Polymerase chain reaction

聚合酶链反应
  • 文章类型: Journal Article
    眼科医生对棘阿米巴角膜炎的诊断和治疗能力差异很大。这个调查员发起的,回顾性,单中心图表回顾检查了有关PCR阳性棘阿米巴角膜炎的电子患者档案.我们包括角膜和隐形眼镜评估。我们进一步回顾了患者的病史,关于病毒或真菌共感染的角膜刮擦结果,以及从症状发作到最终诊断的持续时间。从2010年2月至2023年2月,我们确定了52例患者的59只眼,52例患者中有31例(59.6%)是女性患者。中位数(IQR,范围)患者年龄为33(25.3至45.5[13至90])岁,和平均值(SD,范围)症状发作后诊断时间为18(10.5至35[3至70])天。总的来说,52例患者中有7例(7.7%)出现双侧棘阿米巴感染,48人(92.3%)在症状发作时使用隐形眼镜。关于其他微生物共感染,我们发现52例患者中有45例(86.5%)进行了病毒学PCR检测,3例(6.7%)角膜刮片阳性。52例患者中有49例(94.2%)进行了真菌培养,角膜刮片阳性5例(10.2%)。医疗成功率为45/46(97.8%)。这项研究提高了患者对隐形眼镜处理和筛查可疑棘阿米巴病例中进一步微生物共感染的认识。
    Ophthalmologists\' diagnostic and treatment competence in Acanthamoeba keratitis varies widely. This investigator-initiated, retrospective, single-center chart review examined the electronic patient files regarding PCR-positive Acanthamoeba keratitis. We included corneal and contact lens assessments. We further reviewed the patient\'s medical history, corneal scraping results regarding viral or fungal co-infections, and the duration from symptom onset to final diagnosis. We identified 59 eyes of 52 patients from February 2010 to February 2023, with 31 of 52 (59.6%) being female patients. The median (IQR, range) patient age was 33 (25.3 to 45.5 [13 to 90]) years, and the mean (SD, range) time to diagnosis after symptom onset was 18 (10.5 to 35 [3 to 70]) days. Overall, 7 of 52 (7.7%) patients displayed a bilateral Acanthamoeba infection, and 48 (92.3%) used contact lenses at symptom onset. Regarding other microbiological co-infections, we found virologic PCR testing in 45 of 52 (86.5%) patients, with 3 (6.7%) positive corneal scrapings. Fungal cultures were performed in 49 of 52 (94.2%) patients, with 5 (10.2%) positive corneal scrapings. The medical treatment success rate was 45/46 (97.8%). This study raises awareness of patient education in contact lens handling and screens for further microbial co-infections in suspected Acanthamoeba cases.
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  • 文章类型: Journal Article
    背景:这项荟萃分析检查了聚合酶链反应(PCR)在不同呼吸道样本上诊断肺孢子虫肺炎(PCP)的比较诊断性能,在人类免疫缺陷病毒(HIV)和非HIV人群中。
    方法:共有55篇文章符合纳入标准,包括对7835例有PCP风险的患者的呼吸道标本进行的11434PCR检测。QUADAS-2工具表明所有研究的偏倚风险较低。使用双变量和随机效应荟萃回归分析,针对欧洲癌症研究和治疗组织-真菌病研究小组对已证实的PCP的定义,研究了PCR的诊断性能.
    结果:支气管肺泡灌洗液的定量PCR(qPCR)提供了98.7%的最高合并灵敏度(95%置信区间[CI],96.8%-99.5%),足够的特异性为89.3%(95%CI,84.4%-92.7%),负似然比(LR-)为0.014,正似然比(LR+)为9.19。对诱导痰的qPCR提供了类似的99.0%(95%CI,94.4%-99.3%)的高灵敏度,但降低了81.5%(95%CI,72.1%-88.3%)的特异性,LR-为0.024,LR+为5.30。qPCR对上呼吸道样本的灵敏度较低,为89.2%(95%CI,71.0%-96.5%),高特异性90.5%(95%CI,80.9%-95.5%),LR-为0.120,LR+为9.34。根据患者的HIV状况,PCR的敏感性和特异性没有显着差异。
    结论:在较深的呼吸道标本上,PCR阴性可用于可靠地排除PCP,但PCR阳性可能需要临床解释来区分定植和活动性感染,部分取决于PCR信号的强度(指示真菌负荷),试样类型,和患者人群测试。
    BACKGROUND: This meta-analysis examines the comparative diagnostic performance of polymerase chain reaction (PCR) for the diagnosis of Pneumocystis pneumonia (PCP) on different respiratory tract samples, in both human immunodeficiency virus (HIV) and non-HIV populations.
