感染性肺炎新生儿的血清25-羟基维生素D与炎症因子的相关性分析

目的分析感染性肺炎新生儿的血清25-羟基维生素D[25-hydroxyvitamin D,25(OH)D]与炎症因子干扰素-γ(infectious pneumonia-γ,IFN-γ)、C-反应蛋白(C-reactive protein,CRP)、白细胞介素-2(interleukin-2,IL-2)水平的相关性。方法选取河北省秦皇岛市第一医院2018年12月至2020年12月收治的100例感染性肺炎新生儿,根据血清25(OH)D水平分为缺乏组(≤15.0μg/L,18例)、不足组(15.1~20.0μg/L,42例)、充足组(>20.0μg/L,40例)。统计3组性别、胎龄、血清25(OH)D水平、出生体重、分娩方式等一般资料和临床资料,比较3组IFN-γ、CRP及IL-2水平,分析新生儿血清25(OH)D水平与IFN-γ、CRP、IL-2水平的相关性。结果3组胎龄、性别、出生体质量、分娩方式比较差异无统计学意义(P>0.05);缺乏组、不足组、充足组新生儿的血清25(OH)D水平逐渐升高,差异有统计学意义(P<0.05)。缺乏组IFN-γ、CRP及IL-2水平均高于不足组、充足组(P<0.05),不足组IFN-γ、CRP及IL-2水平高于充足组(P<0.05)。经Pearson相关分析,感染性肺炎新生儿血清25(OH)D水平与IFN-γ、CRP、IL-2水平呈负相关(P<0.05)。结论新生儿血清25(OH)D越低,维生素D越缺乏,IFN-γ、CRP及IL-2水平越高,感染性肺炎的风险越高。

HDP患者血清PLGF、IFI16、ANGPTL2与不良妊娠结局关系及预测价值

目的:研究妊娠高血压疾病(HDP)患者血清胎盘生长因子(PLGF)、γ干扰素诱导蛋白16(IFI16)、血管生成素样蛋白2(ANGPTL2)水平及预测妊娠结局价值。方法:选取2020年1月-2023年1月在本院就诊并分娩的126例HDP孕妇,根据病情严重程度分为轻度组(58例)、重度组(68例),依据妊娠结局分为不良组(52例)、良好组(74例)。同期在本院健康分娩的孕妇60例为健康组。检测血清甘油三酯(TG)、总胆固醇(TC)、糖化血红蛋白(HbA1c)、PLGF、IFI16、Angptl2蛋白表达和尿液中24 h尿蛋白、尿酸(UA)、肌酐(Cr)水平,血清。Spearman分析PLGF、IFI16、Angptl2与HDP严重程度和妊娠结局的相关性;logistic分析不良妊娠结局影响因素;受试者工作特征(ROC)曲线分析PLGF、IFI16、ANGPTL2对HDP孕妇不良妊娠结局的预测价值。结果:健康组、轻度组、重度组血清PLGF水平依次降低,IFI16、ANGPTL2依次升高;良好组舒张压、收缩压、24 h尿蛋白、IFI16、ANGPTL2低于不良组,PLGF高于不良组(均P<0.05)。HDP严重程度、不良妊娠结局与PLGF呈负相关,与IFI16、ANGPTL2呈正相关(P<0.05)。舒张压、收缩压、IFI16、ANGPTL2异常升高是影响HDP患者不良妊娠结局的危险因素,PLGF升高是保护因素(P<0.05)。PLGF、IFI16、ANGPTL2联合检测预测HDP患者不良妊娠结局的曲线下面积为0.905,灵敏度98.1%、特异度73.0%,价值高于单独指标检测(P<0.05)。结论:HDP患者血清PLGF较低,IFI16、ANGPTL2较高,PLGF、IFI16、ANGPTL2联合检测可提高不良妊娠结局预测价值。

