Streptococcus equi

马链球菌
  • 文章类型: Journal Article
    这项研究描述了马氏链球菌亚种动物流行病感染的爆发,该感染引起了与巴西东北部未经巴氏消毒的牛奶消费有关的脑膜脑炎和菌血症。进行了流行病学调查和简短的文献综述。在这些病例之前,巴西尚未发现可能具有神经嗜性的菌株;然而,2023年,又一例由马链球菌引起的脑膜脑炎。描述了动物流行病,揭示了维持监测的必要性,并强调这些嗜神经菌株继续在环境中传播。
    This study describes an outbreak of Streptococcus equi subspecies zooepidemicus infections that caused meningoencephalitis and bacteremia related to unpasteurized milk consumption in northeastern Brazil. Epidemiological investigations and a brief literature review were conducted. Strains with possible neurotropism had not been identified in Brazil before these cases; however, in 2023, another case of meningoencephalitis caused by Streptococcus equi sp. zooepidemicus was described, revealing the need to maintain surveillance and highlighting that these neurotropic strains continue to circulate in the environment.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    勒死,由马氏链球菌亚种引起的,是一种高度传染性的马病,会导致重大的健康问题和经济损失。该研究的目的是检测从怀疑有strangles的马分离出的马链球菌中SeM基因的存在。2022年7月至12月,在Gondar中部地区的五个地区进行了横断面研究设计,埃塞俄比亚。从临床怀疑的动物身上采集了160个拭子样本。使用聚合酶链反应检测SeM基因,采用Kirby-Bauer椎间盘扩散法进行抗菌药物敏感性试验。采用二元logistic回归模型进行统计显著性检验。在31.87%(51/160)的样品中,分离出马链球菌,31.37%(16/51)的物种携带SeM基因。有大量的四环素(81.5%),红霉素(81.5%),16株对万古霉素(75.5%)耐药。在共用饲料容器的动物中更可能存在绞死(AOR=7.59;95%CI=1.44-39.93),从相同的水槽中饮用(AOR=7.74;95%CI=1.44-41.01),并在一起过夜(AOR=5.97;95%CI1.41-25.37)。这项研究的结果表明,研究区域含有马氏链球菌亚种。共享饲料容器和水槽是扼杀感染的潜在来源;因此,这些容器需要定期清洁。
    Strangles, caused by Streptococcus equi subspecies equi, is a highly infectious disease of equines causing major health issues and financial losses. The aim of the study was to detect the presence of the SeM gene in Streptococcus equi isolated from equine suspected of having strangles. A cross-sectional study design was conducted from July to December 2022 in five districts of the central Gondar zone, Ethiopia. One-hundred sixty swab samples were taken from animals that had been clinically suspected. The SeM gene was detected using polymerase chain reaction, and the antimicrobial susceptibility test was performed using the Kirby-Bauer disc diffusion method. The binary logistic regression model was employed to test for statistical significance. In 31.87% (51/160) of the samples, Streptococcus equi species were isolated, and 31.37% (16/51) of these species carried the SeM gene. There was a significant amount of tetracycline (81.5%), erythromycin (81.5%), and vancomycin (75.5%) resistance among the 16 isolates. Strangles were more likely to be present in animals who shared feed containers (AOR = 7.59; 95% CI = 1.44-39.93), drank from the same water troughs (AOR = 7.74; 95% CI = 1.44-41.01), and spent the night together (AOR = 5.97; 95% CI 1.41-25.37). The findings of this study showed that the research areas harboured Streptococcus equi subspecies equi. Sharing feed containers and water troughs were potential sources of strangles infection; thus, these containers need to be cleaned regularly.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    马链球菌亚种。动物流行病(S.动物流行病,SEZ)是一种重要的人畜共患细菌病原体,可引起各种炎症,比如脑膜炎,心内膜炎,和肺炎。UDP-葡萄糖脱氢酶(hasB)在合成SEZ毒力因子透明质酸胶囊中是必不可少的。我们的研究通过使用构建的胶囊缺陷型突变株(称为ΔhasB株)研究了hasB对小鼠对SEZ的反应的感染。这种缺陷与感染后小鼠血液和腹膜灌洗液(PLF)中SEZ细菌负荷减少有关。此外,与野生型(WT)SEZ菌株相比,ΔhasBSEZ菌株在体内表现出更高的中性粒细胞浸润和释放无细胞DNA(cfDNA)的倾向。体外实验进一步表明,ΔhasBSEZ更有效地诱导了含有组蛋白3(H3)的中性粒细胞胞外陷阱(NETs)的形成,中性粒细胞弹性蛋白酶(NE),和DNA,比它的WT对应的。此外,在感染过程中,NETs的释放被确定为GasderminD(GSDMD)依赖性。一起来看,这些发现强调了SEZ中hasB基因的缺乏导致嗜中性粒细胞的GSDMD依赖性NET释放增强,从而降低SEZ在感染期间抵抗NET介导的根除的能力。我们的发现为针对经济特区的创新治疗策略的发展铺平了道路。
