Hypermethylation of thymosinβ4 predicts a poor prognosis for patients with acute-on-chronic hepatitis B liver failure

Background:It has been demonstrated that thymosinβ4(Tβ4)could inflect the severity of acute-on-chronic hepatitis B liver failure(ACHBLF),but the relationship between its methylation status and the prognosis of liver failure is not clear.This study aimed to determine Tβ4 promoter methylation status in patients with ACHBLF and to evaluate its prognostic value.Methods:The study recruited 115 patients with ACHBLF,80 with acute-on-chronic hepatitis B pre-liver failure(pre-ACHBLF),and 86 with chronic hepatitis B(CHB).In addition,there were 36 healthy controls(HCs)from the Department of Hepatology,Qilu Hospital of Shandong University.The 115 patients with ACHBLF were divided into three subgroups:33 with early stage ACHBLF(E-ACHBLF),42 with mid-stage ACHBLF(M-ACHBLF),and 40 with advanced stage ACHBLF(A-ACHBLF).Tβ4 promoter methylation status in peripheral blood mononuclear cells(PBMCs)was measured by methylation-specific polymerase chain reaction,and mRNA was detected by quantitative real-time polymerase chain reaction.Results:Methylation frequency of Tβ4 was significantly higher in patients with ACHBLF than in those with pre-ACHBLF,CHB or HCs.However,expression of Tβ4 mRNA showed the opposite trend.In patients with ACHBLF,Tβ4 promoter methylation status correlated negatively with mRNA levels.The 3-month mortality of ACHBLF in the methylated group was significantly higher than that in the unmethylated group.Also,Tβ4 promoter methylation frequency was lower in survivors than in non-survivors.When used to predict the 1-,2-,and 3-month incidence of ACHBLF,Tβ4 methylation status was better than the model for end-stage liver disease(MELD)score.The predictive value of Tβ4 methylation was higher than that of MELD score for the mortality of patients with E-ACHBLF and M-ACHBLF,but not for A-ACHBLF.Conclusions:Tβ4 methylation might be an important early marker for predicting disease incidence and prognosis in patients with ACHBLF.

血府逐瘀口服液治疗狼疮性肾炎的临床疗效及对免疫学指标的影响

目的探讨血府逐瘀口服液治疗狼疮性肾炎的临床疗效,以及对患者免疫学指标的影响。方法选取河北北方学院附属第二医院2021年10月至2022年10月收治的狼疮性肾炎患者60例,按随机数字表法分为对照组和观察组,各30例。两组患者均予常规药物治疗,观察组患者加用血府逐瘀口服液,两组患者均治疗3个月。结果治疗后,观察组患者的系统性红斑狼疮疾病活动指数(SLEDAI)评分、白细胞计数(WBC)、红细胞沉降率(ESR)、C反应蛋白(CRP)、24 h尿蛋白定量、尿红细胞计数、血肌酐(SCr)、血尿素氮(BUN)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)水平及CD_(8)^(+)T细胞、滤泡辅助性T细胞(Tfh)百分率均显著低于对照组(P<0.05),补体C3、白蛋白(Alb)、胸腺肽β4、活性E玫瑰花结形成细胞百分数、淋巴细胞计数、CD_(4)^(+)T细胞、CD_(4)^(+)/CD_(8)^(+)均显著高于对照组(P<0.05);两组患者的抗双链DNA抗体阳性发生率均较治疗前显著降低(P<0.05),但组间无显著差异。观察组患者的感染发生率显著低于对照组(3.33%比26.67%,P<0.05),总有效率显著高于对照组(86.67%比63.33%,P<0.05),不良反应发生率与对照组相当(6.67%比13.33%,P>0.05)。Pearson相关性分析结果显示,胸腺肽β4水平与SLEDAI评分、WBC、ESR、CRP、24 h尿蛋白定量、尿红细胞计数、SCr、BUN、IgA、IgG、CD_(8)^(+)T细胞、Tfh细胞百分率均呈负相关(r=-0.729,-0.418,-0.674,-0.785,-0.651,-0.512,-0.684,-0.676,-0.499,-0.513,-0.624,-0.631,P<0.05),与补体C3、Alb、活性E玫瑰花结形成细胞百分数、淋巴细胞计数、CD_(4)^(+)T细胞、CD_(4)^(+)/CD_(8)^(+)均呈正相关(r=0.475,0.452,0.619,0.626,0.633,0.645,P<0.05)。结论血府逐瘀口服液治疗狼疮性肾炎的临床疗效良好,可提升患者机体的免疫水平,改善肾功能,减少感染的发生,促进病情恢复,其作用机制可能与血府逐瘀口服液能促进胸腺肽β4的合成相关。

