78) and 2.48(0.94-6.52),respectively].In the current study,the patients who did not achieve ETR or SVR had a lower prevalence of rs12979860 CC(17.4% and 23.3%,respectively) than individuals who had ETR or SVR(47.9% and 47.2%,respectively).Individuals with rs12979860 CC genotype had approximately 6 times the odds of SVR compared to individuals with TT genotype(aOR = 5.92;95%CI:2.09-16.76).Similarly,patients with CT genotype had SVR more often than patients with TT genotype(aOR = 2.48;95%CI:0.94-6.52).Carrying at least one copy of the C allele(genotypes CT and CC) had almost 8 times the probability of ETR compared to those with genotype rs12979860 TT(aOR = 7.87;95%CI:2.84-21.82),and approximately 3 times the odds of SVR compared to those with genotype rs12979860 TT(aOR = 3.46;95%CI:1.37-8.74).In addition,data were consistent with a significant gene-dose relationship(aOR = 4.05/allele;95%CI:2.27-7.22).The association between rs12979860 genotype and SVR was similar among those who achieved and those who did not achieve SVR.CONCLUSION:In HCV-genotype 4 patients,rs12979860 is a sensitive predictor of viral clearance,independent of viral load,age,gender or fibrosis,with no similar relation to severity of fibrosis.

Design,synthesis and biological activity of some novel benzimidazole derivatives against Coxsackie virus B_3

A series of novel benzimidazole derivatives was synthesized and their anti-Coxsackie virus B3 （CVB3） activity was evaluated in VERO ceils. Compounds 9 and 10 exhibited better inhibitory activity than those of ribavirin （RBV） with IC50 values of 5.30 and 1.06 μg/mL, respectively. ？2009 Xian Jin Luo. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Recognition of a multiple antigen peptide containing sequence from mimotope of the dengue type 3 virus NS4B protein by human antibodies

Objective: To evaluate the recognition of NS4 B mimotope, as multiple antigen peptide(MAP), by dengue antibodies presents in serum samples from patients with different serotype infections. Methods: A MAP containing mimotope sequence was synthesized and used to evaluate the recognition of NS4 B mimotope as MAP by a panel of 66 human sera from dengue cases by an indirect ELISA assay. Results: The MAP differentiated between sera from dengue viruses infected patients and sera from healthy individuals and the best reactivity was shown by serum from dengue type 3 virus patients. The recognition was more intense with serum from patients with secondary infection. Conclusions: The findings suggest the potential use of NS4 B mimotope on the development of a multi-epitope diagnostic tool. These results are important for further immunogenicity studies.

Elevated soluble 4-1BB is associated with serum markers of hepatitis B virus in patients with chronic hepatitis B

BACKGROUND Previous studies have suggested that the costimulatory molecule 4-1BB plays pivotal roles in regulating immunity during chronic viral infection.However,up to now,there are few studies about 4-1BB in chronic hepatitis B(CHB).AIM To clarify this issue,we report our comprehensive study results on the expression levels of 4-1BB in patients with CHB.METHODS From September 2018 to June 2019,a total of 64 patients with CHB were recruited from the Department of Hepatology,The First Hospital of Jilin University.Peripheral blood samples were collected from 52 treatment-naïve and 12 entecavir-treated patients with CHB as well as 37 healthy donors(including 24 healthy adults and 13 healthy children).The levels of soluble 4-1BB(s4-1BB)in plasma were measured by ELISA.4-1BB mRNA expression in peripheral blood mononuclear cells was detected by real-time quantitative PCR.RESULTS The s4-1BB levels in the plasma of patients with CHB were significantly higher than those in healthy adults(94.390±7.393 ng/mL vs 8.875±0.914 ng/mL,P<0.001).In addition,the s4-1BB level in plasma was significantly increased in patients with a higher viral load and a disease flare up.However,there were no significant differences between treatment-naïve and entecavir-treated patients.Interestingly,among treatment-naïve patients with CHB,the levels of s4-1BB in plasma had a significant positive correlation with hepatitis B surface antigen,hepatitis B virus DNA,hepatitis B e antigen,and triglyceride levels(r=0.748,P<0.001;r=0.406,P=0.004;r=0.356,P=0.019 and r=-0.469,P=0.007,respectively).The 4-1BB mRNA expression was higher in the peripheral blood mononuclear cells of patients with CHB than in the peripheral blood mononuclear cells of healthy adults,but the difference was not statistically significant.CONCLUSION These results suggest that the levels of s4-1BB may be associated with pathogenesis of hepatitis B virus and therefore may be a promising biomarker for disease progression.

