隐丹参酮体外抗病毒作用及机制研究

目的 探索隐丹参酮在细胞水平的抗病毒作用,从药物调控宿主抗病毒免疫角度,阐释隐丹参酮抑制病毒复制的分子机制。方法 通过流式细胞术、实时荧光定量PCR（qRT-PCR）、免疫印迹实验（Western blotting）等方法检测隐丹参酮对水疱性口炎病毒（VSV）、甲型流感病毒（H1N1）、脑心肌炎病毒（EMCV）、Ⅰ型单纯疱疹病毒（HSV-1）复制的影响。基于基因集富集分析（GSEA）,结合qRT-PCR、流式细胞术,分析隐丹参酮抗病毒的作用机制。在干扰素受体IFNAR1敲除A549细胞（Ifnar1^-/-A549）中,检测I型干扰素（IFN-I）通路对隐丹参酮抗病毒功能的影响。结果 隐丹参酮显著抑制VSV、H1N1、EMCV和HSV-1病毒的复制,在病毒进入前期和进入后阶段发挥抗病毒作用,对病毒吸附过程无影响;通过GSEA分析结合qRT-PCR验证,证明隐丹参酮促进IFN-I信号通路的激活;在Ifnar1^-/-A549细胞中,隐丹参酮的抗病毒功能受到抑制。结论 隐丹参酮通过促进IFN-I信号通路激活抑制多种病毒复制。     

中东呼吸综合征治疗药物研究进展

摘 要中东呼吸综合征（MERS）是由中东呼吸综合征冠状病毒（MERS-CoV）引起的急性呼吸道传染病。MERS-CoV是继SARS冠状病毒（SARS-CoV）之后发现的一种具有高致死率的新型病毒,可由单峰骆驼传染给人类。目前,临床尚无抗MERS-CoV的特效药物。抗MERS-CoV的药物开发策略主要是借鉴抗SARS-CoV药物的开发经验,包括疫苗,抗体,小分子化合物快速筛选等。本文结合临床治疗及动物模型对MERS治疗药物的研究进展进行综述,旨在为该病新型疫苗和抗病毒药物的研发提供借鉴。     

膦甲酸的抗病毒作用和药代动力学及其在乙型肝炎中的应用

近年来,随着分子生物学的发展,针对病毒酶设计、合成的新抗病毒药日益增多,特别在疱疹病毒抑制剂的研究方面进展最大.抗病毒新药膦甲酸目前在国外已广泛用于治疗免疫功能低下者的巨细胞病毒感染及单纯疱疹病毒感染,对人类免疫缺陷病毒(HIV)、乙型肝炎病毒(HBV)等病毒感染亦有一些研究.国内已开始生产并在临床上试用.本文综述膦甲酸在抗病毒作用、药代动力学、乙型肝炎疗效及毒副作用等方面的研究进展,以供参考.     

千金藤素抗病毒作用的免疫学机制研究

目的 探索千金藤素体内外的抗病毒作用,并基于抗病毒天然免疫通路探究其抗病毒作用的分子机制。方法 CCK-8 法检测千金藤素（0.062 5～64.000 0 μmol·L^-1）对 A549 细胞活力的影响;利用表达绿色荧光蛋白的水疱性口炎病毒（VSV-GFP）感染 A549 细胞模型,结合流式细胞术检测千金藤素对病毒复制的影响并探究预处理、吸附过程及吸附后加药对 VSV-GFP 病毒复制的影响;实时荧光定量 PCR（qRT-PCR）检测千金藤素对甲型流感病毒（H1N1）、脑心肌炎病毒（EMCV）和单纯疱疹病毒 I 型（HSV-1）复制的影响;构建 VSV 感染小鼠模型探究千金藤素的体内抗病毒作用;A549细胞中利用生物信息学方法探究其抗病毒机制;qRT-PCR 检测药物处理 A549 和原代胚胎成纤维细胞（MEF）后 IFNB1 及干扰素刺激基因 （ISGs） 表达变化;免疫印迹法 （Immunoblotting） 检测人源单核细胞白血病细胞（THP-1）中 TBK1 和STAT1 的磷酸化水平。结果 与模型组相比,千金藤素在 A549 细胞中显著抑制 VSV、H1N1、EMCV 和 HSV-1 复制;千金藤素不影响 VSV 的吸附过程,而预处理或吸附后给药可以显著抑制病毒复制;千金藤素提高 VSV 感染小鼠的存活率;千金藤素激活基于IFN-I通路的抗病毒天然免疫应答。结论 千金藤素通过激活基于IFN-I通路的抗病毒天然免疫发挥体内外抗病毒作用。     

