Albumin-quercetin combination offers a therapeutic advantage in the prevention of reduced survival of erythrocytes in visceral leishmaniasis

Visceral leishmaniasis is associated with the reduced survival of erythrocytes, the cause of which remains to be fully explored. Here, we described the mechanism underlying the shortened lifespan of erythrocytes in visceral leishmaniasis and proposed a combination therapy with quercetin and hamster serum albumin towards its rectification. Decreased redox potential in erythrocytes followed by oxidative denaturation of hemoglobin and pathologic association of iron with the cell membrane facilitated premature hemolysis during leishmanial infection. Recently, we have reported the therapeutic efficacy of quercetin in arresting the enhanced destruction of erythrocytes in visceral leishmaniasis. Since serum albumin, the principal carrier protein for quercetin gets depleted in visceral leishmaniasis, the situation may compromise the efficacy of quercetin in this disease. We now report the use of quercetin-hamster serum albumin combination to increase the bioavailability of quercetin. The combination targeted hemoglobin oxidation and produced an effective attenuation of heme degradation. This led to decreased iron decompartmentalization, thereby increasing the life span of erythrocytes during leishmanial infection. Thus, we speculate that suppression of iron decompartmentalization, with the combination of quercetin and serum albumin might be a new approach in the prevention of reduced survival of erythrocytes in visceral leishmaniasis.     

Oxidative damage of erythrocytes infected with Plasmodium falciparum

Summary The extent of reduced glutathione, activity of glutathione peroxidase, amount of membrane lipid peroxidation products, and the extent of hemoglobin release from host erythrocytes during in vitroPlasmodium falciparum growth was studied. Highly synchronized parasite cultures were studied to examine the alterations caused by different growth stages of the parasite. There was a moderate increase in the reduced glutathione content as the parasite matured, which was significant only in schizontrich erythrocyte lysates (p<0.05) whereas the activity of glutathione peroxidase was significantly low in all the parasitized red blood cells (ring-infected RBC,p<0.005; trophozoite- and schizont-infected RBC,p<0.001). The lipid peroxidation product, malonyldialdehyde, of the host red cells increased gradually to more than fourfold in schizont-rich cells as compared with normal erythrocytes (p<0.001). The hemoglobin release from cultured cells was significantly higher in all parasitized red cell cultures as well as in uninfected cells kept in in vitro, as compared with normal erythrocytes. The consequence of such changes induced by the malarial parasites in the host red cells in the pathogenesis of erythrocyte destruction and anemia ofP. falciparum malaria is discussed.     

2018年全国内脏利什曼病疫情分析

目的了解2018年全国内脏利什曼病的疫情状况。方法收集2018年中国疾病预防控制中心传染病报告信息管理系统中全国内脏利什曼病病例信息,剔除疑似病例、重复病例以及皮肤利什曼病病例,建立数据库,采用Microsoft Excel 2016软件对内脏利什曼病病例的三间分布进行描述性流行病学分析。结果2018年全国11个省份的78个县共报告内脏利什曼病180例,病例主要分布于甘肃(66例)、山西(38例)和陕西(27例)等3个省。其中40个县属于流行区,共报告本地感染病例134例,其余38个县属于非流行区,共报告输入性病例46例。其中甘肃省舟曲县(22例)、宕昌县(18例)和武都区(10例)为主要流行县,报告病例占全国总报告病例数的27.8%(50/180)。陕西省渭南市化州区和临渭区再现本地感染病例,为内脏利什曼病复燃流行县。内脏利什曼病发病高峰为4-6月,男女病例比为1∶0.7。农民和儿童是我国内脏利什曼病高风险人群,分别占全部病例数的42.2%(76/180)和34.5%(55/180)。内脏利什曼病病例主要分布于≥15岁年龄组,不同流行类型病例年龄分布明显不同,野生动物源型病例主要分布于0~2岁婴幼儿,人源型与犬源型病例主要分布于≥15岁农民。结论我国内脏利什曼病呈低度流行态势,但流行区范围逐渐蔓延。     

阿图什市2005-2011年内脏利什曼病病例流行病学分析

目的 了解新疆阿图什市内脏利什曼病流行现状.方法 通过疾病疫情监测信息网报告系统获得2005- 2011年阿图什市内脏利什曼病病例信息,使用SPSS12.0软件统计分析病例的时间、空间和人群分布.结果 2005-2011年阿图什市共发生内脏利什曼病病例66例,各年的病例数依次为19、15、11、4、8、7和2例.全市7个乡和市区均有病例发生,但主要发生在阿扎克乡(45.45％)、松塔克乡(19.70％)和市区(15.15％).各年龄组均有病例发生,但以15岁及以下年龄为主(77.37％).男女病例之比为1.54∶1.结论 阿图什市内脏利什曼病发病率呈逐年下降之势     