FibroScan联合GPR、APRI和FIB-4诊断血清ALT正常的慢性HBV感染者肝纤维化价值分析

目的 评价基于4因子的肝纤维化指数(FIB-4)、γ-谷氨酰转肽酶/血小板比值(GPR)、天冬氨酸氨基转移酶/血小板比值(APRI)和FibroScan诊断HBV感染者肝纤维化的价值。方法 2015年2月~2020年12月广东省中医院肝病科诊治的血清ALT正常的慢性HBV感染者154例,纳入人群血清HBV DNA阳性,均接受肝活检,采用Scheuer评分系统评价肝纤维化程度。常规检测血小板计数和血生化指标,计算FIB、APRI和GPR指数,使用FibroScan-502诊断仪行肝硬度检测LSM(kPa)。采用多因素二元Logistic回归分析构建联合预测模型,绘制受试者工作特征(ROC)曲线,计算ROC曲线下面积(AUC),采用Delong法对AUC进行两两比较。结果 肝组织病理学检查显示,S0 34例,S1 47例,S2 48例,S3 22例,S4 3例,其中73例(47.4%)存在显著性肝纤维化(≥S2),25例(16.2%)存在进展性肝纤维化;≥S2患者FIB-4、APRI、GPR和LSM分别为1.0(0.8,1.2)、0.3(0.2,0.4)、0.2(0.2,0.3)和7.0(6.0,9.0)kPa,≥S3期分别为1.3(1.0,1.7)、0.4(0.3,0.5)、0.3(0.2,0.4)和9.0(7.4,11.0)kPa,均显著打于S0~S1期患者【分别为0.7(0.6,1.0)、0.2(0.2,0.3)、0.1(0.1,0.2)和5.0(4.1,6.1)kPa,P<0.05】;应用二元Logistics回归分析构建4种指标联合诊断≥S2和≥S3的模型分别为,Logit(≥S2)=1.303×FIB-4+9.8×GPR-0.684×APRI+0.7×LSM-7.565,其诊断≥S2肝纤维化的AUC为0.858,和Logit(≥S3)=3.307×FIB-4+5.361×GPR-4.394×APRI+0.635×LSM-9.632,其诊断≥S3肝纤维化的AUC为0.914,均显著优于各指标单独诊断(P均<0.001)。结论 在血清ALT水平正常的慢性HBV感染者中,近一半存在显著性肝纤维化。应用无创指标联合诊断肝纤维化具有一定的应用价值,值得临床开展研究。

甘草甜素对HBsAg低表达HepG2.2.15细胞株HBV感染及TLR2、4的影响

目的:探讨甘草甜素(GL)对HepG2.2.15细胞上清HBV DNA、e抗原分泌,Toll样受体2、4(TLR2、4)信号分子的表达及对细胞增殖的影响。方法:实时荧光定量PCR检测HBV DNA表达,ELISA检测HBV所分泌抗原,MTT检测细胞增殖活性,直接免疫荧光流式细胞术(FCM)检测TLR2、4在细胞株上表达的阳性细胞率,并与空白对照组对比。结果:HBsAg在该细胞株低表达,但HBeAg则明显阳性,因此研究了GL对e抗原的影响,给药后第3d e抗原总体均数间差异显著(P<0.01),但只有400μg/ml组与对照组间比较显著降低(P<0.05),800μg/ml组则较其余GL组明显升高(P<0.01)。HBV DNA给药后第3 d总体均数间比较差异有显著性,只有50μg/ml组较对照组降低,但无统计学意义(P>0.05),其余各组均较对照组升高;各组TLR2、4数值总体均数间有显著差异(P<0.01)。除200μg/ml组外,各剂量组与对照组比较显示两者均显著上调(P<0.05),但均无剂量依赖关系;MTT实验显示200μg/ml以下三个剂量组均可促进细胞增殖,但只有200μg/ml组与对照组间差异显著(P<0.05);400、800μg/ml两组均显著抑制细胞增殖(P<0.01);而MTT分别与HBeAg、HBV DNA呈显著负相关(P<0.05)。结论:研究表明,GL对HepG2.2.15变异株的病毒复制及e抗原分泌可能具有双向作用;细胞的存活数与病毒复制及e抗原呈负相关;TLR2在变异株低表达,GL呈非剂量依赖关系上调TLR2、4表达,至少在该细胞株GL影响HBV的机理与TLR2、4信号的改变无关,但有可能在体内通过免疫途径影响HBV。

