Antivenom

抗蛇毒血清
  • 文章类型: Journal Article
    Echisocellatus是尼日利亚最常见的毒蛇之一。抗蛇毒血清是唯一有效的治疗方法,但是该国的有效抗蛇毒血清供应有限。因此,本研究旨在探索有效的可行性,使用尼日利亚的Echisocellatus毒液通过免疫兔子产生单特异性抗体。在免疫中采用世界卫生组织抗蛇毒血清生产指南,并使用蛋白A琼脂糖柱层析纯化所得抗体。通过2个月的免疫接种,抗体滴度达到较高的平台,和血清的SDSPAGE表明存在完整的免疫球蛋白,并伴有重链(50kDa)和轻链(25kDa)。毒液在小鼠中的静脉LD50为0.35mg/kg,并且在2LD50的攻击剂量下的毒液致死性被抗体有效地中和,其效力值为每g抗体0.83mg毒液。抗体还以13±0.66ul的有效剂量(ED)中和了毒液的促凝血活性,支持其用于血液毒性毒害。研究确立了开发有效、针对尼日利亚地毯毒蛇的单特异性抗体。
    Echis ocellatus is one of the commonest snakes responsible for envenomation in Nigeria. Antivenom is the only effective treatment, but the country suffers from a limited supply of effective antivenom. This study therefore aimed to explore the feasibility of effective, mono-specific antibodies production through immunization in rabbits using the venom of Echis ocellatus from Nigeria. The World Health Organization guide on antivenom production was employed in the immunization and the resultant antibodies were purified using protein A agarose column chromatography. Antibody titer reached a high plateau by 2-month immunization, and SDS PAGE of the sera suggests the presence of intact immunoglobulins accompanied with the heavy (50 kDa) and light (25 kDa) chains. The venom has an intravenous LD50 of 0.35 mg/kg in mice, and the venom lethality at a challenge dose of 2 LD50 was effectively neutralized by the antibodies with a potency value of 0.83 mg venom per g antibodies. The antibodies also neutralized the procoagulant activity of the venom with an effective dose (ED) of 13±0.66 ul, supporting its use for hemotoxic envenomation. The study establishes the feasibility of developing effective, mono-specific antibodies against the Nigerian Carpet viper.
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  • 文章类型: Journal Article
    小的单链可变片段(scFv)是有前途的生物分子,可以抑制和中和毒素并充当抗蛇毒血清。在这项工作中,我们的目标是在巴斯德毕赤酵母中产生功能性scFv-6009FV,抑制纯Cn2神经毒素和Centruroidesnoxius的整个毒液。我们能够在烧瓶中获得高达31.6±2mg/L的产量。此外,蛋白质显示6.1%的α-螺旋结构,49.1%β-折叠,和44.8%的无规卷曲由CD。质谱证实了氨基酸序列,并且没有显示该分子的糖基化谱。纯化的scFv-6009FV允许我们在兔子中开发抗scFv,然后将其用于亲和柱中以纯化其他scFvs。测定其半最大抑制浓度值(IC50)比作为对照的由大肠杆菌产生的scFvs好40%。最后,我们发现scFv-6009FV能够体外抑制纯Cn2毒素和小鼠解救实验中来自C.noxius的整个毒液。这些结果表明,在这里分析的条件下,巴斯德毕赤酵母适合生产scFv-6009FV,与大肠杆菌产生的scFvs相比,保持抗体的特性并更有效地中和Cn2毒素。
    Small single-chain variable fragments (scFv) are promising biomolecules to inhibit and neutralize toxins and to act as antivenoms. In this work, we aimed to produce a functional scFv-6009FV in the yeast Pichia pastoris, which inhibits the pure Cn2 neurotoxin and the whole venom of Centruroides noxius. We were able to achieve yields of up to 31.6 ± 2 mg/L in flasks. Furthermore, the protein showed a structure of 6.1 % α-helix, 49.1 % β-sheet, and 44.8 % of random coil by CD. Mass spectrometry confirmed the amino acid sequence and showed no glycosylation profile for this molecule. Purified scFv-6009FV allowed us to develop anti-scFvs in rabbits, which were then used in affinity columns to purify other scFvs. Determination of its half-maximal inhibitory concentration value (IC50) was 40 % better than the scFvs produced by E. coli as a control. Finally, we found that scFv-6009FV was able to inhibit ex vivo the pure Cn2 toxin and the whole venom from C. noxius in murine rescue experiments. These results demonstrated that under the conditions assayed here, P. pastoris is suited to produce scFv-6009FV that, compared to scFvs produced by E. coli, maintains the characteristics of an antibody and neutralizes the Cn2 toxin more effectively.