Interleukin 10 receptor blockade--pentavalent antimony treatment in experimental visceral leishmaniasis

Interleukin 10 (IL-10), a suppressive Th2 cell-type cytokine, promotes disease progression in experimental visceral leishmaniasis. To extend testing the therapeutic effects of applying IL-10 receptor (IL-10R) blockade with antileishmanial chemotherapy, BALB/c mice with established intracellular Leishmania donovani infection were injected once with anti-IL-10R mAb at the time low-dose, daily pentavalent antimony (Sb) therapy was initiated. In this treatment model, simultaneous administration of anti-IL-10R enhanced overall antileishmanial activity in the liver in an interferon-gamma-dependent fashion, and accelerated the kinetics of Sb-associated killing, induced a >10-fold Sb dose-sparing effect and shortened the required duration of Sb treatment. These results suggest the possibility of using mAb-induced IL-10R blockade to develop low-dose and/or short-course immunochemotherapeutic regimens in visceral leishmaniasis.     

Antigenuria in visceral leishmaniasis: detection and partial characterisation of a carbohydrate antigen

The detection of antigen in the urine is increasingly being used for diagnosis of parasitic infections. A urinary antigen has recently been demonstrated in visceral leishmaniasis (VL), using a latex agglutination test. The results of our study show that the detected antigen is: heat-stable, precipitates with acetone and ethanol but not TCA, is sensitive to periodate and acid hydrolysis but not to pronase E, lipase, or neuraminidase. The antigen is a low molecular weight glycoconjugate that can be extracted by phenol-water, partitions into the aqueous phase when extracted with Triton X-114 or chloroform/methanol, and can be labelled by biotin hydrazide. Since this urinary antigen cannot be characterised by conventional SDS-PAGE and Western blotting, we used an affinity transfer blotting system in which antigens were captured onto nitro-cellulose paper previously coated with a specific antibody. Using this system a low molecular weight antigen (LMWA) spanning an area of the nitro-cellulose membrane corresponding to molecular weight of 5-20 kDa was detected in the urine of VL patients (from Nepal, Sudan, Brazil, Yemen and Spain) and of experimentally infected animals. No LMWA was detected in the urine of patients with malaria, schistosomiasis, or other nonparasitic diseases including typhoid and brucellosis. Immunoprecipitation, using antibody-coated latex, followed by immunoblotting showed that the LMWA is the target antigen in the previously described latex agglutination test ('KATEX'). The antigen is detectable in both the promastigote and amastigote stages of the parasite. Monoclonal antibodies (mAbs) against Leishmania glycoconjugates strongly react with this molecule. These results suggest that the detected antigen is highly specific and diagnostic for VL.     

我国非流行省内脏利什曼病病例流行病学分析

目的 了解2004-2010年我国非流行省区输入性内脏利什曼病病例状况.方法 从中国疾病疫情监测信息网获取非流行省报告的内脏利什曼病病例信息,以电话、信函等形式进行个案调查,对获取的资料描述性分析其地区,时问和人群性别、年龄、职业分布情况.结果 2004-2010年间,全国共有13个非流行省区有输入性内脏利什曼病病例报...     

新疆伽师县内脏利什曼病媒介白蛉调查

目的调查新疆伽师县的白蛉种类、季节消长等生态学特性。方法 2009年5月中旬至9月上旬,在伽师县卧里托克拉克乡的居民点和野外采集白蛉,鉴定后计算蛉种组成和性别比例,统计白蛉数量和密度,观察白蛉通宵活动情况。解剖白蛉,分析雌蛉生殖营养周期,检查白蛉体内利什曼原虫自然感染情况;以油纸、捕蛉罩等方法调查白蛉栖息地。结果捕获白蛉4540只,其中99.9%为吴氏白蛉,微小司蛉新疆亚种仅占0.1%。人房处在5月底6月初出现第1个密度高峰,8月中旬出现第2个密度高峰。白蛉的活动主要集中在22:00～次日4:00,午夜0:00达到活动最高峰。生殖营养周期分析表明吴氏白蛉为野生野栖蛉种,主要在户外吸血,有较强的嗜人血习性;在4只白蛉体内查到内脏利什曼原虫前鞭毛体,野外(3只)的自然感染率高于人房内(1只)。栖息地调查发现畜圈内墙壁缝隙等处是白蛉的栖息地。结论新疆伽师县的优势蛉种为吴氏白蛉,其每年有2个世代,主要栖息在各种隐蔽、恒温的洞穴和缝隙中。     