Toll样受体4的表达及miR-TLR4干扰对HBV相关肝癌细胞生物学活性的影响

目的观察Toll样受体4在乙肝病毒相关肝癌细胞系中的表达及对其生物学活性的影响。方法Western blot检测Hep3B、Hep G2.2.15、Hep G2、SMMC7721及Huh7五种肝癌细胞中TLR4蛋白表达情况,选择TLR4表达高的HBV阳性肝癌细胞作为研究对象。构建4个mi R-TLR4质粒和一个阴性对照质粒,选择干扰效果明显的质粒转染TLR4表达高的HBV阳性肝癌细胞。实验分为正常组,阴性对照组,干扰质粒3组及干扰质粒4组。通过MTT法、克隆平板形成实验、流式细胞术分别检测干扰TLR4表达对肝癌细胞增殖、克隆形成、周期及凋亡的影响。结果 TLR4基因在五种肝癌细胞株中均有表达,在Hep3B细胞中表达最高。与正常组和阴性对照组相比,干扰TLR4表达后,Hep3B细胞的增殖能力明显受到抑制(P<0.05),克隆形成率显著下降(P<0.05),周期阻滞于G2/M期,并促进细胞凋亡(P<0.05)。结论 TLR4表达上调促进HBV相关肝癌细胞Hep3B生长增殖、周期再分布以及抑制凋亡,在肝癌发生发展过程中起重要作用。

柯萨奇B4病毒非结构蛋白P_2C重组质粒的构建

目的:构建柯萨奇B4病毒非结构蛋白P2C重组质粒。方法:用RT-PCR技术,从柯萨奇B4病毒中钓取非结构蛋白P2CcDNA片段,克隆入PUCm-T载体,转化大肠杆菌DH5α,构建重组质粒。结果:以柯萨奇B4病毒的总RNA为模板,扩增出987bp大小的片段,克隆入PUCm-T载体,用BamHⅠ、HindⅢ双酶切鉴定,与非结构蛋白P2C的PCR扩增产物片段大小一致。结论:成功地构建了柯萨奇B4病毒非结构蛋白P2C重组质粒。

HH方治疗HBV转基因小鼠抗病毒作用及其对TLR3／TLR4的影响

目的:观察HH方治疗乙肝病毒(HBV)转基因小鼠抗HBV的疗效及其作用机制。方法:HBV转基因小鼠随机分4组,模型组小鼠给予同体积生理盐水,苦参素组小鼠给予同体积含0.15 g/(kg·d)苦参素混悬液,高、低剂量HH方组小鼠分别给予同体积含7、2 g/(kg·d)HH混悬液。5周后,ELISA法检测血清及肝组织乙型肝炎病毒表面抗原(HBs Ag),常规HE染色观察肝组织病理变化,定量PCR法检测血清中乙型肝炎病毒脱氧核糖核酸(HBVDNA)及肝组织中HBVDNA、Toll样受体(TLR)3/4 mRNA,Western blot法检测TLR3/4蛋白表达。结果 :高剂量HH方[7 g/(kg·d)]可以减轻肝细胞轻度脂肪变性、肝细胞轻度水肿及Kuffer细胞增生等病理改变,对肝组织炎细胞浸润未见明显缓解。高剂量HH方可以显著降低HBV转基因小鼠肝脏和血清中的HBs Ag、HBVDNA水平,差异具有统计学意义(P<0.05),与苦参素组比较差异无统计学意义(P>0.05)。高剂量HH方可以显著增强肝组织TLR3、TLR4的mRNA及蛋白表达,差异具有统计学意义(P<0.05)。结论:高剂量HH方有一定抗HBV作用,推测与增强TLR3、TLR4表达进而影响干扰素分泌有关。

