外周血单个核细胞感染乙型肝炎病毒在母婴垂直传播中的作用

目的:探讨HBsAg、HBeAg阳性孕妇外周血单个核细胞(PBMC)内乙型肝炎病毒(HBV)DNA感染状况及其在宫内母婴垂直传播中的作用.方法:对HBsAg/HBeAg双阳性共67对孕妇及其新生儿静脉血分离和提纯PBMC后,经抽提、纯化后的DNA进入PCR扩增反应,引物为HBV C区基因序列.结果:67名HBsAg及HBeAg双阳性的孕妇中有35例(52.2%)PBMC中HBV DNA阳性,25例孕妇在血清及PBMC中均发现HBV DNA.67名新生儿有22例感染HBV DNA,感染率32.8%,其中血清HBV DNA阳性者10例,PBMC HBVDNA阳性者19例,二者均阳性者7例.结论:母亲PBMC内HBV DNA阳性可能导致新生儿PBMC中HBV DNA阳性,PBMC内的HBV DNA可能是HBV母婴垂直传播的一条重要途径,同时,HBsAg及HBeAg阳性母亲若血清HBV DNA为阳性就极大增加了其新生儿感染HBV的危险性.     

Science Letters： Solid lipid nanoparticles loading adefovir dipivoxil for antiviral therapy

Herein, solid lipid nanoparticles (SLN) were proposed as a new drug delivery system for adefovir dipivoxil (ADV). The octadecylamine-fluorescein isothiocynate (ODA-FITC) was synthesized and used as a fluorescence maker to be incorporated into SLN to investigate the time-dependent cellular uptake of SLN by HepG2.2.15. The SLN of monostearin with ODA-FITC or ADV were prepared by solvent diffusion method in an aqueous system. About 15wt% drug entrapment efficiency (EE) and 3wt% drug loading (DL) could be reached in SLN loading ADV. Comparing with free ADV, the inhibitory effects of ADV loaded in SLN on hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B virus (HBV) DNA levels in vitro were significantly enhanced.     

Green tea polyphenol,epigallocatechin-3-gallate,possesses the antiviral activity necessary to fight against the hepatitis B virus replication in vitro

Although several antiviral drugs and vaccines are available for use against hepatitis B virus (HBV), hepatitis caused by HBV remains a major public health problem worldwide, which has not yet been resolved, and new anti-HBV drugs are in great demand. The present study was performed to investigate the anti-HBV activity of epigallocatechin-3-gallate (EGCG), a natural-origin compound, in HepG2 2.2.15 cells. The antiviral activity of EGCG was examined by detecting the levels of HBsAg and HBeAg in the supernatant and extracellular HBV DNA. EGCG effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose- and time-dependent manner, and it showed stronger effects at the level of 0.11–0.44 μmol/ml (50–200 μg/ml) than lamivudine (3TC) at 0.87 μmol/ml (200 μg/ml). EGCG also suppressed the amount of extracellular HBV DNA. The data indicated that EGCG possessed anti-HBV activity and suggested the potential of EGCG as an effective anti-HBV agent with low toxicity.     

乙肝血清学指标与腮腺组织HBsAg、HBcAg和HBVDNA的相关性研究

目的通过观察乙型肝炎病毒感染者腮腺组织中HBsAg、HBcAg和HBVDNA的表达情况,研究乙肝血清学指标与它们的相关性。方法免疫组化检测乙肝血清指标与腮腺组织中HbsAg、HbcAg和HBVDNA的相关性。结果23例腮腺组织中HBsAg阳性11例,阳性率为47．83％;HBcAg阳性10例,阳性率为43．48％。其中HBsAg和HBcAg同时阳性8例,占34．78％。14例免疫组化阳性患者检出HBVDNA9例,阳性率为64．29％,乙肝血清学指标与腮腺组织中HBsAg、HBcAg检测结果高度正相关（x^2=3．305,P〉0．05）,其Kendall等级相关系数rs=0．661（P〈0．05）。结论腮腺组织对HBV有较强的亲和力．唾液中HBV的出现可能源于受染的唾液腺组织,含HBV的唾液是乙型肝炎生活接触性传播的媒介。     

