登革2型病毒E蛋白结构域Ⅲ的表达及多克隆抗体制备

登革病毒(Dengue virus, DENV)属于黄病毒科(Flaviviridae),黄病毒属(Flavivirus),为单股正链RNA病毒,有4个不同的血清型(DENV- 1,2,3,4),主要通过埃及伊蚊(Aedes aegypti)和白纹伊蚊(Aedes albopictus)传播,可引起登革热、登革出血热、登革休克综合征等多种疾病 [1,2].     

VEGF-VEGFR2 信号轴对糖尿病血管生成反应的调控研究进展

血管内皮生长因子(Vacularendothelial growth factor,VEGF)-血管内皮生长因子受体-2(VEGF receptor2,VEGFR-2)信号轴调控血管生成反应。糖尿病病理状态下,氧化应激异常激活、NO等血管活性物质功能受损、以及晚期糖基化终末产物增加,该信号轴功能失调,使得血管生成反应在一些器官组织中呈增强状态,如视网膜和肾;然而在另一些组织中却是受到抑制,如外周血管等。不正常的血管生成反应最终导致糖尿病性心血管并发症的发生。因此,阐明血管生产反应功能障碍,将为糖尿病心血管并发症靶向治疗提供依据。     

虫媒黄病毒嵌合疫苗研究进展

黄病毒科黄病毒属包括70多种病毒,分为几种抗原复合组,大多数黄病毒为节肢动物传播的,其中40％以上的病毒对人有致病性,最重要的疾病有登革热／登革出血热(DEN／DHF),日本脑炎(乙型脑炎,JE),黄热(YF),西尼罗热(WNF)和璧虱传播脑炎(森林脑炎,TBE)等。登革热每年约有数千万至1亿病人,数十万人发生严重的登革出血热。乙脑每年约发生4～5万病例,病死率25％,45％     

鞘氨醇激酶-磷酸鞘氨醇轴在血管生成相关性疾病中的作用

血管生成是指在原有血管的基础上形成新血管的过程。病理性血管生成是癌症、心血管类疾病和视网膜病变等一系列疾病的标志。1-磷酸鞘氨醇(sphingosine-1-phosphate,S1P)是一种信号脂质,由鞘氨醇激酶（sphingosine kinases,SPHK）合成,通过5种G蛋白偶联受体(sphingosine-1-phosphate receptors,S1PR1-5)发挥其不同的生物学和病理生理作用,并通过激活受体启动各种信号级联反应,影响细胞命运、血管张力、内皮功能和完整性以及淋巴细胞的运输等。其产生和信号的失衡与内皮功能障碍和异常血管生成等病理过程密切相关。越来越多的证据表明, SPHK-S1P轴在血管生成中发挥重要作用,尤其在癌症的发生发展与肿瘤微环境、动脉粥样硬化、心肌梗死等心血管类疾病,以及糖尿病和视网膜病变中具有重要意义。研究其相关作用与功能,可为治疗血管生成相关疾病提供新见解和药物治疗靶点。本文就SPHK-S1P轴通过SPHK以及S1PR1-5影响内皮细胞和平滑肌增殖、内皮细胞迁移以及由内皮细胞、周细胞和平滑肌细胞等形成管腔的分子机制进行阐述,同时进一步阐述SPHK-S1P轴如何通过鞘氨醇激酶以及S1PR1-5影响肿瘤、心血管类疾病、糖尿病以及视网膜病变中血管生成,旨在通过理解SPHK-S1P轴在血管生成中的分子机制为相关疾病提供新的治疗思路。     

重组人血管内皮抑制素（恩度）临床机制的研究进展

重组人血管内皮抑制素(Endostar,恩度)是由我国科学家自主研发的血管内皮抑制素类抗肿瘤药物,是一种大肠杆菌工程菌发酵产品,在临床上常被用作血管靶向治疗,具有靶向精准、低毒性、不产生耐药等优点。恩度能够抗血管生成、诱导肿瘤细胞凋亡,然而只应用抗血管生成药物治疗肿瘤患者,并不能使其得到明显的生存获益,恩度联合化疗/放疗具有增敏作用,从而取得显著的疗效。本文对恩度的作用机制进行探讨,以便于为更好的应用于临床提供有益的参考信息。     