病毒学研究与抗病毒治疗药物

人们在与病毒的长期斗争中取得了不少重要成就,一些病毒进入人体细胞的过程及其致病机制相继得到阐明。随着病毒学研究的进步,许多抗病毒治疗药物不断出现并被用于临床,人们治疗艾滋病、病毒性肝炎、流感和疱疹病毒感染等病毒性疾病也由此取得了明显进步。然而,面对众多病毒性疾病,人们现有的有效治疗措施仍然极为有限。加强病毒学研究以加深对病毒致病机制的认识是研发新型抗病毒治疗药物的关键。本文介绍目前对临床常见病毒的研究与治疗进展,强调病毒学研究的进步在促进抗病毒治疗药物开发中的重要作用。     

1例多重耐药铜绿假单胞菌椎间隙感染患者的药学监护

摘 要 目的：分享多重耐药铜绿假单胞菌感染的药物治疗经验。方法: 对临床药师参与的1例多重耐药铜绿假单胞菌椎间隙感染患者的诊治过程进行回顾性分析。结果: 药师全程参与治疗,制定了磷霉素+头孢他啶+环丙沙星的联合抗菌方案,并调整用药时间顺序,获得满意疗效。结论:多重耐药铜绿假单胞菌感染可考虑采用三联方案。临床药师能够在疑难感染治疗中发挥积极作用。     

抗疱疹病毒感染药物的研究进展

疱疹病毒感染类疾病高发,可形成终生感染,给人类健康和财产安全带来了极大的损失。然而,临床上批准常用的治疗疱疹病毒感染的药物仅几种核苷类似物,这些药物的应用往往还会引发耐药性和不良反应。因而,克服抗疱疹病毒药物存在的问题,探索新型的抗疱疹病毒药物迫在眉睫。总结了临床常用抗疱疹病毒感染药物、靶向病原体的抗疱疹病毒抑制剂和靶向宿主的抗疱疹病毒抑制剂,介绍了联合用药抗病毒新型策略,以期为科研人员在抗疱疹病毒药物方面的研究提供参考。     

抗乙型肝炎病毒新药——阿德福韦酯的研究进展

阿德福韦酯(adefovir)是新一代核苷类抗病毒药物,能够有效抑制逆转录病毒、嗜肝病毒和疱疹病毒等各种病毒的复制和表达。实验研究表明阿德福韦酯不仅对野生HBV有效,对拉米夫定耐药的HBV病毒株也有显著的抑制作用,提示其能够作为拉米夫定耐药病毒感染的补充、替代产品,成为解决乙型肝炎病毒核苷类似物耐药问题的有效办法。阿德福韦酯有望成为联合治疗乙型肝炎的药物之一,可用于两种或多种药物的联合或序贯应用等方式治疗慢性乙型肝炎或HBV／HIV-1的合并感染。本文主要从阿德福韦酯的作用机制、药效毒理、临床药物动力学、体内抗病毒作用、耐药监测、临床用药的安全性等作一总结。     

2019—2022年临沧市人民医院血液透析患者导管相关性血流感染病原菌分布及耐药性分析

目的 分析临沧市人民医院2019—2022年肾脏免疫风湿科血液透析患者导管相关性血流感染病原菌分布及耐药性,为临床合理预防及治疗导管相关性血流感染提供参考。方法 搜集2019—2022年临沧市人民医院肾脏免疫风湿科送检的发生导管相关性血流感染的血液透析患者的血标本,对其病原菌分布及耐药性进行统计分析。结果 共检出病原菌160株,其中以革兰阳性菌为主,共91株（占56.9%）,常见的是表皮葡萄球菌和金黄色葡萄球菌;革兰阴性菌共69株（占43.1%）,常见的是铜绿假单胞菌、大肠埃希菌、嗜麦芽窄食单胞菌和阴沟肠杆菌。表皮葡萄球菌中对甲氧西林耐药的菌株检出率及对青霉素耐药率均高于金黄色葡萄球菌,均未检出对万古霉素和利奈唑胺耐药的菌株。大肠埃希菌中产超广谱β-内酰胺酶（ESBL）检出率为33.3%,低于全国水平;阴沟肠杆菌对所监测的抗菌药物的敏感性较好。铜绿假单胞菌对绝大多数抗铜绿假单胞菌药物的敏感性较好;嗜麦芽窄食单胞菌对米诺环素和复方磺胺甲噁唑的敏感性均大于80%,对替卡西林/克拉维酸、米诺环素无耐药菌株。结论 临沧市人民医院血液透析患者导管相关性血流感染细菌以革兰阳性菌为主。各病原菌对临床常用抗菌药物的耐药性各有不同,应根据各病原菌的耐药特点和抗菌药物药物的药动学/药效学选择合适的抗感染药物,并及时送检微生物培养。     