非小细胞肺癌晚期患者白细胞介素-2、白细胞介素-6、肿瘤坏死因子-α和γ干扰素对生存情况的预测价值

目的分析非小细胞肺癌(NSCLC)晚期患者血清中白细胞介素-2(IL-2)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和γ干扰素(IFN-γ)变化与生存率的相关性。方法回顾性分析2021年10月至2022年9月在抚州市第一人民医院肿瘤内科确诊的60例NSCLC晚期患者临床资料,作为观察组,根据患者入院6个月后的预后情况将患者分为生存组(n=41)和死亡组(n=19),同期选择入院体检的66例健康人群为健康组。记录患者入院24 h内和治疗2个周期后的IL-2、IL-6、TNF-α、IFN-γ水平,比较治疗2个周期后的生存组和死亡组患者的IL-2、IL-6、TNF-α、IFN-γ水平,以患者的预后情况作为最终变量绘制NSCLC患者预后的ROC曲线图。结果健康组的IL-6水平低于观察组,IL-2、TNF-α和IFN-γ水平均高于观察组,差异均有统计学意义(P<0.05)。患者入院24 h内,死亡组IL-6水平高于生存组,差异有统计学意义(P<0.05)。两组患者入院治疗2个周期后,患者的IL-2水平较入院24 h内低,IL-6、TNF-α和IFN-γ水平较24 h内的指标水平高,差异均有统计学意义(P<0.05)。治疗2个周期后,生存组的IL-6较死亡组低,差异有统计学意义(P<0.05)。ROC结果显示,IL-6水平鉴别诊断NSCLC患者的预后的ROC曲线下面积为0.738,敏感度为89.47%,特异性为65.85%,此时IL-6水平的截断值为60.731 pg/ml(P<0.05)。结论NSCLC晚期患者的IL-6水平对其预后具有一定的预测效能,可以为临床NSCLC患者的治疗方案提供指导作用。

Context-dependent role of sirtuin 2 in inflammation

Sirtuin 2 is a member of the sirtuin family nicotinamide adenine dinucleotide(NAD~+)-dependent deacetylases, known for its regulatory role in different processes, including inflammation. In this context, sirtuin 2 has been involved in the modulation of key inflammatory signaling pathways and transcription factors by deacetylating specific targets, such as nuclear factor κB and nucleotide-binding oligomerization domain-leucine-rich-repeat and pyrin domain-containing protein 3(NLRP3). However, whether sirtuin 2-mediated pathways induce a pro-or an anti-inflammatory response remains controversial. Sirtuin 2 has been implicated in promoting inflammation in conditions such as asthma and neurodegenerative diseases, suggesting that its inhibition in these conditions could be a potential therapeutic strategy. Conversely, arthritis and type 2 diabetes mellitus studies suggest that sirtuin 2 is essential at the peripheral level and, thus, its inhibition in these pathologies would not be recommended. Overall, the precise role of sirtuin 2 in inflammation appears to be context-dependent, and further investigation is needed to determine the specific molecular mechanisms and downstream targets through which sirtuin 2 influences inflammatory processes in various tissues and pathological conditions. The present review explores the involvement of sirtuin 2 in the inflammation associated with different pathologies to elucidate whether its pharmacological modulation could serve as an effective strategy for treating this prevalent symptom across various diseases.

Expression level of interferon-stimulated genes PKR,OAS1,MX1,and ISG15 in peripheral blood mononuclear cells of COVID-19 patients:A retrospective study

Objective:To explore expression level of interferon-stimulated genes PKR,OAS1,MX1,and ISG15 in peripheral blood mononuclear cells of COVID-19 patients.Methods:In this study,changes in the expression of four interferon-stimulated genes(ISGs),including PKR,OAS1,MX1,and ISG15,in peripheral blood mononuclear cells of 45 COVID-19 patients with different severities were evaluated by real-time PCR method.Results:OAS1,MX1,PKR,and ISG15 were differently expressed in COVID-19 patients with different severity.The results showed that the expression of OAS1,MX1,PKR,and ISG15 genes was significantly(P=0.001)lower in severe patients.Conclusions:Weak and defective IFN response and subsequent disruption of ISGs may be associated with COVID-19 severity.