2005～2009年我国内脏利什曼病报告病例的流行病学特征分析

目的明确我国2005～2009年内脏利什曼病流行病学特征,为当前内脏利什曼病防控提供参考数据。方法以中国疾病预防控制中心疾病监测信息管理系统报告的内脏利什曼病病例资料为基础,描述和分析2005～2009年我国内脏利什曼病的地区分布以及不同流行病学类型的利什曼病发病时间和年龄分布特点。结果 (1)2005～2009年我国内脏利什曼病疫情有上升趋势,报告病例数有所增加,其中新疆喀什地区迦师县于2008年出现暴发疫情,报告病例数较上年上升159%;(2)流行范围仍较为集中,报告病例主要分布在我国西部新疆、甘肃和四川3省,3省报告病例数占全国报告病例总数的97.8%,且3省报告病例分别集中在本省少数县市,但出现报告病例县数增加;(3)犬源型(甘肃)、人源型(喀什市和疏附县)和野生动物源型(迦师县)利什曼病病例年龄分布不同(P0.05),野生动物源型病例79%在1岁以内,人源型病例仅0.7%在1岁以内,3～4岁年龄组为发病高峰,1岁以内为犬源型发病高峰年龄段,随年龄增加报告病例数逐渐减少;(4)2岁及以上病例多发生在每年的3～6月份,2岁以下病例(迦师县除外)发生无明显季节性,二者差异有统计学意义(P0.05);迦师县暴发疫情以当年9月份至次年2月份为发病高峰。结论近年我国内脏利什曼病疫情有上升趋势,不同流行病学类型利什曼病发病特点明显不同。应针对不同流行区疫情特点,采取适宜措施,加强我国西部地区内脏利什曼病防治,控制疫情暴发和流行。同时,有必要进一步研究各地内脏利什曼病流行因素和流行特点。     

快速检测内脏利什曼病特异抗体胶体金免疫层析试条方法的建立与效果评价

目的 建立快速、简便地检测内脏利什曼病特异抗体的胶体金免疫层析试条方法,并评价效果. 方法 采用柠檬酸三钠还原法制备胶体金,用以标记链球菌G蛋白（streptococcal protein G,SPG）,并将其吸附于交联释放垫上;将杜氏利什曼原虫前鞭毛体粗抗原作为包被抗原,包被于硝酸纤维素膜适当位置,制成检测特异抗体的免疫层析试条.用该试条检测病原学确诊的内脏利什曼病（129例）、疟疾（20例）、细粒棘球蚴病（10例）、日本血吸虫病（10例）、并殖吸虫病（5例）、华支睾吸虫病（5例）等患者血清,以及健康者（40例）血清,评价其敏感性和特异性.同时用酶联免疫吸附试验（enzyme-linked immunosorbent assay,ELISA）进行平行检测. 结果 试条法检测129份黑热病患者血清,124份为阳性,敏感性为96.1％;与疟疾患者血清存在10.0％ （2/20）的交叉反应;与日本血吸虫病患者血清存在10.0％ （1/10）的交叉反应;与健康者血清存在2.5％ （1/40）的假阳性反应;与10份细粒棘球蚴病患者血清,5份并殖吸虫病患者血清和5份华支睾吸虫病患者血清均无交叉反应,总特异性为95.6％ （86/90）.该方法与ELISA法的符合率为99.2％,二者阳性检出率之间的差异无统计学意义（x2=0.12,P＞0.05）,二者特异性之间的差异也无统计学意义（x2=0.42,P＞0.05）. 结论 成功建立快速检测内脏利什曼病的胶体金免疫层析试条,该试条敏感性、特异性均较高.     