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  • 文章类型: Journal Article
    已知澳大利亚白斑蛇的毒液具有凝血活性,包括一些具有强的促凝血活性和其他具有抗凝血活性,尽管后者不太为人所知。这项研究调查了澳大利亚蛇毒的抗凝血活性,以及该活性是否被商业蛇毒血清和varespladib(PLA2抑制剂)中和。已完成34种澳大利亚类动物的凝血测定。对五个物种进行了虎蛇蛇毒血清(TSAV)的抗蛇毒血清中和测定,以确定是否存在交叉中和。对于相同的五个物种,也完成了Varespladib中和测定。所有假单胞菌的毒液都有抗凝血活性,除了P.卟啉,是促凝血剂。假单胞菌毒液具有相似的抗凝血效力,从最有效的P.colletti毒液到最不有效的P.butleri毒液。三种Austrelaps(铜头)毒液是第二有效的抗凝剂。还有六条蛇,大头蛇,刺五加,A.南极,Sutasuta,Denisoniadevisi和D.maculata,抗凝血活性较弱,除了与假单胞菌具有相似的抗凝血活性外。虎蛇抗蛇毒血清(1200mU/mL)中和浓度高达1mg/mL的澳大利亚假单胞菌的抗凝血作用。TSAV(1200mU/mL)还中和了P.colletti,D.maculata,A.超级巴士和A.pyrhus毒液的EC50,显示交叉中和。Varespladib中和了5μM的澳大利亚疟原虫毒液的抗凝作用,D.maculata,A.超级巴士和A.pyrhus。我们发现在低浓度的澳大利亚蛇的六个属中存在抗凝血活性,可以被抗蛇毒血清和varespladib完全中和。澳大利亚elapid毒液中的抗凝血活性与具有高PLA2活性而没有促凝血蛇毒丝氨酸蛋白酶的物种有关。
    The venoms of Australasian elapid snakes are known to possess coagulant activity, including some with strong procoagulant activity and others with anticoagulant activity, although the latter are less well known. This study investigates the anticoagulant activity of Australasian elapid snake venoms, and whether this activity is neutralised by commercial snake antivenom and varespladib (PLA2 inhibiting agent). Clotting assays were completed for 34 species of Australasian elapids. Antivenom neutralisation assays with tiger snake antivenom (TSAV) were performed on five species to determine if there was cross-neutralisation. Varespladib neutralisation assays were also completed for the same five species. All Pseudechis species venoms had anticoagulant activity, except P. porphyriacus, which was procoagulant. Pseudechis species venoms had similar anticoagulant potency ranging from the most potent P. colletti venom to the least potent P. butleri venom. The three Austrelaps (copperhead) species venoms were the next most potent anticoagulants. Six further snakes, Elapognathus coronatus, Acanthophis pyrrhus, A. antarcticus, Suta suta, Denisonia devisi and D. maculata, had weaker anticoagulant activity, except for D. maculata which had similar anticoagulant activity to Pseudechis species. Tiger Snake Antivenom (1200mU/mL) neutralised the anticoagulant effect of P. australis for concentrations up to 1 mg/mL. TSAV (1200mU/mL) also neutralised P. colletti, D. maculata, A. superbus and A. pyrrhus venoms at their EC50, demonstrating cross neutralisation. Varespladib neutralised the anticoagulant effect of P. australis venom at 5 μM and for venoms of P. colletti, D. maculata, A. superbus and A. pyrrhus. We found anticoagulant activity to be present in six genera of Australasian snakes at low concentrations, which can be completely neutralised by both antivenom and varespladib. Anticoagulant activity in Australian elapid venoms was associated with species possessing high PLA2 activity without procoagulant snake venom serine proteases.
