EB病毒复燃导致弥漫性肺泡出血1例及文献复习

成人EB病毒(EBV)感染的发病率逐渐增高,特别是老年人,急性感染症状不明显,容易忽视。本文报道1例EBV复燃导致弥漫性肺泡出血患者的诊治过程并复习相关文献。该患者以咳嗽、咳痰、喘憋、发热起病,治疗过程中病情曾一度好转,但随后出现咳血、呼吸困难、血红蛋白降低以及低氧血症。肺部CT弥漫浸润影,诊断为肺泡出血。究其病因,考虑为EBV感染所致,给予持续呼吸机辅助呼吸、激素冲击联合抗病毒治疗,以及血液净化、抗感染等综合治疗后病情好转。该结果提示及时使用足量激素联合免疫抑制剂等综合治疗有利于改善患者预后。临床诊疗过程中应进一步提高对病毒感染的认识和关注程度,辅助疾病的早期诊断及治疗。

艾滋病抗病毒复治12例分析

随着艾滋病在我国的流行增加,国家四免一关怀也逐步落实到各地,高效价抗逆转录病毒治疗（HAART）全面铺开,在强调服药依从性的同时,少部分患者因各种原因停止治疗后一段时间,再次服抗艾滋病病毒药治疗.本文回顾性分析艾滋病抗病毒复治12例,旨在总结经验,以提高治疗水平. 1 资料与方法 1.1 一般资料 12例艾滋病为2005年~2011年在我院HAART患者,全部患者HIV-1抗体（免疫印迹法）检测阳性.同时流式细胞仪检查CD4＋T淋巴细胞绝对计数.艾滋病病毒载量送广西自治区疾病控制中心检查.

新型中药制剂肃毒星对乙型肝炎病毒体外复制的抑制作用研究

目的观察新型中药制剂肃毒星对野生和恩替卡韦(ETV)耐药乙型肝炎病毒(HBV)的抑制作用。方法以不同浓度(0、0.0005﹑0.001﹑0.005﹑0.01mg/ml)的肃毒星分别处理HepG2.2.15细胞(D基因型/ayw亚型野生HBV稳定复制细胞系)和HepG2.A64细胞(C基因型/adr亚型ETV耐药HBV稳定复制细胞系),作用时间4d。采用酶联免疫吸附试验检测细胞培养上清中HBsAg和HBeAg含量的变化;提取细胞内病毒核心颗粒,采用实时荧光定量PCR方法检测两种细胞内HBV DNA复制水平的变化。观察不同作用时间下,药物对HBV DNA的抑制作用。结果肃毒星对HepG2.2.15细胞和HepG2.A64细胞内HBV DNA复制和HBsAg、HBeAg的分泌均有明显抑制作用,且具有药物剂量和时间依赖性,最高浓度(0.01mg/ml)的肃毒星对HepG2.2.15细胞内病毒的抑制率达到80.83%,对HBsAg和HBeAg的抑制率分别达51.00%和64.05%;而对HepG2.A64细胞内HBV DNA抑制率为65.18%,对HBsAg和HBeAg的抑制率分别为32.85%和73.86%。随肃毒星作用时间延长,其对HepG2.2.15细胞HBV DNA的抑制效果增强,但未显示对HepG2.A64细胞内ETV耐药型HBV株病毒抑制率增强的效果。结论体外细胞培养实验中,肃毒星不仅可有效抑制野生HBV复制和抗原分泌,而且对ETV耐药HBV的复制和抗原分泌也具有很好的抑制作用。

