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21.
目的:建立一种应用T7 RNA聚合酶体外转录合成大量siRNA的方法。方法:以萤火虫荧光素酶为靶标,应用T7 RNA聚合酶体外转录合成siRNA,脂质体转染法将其导入HepG2细胞中,通过测定荧光素酶的量,评价siRNA对萤火虫荧光素酶基因表达的抑制作用。结果:合成了以萤火虫荧光素酶为靶标的siRNA,合成的siRNA能特异性地抑制荧光素酶基因的表达,抑制率为80%。结论:建立了一种经济简便的体外合成siRNA的方法。 相似文献
22.
Rui T Marinho Ruth M Pinto Maria L Santos Miguel Carneiro de Moura 《Liver international》2004,24(5):413-418
BACKGROUND: Seroreversion, negativation of anti-hepatitis C virus previously positive, is sometimes found in some chronic hepatitis C-sustained responders (SRs) to antiviral therapy. AIMS: To determine the probability of seroreversion in SR treatment with Interferon and Ribavirin, and lymphocyte T helper (CD4+) reactivity to HCV antigens. METHODS: Thirty SR were followed on average for 54.8 months. Anti-HCV was tested by third generation test. Peripheral blood mononuclear cells (PBMCs) were isolated from venous blood and cultured to evaluate CD4+ proliferation in response to 2 microg/ml of eight HCV recombinant antigens from core, NS3, NS4, NS5 regions. RESULTS: Seroreversion was verified in 23% of patients (7/30), appearing at 47.5+/-24.0 months. The probability of anti-HCV loss in this group was 25% at 56 months after ending therapy. In 57% (4/7), anti-HCV returned to positive. These 7 SR patients with seroreversion also showed weaker CD4+ reactivity in 5% of tests (3/56) than the remaining 23 anti-HCV-positive SRs who showed stronger reactivity in 18% of tests (33/184), P=0.036. CONCLUSIONS: One-quarter of the SR showed seroreversion of anti-HCV and weaker CD4+ specific HCV proliferation than those who remained anti-HCV positive. The data suggest that complete viral eradication is a possible and achievable clinical objective. 相似文献
23.
Interferon-induced depressive illness in hep C patients responds to SSRI antidepressant treatments 下载免费PDF全文
This paper examines the role of selective serotonin reuptake inhibitors (SSRIs) in the treatment of hepatitis-C virus (HCV) patients who have developed interferon-α induced depression. A 2-year data analysis of HCV psychiatric liaison clinic has been undertaken. The diagnosis, treatment, and progress of those patients who were treated with interferon-α (INF-α) are reported. 53 of the 78 patients enrolled at the HCV Clinic and treated with INF-α were referred for psychiatric consultation. Six patients developed major depressive illness following INF therapy. They were all treated with SSRIs and they made full recovery. This is a significant observation and is concordant with other studies. Its biochemical ramifications are presented. It is concluded that INF-induced depression is fully reversible. A hypothesis is proposed that SSRIs modulate the neuro-protective neurotoxic ratio by possibly inhibiting the indole-2,3-dioxygenase induction of the kynurenine pathway. 相似文献
24.
S. Caillard C. Lelong F. Pessione B. Moulin for the French PTLD Working Group 《American journal of transplantation》2006,6(11):2735-2742
Post-transplant lymphoproliferative disorders (PTLD) are a rare but serious complication after organ transplantation. A French Registry of PTLD was set up in a nationwide population of kidney transplant recipients. We prospectively enrolled all adult kidney recipients developing PTLD between January 1, 1998, and December 31, 2003. We analyzed the incidence, risk and prognostic factors of PTLD by Kaplan-Meier and Cox analyses. Totally 230 cases of PTLD were referred to the French Registry. Cumulative incidence was 1.18% after 5 years. Older age (per year, AHR = 2.19, CI = 1.22-3.94) and recipient Epstein-Barr virus seronegativity (AHR = 3.01, CI = 1.57-5.08) were associated with an increased risk of PTLD. Patients with PTLD had a reduced survival rate (61% at 5 years). Graft PTLD had the best prognosis with an 81% survival rate after 5 years. Infection with hepatitis C or B virus (HCV or HBV), late-onset PTLD, multiple sites involvement and high Ann Arbor staging were risk factors for patient death. Use of azathioprine was associated with a poorer survival rate. PTLD incidence and risk factors in French recipients are in line with the international or American PTLD series. We highlighted the role of HBV or HCV in patient mortality and described the relevant prognosis factors for patients with post-transplant lymphoproliferations. 相似文献
25.