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  • 文章类型: Journal Article
    Loxosceles属是巴西具有医学重要性的主要蜘蛛属之一。尽管与劳氏有关的事故很严重,只有少数物种被认为在医学上很重要,只有少数物种经历了全面的毒液表征。Loxoscelesamazonica是潜在危险但未充分研究的Loxosceles物种的显着例子。迄今为止,有关亚马逊乳杆菌事故的报道有限,在巴西的北部和东北部地区,与Loxosceles有关的事故正在增加,据报道,亚马逊L.在这项工作中,我们提供了一个互补的生化和免疫学特征的亚马逊乳杆菌毒液,考虑到其最相关的酶活性及其通过当前治疗性抗蛇毒血清的免疫识别和中和作用。此外,构建了富含来自亚马逊乳杆菌毒腺的磷脂酶D(PLD)序列的cDNA文库,随后进行了测序。结果表明,所有测试的抗体都能很好地识别亚马逊蛇毒。它的毒液也显示出蛋白水解,透明质酸酶,和鞘磷脂酶活性。这些活动至少部分被可用的抗蛇毒血清抑制。随着PLDs的cDNA测序,在亚马逊乳杆菌的毒液中鉴定出七个新的推定同工型。这些结果有助于更好地了解合人症的毒液含量和活动,然而研究不足,Loxosceles种。体内测定是必要的,以确认的医学相关性的亚马逊,以及评估其真正的毒性潜力并阐明其相关的病理生理学。
    The Loxosceles genus represents one of the main arachnid genera of medical importance in Brazil. Despite the gravity of Loxosceles-related accidents, just a handful of species are deemed medically important and only a few have undergone comprehensive venom characterization. Loxosceles amazonica is a notable example of a potentially dangerous yet understudied Loxosceles species. While there have been limited reports of accidents involving L. amazonica to date, accidents related to Loxosceles are increasing in the North and Northeast regions of Brazil, where L. amazonica has been reported. In this work, we provide a complementary biochemical and immunological characterization of L. amazonica venom, considering its most relevant enzymatic activities and its immunorecognition and neutralization by current therapeutic antivenoms. Additionally, a cDNA library enriched with phospholipase D (PLD) sequences from L. amazonica venom glands was built and subsequently sequenced. The results showed that L. amazonica venom is well immunorecognised by all the tested antibodies. Its venom also displayed proteolytic, hyaluronidase, and sphingomyelinase activities. These activities were at least partially inhibited by available antivenoms. With cDNA sequencing of PLDs, seven new putative isoforms were identified in the venom of L. amazonica. These results contribute to a better knowledge of the venom content and activities of a synanthropic, yet understudied, Loxosceles species. In vivo assays are essential to confirm the medical relevance of L. amazonica, as well as to assess its true toxic potential and elucidate its related pathophysiology.
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  • 文章类型: Journal Article
    在蛇毒毒性和抗血清效力测试的研究中,建立人道的终点以最大程度地减少动物的痛苦至关重要。我们的发现表明,瑞士小鼠在暴露于不同的蛇毒以及毒液和抗蛇毒血清的组合后表现出早期温度下降,预测以后的死亡率。评估温度,我们可以在接种后3小时内确定,在48小时内无法存活的动物。在这些研究中,将温度作为标准将大大减少动物的痛苦,而不会影响结果。
    Establishing humane endpoints to minimize animal suffering in studies on snake venom toxicity and antivenom potency tests is crucial. Our findings reveal that Swiss mice exhibit early temperature drop following exposure to different snake venoms and combinations of venoms and antivenoms, predicting later mortality. Evaluating temperature we can identify within 3 h post-inoculation, the animals that will not survive in a period of 48 h. Implementing temperature as a criterion would significantly reduce animal suffering in these studies without compromising the outcomes.