口腔种植体周围炎患者血清干扰素调节因子4、可溶性致癌抑制因子2检测的临床意义

目的探讨口腔种植体周围炎患者血清干扰素调节因子4(IRF4)、可溶性致癌抑制因子2(sST2)检测的临床意义。方法选取2021年1月—2022年12月沧州市人民医院口腔种植体周围炎患者30例(口腔种植体周围炎组)、同一类型种植体的健康口腔种植体者30例(健康口腔种植体组)和体检健康者30名(正常对照组)。检测所有研究对象血清IRF4、sST2水平。采用Pearson相关分析评估血清IRF4、sST2水平与年龄、探诊深度(PD)、龈沟液量、出血指数(SBI)的相关性。采用受试者工作特征(ROC)曲线评估血清IRF4、sST2诊断口腔种植体周围炎的效能。结果口腔种植体周围炎组PD、龈沟液量、SBI均显著高于健康口腔种植体组和正常对照组(P<0.05)。正常对照组、健康口腔种植体组、口腔种植体周围炎组血清IRF4、sST2水平依次升高(P<0.001)。口腔种植体周围炎患者男性、女性之间血清IRF4、sST2水平差异均无统计学意义(P>0.05)。≥60岁的口腔种植体周围炎患者血清IRF4、sST2水平高于<60岁的患者(P<0.001)。血清IRF4、sST2与年龄、PD、龈沟液量、SBI均呈正相关(P<0.05)。血清IRF4、sST2单项检测和联合检测诊断口腔种植体周围炎的曲线下面积(AUC)分别为0.919、0.890、0.978。结论口腔种植体周围炎患者血清IRF4、sST2水平显著升高,与牙周指标密切相关,或可作为口腔种植体周围炎的诊断指标。

重组人干扰素α2b凝胶联合阿奇霉素治疗慢性宫颈炎伴HPV感染患者的临床疗效

【目的】探讨重组人干扰素α2b凝胶联合阿奇霉素治疗慢性宫颈炎伴人乳头瘤病毒(HPV)感染患者的临床疗效。【方法】选取2021年3月至2023年3月本院收治的110例慢性宫颈炎伴HPV感染患者,按随机数字表法分为对照组和观察组,每组55例。对照组采用重组人干扰素α2b凝胶治疗,观察组采用重组人干扰素α2b凝胶联合阿奇霉素治疗,均治疗14 d。比较两组患者临床疗效、乳酸杆菌减少率、白细胞酯酶(LE)阳性率、凝固酶(GADP)阳性率、阴道pH值>4.5占比、炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)]水平、HPV病毒载量、HPV转阴率及疾病复发率。【结果】观察组总有效率高于对照组,差异有统计学意义(P<0.05)。观察组乳酸杆菌减少率、LE阳性率、GADP阳性率、阴道pH值>4.5占比均高于对照组,差异有统计学意义(P<0.05)。治疗前,两组炎症因子水平比较,差异无统计学意义(P>0.05);治疗后,观察组TNF-α、IL-1β、IL-8水平均低于对照组,差异有统计学意义(P<0.05)。治疗后,两组病毒载量均下降,且观察组病毒载量低于对照组,差异有统计学意义(P<0.05)。观察组HPV转阴率高于对照组,复阳率、疾病复发率低于对照组,差异有统计学意义(P<0.05)。【结论】重组人干扰素α2b凝胶联合阿奇霉素治疗慢性宫颈炎伴HPV感染患者的效果显著,可降低炎症因子水平,减少HPV病毒载量,促进HPV转阴,值得临床推广。