内脏利什曼病合并HIV感染患者诊断和治疗研究进展

内脏利什曼病是全球被忽视的传染病之一,危害严重。而利什曼原虫-人类免疫缺陷病毒(human immunodeficiency virus,HIV)合并感染对流行地区造成的威胁更甚。利什曼原虫与HIV存在相互作用,合并感染者在临床表现、诊断及治疗方面具有一定特殊性,其病死率及复发率均高于HIV阴性的内脏利什曼病患者。本文对利什曼原虫-HIV合并感染患者的临床表现、诊断和治疗进展进行综述。     

新疆喀什地区人群内脏利什曼病血清流行病学调查

目的通过对新疆喀什人源型内脏利什曼病流行区人群进行血清流行病学调查,掌握当地人群内脏利什曼病的感染状况和影响因素。方法在喀什地区伯什克拉姆乡的3个大队采用系统抽样的方法抽取住户,采用Ka24单抗抑制ELISA法测定居民血清抗利什曼前鞭毛体可溶性抗原Ka24抗体。结果调查1642人,人群内脏利什曼病血清学阳性302人,阳性率为18．39％;既往利什曼病病人血清学阳性率为48．76％（59／121）,无病史者阳性率为15．98％（（243／1521））,差异有统计学意义（Х^2=80．26,P〈0．01）。距发病时间越近,检测的阳性率越高（Х^2=11．55,P〈0．01）。而不同性别、不同职业和不同年龄利什曼病患者血清学阳性率差异无统计学意义（P〉0．05）。结论新疆喀什人群利什曼病血清学阳性率较高,该地区仍为利什曼病流行较为严重的地区,需加强防治力度。     

Central role of interleukin-15 in human immunodeficiency virus (HIV)-infected patients with visceral leishmaniasis

To evaluate clinical and immunological parameters, interleukin (IL)-15 production and outcome of patients with visceral leishmaniasis (VL), including HIV positive patients, we analyzed 48 cases of VL. Clinical manifestations and response to therapy were similar in VL/HIV- and VL/HIV+ patients. However, relapses were more frequent in patients with HIV infection. Low levels of IL-15 concentrations were found in HIV+ patients without VL. These levels were comparable to concentrations obtained in healthy donors. We found a relationship between response to therapy and IL-15 levels. We found increased levels of IL-15 in VL/HIV- and VL/HIV+ patients with clinical and parasitological response to therapy. Our data demonstrate that VL in HIV-infected patients occurs in subjects with severe immunodeficiency and presents high rate of relapses. Low levels of IL-15 in illness patients and restored production in cured persons suggest that this cytokine could play a central role in immune responses during Leishmania/HIV co-infection.     

河南省1例输入性内脏利什曼病的实验室诊断

目的　分析河南省1例输入性内脏利什曼病病例,探讨内脏利什曼病的实验室诊断方法。方法　分析患者的流行病学资料和临床资料,镜检观察骨髓涂片中杜氏利什曼原虫无鞭毛体;rK39试纸条检测血清中利什曼原虫抗体;用两对引物K13A?K13B和 LITSR?L5.8S分别扩增利什曼原虫动基体DNA和核糖体DNA内转录间隔区基因片段。结果　 该患者曾去过内脏利什曼病流行区,有不规则发热、脾肿大、全血细胞减少、白蛋白/球蛋白比例倒置等症状,骨髓涂片查见杜氏利什曼原虫无鞭毛体,rK39试纸条检测阳性,两对引物K13A?K13B和 LITSR?L5.8S分别扩增出87 bp和285 bp的片段。两片段序列与杜氏利什曼原虫相应序列的相似性分别为94%和100%。结论　结合患者的流行病学资料和临床表现以及实验室检测结果,确诊该病例为内脏利什曼病病例,病原体为杜氏利什曼原虫。     

Drug policy for visceral leishmaniasis: a cost-effectiveness analysis

OBJECTIVE: To facilitate the choice of the best visceral leishmaniasis (VL) treatment strategy for first-line health services in (VL)-endemic areas, we compared in a formal decision analysis the cost and the cost-effectiveness of the different available options. METHODS: We selected four drug regimens for VL on the basis of frequency of use, feasibility and reported efficacy studies. The point estimates and the range of plausible values of effectiveness and cost were retrieved from a literature review. A decision tree was constructed and the strategy minimizing the cost per death averted was selected. RESULTS: Treatment with amphotericin B deoxycholate was the most effective approach in the baseline analysis and averted 87.2% of all deaths attributable to VL. The least expensive and the most cost-effective treatment was the miltefosine regimen, and the most expensive and the least cost-effective was AmBisome treatment. The cost of drug and medical care are the main determinants of the cost-effectiveness ranking of the alternative schemes. Sensitivity analysis showed that antimonial was competitive with miltefosine in the low-resistance regions. CONCLUSION: In areas with >94% response rates to antimonials, generic sodium stibogluconate remains the most cost-effective option for VL treatment, mainly due to low drug cost. In other regions, miltefosine is the most cost-effective option of treatment, but its use as a first-line drug is limited by its teratogenicity and rapid resistance development. AmBisome in mono- or combination therapy is too expensive to compete in cost-effectiveness with the other regimens.     