新型冠状病毒肺炎“复阳”患者中医体质类型分析

目的 基于2022年1月-2022年3月河南省定点医院新型冠状病毒肺炎(coronavirus disease 2019,COVID-19,新冠肺炎)患者病历资料,以探讨新冠肺炎“复阳”患者的中医体质类型,为中医药治疗新冠肺炎提供参考。方法 以感染新冠肺炎后复阳患者为复阳组,感染新冠肺炎正常转阴患者为对照组,收集其症状、体征、舌象等病历资料后,根据《中医体质分类与判定表》对两组进行体质类型辨析,并对比复阳组复阳前后体质类型变化。结果 (1)体质类型占比如下,复阳组:湿热质>平和质>痰湿质>阴虚质>气虚质。对照组:湿热质>平和质>痰湿质>气郁质>阴虚质。(2)复阳组复阳后湿热质由39例变为29例占比下降、痰湿质由7例变为12例占比增加、平和质由12例变为11例占比下降、阴虚质由2例变为5例占比增加、增加3例气郁质。结论 新冠肺炎复阳患者易感体质以湿热质为主。新冠肺炎复阳后体质变化主要表现在:湿热质占比下降、痰湿质、阴虚质、气郁质占比增加。

甘薯病毒病复合体(SPVD)对甘薯产量形成的影响

以高淀粉甘薯品种西成薯007的病毒苗、常规苗和脱毒苗为试验材料,研究了甘薯病毒病复合体(SPVD)对甘薯生长发育和产量形成的影响。结果表明,SPVD可导致甘薯地上部丛生和矮化、叶面积指数降低,"源"变小,同化能力和光合能力降低;"流"变小,最终导致生物产量降低;脱毒后可增大叶"源"和叶面积指数,提高植株的光合能力和"流"的通畅性,从而增加产量。SPVD可使甘薯植株的超氧化物歧酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性降低,丙二醛(MDA)含量上升,抗氧化活性降低,细胞膜受到伤害,最终导致产量降低;脱毒后可增加SOD、POD和CAT在植株体内的活性,从而降低MDA对植株的危害。与正常苗相比,SPVD的脱除可增加3.43%的茎叶产量和2.89%的鲜薯产量,而SPVD侵染可使茎叶和鲜薯分别降低69.94%和50.42%。

人类中流行的流感病毒来自何处

针对我国出现H7N9感染病例的状况．作者从流感病毒演化传播及流感的流行病学规律上展开分析。并提出推行对H7N9大范围预防之依据不充分。预告流感要有流行的证据链。不能单凭实验室的某些病毒检测结果下结论。

SARS-CoV-2核酸复阳患者临床特点及不同标本核酸检测结果分析

目的了解SARS-CoV-2核酸复阳的2019新型冠状病毒肺炎(coronavirus disease 2019,COVID-19)患者临床特征,为实行针对性管理提供参考依据。方法分析SARS-CoV-2核酸复阳COVID-19患者的临床表现和实验室检查结果,对该类患者住院期间和出院后呼吸道及粪便标本病毒核酸持续时间进行分析评估。结果43例患者中有15例在住院期间SARS-CoV-2核酸检测短暂转阴后又出现“复阳”(复阳组),复阳比例为34.9%,均为普通型患者。与病毒核酸未出现复阳患者(未复阳组)相比,复阳组在入院时多表现为轻度发热,咳痰持续时间更短,ESR、CRP、血清淀粉样蛋白A水平更低(P均<0.05);2组患者鼻咽拭子标本病毒核酸初始转阴时间差异无统计学意义,但与未复阳组相比,复阳组鼻咽拭子病毒最长脱落时间显著延长,痰液中病毒脱落时间显著延长,粪便核酸阳性率更高(P均<0.05)。呼吸道标本病毒核酸复阳和粪便标本中病毒核酸持续阳性患者均未发现胸部CT较前改变。结论SARS-CoV-2核酸复阳COVID-19患者多表现为轻症感染和病毒持续阳性的特征,同时该类患者存在痰液标本病毒脱落时间延长和粪便标本病毒核酸持续阳性的特点,因而对病毒核酸复阳患者应给予特别关注和更长时间的医学观察,以预防和控制疫情的再次传播。