NSCLC中低氧诱导因子-1α对血管生成素-2表达的影响

目的探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)中低氧诱导因子-1α(hypoxia-inducible fac-tor-1α,HIF-1α)对血管生成素-2(angiopoietin-2)表达的影响及意义。方法免疫组化SP法检测46例NSCLC组织HIF-1α、血管生成素-2蛋白的表达;培养人肺腺癌细胞株A549细胞,CoCl2模拟缺氧处理6h、12h、24h,以及缺氧条件下不同浓度genistein处理细胞12h后,采用Western Blot和RT-PCR方法分别检测A549细胞HIF-1α蛋白和血管生成素-2 mRNA的表达情况。结果46例NSCLC中HIF-1α、血管生成素-2的阳性表达率分别为73.9%(34/46)、63.0%(29/46),明显高于癌旁肺组织(P<0.05);HIF-1α的表达与血管生成素-2的表达呈正相关。急性缺氧可以诱导A549细胞HIF-1α蛋白和血管生成素-2 mRNA表达增加,分别在6h,12h达到高峰,随着缺氧时间延长,HIF-1α蛋白和血管生成素-2 mRNA表达相对减少;genistein抑制HIF-1α蛋白后,血管生成素-2 mRNA的表达也相应减少,呈浓度依赖。结论缺氧时NSCLC中HIF-1α可参与血管生成素-2的调控,HIF-1α、血管生成素-2表达增加与肿瘤新生血管形成有关,二者共同促进NSCLC的发生发展过程。     

登革病毒包膜蛋白的原核表达及免疫原性研究

登革病毒感染引起的登革热和登革出血热/登革休克综合征是目前流行最为广泛的虫媒病毒病,但其发病机制不明,也无疫苗和特异性抗病毒药物用于防治。登革病毒包膜蛋白(E蛋白)在病毒致病和免疫过程中发挥重要作用。本研究构建了登革病毒2型E蛋白基因的重组质粒E/pGEX-6P-1,并优化表达条件,获得登革病毒E蛋白的高效可溶性表达;用谷胱甘肽琼脂糖凝胶4B亲和层析柱纯化,获得纯度较高的蛋白。该蛋白具有较好的免疫原性,免疫小鼠后可获得高效价抗体。本研究为进一步了解登革病毒E蛋白的功能及其在相关疾病中的应用奠定了基础。     

新型登革疫苗研究进展

登革病毒(Dengue virus,DENV)以伊蚊为主要传播媒介,可感染人,引起登革热(Dengue fever,DF)、登革出血热(Dengue hemorrhagic fever,DHF)及登革休克综合征(Dengue shock syndrome,DSS),其中以登革热最常见,广泛流行于热带和亚     

新型登革疫苗研究进展

登革病毒(Dengue virus,DENV)以伊蚊为主要传播媒介,可感染人,引起登革热(Dengue fever,DF)、登革出血热(Dengue hemorrhagic fever,DHF)及登革休克综合征(Dengue shock syndrome,DSS),其中以登革热最常见,广泛流行于热带和亚     

ESM-1、s E-选择素、s TM、Ang-2对儿科脓毒症的治疗效果研究

为观察脓毒症患儿外周血内皮细胞特异性分子-1 (endothelial cell specific molecule-1, ESM-1)、可溶性E-选择素(soluble E-selectin, s E-selectin)、可溶性血栓调节蛋白(soluble thrombomdulin, s TM)、血管生成素-2 (angiopoietin-2, Ang-2)水平的变化,阐明这些指标与脓毒症严重程度之间的关系,探究这些内皮细胞损伤标志物在儿童脓毒症病程发展及预后中的意义,为临床脓毒症患儿的治疗提供理论依据,本研究根据研究目的、按照中华医学会儿科分会急救学组2005年制定的诊断标准,以选取0～3岁儿童并分为对照组(非感染组)、脓毒症组、严重脓毒症组,每组各30例。通过对血清ESM-1、s E-selection、s TM、Ang-2水平进行检测,比较3组患儿4组指标水平与患儿病情进展的相关性。研究发现脓毒症、严重脓毒症患儿第1天在体温、心率、呼吸、血压及外周血白细胞计数方面均显著高于对照组,差异有统计学意义(p0.05)。但是脓毒症与严重脓毒症比较,差异无统计学意义(p0.05)。脓毒症组患儿周血ESM-1、s E-selection、s TM、Ang-2的浓度在第1天明显高于对照组,差异有显著性(p0.05);严重脓毒症组患儿上述指标进一步增高,与脓毒症患儿及对照组相应指标比较,差异均有统计学意义(p0.05);且此4项指标在脓毒症患儿病程的发展早期,尤其是第1、3天均呈逐渐上升趋势,病程第7天血清s-TM、s E-selection、ang-2水平有所下降,而ESM-1水平在第1、3、7天逐渐持续增高,差异有统计学意义(p0.05)。本研究的结果说明,内皮细胞损伤相关标志物ESM-1、s E-selection、s TM、Ang-2可作为临床脓毒症患儿病程发展及预后的一个参考指标,并且能够成为评价脓毒症治疗效果的评价指标。     