    METHODS: A total of 55 articles met inclusion criteria, including 11 434 PCR assays on respiratory specimens from 7835 patients at risk of PCP. QUADAS-2 tool indicated low risk of bias across all studies. Using a bivariate and random-effects meta-regression analysis, the diagnostic performance of PCR against the European Organisation for Research and Treatment of Cancer-Mycoses Study Group definition of proven PCP was examined.
    RESULTS: Quantitative PCR (qPCR) on bronchoalveolar lavage fluid provided the highest pooled sensitivity of 98.7% (95% confidence interval [CI], 96.8%-99.5%), adequate specificity of 89.3% (95% CI, 84.4%-92.7%), negative likelihood ratio (LR-) of 0.014, and positive likelihood ratio (LR+) of 9.19. qPCR on induced sputum provided similarly high sensitivity of 99.0% (95% CI, 94.4%-99.3%) but a reduced specificity of 81.5% (95% CI, 72.1%-88.3%), LR- of 0.024, and LR+ of 5.30. qPCR on upper respiratory tract samples provided lower sensitivity of 89.2% (95% CI, 71.0%-96.5%), high specificity of 90.5% (95% CI, 80.9%-95.5%), LR- of 0.120, and LR+ of 9.34. There was no significant difference in sensitivity and specificity of PCR according to HIV status of patients.
    CONCLUSIONS: On deeper respiratory tract specimens, PCR negativity can be used to confidently exclude PCP, but PCR positivity will likely require clinical interpretation to distinguish between colonization and active infection, partially dependent on the strength of the PCR signal (indicative of fungal burden), the specimen type, and patient population tested.
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  • 文章类型: Case Reports
    单纯疱疹性脑炎(HSVE)是一种潜在致命的传染性中枢神经系统(CNS)疾病。因此,早期发现是决定案件命运的关键。临床病史和检查,脑计算机断层扫描,动态对比增强磁共振成像(DCE-MRI),腰椎穿刺已经被用来建立诊断。本报告描述了一例HSVE,伴有低细胞脑脊液(CSF)和罕见的记忆障碍。然而,MRI结果与HSVE一致,和CSFPCR检测对治疗有反应的HSV-1DNA呈阳性。我们通常建议患者尽快开始抗病毒治疗,以避免并发症。
    Herpes simplex encephalitis (HSVE) is a potentially fatal infectious central nervous system (CNS) disorder. Thus, early detection is critical in determining the case\'s fate. Clinical history and examination, brain computed tomography, dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), and lumbar puncture have been used to establish a diagnosis. This report describes a case of HSVE with hypocellular cerebrospinal fluid (CSF) and an uncommon form of memory impairment. However, MRI results were consistent with HSVE, and CSF PCR tested positive for HSV-1 DNA that responded to treatment. We routinely advise patients to begin antiviral therapy as soon as possible to avoid complications.
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  • 文章类型: Journal Article
    2022年初,40多个国家报告了猴痘(Mpox)的爆发。准确诊断水痘可能具有挑战性,但历史,临床发现,和实验室诊断可以建立诊断。测试的前分析阶段包括收集,存储,和运输标本。建议用来自两个不同部位的含有涤纶或聚酯羊群拭子的病毒转运培养基(VTM)擦拭病变部位。血,尿液,和精液样本也可以使用。需要及时取样以获得足够量的病毒或抗体。传染病控制的分析阶段涉及诊断工具以确定病毒的存在。虽然聚合酶链反应(PCR)是检测水痘的黄金标准,基因组测序用于识别新的或修饰的病毒。作为这些方法的补充,已经设计了等温扩增方法。ELISA测定法也可用于测定抗体。电子显微镜是另一种有效的病毒组织鉴定诊断方法。废水指纹图谱为社区一级的病毒鉴定提供了一些最有效的诊断方法。进一步讨论了这些方法的优缺点。分析阶段后需要正确解释测试结果,并准备包括相关病史在内的准确患者报告。临床指南,以及后续测试或治疗的建议。
    An outbreak of monkeypox (Mpox) was reported in more than 40 countries in early 2022. Accurate diagnosis of Mpox can be challenging, but history, clinical findings, and laboratory diagnosis can establish the diagnosis. The pre-analytic phase of testing includes collecting, storing, and transporting specimens. It is advised to swab the lesion site with virus transport medium (VTM) containing Dacron or polyester flock swabs from two different sites. Blood, urine, and semen samples may also be used. Timely sampling is necessary to obtain a sufficient amount of virus or antibodies. The analytical phase of infectious disease control involves diagnostic tools to determine the presence of the virus. While polymerase chain reaction (PCR) is the gold standard for detecting Mpox, genome sequencing is for identifying new or modified viruses. As a complement to these methods, isothermal amplification methods have been designed. ELISA assays are also available for the determination of antibodies. Electron microscopy is another effective diagnostic method for tissue identification of the virus. Wastewater fingerprinting provides some of the most effective diagnostic methods for virus identification at the community level. The advantages and disadvantages of these methods are further discussed. Post-analytic phase requires proper interpretation of test results and the preparation of accurate patient reports that include relevant medical history, clinical guidelines, and recommendations for follow-up testing or treatment.