    Streptococcus equi subsp. zooepidemicus (S. zooepidemicus, SEZ) is an essential zoonotic bacterial pathogen that can cause various inflammation, such as meningitis, endocarditis, and pneumonia. UDP-glucose dehydrogenase (hasB) is indispensable in synthesizing SEZ virulence factor hyaluronan capsules. Our study investigated the infection of hasB on mice response to SEZ by employing a constructed capsule-deficient mutant strain designated as the ΔhasB strain. This deficiency was associated with a reduced SEZ bacterial load in the mice\'s blood and peritoneal lavage fluid (PLF) post-infection. Besides, the ΔhasB SEZ strain exhibited a higher propensity for neutrophil infiltration and release of cell-free DNA (cfDNA) in vivo compared to the wild-type (WT) SEZ strain. In vitro experiments further revealed that ΔhasB SEZ more effectively induced the formation of neutrophil extracellular traps (NETs) containing histone 3 (H3), neutrophil elastase (NE), and DNA, than its WT counterpart. Moreover, the release of NETs was determined to be gasdermin D (GSDMD)-dependent during the infection process. Taken together, these findings underscore that the deficiency of the hasB gene in SEZ leads to enhanced GSDMD-dependent NET release from neutrophils, thereby reducing SEZ\'s capacity to resist NETs-mediated eradication during infection. Our finding paves the way for the development of innovative therapeutic strategies against SEZ.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    strangles是一种全球普遍存在的,马链球菌亚种引起的常见和重要的传染病。equi.我们对从日本边境的进口马收集的19种马匹分离株进行了全基因组测序。在这些分离物中,从临床病例中获得15个分离株,从亚临床病例中获得4个分离株。通过与出口国相对应的核心基因组单核苷酸多态性分析,将19个分离株分为3个群体结构贝叶斯分析(BAPS)组,SeM打字,或者是马的出口商。无论进口马的临床状况如何,这19个分离株都具有相同的致病基因,并且没有抗菌抗性基因。马的疾病状态可能反映了亚临床感染的动物先前暴露于马氏链球菌。
    Strangles is a globally widespread, commonly diagnosed and important infectious disease of equids caused by Streptococcus equi subsp. equi. We performed whole genome sequencing of 19 S. equi isolates collected from imported horses at the Japanese border. Of these isolates, 15 isolates were obtained from clinical cases and 4 were from subclinical cases. The 19 isolates were grouped into 3 Bayesian analysis of population structure (BAPS) groups by the core genome single nucleotide polymorphism analysis corresponding to exporting country, SeM typing, or exporter of the horses. The 19 isolates possessed same pathogenic genes regardless of clinical status in imported horses and no antimicrobial resistance genes. The disease status of the horses may rather reflect the prior exposure of animals with sub-clinical infection to S. equi.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Case Reports
    马链球菌亚种兽疫(S.动物流行病)是一种主要感染马的人畜共患病原体,猪,还有狗.虽然罕见,它还被证明会感染食用未经巴氏消毒的乳制品或直接接触马的人。这里,我们介绍了一例没有明确传播方式的患者的人畜疫病菌血症。一名86岁男性,有冠状动脉疾病病史,射血分数降低的心力衰竭,完全心脏传导阻滞状态,起搏器后,高血压,高脂血症,和2型糖尿病因发烧和发冷而提交急诊科。他发烧和僵硬三天,但否认减肥,咳嗽,喉咙痛,或皮疹。在急诊室,生命体征显示发烧101.2华氏度,心率110,其他生命体征稳定。除了心动过速,体检没有什么异常,实验室工作显示没有白细胞增多,但炎症标志物升高和乳酸升高。胸部计算机断层扫描,腹部,骨盆没有发现任何感染源。血液培养物增加了动物疫病,并咨询了传染病小组,病人开始服用青霉素G。由于担心起搏器导线感染性心内膜炎,进行了经胸和经食道超声心动图检查,没有显示瓣膜植被。重复血培养显示感染已清除,患者最终使用阿莫西林出院。虽然我们的患者否认食用未经巴氏消毒的乳制品或与马直接接触,经过进一步的询问,他确实支持偶尔与马互动的家庭成员。这种情况是有价值的,因为它增加了关于特别是在人类中的兽疫杆菌感染的稀疏文献。当明确的感染源不容易识别时,广泛的历史记录至关重要。还需要进一步的研究来更好地了解这种细菌的各种传播方式,以更好地瞄准和警告那些感染风险增加的人。
    Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) is a zoonotic pathogen that primarily infects horses, pigs, and dogs. Although rare, it has also been shown to infect humans who consume unpasteurized dairy food or have direct contact with horses. Here, we present a case of S. zooepidemicus bacteremia in a patient without a clear mode of transmission. An 86-year-old male with a past medical history of coronary artery disease, heart failure with reduced ejection fraction, complete heart block status post pacemaker, hypertension, hyperlipidemia, and type 2 diabetes mellitus presented to the Emergency Department with fever and chills. He had fevers and rigors for three days but denied weight loss, cough, sore throat, or rashes. In the Emergency Department, vital signs revealed a fever of 101.2 degrees Fahrenheit and a heart rate of 110 with other stable vital signs. The physical exam was unremarkable except for tachycardia, and laboratory work revealed no leukocytosis but elevated inflammatory markers and elevated lactate. Computed tomography of the chest, abdomen, and pelvis did not reveal any source of infection. Blood cultures grew S. zooepidemicus and the Infectious Diseases team was consulted, who started the patient on Penicillin G. Due to concern for pacer-lead infective endocarditis, transthoracic and transesophageal echocardiograms were performed, which did not show valvular vegetations. Repeat blood cultures showed clearance of the infection, and the patient was ultimately discharged on amoxicillin. While our patient denied consuming unpasteurized dairy products or having direct contact with horses, upon further questioning, he did endorse family members who occasionally interacted with horses. This case is valuable as it adds to the sparse literature on S. zooepidemicus infections specifically in humans. Extensive history taking is of utmost importance when a clear source of infection is not easily identifiable. Further research is also needed to better understand the various modes of transmission of this bacterium to better target and caution those at an increased risk of infection.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    在这项研究中,我们使用不同的底物和马铃薯皮废物(PPW)作为负担得起的底物,检查了兽疫链球菌菌株MW26985产生的透明质酸(HA)。首先,培养基成分,包括碳源和氮源,对细菌HA生产进行了优化。五种不同的碳源(葡萄糖,蔗糖,乳糖,西米淀粉,和马铃薯淀粉,浓度为30g/L)和三种不同的氮源(蛋白胨,酵母提取物,还有硫酸铵,在10g/L)的浓度下进行了研究。葡萄糖,在碳源中,和酵母提取物,在氮源中,产生的HA最多,测定为1.41g/L。之后,马铃薯皮糖通过稀酸和酶水解提取,然后用作动物疫病链球菌生长的经济有效的碳源。根据结果,发酵过程通过酸水解从马铃薯皮糖中产生0.59g/LHA,通过酶水解释放出0.92g/LHA。用葡萄糖作为额外的碳源补充两种水解产物,使用酸性和酶水解产物将HA产量提高到0.95g/L和1.18g/L。分别。采用十六烷基三甲基溴化铵(CTAB)比浊法对发酵液中HA浓度进行比色法评价。此外,通过傅里叶变换红外(FTIR)光谱观察到的峰证实胞外多糖(EPS)由HA组成。这些观察结果表明,马铃薯皮残留物可以作为一种新型的碳源,用于动物疫病链球菌经济生产HA。