EB病毒感染患儿的诊疗体会

目的：探讨EB病毒（人类疱疹病毒）感染患儿的临床特征以及治疗方法。方法选取我院在2012年1月-2014年1月收治的EB病毒感染患儿120例,对所有患儿的临床资料进行回顾性分析。结果发热、咽狭炎、淋巴结肿大是EB病毒感染患儿常见临床症状,感染可累及全身各个系统,并以呼吸道最为常见;对于EB病毒感染患儿的治疗应在原发疾病治疗的基础上加强抗病毒治疗。结论 EB病毒感染患儿临床表现多样复杂,但预后较好,更昔洛韦是治疗EB病毒感染患儿较为有效的抗病毒药物。     

The DNA helicase–primase complex as a target for herpes viral infection

Introduction: The Herpesviridae are responsible for debilitating acute and chronic infections, and some members of this family are associated with human cancers. Conventional anti-herpesviral therapy targets the viral DNA polymerase and has been extremely successful; however, the emergence of drug-resistant virus strains, especially in neonates and immunocompromised patients, underscores the need for continued development of anti-herpes drugs. In this article, we explore an alternative target for antiviral therapy, the HSV helicase/primase complex.

Areas covered: This review addresses the current state of knowledge of HSV DNA replication and the important roles played by the herpesvirus helicase– primase complex. In the last 10 years several helicase/primase inhibitors (HPIs) have been described, and in this article, we discuss and contrast these new agents with established inhibitors.

Expert opinion: The outstanding safety profile of existing nucleoside analogues for α-herpesvirus infection make the development of new therapeutic agents a challenge. Currently used nucleoside analogues exhibit few side effects and have low occurrence of clinically relevant resistance. For HCMV, however, existing drugs have significant toxicity issues and the frequency of drug resistance is high, and no antiviral therapies are available for EBV and KSHV. The development of new anti-herpesvirus drugs is thus well worth pursuing especially for immunocompromised patients and those who develop drug-resistant infections. Although the HPIs are promising, limitations to their development into a successful drug strategy remain.     

Resistance of herpesviruses to antiviral drugs: clinical impacts and molecular mechanisms.

Nucleoside analogues such as acyclovir and ganciclovir have been the mainstay of therapy for alphaherpesviruses (herpes simplex virus (HSV) and varicella-zoster virus (VZV)) and cytomegalovirus (CMV) infections, respectively. Drug-resistant herpesviruses are found relatively frequently in the clinic, almost exclusively among severely immunocompromised patients receiving prolonged antiviral therapy. For instance, close to 10% of patients with AIDS receiving intravenous ganciclovir for 3 months excrete a drug-resistant CMV isolate in their blood or urine and this percentage increases with cumulative drug exposure. Many studies have reported that at least some of the drug-resistant herpesviruses retain their pathogenicity and can be associated with progressive or relapsing disease. Viral mutations conferring resistance to nucleoside analogues have been found in either the drug activating/phosphorylating genes (HSV or VZV thymidine kinase, CMV UL97 kinase) and/or in conserved regions of the viral DNA polymerase. Currently available second line agents for the treatment of herpesvirus infections--the pyrophosphate analogue foscarnet and the acyclic nucleoside phosphonate derivative cidofovir--also inhibit the viral DNA polymerase but are not dependent on prior viral-specific activation. Hence, viral DNA polymerase mutations may lead to a variety of drug resistance patterns which are not totally predictable at the moment due to insufficient information on specific drug binding sites on the polymerase. Although some CMV and HSV DNA polymerase mutants have been found to replicate less efficiently in cell cultures, further research is needed to correlate viral fitness and clinical outcome.     