miR-140-3p.2靶向PD-L1表达增强CD8^(+)T细胞对肝癌细胞的杀伤作用

目的 探究miR-140-3p.2靶向程序性死亡受体配体1(PD-L1)表达对细胞毒性T细胞(CD8~+T细胞)杀伤肝癌细胞的影响。方法 实时荧光定量PCR(qRT-PCR)、Western blotting检测L-02和HepG2、Hep3B、HuH-7细胞系中miR-140-3p.2和PD-L1 mRNA和蛋白水平;Targetscan数据库预测miR-140-3p.2和PD-L1的结合位点,并利用双荧光素酶实验进行验证;HepG2细胞分为过表达miR-140-3p.2(miR-140-3p.2)组及其对照(miR-NC)组、过表达PD-L1(PD-L1)组及其对照(NC)组、过表达miR-140-3p.2和PD-L1(miR-140-3p.2+PD-L1)组、过表达miR-140-3p.2和PD-L1对照(miR-140-3p.2+NC)组;qRT-PCR、Western blotting实验分别检测各组细胞中miR-140-3p.2和PD-L1 mRNA和蛋白水平。30只裸鼠分为过表达miR-140-3p.2(miR-140-3p.2)组及其对照(miR-NC)组,每组各15只,采用sc过表达miR-140-3p.2及其对照(miR-NC)的HepG2细胞制备移植瘤模型;检测肿瘤质量及体积;分离裸鼠脾脏组织CD8^(+)T细胞,并与各转染HepG2细胞共培养,细胞毒性实验检测细胞裂解率,ELISA法检测细胞培养上清液中肿瘤坏死因子-α(TNF-α)、γ干扰素(IFN-γ)水平;ELISA法检测移植瘤中TNF-α、IFN-γ水平,流式细胞术检测移植瘤中CD8^(+)T细胞浸润情况。结果 与L-02细胞相比,HepG2、Hep3B、HuH-7细胞中miR-140-3p.2水平降低,PD-L1 mRNA和蛋白水平升高(P<0.05)。与miR-NC组相比,miR-140-3p.2组细胞中miR-140-3p.2水平升高,PD-L1 mRNA和蛋白水平降低,细胞裂解率升高,细胞培养上清液中TNF-α、IFN-γ水平升高(P<0.05);与NC组相比,PD-L1组细胞中miR-140-3p.2水平降低,PD-L1 mRNA和蛋白水平升高,细胞裂解率降低,细胞培养上清液中TNF-α、IFN-γ水平降低(P<0.05);过表达PD-L1能部分逆转过表达miR-140-3p.2对上述指标的影响(P<0.05)。裸鼠成瘤4周后,与miR-NC组相比,miR-140-3p.2组移植瘤体积和质量明显减小(P<0.05),移植瘤中miR-140-3p.2水平明显升高、PD-L1 mRNA和蛋白水平明显降低(P<0.05),CD8^(+)T细胞浸润水平明显升高(P<0.05),TNF-α、IFN-γ水平明显升高(P<0.05)。结论 miR-140-3p.2靶向PD-L1表达增强CD8^(+)T细胞对肝癌细胞的杀伤作用。

灯盏乙素通过环状GMP-AMP合酶-干扰素基因刺激因子通路抑制BV-2小胶质细胞介导的神经炎症

目的探讨灯盏乙素对脂多糖(LPS)诱导的BV-2小胶质细胞神经炎症的影响。方法培养BV-2小胶质细胞系,将BV-2小胶质细胞分为对照组(Ctrl)、环状GMP-AMP合酶(cGAS)抑制剂RU320521(RU.521)组、LPS组、LPS+RU.521组、LPS+灯盏乙素预处理(LPS+S)组、LPS+S+RU.521组,共6组。Western blotting及免疫荧光双标染色法检测并观察BV-2小胶质细胞中cGAS、干扰素基因刺激因子(STING)、核因子κB(NF-κB)、磷酸化NF-κB(p-NF-κB)、PYD结构域蛋白3(NLRP3)和肿瘤坏死因子α(TNF-α)的表达变化(n=3)。结果Western blotting和免疫荧光双标染色均显示,与对照组相比,LPS诱导后,BV-2小胶质细胞中cGAS、STING、p-NF-κB、NLRP3和TNF-α蛋白的表达水平显著升高(P<0.05);与LPS组相比,LPS+S组中cGAS、STING、p-NF-κB、NLRP3和TNF-α蛋白的表达水平显著下降(P<0.05)。使用cGAS通路抑制剂RU.521后显示了与灯盏乙素预处理组相似的作用效果。此外,NF-κB在各组的变化不明显(P>0.05)。结论灯盏乙素干预抑制BV-2小胶质细胞介导的神经炎症反应,可能与cGAS-STING信号通路有关。

重组干扰素α-2b联合CO_(2)激光气化治疗宫颈低级别上皮内病变伴HR-HPV感染患者的临床效果及对HPV转阴率的影响

目的分析重组干扰素α-2b联合CO_(2)激光气化治疗宫颈低级别上皮内病变伴高危型人乳头瘤病毒(HR-HPV)感染的临床效果。方法选取宫颈低级别上皮内病变伴HR-HPV感染患者80例,采用随机数表法分为对照组及研究组,每组40例。对照组采用重组干扰素α-2b治疗,研究组采用重组干扰素α-2b联合CO_(2)激光气化治疗。比较两组患者的治疗效果。结果研究组治疗总有效率为90.00%,高于对照组的72.50%(P<0.05)。研究组治疗3、6个月的HPV转阴率均高于对照组(P<0.05)。两组不良反应总发生率比较,差异无统计学意义(P>0.05)。结论采用重组干扰素α-2b联合CO_(2)激光气化治疗宫颈低级别上皮内病变伴HR-HPV感染患者,可提高疗效及HPV转阴率。