COVID-19恢复期MSCT征象及核酸复阳与非复阳MSCT征象对比分析

目的:分析新型冠状病毒肺炎(COVID-19)恢复期多层螺旋CT(MSCT)表现特点及新型冠状病毒核酸检测非复阳患者与复阳患者的MSCT表现差异。方法:回顾性分析深圳市萨米医疗中心收治的131例恢复期COVID-19患者的一般资料及MSCT图像,并将病例分为非复阳组(93例)及复阳组(38例),采用t检验、Mann-Whitney U检验、χ^2检验或Fisher确切概率法进行统计分析。结果:恢复期COVID-19患者男女比例为61:70,平均发病年龄(48.6±16.3)岁。非复阳组和复阳组性别比例差异无统计学意义[(45∶48)vs.(16∶22),χ^2=0.428,P=0.513],非复阳组较复阳组平均发病年龄大[(51.3±14.9)vs.(42.7±18.0),t=2.836,P=0.005]。恢复期COVID-19患者肺叶病灶受累情况如下:右肺下叶(143分)、左肺下叶(140分)、右肺上叶(93分)、右肺中叶(90分)、左肺上叶舌段(84分)、左肺固有上叶(70分)。非复阳组总肺叶、右肺上叶、右肺中叶、右肺下叶及左肺下叶受累情况计分均高于复阳组,差异有统计学意义(P<0.05)。恢复期患者病灶主要位于肺外带,其次弥漫分布。非复阳组病灶外带和中内带分布比例均低于复阳组,差异有统计学意义(χ^2=6.711,P=0.035)。恢复期COVID-19患者MSCT主要征象为:磨玻璃样密度影(GGO)及索条状影,其次是GGO合并实变影,少数征象依次有钙化灶、小结节灶、树芽征、支气管扩张、肺气肿/肺大泡、胸膜增厚伴/不伴钙化、铺石路征、肺不张。其中支气管扩张均为柱状及囊状,支气管扩张主要位于双肺下叶。两组病例MSCT征象出现率无显著差异(χ^2=10.196,P=0.423),GGO与GGO合并实变征象差异无统计学意义(χ^2=0.011,P=0.915)。两组病例支气管扩张位置、数量差异均无统计学意义。结论:COVID-19恢复期MSCT表现有其特征性,新型冠状病毒核酸检测非复阳与复阳患者年龄及MSCT病灶的累及程度、分布范围均有差异,MSCT可以反映COVID-19的进展情况,对疾病的诊断、预防起到积极作用。

英研制通用流感疫苗 一针可预防各种类型流感

英国科学家研制的一款新型流感疫苗近期通过人体测试,效果良好。这一新型流感疫苗属全流感疫苗,或称通用流感疫苗,可促使人体对各型流感产生免疫力。如果这款疫苗顺利通过临床试验和安全测试,流感免疫学科可望发生革命性转变,未来的流感疫苗可不必随流感病毒的变异而不断更新。打"共性"作为这款新型疫苗的"制造者"。

转型有机遇也有挑战

点评1这是一个企业转型的时代,传统管理软件厂商已加速进入网络时代,我们期待国内软件厂商能够在互联网浪潮中再创当年神话。这是一个互联网的时代,几乎所有的企业都在谈转型,都在向互联网靠拢,仿佛只要沾了互联网的边儿,企业一下子就有了发展,公司的市值就能大幅提升,特别是从2013年起,以余额宝为代表的互联网理财产品引起了互联网金融的发展高潮。