    In this research, we examined the production of hyaluronic acid (HA) by Streptococcus zooepidemicus strain MW26985 using different substrates and potato peel waste (PPW) as an affordable substrate. First, culture medium components, including carbon and nitrogen sources, were optimized for bacterial HA production. Five different carbon sources (glucose, sucrose, lactose, sago starch, and potato starch, at a concentration of 30 g/L) and three distinct nitrogen sources (peptone, yeast extract, and ammonium sulfate, at a concentration of 10 g/L) were investigated. Glucose, among the carbon sources, and yeast extract, among nitrogen sources, produced the most HA which was determined as 1.41 g/L. Afterward, potato peel sugars were extracted by dilute acid and enzymatic hydrolysis and then employed as a cost-effective carbon source for the growth of S. zooepidemicus. Based on the results, the fermentation process yielded 0.59 g/L HA from potato peel sugars through acid hydrolysis and 0.92 g/L HA from those released by enzymatic hydrolysis. The supplementation of both hydrolyzates with glucose as an additional carbon source enhanced HA production to 0.95 g/L and 1.18 g/L using acidic and enzymatic hydrolyzates, respectively. The cetyltrimethylammonium bromide (CTAB) turbidimetric method was used to evaluate the concentration of HA in the fermentation broth using the colorimetric method. Also, the peaks observed by Fourier transform infrared (FTIR) spectroscopy confirmed that the exopolysaccharide (EPS) was composed of HA. These observations demonstrate that potato peel residues can be a novel alternative as a carbon source for the economical production of HA by S. zooepidemicus.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    勒死,一种由马氏链球菌亚种感染引起的疾病(S.equi),是全球特有的,也是最常见的马传染病之一。最近的工作提高了我们对变速器动力学关键参数的认识,但是重要的知识差距仍然存在。我们的目标是应用S.equi传输动力学的数学模型来优先考虑未来的研究领域,并增加传输参数的估计精度,从而提高对S.equi流行病学的理解并量化所需的控制努力。建立了房室确定性模型。在可能的情况下,从当前文献中估算参数值。我们评估了人口规模上基本繁殖数的估计值对以下未知或不确定参数的不同假设的敏感性:(平均)运输持续时间(1/γC),携带者的相对传染性(f),导致运输的感染比例(P),和(平均)自然感染后的免疫持续时间(1/γR)。将可用的发病率和(血清)患病率数据与模型输出进行比较,以改善这些当前未知或不确定的传播相关参数的点估计和范围。估计了在一系列控制情况下预防重大疫情的理想疫苗所需的疫苗接种覆盖率,并比较了现有疫苗的可用数据。承运人的相对传染性(与急病马相比)和承运人的持续时间被确定为关键的知识差距。确定性隔室模拟,结合血清阳性率数据,这表明0.05 Strangles, a disease caused by infection with Streptococccus equi subspecies equi (S. equi), is endemic worldwide and one of the most frequently diagnosed infectious diseases of horses. Recent work has improved our knowledge of key parameters of transmission dynamics, but important knowledge gaps remain. Our aim was to apply mathematical modelling of S. equi transmission dynamics to prioritise future research areas, and add precision to estimates of transmission parameters thereby improving understanding of S. equi epidemiology and quantifying the control effort required. A compartmental deterministic model was constructed. Parameter values were estimated from current literature wherever possible. We assessed the sensitivity of estimates for the basic reproduction number on the population scale to varying assumptions for the unknown or uncertain parameters of: (mean) duration of carriership (1∕γC), relative infectiousness of carriers (f), proportion of infections that result in carriership (p), and (mean) duration of immunity after natural infection (1∕γR). Available incidence and (sero-)prevalence data were compared to model outputs to improve point estimates and ranges for these currently unknown or uncertain transmission-related parameters. The required vaccination coverage of an ideal vaccine to prevent major outbreaks under a range of control scenarios was estimated, and compared available data on existing vaccines. The relative infectiousness of carriers (as compared to acutely ill horses) and the duration of carriership were identified as key knowledge gaps. Deterministic compartmental simulations, combined with seroprevalence data, suggest that 0.05