2012—2019年上海市某三甲医院境外输入性传染病病例特征分析

本研究分析近年来上海市境外输入性传染病的病例特点,为上海市输入性传染病的防控提供依据。对上海市某三甲医院2012—2019年报告的所有法定及重点监测传染病中确定为境外感染的305例传染病病例进行回顾性分析。采用WPS Office 2019和 SPSS17.0软件对所收集病例的病种、数量、输入时间、病例人口学特征以及输入地区等进行整理分析。结果显示,2012—2019年该院报告境外输入性传染病11种,分别为疟疾、登革热、肺结核、麻疹、甲型肝炎、戊型肝炎、伤寒、人类免疫缺陷病毒(human immunodeficiency virus,HIV)感染、黄热病、白喉、隐性梅毒。确诊的305例病例中,疟疾226例,占74.10%;登革热68例,占22.30%;其他9种疾病共11例,占3.6%。疟疾、登革热全年均有输入,其中6月、9月和11月为疟疾输入高峰,6～8月为登革热输入高峰,肺结核、麻疹、甲型肝炎、戊型肝炎、伤寒、HIV感染、黄热病、白喉、隐性梅毒无明显季节性分布特征。输入性病例中,中国籍多于外籍,男性多于女性,以青壮年为主;主要职业为商业服务和工人;主要输入地为非洲。本研究结果提示境外输入性传染病以疟疾、登革热等虫媒传染病为主,病种呈现多样化的变化趋势,输入高峰主要在夏秋季。应加强非洲、东南亚劳务和旅游人群的健康教育,做好病例监管和防蚊、灭蚊,减少二代病例。     

Enhanced passive surveillance dengue infection among febrile children: Prevalence,co-infections and associated factors in Cameroon

Dengue virus (DENV) causes a spectrum of diseases ranging from asymptomatic, mild febrile to a life-threatening illness: dengue hemorrhagic fever. The main clinical symptom of dengue is fever, similar to that of malaria. The prevalence of dengue virus infection, alone or in association with other endemic infectious diseases in children in Cameroon is unknown. The aim of this study was to determine the prevalence of dengue, malaria and HIV in children presenting with fever and associated risk factors.Dengue overall prevalence was 20.2%, Malaria cases were 52.7% and HIV cases represented 12.6%. The prevalence of dengue-HIV co-infection was 6.0% and that of Malaria-dengue co-infection was 19.5%. Triple infection prevalence was 4.3%. Dengue virus infection is present in children and HIV-Dengue or Dengue- Malaria co-infections are common. Dengue peak prevalence was between August and October. Sex and age were not associated with dengue and dengue co-infections. However, malaria as well as HIV were significantly associated with dengue (P = 0.001 and 0.028 respectively). The diagnosis of dengue and Malaria should be carried out routinely for better management of fever.     

Virus-induced decline in soluble vascular endothelial growth receptor 2 is associated with plasma leakage in dengue hemorrhagic Fever

Some individuals infected with dengue virus develop dengue hemorrhagic fever (DHF), a viral hemorrhagic disease characterized by a transient period of localized plasma leakage. To determine the importance of vascular endothelial growth factor A (VEGF-A) in this syndrome, we compared plasma levels of VEGF-A and the soluble forms of its receptors in patients with DHF to patients with dengue fever (DF), a milder form of dengue virus infection without plasma leakage. We observed a rise in the plasma levels of free, but not total VEGF-A in DHF patients at the time of plasma leakage. This was associated with a decline in the soluble form of VEGF receptor 2 (VEGFR2) and VEGF-soluble VEGFR2 complexes, but not the soluble form of VEGFR1. The severity of plasma leakage in patients inversely correlated with plasma levels of soluble VEGFR2. In vitro, dengue virus suppressed soluble VEGFR2 production by endothelial cells but up-regulated surface VEGFR2 expression and promoted response to VEGF stimulation. In vivo, plasma viral load correlated with the degree of decline in plasma soluble VEGFR2. These results suggest that VEGF regulates vascular permeability and its activity is controlled by binding to soluble VEGFR2. Dengue virus-induced changes in surface and soluble VEGFR2 expression may be an important mechanism of plasma leakage in DHF.     