RNA interference against interleukin-5 attenuates airway inflammation and hyperresponsiveness in an asthma model

Interleukin-5 (IL-5) accompanies the development of airway inflammation and hyperresponsiveness through the activation of eosinophils. Therefore, interference of IL-5 expression in lung tissue seems to be an accepted approach in asthma therapy. In this study, we designed a small interfering RNA (siRNA) to inhibit the expression of IL-5. The siRNAs against IL-5 were constructed in a lentivirus expressing system, and 1.5×106 IFU (inclusion-forming unit) lentiviruses were administered intratracheally to ovalbumin (OVA)-sensitized murine asthmatic models. Our results show that lentivirus-delivered siRNA against IL-5 efficiently inhibited the IL-5 messenger ribonucleic acid (mRNA) expression and significantly attenuated the inflammation in lung tissue. Significant decrease of eosinophils and inflammatory cells were found in peripheral blood, bronchoalveolar lavage fluid (BALF), and lung tissue. In addition, significant inhibition of airway hyperresponsiveness (AHR) was found in the mice treated with siRNA against IL-5. These observations demonstrate that siRNA delivered by means of the lentivirus system is possibly an efficacious therapeutic approach for asthma.     

基于支持向量机的siRNA降解效率预测

为了辅助siRNA的设计,从已发表文献中共收集到573个siRNA的实验数据,使用基于统计学习理论的支持向量机(SVM)方法,提取了siRNA序列的碱基对关联性(BBC)特征,然后使用十倍交叉验证方法,对siRNA沉默目标基因的效率进行了预测.结果表明,基于支持向量机,选用多项式核作为核函数的算法具有最高的AUC值(0.73,ROC曲线图)和最高的r值(0.43,Pearson相关系数分析),优于以前基于打分的算法.结果说明,在以后的siRNA的设计中应该更多关注碱基之间的关联信息.     

Effect of siRNA-mediated knockdown of eIF3c gene on survival of colon cancer cells

Eukaryotic initiation factor subunit c(eIF3c) has been identified as an oncogene that is over-expressed in tumor cells and,therefore,is a potential therapeutic target for gene-based cancer treatment.This study was focused on investigating the effect of small interfering RNA(siRNA)-mediated eIF3c gene knockdown on colon cancer cell survival.The eIF3c gene was observed to be highly expressed in colon cancer cell models.The expression levels of the gene in eIF3c siRNA infected and control siRNA infected cells were compared via real-time polymerase chain reaction(PCR) and western blotting analysis.Cell proliferation levels were analyzed employing 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide(MTT) and colony formation assays.Furthermore,the effects of eIF3c gene knockdown on the cell cycle and apoptosis were analyzed using flow cytometry.The results showed that suppression of eIF3c expression significantly(P<0.001) reduced cell proliferation and colony formation of RKO colon cancer cells.The cell cycle was arrested by decreasing the number of cells entering S phase.Further,apoptosis was induced as a result of eIF3c knockdown.Collectively,eIF3c deletion effectively reduced the survival of colon cancer cells and could be used as a therapeutic tool for colon cancer therapy.     

Construction and identification of Fas-targeting siRNA-expressing plasmid

Objective: To study the therapeutic potential of Fas inhibition in different diseases, a Fas-targeting siRNA (small interfering)-expressing plasmid was constructed. Methods: The U6 promoter cassette and siFas (small interfering RNA that inhibit Fas expression) template sequence were obtained by PCR method. They were cloned into modified pcDNA3.1. The resultant plasmid pU6-siFas was transfected into P815 cells with lipofectin2000 and selected under G-418-containing culture medium. Fas inhibition in stably transfected cells was detected by immunocytochemistry. Results: The plasmid pU6-siFas efficiently reduced the expression of Fas and conferred G-418 resistance in P815 cells. Conclusion: The successful construction of the siRNA expressing plasmid will facilitate the application of RNA interference technique and lay the foundation for further study of Fas inhibition in the treatment of different diseases such as aplastic anemia and acute liver failure.     