Assessment of natural and interleukin-2-induced production of interferon-gamma in patients with liver diseases

AIMS To clarify whether the lower interferon gamma (IFNγ) production by lymphocytes in patients with liver diseases is due to defects of lymphocytes themselves or of other cofactors such as interleukin-2(IL-2). METHODS Peripheral blood mononuclear cells (PBMCs) from patients with various liver diseases were cultured with or without PHA and IL-2. The cells were harvested and counted and the su- pernatants were tested for IFNγ by a sensitive and quantitative ABC-ELISA. RESULTS IFNγ was not round in serum samples from patients as well as normal individuals. However,in supernatants of non-in- duced and induced PBMCs,IFN7 was detected by ABC-ELISA. In non-induced PBMCs (group 1),the content of IFNγ in super- natants from control,CAH,CPH and HCC was 8.72 μg/L, 5.03 μg/L,6.02 μg/L and 4.91 μg/L respectively. The pro- duction of IFNγ in liver disease was significantly decreased,com- pared to control. In group 2 in which PBMCs were stimulated with PHA,the content of IFNγ was 22.71,17.12,14.54 and 17.63 μg/L respectively. In group 3 in which PBMCs were in- duced by IL-2,the amount of IFN7 in supernatant from control (60.67 μg/L) was much larger than those from CAH (21.70 μg/ L),CPH (24.00 μg/L) and HCC (19.15 μg/L) (P<0.01). Comparing the amount of IFNγ in group 3 (IL-2-induced) with that in group 1 (non-induced),we found that IFNγ production was en- hanced by nearly 4 folds in liver diseases and by over 7 folds in control,Whereas the number of PBMCs,whether from liver dis- eases or from control,was increased by only approximately 3 folds. CONCLUSIONS The decreased production of IFNγ in liver dis- eases including HCC is mainly due to endogenous defects of lym- phocytes though the defects of stimulating cofactors such as IL-2 may also be involved.

Cloning and Sequence Analysis of Interferon γ-2β Full-length cDNA in Cyprinus carpio L.

[Objective] The research aimed to carry out the cloning, identification and sequence analysis of full-length cDNA of carp interferon γ-2β （IFNγ-2β）. [Method] The cDNA library of peripheral blood leucocytes which were separated from carp and stimulated with mitogen was screened by a probe labeled with DIG. The IFNγ- 2β EST sequence was picked out from the constructed cDNA library of peripheral blood leucocyte, and the full length of carp interferon γ-2β was cloned. In addition, the sequence analysis was carried out. [Result] Three positive clones were obtained. Sequence analysis indicated that the sequence had a 119 bp 5’-UTR and a 218 bp 3’-UTR, and the open reading frame （ORF）of this gene was 537 bp which putatively coded 178 amino acids and there were several instable motifs for mRNA （ATTTA） in the 3’-untranslated region. Its homology with IFN from GenBank was up to 97% . Analysis on protein sequence and structure showed that the predicted protein sequence was identified as an IFN family signature. [Conclusion] The research laid the foundation for further studying the expression manner, function characteristic and regulation mechanism of IFNγ-2β in vivo and the action mechanism in the inflammatory reaction, emergency reaction and immune response.