家蝇Thymosin(THY)基因的克隆及原核表达

采用EST测序技术从构建的家蝇(Musca domestica)幼虫c DNA质粒文库中筛选到胸腺肽(Thymosin,THY)基因,以该基因的c DNA文库质粒为模板,设计引物,通过PCR扩增,测序鉴定,获得THY基因完整编码序列。运用生物信息学方法对该基因及其编码蛋白的基本理化性质、信号肽和亚细胞定位等方面进行预测和分析。构建p ET-28a(+)-THY重组质粒,转化到大肠杆菌BL21(DE3)中进行诱导表达。研究结果表明,THY基因ORF全长384 bp,编码127个氨基酸,理论分子量14.3 k D,等电点为5.22,具有THY家族的蛋白保守结构域。构建重组原核质粒p ET-28a(+)-THY,经IPTG诱导,蛋白在大肠杆菌中获得表达,经亲和层析柱纯化获得目的蛋白,Western blot检测发现纯化的目的蛋白大小正确。

基于改进YOLOv5的金属表面缺陷分割

针对当前工业图像表面缺陷检测算法定位精度差等问题,提出了一种基于改进YOLOv5缺陷分割算法。首先,在骨干网络的前两层使用ODConv替换原有的Conv模块,使图片的下采样信息更好地保存;其次,使用Meta-ACON激活函数替代SiLU激活函数,能够通过学习自动使用性能更好地激活函数来提高特征提取能力;然后,在下层特征提取部分以及Neck层引入SimAM注意力机制,增强特征提取能力;最后,引入Alpha-IoU作为损失函数,提升了边界框回归的精确度。实验结果表明,改进的分割模型检测精度(map)为86.7%,比原YOLOv5网络提升了20.1%,比最新的检测模型YOLOv8高出2%。改进的模型不仅具有较高的检测精度,而且分割检测算法可以更加精确定位缺陷位置。

胸腺肽α1对衰老小鼠肌肉减少症的调控机制

目的 探讨胸腺肽α1对自噬接头蛋白SQSTM1/p62和衰老小鼠肌肉减少症的调节作用。方法 将小鼠成肌细胞(C2C12)分为对照组、地塞米松组、地塞米松+胸腺肽α1组、地塞米松+胸腺肽α1+p62沉默组。采用细胞计数试剂盒-8检测各组细胞增殖活性(450 nm波长的OD值)的变化,并检测各组C2C12的肌管细胞形成情况。另外,将30只SAMP8快速衰老小鼠用随机数表法分为SAMP8组、胸腺肽α1+SAMP8组及胸腺肽α1+p62沉默+SAMP8组,每组小鼠10只。检测各组小鼠的瘦体质量(LBM)与体质量(BM)的比值[LBM/BM(%)]。采用Western blotting法检测各组C2C12和小鼠肌肉组织中p62、肌球蛋白原D(MyoD)、肌原细胞转录因子(MyoG)、肌球蛋白重链(MyHC)、肌肉RING-指蛋白1(MuRF1)和肌肉萎缩相关蛋白(MAFbx)的表达。采用GraphPad PRISM 5.01软件进行数据分析。根据数据类型,组间比较采用t检验。结果 与对照组比较,地塞米松组细胞的增殖活性和肌管细胞形成能力均显著下调(均P<0.05),p62、MyoD、MyoG、MyHC的表达量都减少(均P<0.05),但MuRF1和MAFbx的表达量均增加(均P<0.05)。与地塞米松组比较,地塞米松+胸腺肽α1组的增殖活性和肌管细胞形成能力均显著上调(均P<0.05),而且p62、MyoD、MyoG、MyHC的表达量也都增加(均P<0.05),MuRF1和MAFbx的表达量均减少(均P<0.05)。与地塞米松+胸腺肽α1组比较,地塞米松+胸腺肽α1+p62沉默组的肌管细胞形成能力显著降低(均P<0.05),而且p62、MyoD、MyoG、MyHC的表达量也都减少(均P<0.05),但是MuRF1和MAFbx的表达量均增加(均P<0.05)。与SAMP8组比较,胸腺肽α1+SAMP8组的LBM/BM比值显著上调(P<0.05),p62的表达量增加(均P<0.05),但MuRF1和MAFbx的表达量均减少(均P<0.05)。与胸腺肽α1+SAMP8组比较,胸腺肽α1+p62沉默+SAMP8组的LBM/BM比值显著下调(P<0.05),p62的表达量减少(均P<0.05),但MuRF1和MAFbx的表达量均增加(均P<0.05)。结论 胸腺肽α1通过激活SQSTM1/p62信号从而缓解衰老小鼠的肌肉减少症。