河南省1例输入性内脏利什曼病的实验室诊断

目的　分析河南省1例输入性内脏利什曼病病例，探讨内脏利什曼病的实验室诊断方法。方法　分析患者的流行病学资料和临床资料，镜检观察骨髓涂片中杜氏利什曼原虫无鞭毛体；rK39试纸条检测血清中利什曼原虫抗体；用两对引物K13A?K13B和 LITSR?L5.8S分别扩增利什曼原虫动基体DNA和核糖体DNA内转录间隔区基因片段。结果　 该患者曾去过内脏利什曼病流行区，有不规则发热、脾肿大、全血细胞减少、白蛋白/球蛋白比例倒置等症状，骨髓涂片查见杜氏利什曼原虫无鞭毛体，rK39试纸条检测阳性，两对引物K13A?K13B和 LITSR?L5.8S分别扩增出87 bp和285 bp的片段。两片段序列与杜氏利什曼原虫相应序列的相似性分别为94%和100%。结论　结合患者的流行病学资料和临床表现以及实验室检测结果，确诊该病例为内脏利什曼病病例，病原体为杜氏利什曼原虫。     

Spatial analysis for identification of priority areas for surveillance and control in a visceral leishmaniasis endemic area in Brazil

Spatial analysis of epidemiological data may be used to assist in the implementation of surveillance and control measures against visceral leishmaniasis (VL) in endemic areas. This ecological study aimed to identify priority areas for surveillance and control of VL in São Luís, the capital of the state of Maranhão in northeast Brazil, a highly endemic area for the disease. We evaluated the spatial structure of the incidence rates of human VL and of the mean number of human and canine cases occurring between 2005 and 2007 in 355 neighborhoods (aggregated into 203 geographical analytical units) within the municipality. The presence of spatial autocorrelation was explored using global and local Moran's I statistics. A local indicator of spatial autocorrelation was used to generate maps for the identification of VL clusters. The global Moran's I index revealed a weak, but statistically significant spatial autocorrelation for human VL incidence rates (I = 0.138). A total of 43 geographical analytical units, encompassing 121 neighborhoods, were identified as priority areas for implementing surveillance and control actions. For the purpose of defining an action plan for the delivery of these measures, those 16 geographical analytical units (encompassing 54 neighborhoods) identified as clusters with high incidence rates of human VL should receive the highest priority. An additional nine geographical analytical units (comprising 28 neighborhoods) showed non-significant clustering of high rates of human, and might be considered as the next priority for VL management. Finally, a further 18 geographical analytical units (covering 39 neighborhoods) had records of coexisting human and canine VL cases during the study period, and these should receive priority attention when resources become available. Spatial data analysis is a valuable tool for defining priority areas for VL surveillance in high transmission areas contributing to a more effective management of financial and technical resources, increasing the sustainability and efficiency of control efforts.     

Relapse of visceral leishmaniasis in an HIV-infected patient successfully treated with a combination of miltefosine and amphotericin B

The present report documents a 49-year-old HIV-infected man receiving antiretroviral therapy with a suboptimal immune response and a CD4 count of 95 cells/mm³, despite virological suppression. Investigation of bone marrow was conducted and yielded a diagnosis of visceral leishmaniasis. The clinical course was complicated by gastrointestinal involvment and relapse occurred after amphotericin B therapy. With the addition of miltefosine, the patient no longer presented with bone marrow amastigotes, and displayed an increased CD4 count and negative Leishmania polymerase chain reaction results. The present case highlights atypical presentation of visceral leishmaniasis, including poor immune reconstitution and gastrointestinal involvement. The high likelihood of relapse and response to combination therapy are illustrated.     

Entomological investigation following the resurgence of human visceral leishmaniasis in southern Algeria

Visceral and cutaneous leishmaniasis are the main endemic vector born diseases in Algeria. In the Hoggar region (extreme south of the country) human visceral leishmaniasis (HVL) is known to be sporadic but during the last decade the number of cases has increased significantly. In 2010, a peak of HVL cases was registered mostly among children. Therefore an entomological survey and a retrospective study on HVL cases were carried out in order to explore the transmission of the disease. Among the sand fly caught Phlebotomus bergeroti was the most frequent species (68%) followed by Sergentomyia schwetzi (22%). In this work we describe the presence of Phlebotomus (Paraphlebotomus) kazeruni for the first time in the Hoggar region.