慢加急性乙型肝炎肝衰竭患者外周血单个核细胞IP-10和COX-2水平变化及其临床意义探讨

目的 探讨乙型肝炎病毒相关慢加急性肝衰竭(HBV-ACLF)患者外周血单个核细胞(PBMC)干扰素诱导蛋白-10(IP-10)和环氧化酶-2(COX-2)水平变化及其临床意义。方法 2019年1月~2021年3月我院收治的73例HBV-ACLF患者和95例慢性乙型肝炎(CHB)患者,分离PBMCs,采用RT-PCR法检测PBMC IP-10和COX-2mRNA水平,采用化学发光法检测血清HBsAg水平,采用ELISA法检测血清IFN-γ水平。结果 HBV-ACLF组血清IFN-γ及PBMC IP-10 mRNA和COX-2 mRNA水平分别为(59.3±7.9)ng/L、(0.9±0.1)和(1.6±0.2),显著高于CHB组【分别为(35.8±6.2)ng/L、(0.6±0.1)和(0.5±0.1),P<0.05】;HBV-ACLF患者PBMC IP-10和COX-2水平与血清HBsAg水平呈负相关(r=-0.828,P<0.05;r=-0.795,P<0.05);29例死亡组血清IFN-γ及PBMC IP-10 mRNA和COX-2 mRNA水平分别为(67.5±8.1)ng/L、(1.2±0.2)和(1.9±0.4),显著高于44例生存组【分别为(53.9±7.4)ng/L、(0.7±0.1)和(1.4±0.2),P<0.05】;应用PBMC IP-10和COX-2水平评估HBV-ACLF患者预后的AUC分别为0.835和0.828,两者差异无统计学意义(P>0.05);32例PBMC IP-10 mRNA≥0.9患者90 d生存率为46.9%,显著低于41例IP-10 mRNA<0.9患者的70.7%(P<0.05),38例COX-2 mRNA≥1.7患者90 d生存率为47.4%,显著低于35例COX-2 mRNA<1.7患者的74.3%(P<0.05)。结论 检测HBV-ACLF患者PBMC IP-10和COX-2水平可能能帮助预测其短期预后,值得进一步研究。

弥漫大B细胞淋巴瘤患者血浆IFN-γ、IL-2及外周血T淋巴细胞亚群、NK细胞与其临床分期的关系

目的探讨弥漫大B细胞淋巴瘤(DLBCL)患者血浆干扰素γ(IFN-γ)、白细胞介素-2(IL-2)水平及外周血T淋巴细胞亚群、自然杀伤(NK)细胞与临床分期的关系。方法纳入河南中医药大学第三附属医院2020年12月至2022年12月收治的DLBCL患者198例,根据临床分期分成Ⅰ期组(31例)、Ⅱ期组(56例)、Ⅲ期组(59例)、Ⅳ期组(52例)。比较4组血浆IFN-γ、IL-2及外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平,分析各指标与DLBCL分期的相关性。结果Ⅲ期组、Ⅳ期组的血浆IFN-γ、IL-2水平低于Ⅰ期组、Ⅱ期组(P<0.05)。Ⅲ期组、Ⅳ期组外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平低于Ⅰ期组、Ⅱ期组(P<0.05)。DLBCL患者IFN-γ、IL-2与CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平呈正相关(P<0.05)。DLBCL患者的血浆IFN-γ、IL-2及外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平与临床分期呈负相关(P<0.05)。IFN-γ、IL-2、CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平增高是控制临床分期增高的保护因素(P<0.05)。结论DLBCL患者的临床分期与血浆IFN-γ、IL-2及外周血T淋巴细胞亚群、NK细胞水平相关,且血浆IFN-γ、IL-2水平与T淋巴细胞亚群、NK细胞存在相关性,上述指标有望对DLBCL进展风险进行预测。

PLP2, a potent deubiquitinase from murine hepatitis virus, strongly inhibits cellular type I interferon production

Infections by coronaviruses such as severe acute respiratory syndrome （SARS） coronavirus （SCoV） and mouse hepatitis virus A59 （MHV-A59） result in very little type I interferon （IFN） production by host cells, which is potentially responsible for the rapid viral growth and severe immunopathology associated with SARS. However, the molecular mechanisms for the low IFN production in cells infected with coronaviruses remain unclear. Here, we provide evidence that Papain-like protease domain 2 （PLP2）, a catalytic domain of the nonstructural protein 3 （nsp3） of MHV-A59, can bind to IRF3, cause its deubiquitination and prevent its nuclear translocation. As a consequence, co-expression of PLP2 strongly inhibits CARDIF-, TBK1- and IRF3-mediated IFNp reporter activities. In addition, we show that wild-type PLP2 but not the mutant PLP2 lacking the deubiquitinase （DUB） activity can reduce IFN induction and promote viral growth in cells infected with VSV. Thus, our study uncovered a viral DUB which coronaviruses may use to escape from the host innate antiviral responses.