七彩神仙鱼β-胸腺素基因克隆及表达分析

为了探究胸腺素基因在七彩神仙鱼(Symphysodon aquifasciatus)中的特征、进化和功能,基于转录组序列比对及生物信息学的方法对七彩神仙鱼的胸腺素进行了鉴定、基因与蛋白结构分析、系统发生分析,通过RACE技术克隆获得了七彩神仙鱼胸腺素基因全长,利用qRT-PCR技术研究其在各组织中的表达规律,通过病原感染实验,研究了七彩神仙鱼β-胸腺素基因在病原胁迫下的表达规律。结果显示,七彩神仙鱼的胸腺素基因鉴定为胸腺素β1型,命名为dfTβ。克隆结果显示,dfTβ的全长为779 bp,ORF区序列长度为303 bp,编码100个氨基酸,其5′-UTR为43 bp,3′-UTR为433 bp,含有22个poly-A。系统发育进化分析结果显示,七彩神仙鱼β-胸腺素同尼加拉瓜湖始丽鱼β-胸腺素聚为一簇。qPCR结果显示,β-胸腺素在七彩神仙鱼各组织中均有表达,其中在肠、皮肤、鳃中表达最高,在头肾和脾中表达最低。嗜水气单胞菌(Aeromonas hydrophila)感染后,七彩神仙鱼皮肤、头肾、脾和肠中dfTβ均在短时间内显著上调,并随着时间出现先上升后下降的趋势,但各组织中上调倍数最高峰的时间不同。研究结果表明,dfTβ基因参与了七彩神仙鱼受病原刺激后的免疫应答,且表达模式不同,表明其可能在不同组织、免疫的不同阶段发挥不同的作用。

脓毒症免疫抑制机制及治疗策略

脓毒症是机体对感染反应失调引起的危及生命的器官功能障碍。随着研究的深入,对脓毒症的理解已经从单纯的炎症反应拓展到涉及免疫抑制和免疫失衡。本文综述了脓毒症免疫抑制的生物学机制和临床影响,评估了针对脓毒症免疫抑制的多种潜在治疗策略,包括免疫检查点抑制剂、免疫刺激性细胞因子、免疫球蛋白、胸腺素α1和间充质干细胞等。旨在为临床医生和研究者提供关于脓毒症免疫抑制的最新见解,并探索改善患者长期预后的新兴治疗方法。