南非HIV-1感染患儿接受补锌疗法的安全性与疗效研究:随机双盲、安慰剂对照试验

Background: Zinc deficiency is associated with impaired immune function and an increased risk of infection. Supplementation can decrease the incidence of diarrhoea and pneumonia in children in resource-poor countries. However, in children with HIV- 1 infection, the safety of zinc supplementation is uncertain. We aimed to assess the role of zinc in HIV- 1 replication before mass zinc supplementation is recommended in regions of high HIV- 1 prevalence. Methods: We did a randomised double-blind placebo-controlled equivalence trial of zinc supplementation at Grey’s Hospital in Pietermaritzburg, South Africa. 96 children with HIV- 1 infection were randomly assigned to receive 10 mg of elemental zinc as sulphate or placebo daily for 6 months. Baseline measurements of plasma HIV- 1 viral load and the percentage of CD4+ T lymphocytes were established at two study visits before randomisation, and measurements were repeated 3, 6, and 9 months after the start of supplementation. The primary outcome measure was plasma HIV- 1 viral load. Analysis was per protocol. Findings: The mean log10 HIV- 1 viral load was 5.4 (SD 0.61) for the placebo group and 5.4 (SD 0.66) for the zinc-supplemented group 6 months after supplementation began (difference 0.0002, 95% CI - 0.27 to 0.27). 3 months after supplementation ended, the corresponding values were 5.5 (SD 0.77) and 5.4 (SD 0.61), a difference of 0.05 (- 0.24 to 0.35). The mean percentage of CD4+ T lymphocytes and median haemoglobin concentrations were also similar between the two groups after zinc supplementation. Two deaths occurred in the zinc supplementation group and seven in the placebo group (p=0.1). Children given zinc supplementation were less likely to get watery diarrhoea than those given placebo. Watery diarrhoea was diagnosed at 30 (7.4% ) of 407 clinic visits in the zinc-supplemented group versus 65 (14.5% ) of 447 visits in the placebo group (p=0.001). Interpretation. Zinc supplementation of HIV- 1- infected children does not result in an increase in plasma HIV- 1 viral load and could reduce morbidity caused by diarrhoea. Relevance to Practice: Programmes to enhance zinc intake in deficient populations with a high prevalence of HIV- 1 infection can be implemented without concern for adverse effects on HIV- 1 replication. In view of the reductions in diarrhoea and pneumonia morbidity, zinc supplementation should be used as adjunct therapy for children with HIV- 1 infection.

Cyclosporine versus tacrolimus in patients with HCV infection after liver transplantation:Effects on virus replication and recurrent hepatitis

AIM： To determine the effects of the calcineurin inhibitors, cyclosporine and tacrolimus, on hepatitis C virus （HCV） replication and activity of recurrent hepatitis C in patients post liver transplantation. METHODS： The data of a cohort of 107 patients who received liver transplantation for HCV-associated liver cirrhosis between 1999 and 2003 in our center were retrospectively analyzed. The level of serum HCV-RNA and the activity of recurrent hepatitis were compared between 47 patients who received either cyclosporine or tacrolimus as the primary immunosuppressive agent and an otherwise similar immunosuppressive regimen which did not lead to biliary complications within the first 12 mo after transplantation. RESULTS： HCV-RNA increased within 3 mo after transplantation but the differences between the cyclosporine group and the tacrolimus group were insignificant （P=0.49 at 12 too）. In addition, recurrent hepatitis as determined by serum transarninases and histological grading of portal inflammation and fibrosis showed no significant difference after 12 mo （P= 0.34）.CONCLUSION： Cyclosporine or tacrolimus as a primary immunosuppressive agent does not influence the induction or severity of recurrent hepatitis in HCV- infected patients after liver transplantation.