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  • 文章类型: Journal Article
    21世纪死亡率最高的疾病之一是癌症。全球范围内,分子方法被广泛用于治疗癌症相关疾病,关于这个主题的研究机构每年都在增长。需要对支持与癌症有关的疾病的分子方法的数据进行全面而重要的总结。
    为了指导临床实践和未来的研究,重要的是检查和总结评估分子方法治疗癌症相关疾病的有效性和安全性的系统评价(SRs).
    我们开发了一种全面的搜索策略,可以从PubMed等电子数据库中查找相关文章,谷歌学者,WebofScience(WoS),或者Scopus.我们浏览了文献,并确定了癌症遗传学中哪些诊断方法特别可靠。我们使用了像“癌症遗传学”这样的短语,遗传易感性,遗传性癌症,癌症风险评估,\'癌症诊断工具\',癌症筛查,生物标志物,和分子诊断,综述和荟萃分析评估分子疗法对癌症相关疾病的疗效和安全性。仅考虑不需要遗传或分子诊断的治疗方式的研究属于排除标准。
    这篇综合综述的结果清楚地证明了分子方法在癌症遗传学领域的转化作用。这篇综述强调了这些技术如何使研究人员和临床医生能够识别和理解导致恶性肿瘤的关键遗传改变。从点突变到结构变异。这些见解有助于确定关键的致癌驱动因素和潜在的治疗目标,从而为能够显著改善患者预后的精准医学方法打开了大门。
    搜索没有指定包含出版物的时间表,它可能错过了癌症分子方法领域的最新进展或变化。因此,它可能没有包括该领域的最新发展。
    在对癌症遗传学中的分子方法进行了深入研究之后,很明显,这些尖端技术彻底改变了肿瘤学领域,为研究人员和临床医生提供强大的工具,在基因水平上解开癌症的复杂性。分子方法技术的整合不仅增强了我们对癌症病因的理解,programming,和治疗反应,但也为个性化医疗和靶向治疗开辟了新的途径,改善患者预后。
    UNASSIGNED: One of the ailments with the greatest fatality rates in the 21st century is cancer. Globally, molecular methods are widely employed to treat cancer-related disorders, and the body of research on this subject is growing yearly. A thorough and critical summary of the data supporting molecular methods for illnesses linked to cancer is required.
    UNASSIGNED: In order to guide clinical practice and future research, it is important to examine and summarize the systematic reviews (SRs) that evaluate the efficacy and safety of molecular methods for disorders associated to cancer.
    UNASSIGNED: We developed a comprehensive search strategy to find relevant articles from electronic databases like PubMed, Google Scholar, Web of Science (WoS), or Scopus. We looked through the literature and determined which diagnostic methods in cancer genetics were particularly reliable. We used phrases like \'cancer genetics\', genetic susceptibility, Hereditary cancer, cancer risk assessment, \'cancer diagnostic tools\', cancer screening\', biomarkers, and molecular diagnostics, reviews and meta-analyses evaluating the efficacy and safety of molecular therapies for cancer-related disorders. Research that only consider treatment modalities that don\'t necessitate genetic or molecular diagnostics fall under the exclusion criteria.
    UNASSIGNED: The results of this comprehensive review clearly demonstrate the transformative impact of molecular methods in the realm of cancer genetics.This review underscores how these technologies have empowered researchers and clinicians to identify and understand key genetic alterations that drive malignancy, ranging from point mutations to structural variations. Such insights are instrumental in pinpointing critical oncogenic drivers and potential therapeutic targets, thus opening the door for methods in precision medicine that can significantly improve patient outcomes.
    UNASSIGNED: The search does not specify a timeframe for publication inclusion, it may have missed recent advancements or changes in the field\'s landscape of molecular methods for cancer. As a result, it may not have included the most recent developments in the field.
    UNASSIGNED: After conducting an in-depth study on the molecular methods in cancer genetics, it is evident that these cutting-edge technologies have revolutionized the field of oncology, providing researchers and clinicians with powerful tools to unravel the complexities of cancer at the genetic level. The integration of molecular methods techniques has not only enhanced our understanding of cancer etiology, progression, and treatment response but has also opened new avenues for personalized medicine and targeted therapies, leading to improved patient outcomes.