Antiviral therapy for adenovirus infections

The treatment of severe adenovirus keratoconjunctivitis and life-threatening adenovirus infections in immunocompromised patients is still unsatisfactory. We here review the mode of action and antiviral data for cidofovir and ribavirin, obtained in cell culture, animal models or patients. Several nucleoside or nucleotide analogues have been described that target the adenovirus polymerase, whereas other antiviral targets have been poorly investigated. Furthermore, optimal therapeutic response may be achieved by combining antiviral therapy with immunotherapeutic approaches, as currently being explored.     

Prophylaxis against herpesvirus infections in transplant recipients

Herpesvirus infections are important after stem cell and organ transplant. During the last decades several antiviral agents have been introduced with efficacy against herpesviruses. These agents are the nucleoside analogues aciclovir, valaciclovir, famciclovir, and ganciclovir; the nucleotide analogue cidofovir; and the pyrophosphate analogue foscarnet. Several studies have been performed with antiviral agents with the aim to reduce morbidity and mortality associated with herpesvirus infections in transplant recipients. Aciclovir and valaciclovir have been examined in randomised, controlled trials in both solid organ and stem cell transplant patients, and were shown to be very effective for the prevention of herpes simplex virus (HSV) and varicella-zoster virus infections. In addition, these drugs were shown to reduce cytomegalovirus (CMV) infection and improve survival in allogenic stem cell transplant patients and to reduce CMV infection, CMV disease (aciclovir and valaciclovir), and acute rejection (valaciclovir) in renal transplant patients. Ganciclovir is very effective for the prevention of CMV infection and disease in both stem cell and solid organ transplant recipients. It can also be used in preemptive strategies in which the aim is to prevent CMV disease in patients who have ongoing CMV infection documented by antigenaemia or detection of CMV DNA. The latter strategy has the advantage of reducing the exposure to the drug and thereby the risk for toxicity. Foscarnet has also been shown to be effective as preemptive therapy for CMV in allogenic stem cell transplant patients and as therapy for aciclovir-resistant HSV infections. Finally cidofovir is an interesting agent with broad spectrum antiherpesvirus efficacy. However, because of the drug's toxicity profile, further studies are needed.     

抗单纯疱疹病毒活性的海洋化合物研究进展

单纯疱疹病毒(herpes simplex virus, HSV)是全球高度流行的病原体。目前核苷类似物如阿昔洛韦被用于单纯疱疹病毒的治疗,尽管其具有安全性和有效性,但这些药物的广泛使用导致耐药病毒株的出现,尤其是对于免疫功能低下的患者。为了解决这些问题,已有来自海洋的活性物质作为抗HSV药物的报道,且人们对海洋生物如藻类,海绵,被囊类动物,棘皮动物,软体动物,细菌以及真菌等作为抗HSV药物的来源产生了更大的兴趣。本综述重点介绍和总结了作为潜在抗HSV药物最有希望的海洋生物提取物,组分等, 以期为新型抗HSV药物的研发提供参考。     

Acyclovir. An updated review of its antiviral activity, pharmacokinetic properties and therapeutic efficacy

Acyclovir (aciclovir) is a nucleoside antiviral drug with antiviral activity in vitro against members of the herpes group of DNA viruses. As an established treatment of herpes simplex infection, intravenous, oral and to a lesser extent topical formulations of acyclovir provide significant therapeutic benefit in genital herpes simplex and recurrent orofacial herpes simplex. The effect of acyclovir therapy is maximised by early initiation of treatment, especially in non-primary infection which tends to have a less protracted course than the primary episode. Long term prophylactic oral acyclovir, in patients with frequent episodes of genital herpes simplex, totally suppresses recurrences in the majority of subjects; as with other infections responding to acyclovir, viral latency is not eradicated and pretreatment frequencies of recurrence return after discontinuation of treatment. Caution should accompany the prophylactic use of acyclovir in the general population, due to the theoretical risk of the emergence of viral strains resistant to acyclovir and other agents whose mechanism of action is dependent on viral thymidine kinase. Intravenous acyclovir is the treatment of choice in biopsy-proven herpes simplex encephalitis in adults, and has also been successful in the treatment of disseminated herpes simplex in pregnancy and herpes neonatorium. Intravenous and oral acyclovir protect against dissemination and progression of varicella zoster virus infection, but do not protect against post-herpetic neuralgia. In immunocompromised patients, intravenous, oral and topical acyclovir shorten the clinical course of herpes simplex infections while prophylaxis with oral or intravenous dosage forms suppresses reactivation of infection during the period of drug administration. Ophthalmic application of 3% acyclovir ointment rapidly heals herpetic dendritic corneal ulcers and superficial herpetic keratitis. Thus, despite an inability to eradicate latent virus, acyclovir administered in therapeutic or prophylactic fashion is now the standard antiviral therapy in several manifestations of herpes simplex virus infection, and indeed represents a major advance in this regard. With the exception of varicella zoster virus infections, early optimism concerning the use of the drug in diseases due to other herpes viruses has generally not been supported in clinical investigations.     