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    马链球菌。动物流行病(SEZ)是一种重要的病原体,并有助于许多动物物种的各种感染。近年来,一些国家已经报道了由经济特区引起的猪链球菌病暴发。SzM蛋白是一种细胞膜锚定蛋白,它是经济特区的重要毒力因子。SzM蛋白对宿主固有免疫的影响有待进一步研究。这里,获得SEZ的重组SzM(rSzM)蛋白,小鼠腹腔注射rSzM蛋白。我们发现rSzM蛋白可以招募嗜中性粒细胞进入注射部位。在进一步的研究中,分离中性粒细胞并用rSzM蛋白处理,NETs释放由rSzM蛋白独立触发,GSDMD蛋白被促进表达和激活。为了研究GSDMD在NETs形成中的作用,用rSzM蛋白处理从WT小鼠和GSDMD-/-小鼠分离的嗜中性粒细胞。结果表明,GSDMD缺乏抑制了NETs的释放。总之,SEZ的SzM蛋白可以以GSDMD依赖的方式触发NETs释放。
    Streptococcus equissp.zooepidemicus (SEZ) is a crucial pathogen and contributes to various infections in numerous animal species. Swine streptococcicosis outbreak caused by SEZ has been reported in several countries in recent years. SzM protein is a cell membrane-anchored protein, which exhibits as an important virulence factor of SEZ. Effects of SzM protein on host innate immune need further study. Here, recombinant SzM (rSzM) protein of the SEZ was obtained, and mice were intraperitoneally injected with rSzM protein. We discovered that rSzM protein can recruit neutrophils into the injected site. In further study, neutrophils were isolated and treated with rSzM protein, NETs release were triggered by rSzM protein independently, and GSDMD protein was promoted-expressed and activated. In order to investigate the role of GSDMD in NETs formation, neutrophils isolated from WT mice and GSDMD-/- mice were treated with rSzM protein. The results showed that GSDMD deficiency suppressed the NETs release. In conclusion, SzM protein of SEZ can trigger the NETs release in a GSDMD-depending manner.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    绞死是由马链球菌亚种引起的高度传染性的马上呼吸道疾病。马链球菌亚种。equi(S.equi)和马氏链球菌亚种。动物流行病(S.动物流行病)被隔离,作为本地,热,和田间菌株,从临床上患有呼吸窘迫的马。使用细菌学和分子技术鉴定了分离的链球菌。开发并评价了四种灭活马氏链球菌疫苗的制剂。第一个制剂是使用S.equi分离株制备的,用MONTANIDE凝胶佐剂佐剂,而第二种制剂用MONTANIDEISA-70佐剂佐剂。其他2种制剂是从马氏链球菌和兽疫链球菌分离株制备的灭活联合疫苗。第三制剂是用MONTANIDE凝胶佐剂化的组合分离物,而第四制剂是用MONTANIDEISA-70佐剂化的组合分离物。开发的疫苗的物理性质,纯度,不育,安全,和效力得到了保证。使用被动血凝试验在等基因BALB/c小鼠和新西兰白兔中测定免疫效力。此外,采用间接酶联免疫吸附试验,对马驹用MONTANIDEISA-70佐剂化马驹和兽疫链球菌联合疫苗的抗体滴度进行了追踪.在实验室和野外动物模型中使用攻击测试确定开发的疫苗的保护效力。达到75%的保护率。联合疫苗被证明比单价疫苗更有效。此外,与Montanide凝胶相比,MontanideISA-70佐剂具有显著的保护功效.当前的工作正在为strangles引入一种非常有前途的缓解和战略控制解决方案。
    Strangles is a highly contagious disease of the equine upper respiratory tract caused by Streptococcus equi subspecies. Streptococcus equi subsp. equi (S. equi) and Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) was isolated, as local, hot, and field strains, from horses clinically suffering from respiratory distress. The isolated Streptococci were identified using bacteriological and molecular techniques. Four formulations of inactivated S. equi vaccines were developed and evaluated. The first formulation was prepared using the S. equi isolates, adjuvanted with MONTANIDE GEL adjuvant, while the second formulation was