慢性肝炎病毒感染致病机制相关新基因克隆化研究策略

乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)的慢性感染,与慢性病毒性肝炎、肝硬化、肝细胞癌(HCC)的发病密切相关,其发病机理涉及到很多基因的共同参与.病毒基因的复制和表达受到肝细胞中蛋白质因子的调节,肝炎病毒基因编码的蛋白与肝细胞中的蛋白能够结合,肝炎病毒蛋白在肝细胞中的表达对于肝细胞的基因表达谱产生影响,也可能是病毒性肝炎、肝硬化、肝细胞癌发病机制的重要机制.酵母单杂交技术、酵母双杂交技术、基因芯片技术、抑制性消减杂交技术、噬菌体展示技术、蛋白质分离纯化与基因克隆化的反向遗传学技术等,在肝炎病毒基因调控、肝炎病毒蛋白结合蛋白的研究、肝炎病毒蛋白反式激活靶基因的研究中都有重要的应用,是促进慢性病毒性肝炎发病机理研究,探索病毒性肝炎新型治疗技术和治疗方法的有效途径.     

复方甘草酸苷联合拉米夫定治疗慢性乙型肝炎的临床研究

[目的]观察复方甘草酸苷对慢性乙型肝炎患者血清病毒学标志的影响．[方法]采用复方甘草酸苷联合拉米夫定治疗30例慢性乙肝患者,并与30例拉米夫定单药治疗患者比较,观察治疗后血清病毒学标志及肝功能指标的变化．[结果]治疗12周时联合治疗组血清HBeAg／抗-HBe转换率明显高于对照组；治疗12周和24周时联合治疗组肝功能指标复常率明显高于对照组．[结论]复方甘草酸苷治疗慢性乙型肝炎可能有助于患者在拉米夫定治疗过程中早期发生HBeAg／抗-HBe血清转换．     

Enhancing cellular immune response to HBV M DNA vaccine in mice by codelivery of interleukin-18 recombinant

INTRODUCTIONChronicinfectionwithHBVaffectsmorethan250millionpeopleworldwide.Therearemorethan120millionchronicHBVcarriersinChina;appro-ximately10percentofthemremaininstateofchronichepatitisandhaveahighriskofdevelop-mentofcirrhosisandhepatocellularcarcinoma.ButthereisnoeffectivemethodtocontrolchronicHBVinfectionatpresent.Recentdataindicatedthatim-munotherapeuticstrategiesstimulatingbothcellularandhumoralimmuneresponsestoHBVantigensareessentialforcuringchronicHBVinfection(ChisariandFe…     

Enhancing cellular immune response to HBV M DNA vaccine in mice by codelivery of interleukin-18 recombinant

Objective: To investigate the effect of interleukin-18 (IL-18) on immune response induced by plasmid encoding hepatitis B virus middle protein antigen and to explore new strategies for prophylactic and therapeutic HBV DNA vaccines. Methods: BALB/c mice were immunized with pCMV-M alone or co-immunized with pcDNA3-18 and pCMV-M and then their sera were collected for analysing anti-HBsAg antibody by ELISA; splenocytes were isolated for detecting specific CTL response and cytokine assay in vitro. Results: The anti-HBs antibody level of mice co-immunized with pcDNA3-18 and pCMV-M was slightly higher than that of mice immunized with pCMV-M alone, but there was not significantly different (P>0.05). Compared with mice injected with pCMV-M, the specific CTL cytotoxity activity of mice immunized with pcDNA3-18 and pCMV-M was significantly enhanced (P<0.05) and the level of IFN-γ in supernatant of splenocytes cultured with HBsAg in vitro was significantly elevated (P<0.05) while the level of IL-4 had no significant difference (P>0.05). Conclusion: The plasmid encoding IL-18 together with HBV M gene DNA vaccines may enhance specific TH1 cells and CTL cellular immune response induced in mice, so that IL-18 is a promising immune adjuvant.     