Combination of "low-dose" ribavirin and interferon alfa-2a therapy followed by interferon alfa-2a monotherapy in chronic HCV-infected nonresponders and relapsers after interferon alfa-2a monotherapy

AIM To report on the efficacy, safety and tolerability of interferon alfa-2a combined with a "low dose" of ribavirin for relapsers and non responders to alpha interferon monotherapy.METHODS Thirty-four chronic hepatitis C virus-infected non-responders to interferon alfa2a monotherapy (a course of at least 3 months treatment) and 13 relapsers to interferon alfa 2a monotherapy (a dose of 3 to 6 million units three times per week for at least 20 weeks but not more than 18 months) were treated with the same dose of interferon alfa-2a used before (3 to 6 million units three times per week) and ribavirin (10 mg/ kg daily) for 6 months. In complete responders, interferon alfa-2a was administered for further 6 months at the same dose used before as monotherapy.RESULTS Seven (20.6%) of 34 non-responders stopped the combined therapy due to adverse events, including two patients with histological and clinical Child A cirrhosis. In 17/27 (63%)non-responders, the combined therapy was stopped after three months because of non-response. Ten of the 27 non-responders completed the 1;2-month treatment course. At a mean follow up of 28 months (16- 37 months)after the treatment, 4/10 (15%) previous non-responders still remained complete responders,All 13 previous relapsers completed the 12-month treatment course. At a mean follow up of 22months (9 - 36 months) after treatment, 6/13(46%) the previous relapsers were stillsustained complete responders.CONCLUSION Our treatment schedule of the combined therapy for 6 months of interferon alfa2a with a low dose of ribavirin (10 mg/kg/day)followed by 6 months of interferon alfa-2amonotherapy is able to induce a sustainedcomplete response rate in 15% of non-responders and 46% of relapsers with chronic hepatitis C virus-related liver diseases comparable to those obtained with the standarddoses of ribavirin 1000 - 1200 mg/day.Randomized prospective controlled trials using lower total amounts of ribavirin in combination with interferon should be performed.

Pegylated interferon α-2b up-regulates specific CD8+ T cells in patients with chronic hepatitis B

AIM:To investigate the effect of pegylated interferon (IFN) α-2b on specific CD8+ T lymphocytes in patients with chronic hepatitis B (CHB). METHODS:Twenty-one patients with CHB were treated with pegylated IFN α-2b. Periphery blood mononuclear cells were isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation (density:1.077 g/L,Pharmingen) at weeks 0,4,8,12,and 24,respectively. Frequency of circulating hepatitis B virus (HBV) epitope-specific CD8 T cells was detected by flow cytometry. Cytokines were detected by cytometric bead assay. RESULTS:The frequency of circulating HBV core or env-specific CD8 T cells was higher (P < 0.05),the number of HBV core specific CD8 T cells was greater at week 24 (P < 0.05),the level of Th1-type cytokines [interleukin (IL)-12,tumor necrosis factor-α,and IFN-γ] was higher,while that of Th2-type cytokines (IL-4,IL-6,and IL-10) was lower in responders than in nonresponders (P < 0.05) after pegylated IFN α-2b treatment. The IL-6 level was correlated with HBV DNA (r = 0.597,P = 0.04),while the inducible protein-10 (IP-10) level was correlated with serum alanine aminotransferase (ALT) (r = 0.545,P = 0.005). The IP-10 level at week 8 after pegylated IFN α-2b treatment could predict the normalization of ALT in CHB patients (positive predict value = 56%,negative predict value = 92%). CONCLUSION:Pegylated IFN α-2b can enhance the immune response of CHB patients by increasing the frequency of HBV specific CD8+ T cells and regulating the Th1/Th2 cytokines.