低温胁迫对凡纳滨对虾胸腺肽基因功能的影响

低温是影响凡纳滨对虾养殖的重要因素。为探明凡纳滨对虾在应对低温胁迫下的分子机制,通过RACE方法对凡纳滨对虾胸腺肽(Lvthy)基因进行克隆,并对其序列特征进行分析,并通过荧光定量PCR对其组织分布表达及低温胁迫下的表达谱进行分析,最后通过siRNA干扰的方法对其表达量进行敲降,对其在凡纳滨对虾低温胁迫下的功能进行验证。Lvthy基因cDNA全长1765 bp,开放阅读框长度729 bp,编码242个氨基酸。组织分布分析结果显示,Lvthy基因在对虾不同组织中均有表达,在胃组织中表达量最高,其次是在肠道、心脏和脑组织。低温胁迫试验结果显示:Lvthy基因在低温刺激1 h后表达量开始上调;3 h后表达量达到最高,是正常状态下的2.7倍;随后Lvthy基因表达量开始下降。RNAi干扰试验结果显示,敲降Lvthy基因后,凡纳滨对虾在低温胁迫下的存活率仅为31%,显著低于对照组存活率(83%)。试验结果表明,胸腺肽在凡纳滨对虾响应低温胁迫的生理过程中发挥重要的作用,本试验结果可为胸腺肽在甲壳动物应对低温胁迫下的适应机制研究奠定基础。

维生素D联合胸腺肽辅助治疗急性加重期COPD患者的临床研究

目的探讨维生素D联合胸腺肽辅助治疗急性加重期慢性阻塞性肺疾病(COPD)患者的临床效果。方法80例急性加重期COPD患者随机分为两组。在常规治疗基础上,对照组给予胸腺肽辅助治疗,观察组给予维生素D联合胸腺肽辅助治疗。比较两组的治疗效果、动脉血气指标及血清炎性因子。结果观察组的治疗总有效率为97.50%,明显高于对照组的85.00%(P<0.05)。治疗后,观察组的PaO_(2)水平明显高于对照组,PaCO_(2)、IL-6、TNF-α水平均明显低于对照组(P<0.05)。结论维生素D联合胸腺肽辅助治疗急性加重期COPD效果显著,可明显改善患者的动脉血气指标,减轻患者的炎性反应。

胸腺肽联合痰热清注射液辅助治疗老年慢阻肺合并肺部感染的临床效果

目的探究胸腺肽联合痰热清注射液辅助治疗老年慢阻肺合并肺部感染(COPD-PI)的临床效果。方法通过随机数字表法将本院2019年9月-2020年9月接收的老年COPD-PI患者(126例)分为研究组(63例,胸腺肽治疗)和对照组(63例,痰热清注射液治疗)。比较2组治疗总有效率、治疗前后肺功能指标、免疫功能指标、炎性细胞因子水平、氧化应激指标。结果治疗后,研究组治疗总有效率92.06%明显高于对照组77.78%(P<0.05)。治疗后2组FVC%、FEV1%、FEV1%/FVC%、CD4+、CD4+/CD8+水平较治疗前明显升高,2组血清TNF-α、IL-8、TGF-β1水平均明显降低(P<0.05),且研究组明显优于对照组(P<0.05)。结论胸腺肽与痰热清注射液对老年COPD-PI进行联合治疗的效果较好,可使炎性因子水平降低,改善肺功能及免疫功能。

ALA-PDT、卡介菌多糖核酸注射液联合胸腺肽肠溶片治疗尖锐湿疣的效果

目的研究艾拉光动力治疗(ALA-PDT)、卡介菌多糖核酸注射液联合胸腺肽肠溶片治疗尖锐湿疣的效果。方法选取2020年2月至2022年6月郑州大学附属郑州中心医院收治的82例尖锐湿疣患者为研究对象,以随机双盲法分为联合组、对照组,各41例。对照组接受胸腺肽肠溶片联合ALA-PDT治疗,联合组在对照组基础上接受卡介菌多糖核酸注射液治疗。比较两组临床疗效、治疗前和治疗3个月后疣体数、皮损面积、免疫功能指标(CD3^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))、血清细胞因子[脂质过氧化物(Lpo)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)、超敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)]水平、6个月复发率。结果联合组临床总有效率高于对照组(P<0.05);治疗3个月后,联合组疣体数、皮损面积小于对照组(P<0.05);治疗3个月后与对照组相比,联合组CD3^(+)、CD4^(+)/CD8^(+)、CD4^(+)水平较高,CD8^(+)水平较低(P<0.05);治疗3个月后,联合组血清GM-CSF、TNF-α、Lpo、hs-CRP水平低于对照组(P<0.05);治疗后随访6个月,联合组复发率(2.56%)低于对照组(21.05%)(P<0.05)。结论采用ALA-PDT、卡介菌多糖核酸注射液与胸腺肽肠溶片联合治疗可促进尖锐湿疣患者免疫功能改善,抑制炎症反应,减轻临床症状,提高临床疗效,且复发风险较低。