HBV相关慢加急性肝衰竭前期外周血中Toll样受体3、4的表达

目的探讨HBV相关慢加急性肝衰竭前期(pre-ACLF)外周血单个核细胞中Toll样受体3(TLR3)和Toll样受体4(TLR4)表达水平的变化及其意义。方法采用流式细胞仪检测HBV相关的pre-ACLF患者30例、慢性乙型肝炎患者30例以及健康对照者15名外周血单个核细胞TLR3、TLR4的表达水平,绘制ROC曲线,评价TLR3和TLR4对pre-ACLF的诊断价值。结果pre-ACLF组患者外周血中TLR3(20.81±5.15)明显升高,分别与慢性乙型肝炎组、健康对照组比较,均差异有统计学意义(均P<0.05)。同时,pre-ACLF组患者外周血中TLR4(13.73±3.56)也明显上升,分别与慢性乙型肝炎组、健康对照组比较,也均差异有统计学意义(均P<0.05)。根据TLR3和TLR4检测结果分别绘制ROC曲线,外周血TLR3诊断pre-ACLF的ROC曲线下的面积(AUC)为0.830(95%CI为0.728~0.932);外周血TLR4诊断pre-ACLF的ROC曲线下面积为0.977(95%CI为0.948~1.000)。结论通过对pre-ACLF患者TLR3和TLR4水平的检测,有助于对pre-ACLF的诊断。

肝癌组织HBx和ANGPTL4表达及与临床病理、预后的关系

目的探究肝癌组织乙型肝炎病毒(HBV)x蛋白(HBx)、血管生成素样蛋白4(ANGPTL4)表达与临床病理、预后的关系。方法选择2014年12月—2017年3月手术切除的HBV相关性肝癌组织98例,另择50例癌旁组织。分析肝癌及癌旁组织中HBx、ANGPTL4的表达及与肝癌患者临床病理、预后的关系。结果与癌旁组织比较,肝癌组织HBx表达阳性率降低,ANGPTL4表达阳性率升高(P<0.05)。肝癌HBx和ANGPTL4表达与肝癌患者包膜侵犯、门静脉浸润、远处转移相关(P<0.05,P<0.01)。肝癌包膜侵犯、门静脉浸润、临床分期、远处转移、HBx、ANGPTL4表达是影响预后的因素(P<0.05,P<0.01);HBx、ANGPTL4阳性肝癌患者生存率低于阴性患者(P<0.05)。结论肝癌组织HBx和ANGPTL4表达与临床病理及预后相关,临床可通过检测其表达情况判断患者病情及预后。

干扰素调节因子4基因在慢性HBV携带者外周血中的表达及临床意义

目的:从mRNA和蛋白水平两个层面探讨健康人群和慢性HBV携带者外周血干扰素调节因子4(IRF-4)基因表达情况。方法:选取慢性HBV携带者19例(病例组),健康对照者22例(对照组),采集外周血分离PBMC,利用流式细胞技术观测CD4+T细胞中IRF-4+细胞比例,利用实时定量PCR技术检测IRF-4mRNA水平,比较两组人群IRF-4表达差异。结果:通过FACS分析在CD4+T细胞群中明显出现IRF-4+细胞群;病例组和对照组IRF-4+细胞比例分别为(3.29±0.83)%、(2.26±0.82)%,差异具有统计学意义(t=3.98,P=0.000 3);病例组IRF-4 mRNA表达水平显著高于对照组(10.66±5.15 vs 5.53±2.74,t=4.06,P=0.000 2)。结论:慢性HBV携带者IRF-4表达在mRNA和蛋白水平均高于正常人群,可以作为免疫干预的潜在靶点。