Interferon alpha plus ribavirin combination treatment of Japanese chronic hepatitis C patients with HCV genotype 2:A project of the Kyushu University Liver Disease Study Group

AIM： To determine the efficacy of an interferon alpha and ribavirin combination treatment for Japanese patients infected with hepatitis C virus （HCV） of genotype 2, a multi-center study was retrospectively analyzed. METHODS： In total, 173 patients with HCV genotype 2 started to receive interferon-alpha subcutaneously thrice a week and 600-800 mg of ribavirin daily for 24 wk. RESULTS： The overall sustained virological response （SVR）, defined as undetectable HCV RNA in serum, 24 wk after the end of treatment, was remarkably high by 84.4%, （146/173） by an intention-to-treat analysis. A significant difference in SVR was found between patients with and without the discontinuation of ribavirin （46.9% vs 92.9 %）, but no difference was found between those with and without a dose reduction of ribavirin. A significant difference in SVR was also found between patients with less than 16 wk and patients with 16 or more weeks of ribavirin treatment （34.8 % vs 92.0 %）. CONCLUSION： The 24-wk interferon and ribavirin treatment is highly effective for Japanese patients with HCV genotype 2. The significant predictor of SVR is continuation of the ribavirin treatment for up to 16

The Predictive Value of Baseline HBs Ag Level and Early Response for HBs Ag Loss in Patients with HBe Ag-positive Chronic Hepatitis B during Pegylated Interferon Alpha-2a Treatment

Objective To explore the predictive value of baseline HBs Ag level and early response for HBs Ag loss in patients with HBe Ag-positive chronic hepatitis B during pegylated interferon alpha-2a treatment. Methods A total of 121 patients with HBe Ag-positive chronic hepatitis B who achieved HBs Ag loss were enrolled; all patients were treated with PEG-IFNα-2a 180 μg/week. Serum HBV DNA and serological indicators （HBs Ag, anti-HBs, HBe Ag, and anti-HBe） were determined before and every 3 months during treatment. Results The median treatment time for HBs Ag loss was 84 weeks （7-273 weeks）, and 74.38% （90 cases） of the patients needed extended treatment （〉 48 weeks）. The correlation between baseline HBs Ag levels and the treatment time of HBs Ag loss was significant （B = 14.465, t = 2.342, P = 0.021）. Baseline HBs Ag levels together with the decline range of HBs Ag at 24 weeks significantly correlated with the treatment time of HBs Ag loss （B = 29.862, t = 4.890, P = 0.000 and B = 27.993, t = 27.993, P = 0.005）. Conclusion Baseline HBs Ag levels and extended therapy are critical steps toward HBs Ag loss. Baseline HBs Ag levels together with early response determined the treatment time of HBs Ag loss in patients with HBe Ag-positive chronic hepatitis B during pegylated interferon alpha-2a treatment.

Mutations in carboxy-terminal part of E2 including PKR/eIF2αphosphorylation homology domain and interferon sensitivity determining region of nonstructural 5A of hepatitis C virus 1b:Their correlation with response to interferon monotherapy and viral load

AIM： To study the amino acid substitutions in the carboxy （C）-terminal part of E2 protein and in the interferon （IFN） sensitivity determining region （ISDR） and their correlation with response to IFN and viral load in 85 hepatitis C virus （HCV）-lb-infected patients treated with IFN. METHODS： The C-terminal part of E2 （codons 617-711） including PKR/eIF2α phosphorylation homology domain （PePHD） and ISDR was sequenced in 85 HCV-1b-infected patients treated by IFN monotherapy. RESULTS： The amino acid substitutions in PePHD detected only in 4 of 85 patients were not correlated either with response to iFN or with viral load. The presence of substitutions in a N-terminal variable region （codons 617-641） in the C-terminal part of E2 was significantly correlated with both small viral load （33.9% vs 13.8%, P = 0.0394） and sustained response to iFN （25.0% vs 6.9 %, P = 0.0429）. Four or more substitutions in ISDR were significantly correlated with both small viral load （78.6% vs 16.2%, P 〈 0.0001） and sustained response to iFN （85.7% vs 2.9%, P 〈 0.0001）. In multivariate analysis, ISDR in nonstructural （NS） 5A （OR = 0.39, P 〈 0.0001） and N-terminal variable region （OR = 0.51, P = 0.039） was selected as the independentpredictors for small viral load, and ISDR （OR = 39.0, P 〈 0.0001） was selected as the only independent predictor for sustained response. CONCLUSION： The N-terminal variable region in the C-terminal part of E2 correlates with both response to IFN monotherapy and viral load and is one of the factors independently associated with a small viral load.