A randomized controlled clinical trial on the treatment of Thymosin-a1 versus interferon-α in patients with hepatitis B

INTRODUCTIONChronic hepatitis B virus (HBV) infection is a serious problem because of its world wide distribution and possible adverse sequelae ,such as cirrhosis and hepatocellular carcinoma .The World Health Organization estimates that HBV has infected mord than 350 million people worldwide ,and up to 20% of them will become chromic carricrs and will be at significant risk for cirrhosis and HCC .The ultimate goal of the therapy for chronic hepatitis B is to prevent progression to cirrhosis and to prevent development of HCC.

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.

Recombinant adeno-associated virus carrying thymosin β4 suppresses experimental colitis in mice

AIM To investigate the protective effect of a recombinant adeno-associated virus carrying thymosin β4(AAV-Tβ4) on murine colitis via intracolonic administration.METHODS AAV-Tβ4 was prepared and intracolonically used to mediate the secretory expression of Tβ4 in mouse colons. Dextran sulfate sodium(DSS) was applied to induce the murine ulcerative colitis, and 2,4,6-trinitrobenzene sulfonic acid(TNBS) was used to establish a mouse colitis model resembling Crohn's disease. The disease severity and colon injuries were observed and graded to reveal the effects of AAV-Tβ4 on colitis. The activities of myeloperoxidase(MPO) and superoxide dismutase(SOD) and the content of malondialdehyde(MDA) were determined using biochemical assays. Colonic levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-1β and IL-10 were measured using ELISA, and mucosal epithelial cell apoptosis andproliferation were detected by TUNEL assay and immunochemistry, respectively.RESULTS Recombinant AAVs efficiently delivered Lac Z and Tβ4 into the colonic tissues of the mice, and AAV-Tβ4 led to a strong expression of Tβ4 in mouse colons. In both the DSS and TNBS colitis models, AAV-Tβ4-treated mice displayed distinctly attenuated colon injuries and reduced apoptosis rate of colonic mucosal epithelia. AAV-Tβ4 significantly reduced inflammatory cell infiltrations and relieved oxidative stress in the inflamed colons of the mice, as evidenced by decreases in MPO activity and MDA content and increases in SOD activity. AAV-Tβ4 also modulated colonic TNF-α, IL-1β and IL-10 levels and suppressed the compensatory proliferation of colonic epithelial cells in DSS- and TNBS-treated mice.CONCLUSION Tβ4 exerts a protective effect on murine colitis, indicating that AAV-Tβ4 could potentially be developed into a promising agent for the therapy of inflammatory bowel diseases.

Thymosin alpha 1:A comprehensive review of the literature

Thymosin alpha 1 is a peptide naturally occurring in the thymus that has long been recognized for modifying,enhancing,and restoring immune function.Thymosin alpha 1 has been utilized in the treatment of immunocompromised states and malignancies,as an enhancer of vaccine response,and as a means of curbing morbidity and mortality in sepsis and numerous infections.Studies have postulated that thymosin alpha 1 could help improve the outcome in severely ill corona virus disease 2019 patients by repairing damage caused by overactivation of lymphocytic immunity and how thymosin alpha 1 could prevent the excessive activation of T cells.In this review,we discuss key literature on the background knowledge and current clinical uses of thymosin alpha 1.Considering the known biochemical properties including antibacterial and antiviral properties,timehonored applications,and the new promising findings regarding the use of thymosin,we believe that thymosin alpha 1 deserves further investigation into its antiviral properties and possible repurposing as a treatment against severe acute respiratory syndrome coronavirus-2.