Early warning systems (EWSs) for chikungunya,dengue, malaria,yellow fever,and Zika outbreaks: What is the evidence? A scoping review

BackgroundEarly warning systems (EWSs) are of increasing importance in the context of outbreak-prone diseases such as chikungunya, dengue, malaria, yellow fever, and Zika. A scoping review has been undertaken for all 5 diseases to summarize existing evidence of EWS tools in terms of their structural and statistical designs, feasibility of integration and implementation into national surveillance programs, and the users’ perspective of their applications.MethodsData were extracted from Cochrane Database of Systematic Reviews (CDSR), Google Scholar, Latin American and Caribbean Health Sciences Literature (LILACS), PubMed, Web of Science, and WHO Library Database (WHOLIS) databases until August 2019. Included were studies reporting on (a) experiences with existing EWS, including implemented tools; and (b) the development or implementation of EWS in a particular setting. No restrictions were applied regarding year of publication, language or geographical area.FindingsThrough the first screening, 11,710 documents for dengue, 2,757 for Zika, 2,706 for chikungunya, 24,611 for malaria, and 4,963 for yellow fever were identified. After applying the selection criteria, a total of 37 studies were included in this review. Key findings were the following: (1) a large number of studies showed the quality performance of their prediction models but except for dengue outbreaks, only few presented statistical prediction validity of EWS; (2) while entomological, epidemiological, and social media alarm indicators are potentially useful for outbreak warning, almost all studies focus primarily or exclusively on meteorological indicators, which tends to limit the prediction capacity; (3) no assessment of the integration of the EWS into a routine surveillance system could be found, and only few studies addressed the users’ perspective of the tool; (4) almost all EWS tools require highly skilled users with advanced statistics; and (5) spatial prediction remains a limitation with no tool currently able to map high transmission areas at small spatial level.ConclusionsIn view of the escalating infectious diseases as global threats, gaps and challenges are significantly present within the EWS applications. While some advanced EWS showed high prediction abilities, the scarcity of tool assessments in terms of integration into existing national surveillance systems as well as of the feasibility of transforming model outputs into local vector control or action plans tends to limit in most cases the support of countries in controlling disease outbreaks.     

Activation of endothelial cells via antibody-enhanced dengue virus infection of peripheral blood monocytes.

Although endothelial cells have been speculated to be a target in the pathogenesis of dengue hemorrhagic fever (DHF), there has been little evidence linking dengue virus infection to any alteration in endothelial cell function. In this study, we show that human umbilical vein endothelial cells become activated when exposed to culture fluids from dengue virus-infected peripheral blood monocytes. Maximum activation was achieved with culture fluids from monocytes in which virus infection was enhanced by the addition of dengue virus-immune serum, thus correlating with epidemiological evidence that prior immunity to dengue virus is a major risk factor for DHF. Activation was strongest for endothelial cell expression of VCAM-1 and ICAM-1. In contrast, activation of endothelial cell E-selectin expression appeared to be more transient, as indicated by its detection at 3 h, but not at 16 h, of treatment. Treatment of monocyte culture fluids with anti-tumor necrosis factor alpha (TNF-alpha) antibody largely abolished the activation effect (as measured by endothelial cell expression of ICAM-1), whereas treatment with IL-1beta receptor antagonist had a much smaller inhibitory effect on activation. Endothelial cells inoculated directly with dengue virus or with virus-antibody combinations were poorly infectable (compared to Vero cells or peripheral blood monocytes), and virus-inoculated endothelial cells showed no increased expression of VCAM-1, ICAM-1, or E-selectin. Taken together, the results strongly indicate that dengue virus can modulate endothelial cell function by an indirect route, in which a key intermediary is TNF-alpha released from virus-infected monocytes.     

Dengue and Chikungunya Fever among Viral Diseases in Outpatient Febrile Children in Kilosa District Hospital,Tanzania

Introduction

Viral etiologies of fever, including dengue, Chikungunya, influenza, rota and adeno viruses, cause major disease burden in tropical and subtropical countries. The lack of diagnostic facilities in developing countries leads to failure to estimate the true burden of such illnesses, and generally the diseases are underreported. These diseases may have similar symptoms with other causes of acute febrile illnesses including malaria and hence clinical diagnosis without laboratory tests can be difficult. This study aimed to identify viral etiologies as a cause of fever in children and their co-infections with malaria.