HBV DNA与HBV-m对比分析

目的：探讨荧光定量PCR检测HBV DNA与HBVm的关系。方法；采用荧光定量PCR法和酶联免疫吸附试验（ELISA）对159份HBVm8种不同阳性模式及67份HBVm全阴模式血清进行HBV DNA检测。结果；HBsAg、HBcAb、HBeAe阳性者HBV DNA阳性率为89．83％；HBsAg、HBcAb、HBeAb阳性者HBV DNA阳性率为28．81％；HBsAb、HBcAb、HBeAb阳性者HBV DNA阳性率为16．66％；HBVm全阴性者HBV DNA阳性率为17．91％。结论：荧光定量PCR法检测HBV DNA是乙型肝炎病毒复制最直接可信的指标．可反映HBV真实感染和复制状态．特别是低复制状态，明显优于ELISA法检测HBVm，具有重要临床意义。     

Inhibition of chemotherapy-related breast tumor EMT by application of redox-sensitive siRNA delivery system CSO-ss-SA/siRNA along with doxorubicin treatment

Metastasis is one of the main reasons causing death in cancer patients. It was reported that chemotherapy might induce metastasis. In order to uncover the mechanism of chemotherapy-induced metastasis and find solutions to inhibit treatment-induced metastasis, the relationship between epithelial-mesenchymal transition (EMT) and doxorubicin (DOX) treatment was investigated and a redox-sensitive small interfering RNA (siRNA) delivery system was designed. DOX-related reactive oxygen species (ROS) were found to be responsible for the invasiveness of tumor cells in vitro, causing enhanced EMT and cytoskeleton reconstruction regulated by Ras-related C3 botulinum toxin substrate 1 (RAC1). In order to decrease RAC1, a redox-sensitive glycolipid drug delivery system (chitosan-ss-stearylamine conjugate (CSO-ss-SA)) was designed to carry siRNA, forming a gene delivery system (CSO-ss-SA/siRNA) down-regulating RAC1. CSO-ss-SA/siRNA exhibited an enhanced redox sensitivity compared to nonresponsive complexes in 10 mmol/L glutathione (GSH) and showed a significant safety. CSO-ss-SA/siRNA could effectively transmit siRNA into tumor cells, reducing the expression of RAC1 protein by 38.2% and decreasing the number of tumor-induced invasion cells by 42.5%. When combined with DOX, CSO-ss-SA/siRNA remarkably inhibited the chemotherapy-induced EMT in vivo and enhanced therapeutic efficiency. The present study indicates that RAC1 protein is a key regulator of chemotherapy-induced EMT and CSO-ss-SA/siRNA silencing RAC1 could efficiently decrease the tumor metastasis risk after chemotherapy.

     

二对半e抗原假阳性原因分析

目的:探讨导致二对半e抗原假阳性的原因.方法:用ELISA法测定45份离心与不离心抗凝标本的HBeAg.结果:不离心标本的HBeAg单独阳性很高,离心标本基本无此情况.结论:假阳性与不离心有关,全血中有许多与HBeAg有共同抗原颗粒物质,在酶免过程中出现非特异性结合,而造成假阳性,洗板不清,可能也是造成假阳性的原因之一.故要求对单HBeAg阳性标本进行离心,以提高测定结果的准确性.     

大学新生乙型肝炎病毒感染调查

本文报道近三年安徽农业技术师范学院新生动型肝炎病毒感染调查结果。受检学生1628人，乙型肝炎表面抗原阳性率为10～12％，谷丙转氨酶异常率为4～6％，西指标均异常为0．9～1．5％。     

我校体育系学生乙肝表面抗原携带情况调查

调查体育系学生HBsAg阳性率 17.1%高于全国平均水平 .目的是为了更有效地防治乙肝病毒的传播 ,有益于学生的身心健康 .