Thymosin <i>β</i>4 Improves Neurological Outcome and Enhances Induced Oligodendrogenesis in the Rat after ICH

Thymosin β4 (Tβ4), a G-actin binding protein, has diverse biological functions. This study tested the effects of Tβ4 on oligodendrogenesis in a rat model of intracerebral hemorrhage (ICH). ICH was induced by stereotactic injection of 100 μm of autologous blood into the striatum in 32 male Wistar rats. The rats were randomly divided into four groups: 1) saline control group (n = 8);2) 3 mg/kg Tβ4-treated group (n = 8);3) 6 mg/kg Tβ4-treated group (n = 8);and 4) 12 mg/kg Tβ4treated group (n = 8). Tβ4 or saline was administered intraperitoneally starting at 24 h post ICH and then every 3 days for 4 additional doses. The neurological functional outcome was evaluated by behavioral tests (i.e., modified Neurological Severity Score and corner turn test) at multiple time points after ICH. Animals were sacrificed at 28 days post ICH, and histological studies were completed. Tβ4 treatment improved neurological functional recovery significantly and increased actively proliferating oligodendrocytic progenitor cells and myelinating oligodendrocytes in the ICH-affected brain tissue, compared with the saline-treated group. The high-dose treatment of Tβ4 showed better restorative effects compared with the low-dose treatment. Tβ4 treatment enhanced ICH-induced oligodendrogenesis that may contribute to the enhanced functional recovery after ICH. Further investigation is warranted to determine the associated underlying mechanisms of Tβ4 treatment for ICH.

Efficacy of thymosin alpha-1 and interferon alpha in treatment of chronic viral hepatitis B:A randomized controlled study

AIM: To observe the efficiency and safety of thymosin-α1 treatment in patients with hepatitis B e antigen (HBeAg) and HBV DNA positive chronic hepatitis. METHODS: Sixty-two patients were randomly divided into groups A and B. The patients in group A received subcutaneous injection of 1.6 mg thymosin-α1, twice a week (T-α1 group) for six months, and the patients in group B received 5 MU interferon alpha (IFN-α) each day for fifteen days, then three times weekly (IFN-α group) for six months. The results between two groups treated with and the group untreated with IFN-α which was followed up for 12 mo (historical control group consisting of 30 patients) were compared, and three groups were comparable between each other (P > 0.05) at baseline (age, sex, clinical history, biochemical, and serological parameters). RESULTS: At the end of treatment, complete response, which was defined as alanine aminotransferase (ALT) normalization and HBV DNA and HBeAg loss, occurred in 9 of 29 (31.0%) patients in the T-α1 group and in 15 of 33 (45.5%) patients in the IFN-α group (c2 = 1.36, P >0.05). After a follow-up period of six months, a complete response was observed in 14 of 29 (48.3%) patients in the T-α1 group and in 9 of 33 (27.3%) patients in the IFN-α group (c2 = 2.93, P > 0.05). Compared with the results observed in the historical control (HC) group untreated with IFN-α which was followed up for 12 mo, the rate of complete response was significantly higher in IFN-α group at the end of therapy (1 of 30 vs 15 of 33, c2 = 14.72, P < 0.001) and in the T-α1 group at the end of follow-up (1 of 30 vs 14 of 29, c2 = 15.71, P < 0.001). In T-α1 and IFN-α treatment groups, the area under (the plasma concentration time) curve (AUC) of negative HBV DNA and HBeAg was 34%, 17%, 31% and 19% smaller than that in the HC group. By the end of the follow- up period, the proportions of ALT normalization and negative HBV DNA in the T-α1 group were significantly higher than those in the IFN-α and HC groups. The odds of ALT normalization and negative HBV DNA at the end of the follow-up was three-fold higher in the T-α1 group than in the IFN-α group. Unlike IFN-α, T-α1 was well tolerated by all patients, and no side effects appeared in T-α1 group.CONCLUSION: The results suggest that a 6-mo course of T-α1 therapy is effective and safe in patients with chronic hepatitis B. T-α1 is able to reduce HBV replication in patients with chronic hepatitis B. Furthermore, T-α1 is better tolerated than IFN-α and can gradually induce more sustained ALT normalization and HBV DNA and HBeAg loss. However, a response rate of 48.3% is still less ideal. A more effective therapeutic approach warrants further study.