Hepatitis B virus replication

Hepadnaviruses, including human hepatitis B virus （HBV）, replicate through reverse transcription of an RNA intermediate, the pregenomic RNA （pgRNA）. Despite this kinship to retroviruses, there are fundamental differences beyond the fact that hepadnavirions contain DNA instead of RNA. Most peculiar is the initiation of reverse transcription： it occurs by protein-priming, is strictly committed to using an RNA hairpin on the pgRNA, ε, as template, and depends on cellular chaperones; moreover, proper replication can apparently occur only in the specialized environment of intact nucleocapsids. This complexity has hampered an in-depth mechanistic understanding. The recent successful reconstitution in the test tube of active replication initiation complexes from purified components, for duck HBV （DHBV）, now allows for the analysis of the biochemistry of hepadnaviral replication at the molecular level. Here we review the current state of knowledge at all steps of the hepadnaviral genome replication cycle, with emphasis on new insights that turned up by the use of such cellfree systems. At this time, they can, unfortunately, not be complemented by three-dimensional structural information on the involved components. However, at least for the ~ RNA element such information is emerging, raising expectations that combining biophysics with biochemistry and genetics will soon provide a powerful integrated approach for solving the many outstanding questions. The ultimate, though most challenging goal, will be to visualize the hepadnaviral reverse transcriptase in the act of synthesizing DNA, which will also have strong implications for drug development.

Inhibition of hepatitis B virus expression and replication by RNA interference in HepG2.2.15

AIM： To observe the inhibition of hepatitis B virus replication and expression by transfecting vector-based small interference RNA （siRNA） pGenesiI-HBV X targeting HBV X gene region into HepG2.2.15 cells. METHODS：pGenesil-HBV X was constructed and transfected into HepG2.2.15 cells via lipofection. HBV antigen secretion was determined 24, 48, and 72 h after transfection by time-resolved immunofluorometric assays （TRFIA）. HBV replication was examined by fluorescence quantitative PCR, and the expression of cytoplasmic viral proteins was determined by immunohistochemistry. RESULTS： The secretion of HBsAg and HBeAg into the supernatant was found to be inhibited by 28.5% and 32.2% （P 〈 0.01）, and by 38.67% （P 〈 0.05） and 42.86% （P 〈 0.01） at 48 h and 72 h after pGenesil-HBV X transfection, respectively. Immunohistochemical staining for cytoplasmic HBsAg showed a similar decline in HepG2.2.15 cells 48 h after transfection. The number of HBV genomes within culture supernatants was also significantly decreased 48 h and 72 h post-transfection as quantified by fluorescence PCR （P 〈 0.05）. CONCLUSION： In HepG2.2.15 cells, HBV replication and expression is inhibited by vector-based siRNA pGenesil- HBV X targeting the HBV X coding region.

Inhibitory effect of emodin and Astragalus polysaccharide on the replication of HBV

AIM: To evaluate the anti-viral effect of emodin plus Astragalus polysaccharide (APS) in hepatitis B virus (HBV) transgenic mice.METHODS: Sixty HBV transgenic mice (HBV TGM) whose weight varied between 18 and 24 g were randomly divided into 3 groups, with 20 mice in each group. Group A was the normal control, where the mice were treated with physiological saline; group B was the positive control where the mice were treated with lamivudine solution (100 mL/kg per day). Group C was the experimental group where the mice were treated with physiological saline containing emodin and APS (57.59 mg/kg per day and 287.95 mg/kg per day, respectively). The mice were treated daily for 3 wk. After 1 wk recovery time, the mice were sacrifi ced and serum as well as liver tissues were collected for ELISA and histological examination.RESULTS: After 21 d treatment, HBV DNA levels in group B and group C significantly declined when compared with group A (P < 0.05). However, a signif icant increase in HBV DNA content was observed in group B, whereas this phenomenon was not observed in group C. A reduction in the contents of HBsAg, HBeAg and HBcAg in the mice from group B and C was observed when compared with group A.CONCLUSION: Emodin and APS have a weak but persistent inhibitory effect on HBV replication in vivo, which may function as a supplementary modality in the treatment of hepatitis B infection.