adjuvanted with MONTANIDE ISA-70 adjuvant. The other 2 formulations were inactivated combined vaccines prepared from both S. equi and S. zooepidemicus isolates. The 3rd formulation was the combined isolates adjuvanted with MONTANIDE GEL while the 4th formulation was the combined isolates adjuvanted with MONTANIDE ISA-70. The developed vaccines\' physical properties, purity, sterility, safety, and potency were ensured. The immunizing efficacy was determined in isogenic BALB/c mice and white New Zealand rabbits using the passive hemagglutination test. Also, the antibodies\' titer of the combined S. equi and S. zooepidemicus vaccine adjuvanted with MONTANIDE ISA-70 in foals was tracked using an indirect enzyme-linked immunosorbent assay. The protective efficacy of the developed vaccines was determined using a challenge test in both laboratory and field animal models, where a 75% protection rate was achieved. The combined vaccine proved to be more efficacious than the monovalent vaccine. Also, the MONTANIDE ISA-70 adjuvant provided significant protective efficacy than the MONTANIDE GEL. The current work is introducing a very promising mitigative and strategic controlling solution for strangles.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    马链球菌亚种。动物流行病(S.兽疫)是马下生殖道的粘膜共生菌,是引起母马子宫内膜炎的最孤立的细菌。这项研究的目的是确定从布宜诺斯艾利斯省的母马子宫内膜炎中获得的S.ezoopempopicus的分子多样性,阿根廷。研究了2005年和2017年从母马子宫获得的30株分离株。MLST方案用于鉴定阿根廷基因型,并使用eBURST算法-goeBURST鉴定了进化下降的克隆关系和模式。确定了26种不同的序列类型(ST),以前只有11个在马中报道过,来自几种宿主物种和组织。在我们的研究中,其他15种ST是首次在阿根廷的母马繁殖品系中报道的。当分析所有已发表的动物疫病链球菌时,从阿根廷子宫获得的基因型分布并不均匀,因此,不可能确定相同的谱系在我们的马种群中循环。其他国家也报道了确定的基因型,不同的样本和寄主物种表明没有寄主,和解剖学生态位适应。最后,在同一母马的阴道/阴蒂和子宫中分离出相同的基因型,这凸显了人疫链球菌的多功能性及其作为机会病原体的作用.
    Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a mucosal commensal of the lower genital tract in horses and is the most isolated bacterium causing endometritis in mares. The aim of this study was to determine the molecular diversity of S. zooepidemicus obtained from endometritis in mares in Buenos Aires province, Argentina. Thirty isolates obtained from the uterus of mares in 2005 and 2017 were studied. The MLST scheme was applied to identify the Argentinian genotypes and the clonal relationships and patterns of evolutionary descent were identified using the eBURST algorithm - goeBURST. Twenty six different Sequence types (STs) were identified, being only 11 of them previously reported in horses and also, from several host species and tissues. The other 15 STs were reported in Argentinian reproductive strains of mares in our study for the first time. The genotypes obtained from uterus in Argentina were not evenly distributed when all the published S. zooepidemicus STs were analysed, thus, it was not possible to establish that the same lineage circulates in our equine population. The fact that the identified genotypes were also reported in other countries, diverse samples and host species suggest that there is not a host, and an anatomical niche adaptation. Finally, the isolation of the same genotype in the vagina/clitoris and the uterus of the same mare highlights the versatility of S. zooepidemicus and its role as an opportunistic pathogen.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

公众号