Preliminary results of Thymosin-a1 versus interferon-α treatment in patients with HBeAg negative and serum HBV DNA positive chronic hepatitis B

INTRODUCTIONIn China ,the incidence and mortality of gastric cancer rank the second among all cancers. Recent development of cancer [1-20].The aim of this study was investigat the insight of apoptosis and bcl-2, p53 and C-myc protein expression in the development of gastric cancer .

Molecular characteristics of three thymosin-repeat proteins from Marsupenaeus japonicus and their responses to WSSV infection

β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis,antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjthm2, were cloned from Marsupenaeus japonicus using expressed sequence tags（EST） from suppression subtractive hybridization. The mjthm4, mjthm3 and mjthm2 c DNAs possessed open reading frames that encoded166, 128 and 90 amino acid residue polypeptides and contained four, three and two β-thymosin actin binding modules, respectively. Blast analysis demonstrated that mjthm4, mjthm3 and mjthm2 shared high homology with known invertebrate multi-repeat β-thymosins. These proteins are ubiquitously expressed in all of the examined tissues, and the transcriptional levels were highest in the intestine. Further investigation revealed that mjthm4,mjthm3 and mjthm2 were remarkably up-regulated 6 h after WSSV infection. Moreover, while mjthm4 transcriptional levels displayed no changes, mjthm3 and mjthm2 levels decreased in the virus-resistant shrimps.The results indicate that mjthm4, mjthm3 and mjthm2 are novel multi-repeat β-thymosin homologues, have a close relationship with WSSV infection, and might contribute to a better understanding of host defense and/or virus invasion interactions in shrimps.

Construction, expression and characterization of human interferon α2b-(G4S)n-thymosin α1 fusion proteins in Pichia pastoris

AIM: Interferon α2b (IFNα2b) and thymosin α1 (Tα1) exhibit synergic effects in the treatment of hepatitis B and hepatitis C when used together. For developing a fusion protein drug, fusion proteins of IFNα2b and Tα1 linked by different lengths of (G4S)n (n = 1-3) were constructed and expressed in Pichia pastoris.METHODS: Using PCR and molecular clone techniques, the fusion genes of IFNα2b-(G4S)n-Tα1 (n = 1-3) were constructed and subcloned into the eukaryotic expression vector pPIC9. After transformation of these plasmids into P. pastoris, the expressed fusion proteins IFNα2b-(G4S)n-Tα1 (n = 1-3) were obtained. These proteins were purified through diethylaminoethyl (DEAE) affinity chromatography and SuperdexTM 75 gel filtration and analyzed by SDSPAGE and Western blot. Antiviral and E-rosette assays were used to investigate the bioactivities of these fusion proteins.RESULTS: DNA sequencing confirmed that the fusion genes of IFNα2b-(G4 S)n-Tα1 (n = 1-3) were correctly cloned to the pPIC9 vector. The recombinant IFNα2b(G4 S)n-Tα1 (n = 1-3) fusion proteins expressed in P. pastoris were purified with DEAE and SuperdexTM 75 gel filtration chromatography. The fusion proteins could be observed on sodium dodecylsulfate-polyacrylamide gel electrophoresis with molecular weight (MW) of 23.2, 22.9, and 22.6 ku, respectively, and reacted to the IFNα2b monoclonal antibody and Tα1 polyclonal antibody. The purified fusion proteins exhibit antiviral activity and can enhance the percentage of E-rosette-forming-cell in E-rosette assay.CONCLUSION: The recombinant IFNα2b-(G4S)n-Tα1 (n = 1-3) fusion proteins were successfully expressed in P. pastoris. Purified fusion proteins exhibit both antiviral activity of IFNα2b and immunomodulatory activity of Tα1 in vitro. These results will be the basis for further evaluation of the fusion proteins' function in vivo.