柯萨奇B_4病毒对小鼠胰岛的影响及干扰素的保护作用

采用我国流行的柯萨奇B4病毒 (CB4V)感染swiss小鼠 ,建立I型糖尿病的动物模型 ,并观察干扰素对CB4V引起的I型糖尿病的保护作用。结果表明 ,80 %的小鼠在感染后 72h出现低血糖 ,血糖在 (90 .87±8.2 6 )mg/L范围之内 ,6～ 8周全部发展为高血糖。糖刺激的胰岛素释放量在感染后 72h增加 ,但至 3～ 8周明显低于正常对照组。在用CB4V感染小鼠的同时给予干扰素 ,目的在于观察干扰素对CB4V攻击胰岛 β细胞的保护作用 ,结果经干扰素治疗后的感染小鼠 ,当给干扰素到 6周 ,80 %小鼠血糖在 (118.6 6± 5 .73)mg/L范围之内 ,而低糖与高糖刺激下的胰岛素释放量分别在 (18.34± 3.5 0 )uIU/mL ,(31.5 1± 4 .71)uIU/mL范围之内 ,均与正常对照组无差异 (p >0 .0 5 )。

益肾固本法对慢性HBV携带者外周血干扰素调节因子-4表达的影响

目的:从mRNA和蛋白水平两个层面探讨益肾固本方对慢性HBV携带者外周血IRF(干扰素调节因子)-4表达的影响。方法:选取慢性HBV携带者46例,其中27例服用益肾固本方进行中药干预,19例作为对照组不予任何药物治疗,动态随访24周后,采集外周血分离外周血单个核细胞(PBMC),利用流式细胞技术(FACS)观测CD4+T细胞中IRF-4+细胞比例,利用实时定量PCR(聚合酶链反应)技术检测两组人群IRF-4 mRNA水平,比较其IRF-4表达差异。结果:FACS可以有效鉴定IRF-4+细胞群,中药组IRF-4+细胞群峰值偏移程度小于对照组;中药组和对照组IRF-4+细胞比例分别为(2.61±0.19)%、(3.29±0.18)%,差异具有统计学意义(t=2.48,P=0.02);中药组IRF-4 mRNA表达量显著低于对照组[(6.78±0.89)vs(10.66±1.18),t=2.67,P=0.01]。结论:益肾固本方可以有效降低慢性HBV携带者IRF-4表达水平,对于纠正Th1/Th2细胞失衡、打破免疫耐受具有重要意义。

CHB并EBV感染患者外周血单个核细胞中TLR4、TLR9表达及临床意义

目的研究慢性乙肝(CHB)合并EB病毒(EBV)感染患者外周血单个核细胞中Toll样受体4(TLR4)、Toll样受体9(TLR9)表达水平及临床意义。方法选取2017年3月至2020年3月江苏省中医院收治的54例CHB合并EBV感染患者作为研究对象,另选取本院同期收治的25例CHB未合并EBV感染患者作为对照组1,25例仅感染EBV病毒患者作为对照组2。检测所有患者丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(TBIL)、直接胆红素(DBIL)、间接胆红素(IBIL)、碱性磷酸酶(ALP)水平,采用流式细胞术检测TLR4、TLR9阳性表达情况,采用Spearman分析数据间相关性,采用Kaplan-Meier生存曲线对比两组患者生存情况。结果研究组患者外周血单个核细胞TLR4、TLR9阳性表达情况显著高于两组对照组,差异有统计学意义(P<0.05);研究组患者的血ALT、AST、TBIL、DBIL、IBIL、ALP水平均明显高于两组对照组,差异有统计学意义(P<0.05);TLR4阳性表达与血ALT、AST、DBIL及ALP水平呈正相关,TLR9阳性表达与血ALT、AST、IBIL及ALP水平呈正相关;TLR4、TLR9高表达患者疾病进展率高于低表达患者,差异有统计学意义(χ^(2)=6.135、5.684,P<0.05);TLR4、TLR9高表达患者15个月生存率较低表达患者比较,差异无统计学意义(P>0.05)。结论CHB合并EBV感染患者外周血单个核细胞TLR4、TLR9阳性表达增多,与肝功能损伤加重和疾病进展相关。