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  • 文章类型: Journal Article
    氯化钌(RuCl3)在学术界和工业界广泛用于许多化合物的合成和催化,并且在具有医学应用的各种化合物中被用作关键分子。有趣的是,已证明RuCl3可调节人的血浆凝血,并可作为复合无机抗蛇毒血清的组成部分,在体内和体外中和蛇毒的凝血效应。使用血栓弹力图,这项研究试图确定RuCl3对Crotalusatrox毒液纤溶作用的抑制作用是否可以通过人血浆中的载体成分来调节。毒液在0.9%NaCl中暴露于RuCl3,磷酸盐缓冲盐水(PBS),或含1%二甲基亚砜(DMSO)的0.9%NaCl。RuCl3抑制了毒液介导的血栓形成延迟,凝块生长速度降低,凝块强度降低。PBS和DMSO增强了RuCl3的作用。结论是,虽然Ru基阳离子对毒液活性有显著的抑制作用,含有磷酸根和DMSO的Ru基离子的组合增强RuCl3介导的毒液抑制。指出了进一步的研究,以确定哪些特定的含Ru分子会引起毒液抑制,以及哪些其他无机/有机化合物的组合可能会增强RuCl3的抗蛇毒作用。
    Ruthenium chloride (RuCl3) is widely utilized for synthesis and catalysis of numerous compounds in academia and industry and is utilized as a key molecule in a variety of compounds with medical applications. Interestingly, RuCl3 has been demonstrated to modulate human plasmatic coagulation and serves as a constituent of a compounded inorganic antivenom that neutralizes the coagulopathic effects of snake venom in vitro and in vivo. Using thrombelastography, this investigation sought to determine if RuCl3 inhibition of the fibrinogenolytic effects of Crotalus atrox venom could be modulated by vehicle composition in human plasma. Venom was exposed to RuCl3 in 0.9% NaCl, phosphate-buffered saline (PBS), or 0.9% NaCl containing 1% dimethyl sulfoxide (DMSO). RuCl3 inhibited venom-mediated delay in the onset of thrombus formation, decreased clot growth velocity, and decreased clot strength. PBS and DMSO enhanced the effects of RuCl3. It is concluded that while a Ru-based cation is responsible for significant inhibition of venom activity, a combination of Ru-based ions containing phosphate and DMSO enhances RuCl3-mediated venom inhibition. Additional investigation is indicated to determine what specific Ru-containing molecules cause venom inhibition and what other combinations of inorganic/organic compounds may enhance the antivenom effects of RuCl3.
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  • 文章类型: Journal Article
    目的:分析我国蛇咬伤患者的易感因素。
    方法:以多阶段随机抽样为主要抽样方法,以滚雪球抽样为辅助抽样方法。知识,调查我国居民对蛇咬伤的态度和行为。非参数检验用于比较居民知识的百分比差异,毒蛇咬伤的态度和行为,并采用广义线性回归分析方法对影响因素进行分析,综合分析蛇咬伤患者的易感因素。
    结果:本研究共纳入6338名受试者,其中68.4%是男性,58.6%是农民,工人和服务人员。知识总分中位数,态度,行为为26(22,36)。受伤后治疗不当的患者在患处近端结扎(23.43%),挤压(21.82%),口腔和吸伤(8.74%)。因贫困未住院(1351例),未接受抗蛇毒血清(2068例)。分别为21.32%和32.63%,分别。在4270例注射抗蛇毒血清的患者中,有30.7%在2h内接种疫苗。在去医院治疗的患者中(4987),75.0%在6小时内到达医院;在4,761例拨打急救电话的患者中,37.4%在0.5h内处理。
    结论:中国蛇咬伤患者对蛇咬伤的认识薄弱,医疗意识低,缺乏正确的预防和紧急处理措施,对民间疗法的依赖,住房差等等。此外,我国部分地区存在抗蛇毒血清利用率低、医疗资源分布不合理等问题。应发展多部门和多学科合作,预防和控制蛇咬伤,以减轻蛇咬伤造成的负担。
    OBJECTIVE: To analyze the vulnerability factors of snakebite patients in China.
    METHODS: Multi-stage random sampling was used as the main sampling method and snowball sampling as the auxiliary sampling method. The knowledge, attitude and behavior of snakebite among Chinese residents were investigated. Non-parametric test was used to compare the percentage differences in residents\' knowledge, attitude and behavior of snakebite, and generalized linear regression analysis was used to analyze the influencing factors, and the vulnerability factors of snakebite patients were comprehensively analyzed.