A case-control study of the relationship between hepatitis B virus DNA level and risk of hepatocellular carcinoma in Qidong,China

AIM:To investigate the role of hepatitis B virus (HBV) replication in the development of hepatocellular carcinoma (HCC), a nested case-control study was performed to study the relationship between HBV DNA level and risk of HCC. METHODS:One hundred and seventy cases of HCC and 276 control subjects free of HCC and cirrhosis were selected for this study. Serum HBV DNA level was measured using fluorescein quantitative polymerase chain reaction at study entry and the last visit. RESULTS:In a binary unconditional logistic regression analysis adjusted for age, cigarette smoking, alcohol consumption and family history of chronic liver diseases, the adjusted odds ratios (95% confidence intervals) of HCC in patients with increasing HBV DNA level were 2.834 (1.237-6.492), 48.403 (14.392-162.789), 42.252 (14.784-120.750), and 14.819 (6.992-31.411) for HBV DNA levels ≥ 104 to < 105; ≥ 105 to < 106; ≥ 106 to < 107; ≥ 107 copies/mL, respectively. Forty-six HCC cases were selected to compare the serums viral loads of HBV DNA at study entry with those at the last visit. The HBV DNA levels measured at the two time points did not differ significantly.CONCLUSION:The findings of this study provide strong longitudinal evidence of an increased risk of HCC associated with persistent elevation of serum HBV DNA level in the 104-107 range.

Inhibition of hepatitis B virus DNA replicative intermediate forms by recombinant interferon-γ

AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-γ, alone and in combination with lamivudine. METHODS: A recombinant baculovirus-HBV/HepG2 culture system was developed which could support productive HBV infection in vitro. Expression of HBsAg and HBeAg in infected HepG2 culture medium was detected by commercial enzyme immunoassays. HBV DNA replication intermediates were detected in infected cells by Southern hybridization and viral DNA load was determined by dot hybridization. RESULTS: IFN-γat 0.1 to 5μg/L efficiently down regulated HBsAg expression in transduced HepG2 cells. At 5μg/L, IFN-γalso suppressed HBV DNA replication in these cells. While treatment with a combination of lamivudine and IFN-γshowed no additive effect, sequential treatment first with lamivudine and then IFN-γwas found to be promising. In this culture system the best HBV suppression was observed with a pulse of 2μmol/L lamivudine for two days, followed by 1μg/L IFN-γfor another four days. Compared to treatment with lamivudine alone, the sequential use of 0.2μmol/L lamivudine for two days, followed by 5μg/L IFN-γfor six days showed a 72% reduction in HBV cccDNA pool. CONCLUSION: This in vitro study warrants further evaluation of a combination of IFN-γand lamivudine, especially in IFN-αnon-responder chronic hepatitis B patients. A reduced duration of lamivudine treatment would also restrict the emergence of drug-resistant HBV mutants.