Penciclovir cream--improved topical treatment for herpes simplex infections

Human herpesviruses can be found worldwide and cause many viral infections in immunocompetent as well as in immunocompromised patients. Herpes simplex virus (HSV) diseases can be the cause of life-threatening disease, especially in neonates. After initial infection, HSV persists latently in host neurons with the risk of periodical reactivation over a lifetime. The development of acyclovir, a potent and specific nucleoside inhibitor of the herpes DNA polymerase, was a milestone in the history of antiviral drugs in the late 1970s. During the last decades a better understanding of the replication and disease-causing state of HSV types 1 and 2 has been achieved enabling the development of new and potent antiviral compounds. In the mid-1990s, for example, valacyclovir and famciclovir were launched as prodrugs of acyclovir with improved bioavailability. Despite the numerous drugs available for the systemic treatment of HSV infections, the topical application of a cream containing an antiviral agent is still the most convenient method of treating herpes simplex labialis/facialis in the general population. For some time, the topical standard treatment for recurrent HSV infections has been acyclovir cream, despite the fact that the evidence for efficacy in recurrent episodes has been equivocal. Penciclovir, a novel acyclic nucleoside analogue, has demonstrated efficacy against HSV types 1 and 2 and seems to have a pharmacological advantage due to a prolonged half-life of its active form in HSV-infected cells. This review discusses and compares the topical treatment modalities available for HSV infections. As a conclusion, different studies are available that have shown that it is possible to reduce viral replication and hasten lesion resolution with 1% penciclovir treatment beyond the prodromal phase of the HSV infection. Comparing data of topical treatment with acyclovir and penciclovir revealed a superiority for penciclovir cream showing a significant decrease in time to lesion healing, lesion area and pain. While systemic acyclovir or valacyclovir may be valid drugs especially for HSV prophylaxis, 1% penciclovir cream should be preferred as topical treatment since there are good therapeutic results independent of the phase of development of herpetic eruptions.     

Cellular factors involved in the induction of resistance of HIV to antiretroviral agents

Long-term treatment of HIV-1 infected patients with antiretroviral agents may result in failure of therapy due to the emergence of resistant virus mutants with decreased susceptibility to the therapeutic agents. Several authors have asked whether cellular factors, other than viral mutation may contribute to the declining efficiency of chemotherapy including nucleoside analogues and protease inhibitors (PI). Prolonged treatment with AZT may induce a defect of thymidine kinase activity in vitro and in vivo. Long-term treatment with other nucleoside analogues, such as d4T and 3TC, is also able to induce in host cells, a decreased sensitivity to the antiviral activity of these compounds. It is suggested that antiviral activity of PI could be modified by the expression of a protein P-gp that has been demonstrated to be able to bind PI and is involved in extrusion of anticancer agents.     

Current status of anti-HBV chemotherapy

In the past decade, significant progress has been achieved in the battle against hepatitis B virus. In addition to the immunomodulating agents such as interferon-α and thymosin, many novel antiviral agents have been discovered, among which nucleoside analogues are the main-stay. New-generation compounds such as 3TC and famciclovir have shown promise in the treatment of patients chronically infected by this virus, and are on the line for approval. However, viral rebound after cessation of therapy still remains a major problem. Additionally, the reports on the drug resistance to these antiviral agents suggest that combination therapy will be the eventual strategy (Bartholomewet al., 1997; Tippleset al., 1996). Therefore, developments of safe and effective antiviral agents which do not cross-resist with currently available antiviral drugs are still much needed.