    RESULTS: A total of 6338 subjects were included in this study, of which 68.4% were males, and 58.6% were farmers, workers and service personnel. The median total score of knowledge, attitude, and behavior was 26 (22,36). The patients who were improperly treated after injury were ligation proximal to the affected area (23.43%), squeezing (21.82%), and oral and suction wounds (8.74%). Did not go to hospital due to poverty (1351 cases) and did not receive antivenom (2068 cases). There were 21.32% and 32.63%, respectively. Among 4270 patients injected with antivenom 30.7% were vaccinated within 2 h. Among the patients who went to the hospital for treatment (4987), 75.0% arrived at the hospital within 6 h; Among the 4,761 patients who made emergency calls, 37.4% were treated within 0.5 h.
    CONCLUSIONS: Snakebite patients in China have weak knowledge about snakebite, low awareness of medical treatment, lack of correct prevention and emergency treatment measures, dependence on folk remedies, poor housing and so on. In addition, there are low availability of antivenoms and unreasonable distribution of medical resources in some areas of China. Multisectoral and multidisciplinary cooperation should be developed to prevent and control snakebites in order to reduce the burden caused by snakebites.
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  • 文章类型: Journal Article
    Vipera属包括欧洲大多数具有医学意义的有毒蛇,意大利藏有其中的四个。欧洲毒蛇的毒害会导致严重后果,但是漏报和缺乏标准化的临床方案阻碍了有效的蛇咬伤管理。这项研究提供了最新的,为意大利临床医生量身定制的Vipera蛇咬伤的管理和治疗指南。它包括用于蛇识别的分类键,对毒蛇毒液组成的见解,和临床管理建议。重点是快速可靠地识别与医学相关的蛇种,采取适当的急救措施。概述了抗蛇毒血清管理标准,以及管理潜在副作用的适应症。虽然协议是专门针对意大利的,它的方法可能适用于其他欧洲国家,取决于当地资源。提倡在毒物控制中心之间促进全面的数据收集和合作,以优化毒物管理协议,并改善国家一级有关蛇咬伤的流行病学数据的报告。
    The genus Vipera encompasses most species of medically significant venomous snakes of Europe, with Italy harbouring four of them. Envenomation by European vipers can result in severe consequences, but underreporting and the absence of standardised clinical protocols hinder effective snakebite management. This study provides an updated, detailed set of guidelines for the management and treatment of Vipera snakebite tailored for Italian clinicians. It includes taxonomic keys for snake identification, insights into viper venom composition, and recommendations for clinical management. Emphasis is placed on quick and reliable identification of medically relevant snake species, along with appropriate first aid measures. Criteria for antivenom administration are outlined, as well as indications on managing potential side effects. While the protocol is specific to Italy, its methodology can potentially be adapted for other European countries, depending on local resources. The promotion of comprehensive data collection and collaboration among Poison Control Centres is advocated to optimise envenomation management protocols and improve the reporting of epidemiological data concerning snakebite at the country level.