目的构建并筛选大鼠胶质原纤维酸性蛋白(GFAP)表达抑制短发夹样RNA(shRNA)真核表达载体。方法针对GFAP基因全编码序列设计并合成三对9bp茎环结构、19bp干扰序列特异性shRNA模板,体外定向克隆构建特异性重组质粒真核表达载体;通过体外大鼠脊髓源星形胶质细胞GFAP表达抑制模型,脂质体介导RNA干扰分子转染,实时荧光定量RT—PCR及Wesem blot技术观察RNA干扰后原代星形胶质细胞GFAP表达抑制效果.筛选最佳GFAP表达干扰抑制真核表达载体。结果序列测定证实GFAP—shRNA重组质粒真核表达载体构建成功,三对shRNA模板在mRNA及蛋白表达水平抑制靶基因表达效率分别为81%、63%、56%。结论高效率的GFAP—shRNA真核表达载体在大鼠原代星形胶质细胞GFAP表达抑制模型中能高效抑制GFAP基因表达,为后续多靶点RNA干扰技术在脊髓损伤胶质瘢痕抑制基因治疗中的应用奠定了前期基础。 相似文献
26.
J. A. Doebler W. R. Markesbery A. Anthony S. W. Scheff R. E. Rhoads 《Acta neuropathologica》1988,75(3):272-276
Summary A new double-staining procedure, in which the techniques of immunocytochemistry of glial fibrillary acidic protein (GFAP) and quantitative microdensitometry of azure B-RNA were combined, was used to study nucleic acid alterations in fibrous astrocytes in Alzheimer's disease (AD). RNA contents of GFAP-positive cells of the hippocampal endplate (Rose's H3-H5 fields) and the dentate gyrus molecular layer were determined in ten autopsy-proven AD patients (ages 51–88) and ten age-matched, non-demented control. In addition, RNA contents of pyramidal neurons of the endplate were examined. While there were no differences in RNA contents of astrocytes of either region between AD patients and controls, neuronal RNA was markedly depleted. These data suggest that astrocytes maintain protein synthetic capabilities in AD and that RNA loss is limited to the neuronal compartment.Supported by Grants 1P01-AG05119 and 1P50-AG05144 from the National Institutes of Health and by a Small Research Project Award from the University of Kentucky Medical Center 相似文献
27.
丙型肝炎患者外周血单核细胞中丙型肝炎病毒复制的研究 总被引:2,自引:0,他引:2
9例临床诊断为丙型肝炎患者,研究其外周血单核细胞中HCVRNA的存在及复制。9例患者血清标本抗-HCV及HCVRNA均为阳性,采用高敏感的逆转录一套式PCR法测定其外周血单核细胞中HCV正、负链RNA,结果9例患者外周血单核细胞中7例HCV正链RNA阳性,3例HCV负链RNA阳性,证实部分丙肝患者外周血单核细胞中存在HCV的复制,表明肝细胞并非为HCV感染与复制的唯一场所。 相似文献
28.
29.
Soonmyoung Paik M.D. 《Breast cancer research and treatment》1992,22(1):31-38
Summary Expression of IGF-I and IGF-II was studied in human breast cancer tissues by in situ hybridization. IGF-I mRNA was detected only in stromal cells adjacent to normal breast epithelial cells. Stromal cells associated with the tumor cells did not contain IGF-I, nor did malignant or benign breast epithelial cells. In contrast, IGF-II mRNA was found in both the malignant epithelial cells and their adjacent stromal cells. These data imply that stromal cells associated with breast epithelium may switch expression from IGF-I to IGF-II during breast cancer evolution. This appearance of IGF-II expression may identify cancer-associated stromal cells that have a fetal phenotype. 相似文献
30.
性传播疾病危险人群乙型肝炎和丙型肝炎病毒感染的血清流行病学调查 总被引:1,自引:0,他引:1
通过对431名卖淫妇女和109名正常妇女HBV和HCV感染的血清学指标的检测、性传播有关因素的调查,发现卖淫妇女HBV和HCV感染中的抗-Hkc为43.15%,抗-HBs为36.89%,HBsAg为10.44%,抗-HCV为9.51%,明显高于正常妇女;有性传播疾病者HBV和HCV的各项感染指标也明显高于正常妇女。卖淫妇女的抗-HCV感染率有随年龄增长而增高的趋势,与卖淫时限长、卖淫次数多有明显差别,这表明卖淫妇女中有较严重的HBV和HCV感染。 相似文献