Anti-HBV effect of liposome-encapsulated matrine in vitro ana in vivo

AIM: To study the anti-HBV effect of liposome-encapsulated matrine (Lip-M) in vitro and in vivo. METHODS: 2.2.15 cell line was cultured in vitro observe the effect of Lip-M and matrine on the secretion of HBsAg and HBeAg. The toxicity of Lip-M and matrine to 2.2.15 cell line was also studied by MTT method. In in vivo study, drug treatment experiment was carried out on the 13th day after ducks were infected with duck hepatitis B virus (DHBV). The ducks were randomly divided into 4 groups with 5-6 ducks in each group. Lip-M and matrine were given to DHBV-infected ducks respectively by gastric perfusion. Four groups were observed: group of Lip-M (20 mg/kg), group of Lip-M (10 mg/kg), group of matrine (20 mg/kg) and group of blank model. The drug was given once daily for 20 d continuously, and normal saline was used as control. The blood was drawn from the posterior tibial vein of all ducks before treatment (T0), after the medication for 5 (T5), 10 (T10), 15 (T15), 20 (T20) d and withdrawl of the drug for 3 d (P3). The serum samples were separated and stored at -70 ℃, DHBV-DNA was detected by the dot-blot hybridization. RESULTS: After addition of Lip-M and matrine to 2.2.15 cell line for eleven d, the median toxic concentration (TC50) of Lip-M and matrine was 7.29 mg/mL and 1.33 mg/mL respectively. The median concentration (IC50) of Lip-M to inhibit HBsAg and HBeAg expression was 0.078 mg/mL and 3.35 mg/mL respectively. The treatment index (TI) value of Lip-M for HBsAg and HBeAg was 93.46 and 2.17 respectively, better than that of matrine. The DHBV-infected duck model treatment test showed that the duck serum DHBV-DNA levels were markedly reduced in the group of Lip-M (20 mg/kg) after treated by gastric perfusion for 10, 15 and 20 d (0.43±0.22 vs 0.95±0.18, t = 4.70, P= 0.001<0.01.0.40±0.12 vs 0.95±0.18, t = 6.34, P= 0.000<0.01. 0.22±0.10 vs 0.95±0.18, t = 8.30, P= 0.000<0.01), compared to the group of matrine (20 mg/kg) (0.43±0.22 vs 0.79±0.19, t = 3.17, P= 0.01<0.05. 0.40±0.12 vs 0.73±0.24, t = 3.21, P= 0.009<0.05. 0.22±0.10 vs0.55±0.32, t = 2.27, P= 0.046<0.05.), and the control (0.43±0.22 vs50.98±0.29, t = 3.68, P = 0.005<0.01. 0.40±0.12 vs 0.97±0.30, t = 4.26, P= 0.002<0.01. 0.22±0.10 vs 0.95±0.27, t = 5.76, P= 0.000<0.01). After the treatment for 20 d and withdrawl of the drug for 3 d, duck serum DHBV-DNA level in the group of Lip-M (10 mg/kg) markedly reduced (0.56±0.26 vs0.95±0.38, t = 5.26, P= 0.003<0.05. 0.55±0.25 vs 0.95±0.38, t = 5.52, P= 0.003<0.05), and the difference was significant as compared with the control (0.56±0.26 vs 0.95±0.27, t = 2.37, P = 0.042<0.05. 0.55±0.25 vs 0.89±0.18, t = 2.55, P= 0.031<0.05), but not significant as compared with the group of matrine (20 mg/kg). After withdrawl of the drug for 3 d, the levels of DHBV-DNA did not relapse in both groups of Lip-M. CONCLUSION: Lip-M can evidently inhibit the replication of hepatitis B virus In vitro and in viva, its anti-HBV effect is better than that of matrine.

Combination of small interfering RNAs mediates greater inhibition of human hepatitis B virus replication and antigen expression

Objectives: To evaluate the inhibitory effect mediated by combination of small interfering RNAs (siRNAs) targeting different sites of hepatitis B virus (HBV) transcripts on the viral replication and antigen expression in vitro. Methods: (1) Seven siRNAs targeting surface (S), polymerase (P) or precore (PreC) region of HBV genome were designed and chemically synthesized. (2) HBV-producing HepG2.2.15 cells were treated with or without siRNAs for 72 h. (3) HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay. (4) Intracellular viral DNA was quantified by real-time PCR (Polymerase Chain Reaction). (5) HBV viral mRNA was reverse transcribed and quantified by real-time PCR. (6) The change of cell cycle and apoptosis was determined by flow cytometry. Results: Our data demonstrated that synthetic small interfering RNAs (siRNAs) targeting S and PreC gene could efficiently and specifically inhibit HBV replication and antigen expression. The ex- pression of HBsAg and HBeAg and the replication of HBV could be specifically inhibited in a dose-dependent manner by siRNAs. Furthermore, our results showed that the combination of siRNAs targeting various regions could inhibit HBV replication and antigen expression in a more efficient way than the use of single siRNA at the same final concentration. No apoptotic change was observed in the cell after siRNA treatment. Conclusion: Our results demonstrated that siRNAs exerted robust and specific inhibi- tion on HBV replication and antigen expression in a cell culture system and combination of siRNAs targeting different regions exhibited more potency.