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  • 文章类型: Journal Article
    这次回顾,观察性研究描述了临床发现,案件管理趋势,以及在大学教学医院环境中暴露于东部珊瑚蛇的83只狗和9只猫的结果。回顾了在珊瑚蛇暴露后接受抗蛇毒血清的狗和猫的医疗记录。收集的数据包括信号,抗蛇毒血清管理时间到了,演示时的物理和实验室特征,住院期间的临床过程,住院时间,生存到出院。从提交到珊瑚蛇抗蛇毒血清管理的平均时间为2.26±1.46小时。不包括所有者拒绝住院护理的情况,狗和猫的平均住院时间为50.8h和34h,分别。抗蛇毒血清小瓶的平均数目为1.29(1-4)。胃肠道症状(呕吐和呕吐)发生在42.2%(35/83)的狗和33.3%(3/9)的猫中。周围神经系统缺陷(共济失调,麻痹至麻痹,无反射,和通气不足)的狗和猫占19.6%(18/92)。溶血在37.9%(25/66)的狗中也很常见,但在猫中没有观察到。12%(10/83)的狗指示机械通气(MV),但没有猫。急性肾损伤(AKI),虽然罕见,是安乐死的常见原因,占20%(2/5),是MV期间最常见的并发症,占44.4%(4/9)。88.9%(8/9)的MV病例和所有AKI病例发生色素尿/溶血。尽管抗蛇毒血清管理延迟了几个小时,接触珊瑚蛇的狗和猫的死亡率较低(6%的狗(5/83)和0%的猫)。胃肠道体征很常见,但不能预测神经系统体征的进展。因此,在神经系统症状出现之前区分珊瑚蛇的暴露和毒液仍然具有挑战性。
    This retrospective, observational study describes the clinical findings, case management trends, and outcomes of 83 dogs and nine cats exposed to eastern coral snakes in a university teaching hospital setting. The medical records of dogs and cats that received antivenom following coral snake exposure were reviewed. Data collected included signalment, time to antivenom administration, physical and laboratory characteristics at presentation, clinical course during hospitalization, length of hospitalization, and survival to discharge. The mean time from presentation to coral snake antivenom administration was 2.26 ± 1.46 h. Excluding cases where the owner declined in-hospital care, the mean hospitalization time for dogs and cats was 50.8 h and 34 h, respectively. The mean number of antivenom vials was 1.29 (1-4). Gastrointestinal signs (vomiting and ptyalism) occurred in 42.2% (35/83) of dogs and 33.3% (3/9) of cats. Peripheral neurologic system deficits (ataxia, paresis to plegia, absent reflexes, and hypoventilation) were noted in 19.6% (18/92) of dogs and cats. Hemolysis was also common in 37.9% (25/66) of dogs but was not observed in cats. Mechanical ventilation (MV) was indicated in 12% (10/83) of dogs but no cats. Acute kidney injury (AKI), while rare, was a common cause of euthanasia at 20% (2/5) and was the most common complication during MV at 44.4% (4/9). Pigmenturia/hemolysis occurred in 88.9% (8/9) of MV cases and in all cases with AKI. Despite delays in antivenom administration by several hours, dogs and cats with coral snake exposure have low mortality rates (6% of dogs (5/83) and 0% of cats). Gastrointestinal signs were common but were not predictive of progression to neurological signs. Thus, differentiating between coral snake exposure and envenomation before the onset of neurological signs remains challenging.
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  • 文章类型: Journal Article
    蛇咬伤在非洲是一个重大的健康问题,特别是由于神经毒性毒液可导致神经肌肉麻痹和呼吸衰竭。在尼日利亚,来自Elapidae家族的蛇是引起毒液感染的主要原因,尽管这些事件被低估了。这篇综述审查了非洲神经毒性毒液的病例报告,强调可用抗蛇毒血清的临床影响和疗效。临床前研究表明,南非医学研究所(SAIMR)的多价抗蛇毒血清对神经毒性非常有效,其保护功效(R)为1346.80mg/mL,而临床评估强调需要大剂量抗蛇毒血清治疗以及机械通气等支持性措施。与出血性毒液不同,抗蛇毒血清迅速解决出血,神经毒性病例通常需要额外的干预措施.该评论强调了在抗蛇毒血清治疗中采用量身定制的方法的必要性,以解决神经毒性蛇咬伤的复杂性并减轻其在非洲的公共卫生负担。
    Snakebite is a significant health concern in Africa, particularly due to neurotoxic envenomation which can lead to neuromuscular paralysis and respiratory failure. In Nigeria, snakes from the Elapidae family are a notable cause of envenomation cases, though these incidents are underreported. This review examined case reports of neurotoxic envenomation in Africa, highlighting the clinical impacts and the efficacy of available antivenoms. Preclinical studies showed that the polyvalent antivenom from the South African Institute for Medical Research (SAIMR) was highly effective against neurotoxicity with a protective efficacy (R) of 1346.80 mg/mL, while clinical assessment emphasized the need for high-dose antivenom therapy along with supportive measures like mechanical ventilation. Unlike hemorrhagic envenomation, where antivenom promptly resolves bleeding, neurotoxic cases often require additional interventions. The review underscores the necessity for tailored approaches in antivenom therapy to address the complexities of neurotoxic snakebites and reduce their public health burden in Africa.
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