Methods

A cross sectional study was conducted for 6 months at Kilosa district hospital, Tanzania. The participants were febrile children aged 2–13 years presented at the outpatient department. Diagnostic tests such as IgM and IgG ELISA, and PCR were used.

Results

A total of 364 patients were enrolled, of these 83(22.8%) had malaria parasites, 76 (20.9%) had presumptive acute dengue infection and among those, 29(38.2%) were confirmed cases. Dengue was more likely to occur in children ≥ 5 years than in <5 years (OR 2.28, 95% CI: 1.35–3.86). Presumptive acute Chikungunya infection was identified in 17(4.7%) of patients. We observed no presenting symptoms that distinguished patients with Chikungunya infection from those with dengue infection or malaria. Co-infections between malaria and Chikungunya, malaria and dengue fever as well as Chikungunya and dengue were detected. Most patients with Chikungunya and dengue infections were treated with antibacterials. Furthermore, our results revealed that 5(5.2%) of patients had influenza virus while 5(12.8%) had rotavirus and 2(5.1%) had adenovirus.

Conclusion

Our results suggest that even though viral diseases are a major public health concern, they are not given due recognition as a cause of fever in febrile patients. Emphasis on laboratory diagnostic tests for proper diagnosis and management of febrile patients is recommended.     

Emerging infectious disease: what are the relative roles of ecology and evolution?

The increasing threat of infectious diseases in humans has renewed interest in factors leading to the emergence of new diseases and the re-emergence of familiar diseases. Examples of seemingly novel diseases currently spreading in human populations include HIV, dengue hemorrhagic fever and Lyme disease; drug-resistant forms of well-known diseases such as tuberculosis are also increasing. The problem of disease emergence also extends to other animal and plant populations. In most current epidemics, ecological factors (e.g. migration, climate, agricultural practices) play a more significant role in disease emergence than evolutionary changes in pathogens or hosts. Evolutionary biologists and ecologists have much to offer to the development of strategies for the control of emerging diseases.     

Serotype-specific entry of dengue virus into liver cells: identification of the 37-kilodalton/67-kilodalton high-affinity laminin receptor as a dengue virus serotype 1 receptor

Dengue virus, the causative agent of dengue fever, dengue shock syndrome, and dengue hemorrhagic fever, infects susceptible cells by initially binding to a receptor(s) located on the host cell surface. Evidence to date suggests that receptor usage may be cell and serotype specific, and this study sought to identify dengue virus serotype 1 binding proteins on the surface of liver cells, a known target organ. By using a virus overlay protein binding assay (VOPBA), in both nondenaturing and denaturing gel systems, a putative dengue virus serotype 1 binding protein of approximately 37 kDa expressed on the surface of liver (HepG2) cells was identified. Mass spectrometry analysis identified a candidate protein, the 37/67-kDa high-affinity laminin receptor. Entry of the dengue virus serotype 1 was significantly inhibited in a dose-dependent manner by both antibodies directed against the 37/67-kDa high-affinity laminin receptor and soluble laminin. No inhibition of virus entry was seen with dengue virus serotypes 2, 3, or 4, demonstrating that the 37/67-kDa high-affinity laminin receptor is a serotype-specific receptor for dengue virus entry into liver cells.     

Role of mitogen-activated protein kinase signaling in the pathogenesis of dengue virus infection

Dengue virus (DENV) infection is a disease that is endemic to many parts of the world, and its increasing prevalence ranks it among the diseases considered to be a significant threat to public health. The clinical manifestations of DENV infection range from mild dengue fever (DF) to more severe dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Increased proinflammatory cytokines and vascular permeability, both of which cause organ injury, are the hallmarks of severe dengue disease. Signs of liver injury were observed in studies using hepatic cell lines, mouse models, and autopsy specimens from DENV-infected patients, and these signs substantiated the effects of inflammatory responses and hepatic cell apoptosis. Mitogen-activated protein kinases (MAPK) are involved in inflammatory responses and cellular stress during viral infections. The roles of MAPK signaling in DENV infection were reviewed, and published data indicate MAPK signaling to be involved in inflammatory responses and hepatic cell apoptosis in both in vitro cultures and in vivo models. Modulation of MAPK signaling ameliorates the inflammatory responses and hepatic cell apoptosis in DENV infection. This accumulation of published data relative to the role of MAPK signaling in inflammatory responses and cell apoptosis in DENV infection is elucidatory, and may help to accelerate the development of novel or repositioned therapies to treat this unpredictable and often debilitating disease.