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  • 文章类型: Systematic Review
    背景:膀胱癌(BC)是全球第七大最常见的癌症。不是所有的感染都以癌症结束,尽管HPV诱导的致癌作用是炎症的复杂过程。探讨人乳头瘤病毒(HPV)与膀胱癌诊断的关系。
    方法:我们根据Cochrane和PRISMA的建议进行了系统评价。我们在EMBASE搜索,Medline(Ovid),和Cochrane中央控制试验登记册(CENTRAL),从开始到现在。我们纳入了病例对照研究。基于QUADAS2进行偏倚风险评估。我们进行了随机效应Meta分析。
    结果:我们纳入了14项定性和定量分析研究。主要存在低偏倚风险。我们最终发现HPV的存在与膀胱癌诊断之间存在很强的相关性(OR4.1895CI2.63至6.66;I2=40%)。
    结论:HPV目前与膀胱癌的诊断相关。
    BACKGROUND: Bladder cancer (BC) is the seventh most common cancer worldwide. Not every infection ends as cancer, although the HPV-induced carcinogenesis is a complex process consequence of inflammation. To determine the association between human papillomavirus (HPV) and the diagnosis of bladder cancer.
    METHODS: We carried out a systematic review according to Cochrane and PRISMA recommendations. We searched in EMBASE, Medline (Ovid), and The Cochrane Central Register of Controlled Trials (CENTRAL), from inception to nowadays. We included case-control studies. The risk of bias assessment was performed based on QUADAS2. We performed a random effect Meta-analysis.
    RESULTS: We included 14 studies in qualitative and quantitative analysis. There was mainly a low risk of bias. We finally found a strong association between the presence of HPV and bladder cancer diagnosis (OR 4.18 95%CI 2.63-6.66; I2 = 40%).
    CONCLUSIONS: HPV is currently associated with the diagnosis of bladder cancer.
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  • 文章类型: Systematic Review
    背景:猫相关的嗜血支原体(血液原虫)被认为通过两种主要机制传播:(1)通过战斗直接传播和(2)由猫蚤(Ctenocephalidesfelis)传播。虽然C.felis的传输效率似乎很低,大多数手稿都集中在野生跳蚤中的血原虫的患病率,并报告了非常低(<3%)或高(>26%)的患病率。因此,我们旨在评估样品处理和PCR方法对C.felis血浆感染患病率的影响。
    方法:对PubMed文章的系统审查确定了13份手稿(1,531只跳蚤/跳蚤池)符合纳入标准(对从猫收集的C.felis上的>1份血浆进行PCR)。使用ROBINS-E工具评估偏倚风险。在这些手稿的R中进行的荟萃分析发现,不洗涤样品和一组常见的16SrRNA引物首次发表在Jensen等人。2001年与血血浆患病率增加有关。为了评估洗涤对新收集的跳蚤的影响,我们评估了20个5个C.felis池的血浆状态,其中一半被洗了,一半没有洗。
    结果:跳蚤冲洗并不影响血血浆的检测,而是扩增了螺血浆。用Jensen等人评估非特异性扩增。2001引物,对67例C.felis样品(34%先前报道的血血浆感染)进行PCR和测序。通过这种方法,仅在3%的样本中检测到血浆.在剩下的“血支原体感染”跳蚤中,PCR扩增螺旋体或其他细菌。
    结论:因此,我们得出的结论是,在C.felis中的血浆感染是罕见的,未来的跳蚤流行研究应该对所有阳性扩增子进行测序以验证PCR特异性。有必要进一步研究猫相关的血血浆传递的替代方法以及C.felis维持血血浆感染的能力。
    BACKGROUND: Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence.
    METHODS: A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed.
    RESULTS: Flea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining \"hemoplasma infected\" fleas, PCR amplified Spiroplasma or other bacteria.
    CONCLUSIONS: Therefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary.
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  • 文章类型: Journal Article
    聚合酶链反应(PCR)在我们对世界的理解中发挥了重要作用,并在广泛的学科中应用。将PCR引入法医学标志着DNA分析新时代的开始。这个时代将PCR推向了极限,并允许从痕量DNA生成遗传数据。痕量样品含有与抑制性化合物和离子相关的非常少量的降解DNA。尽管自首次引入以来,PCR过程取得了重大进展,抑制和降解样品的概况分析挑战仍然存在。这篇综述考察了PCR从20世纪80年代开始的演变,直至其目前在法医学中的应用。讨论了用于DNA分析的PCR进化背后的驱动因素,以及商业PCR试剂盒中使用的循环条件的关键比较。检查了目前在法医实践中使用的较新的PCR方法,并评估了用于DNA谱分析的PCR未来可能的方向。
    The polymerase chain reaction (PCR) has played a fundamental role in our understanding of the world, and has applications across a broad range of disciplines. The introduction of PCR into forensic science marked the beginning of a new era of DNA profiling. This era has pushed PCR to its limits and allowed genetic data to be generated from trace DNA. Trace samples contain very small amounts of degraded DNA associated with inhibitory compounds and ions. Despite significant development in the PCR process since it was first introduced, the challenges of profiling inhibited and degraded samples remain. This review examines the evolution of the PCR from its inception in the 1980s, through to its current application in forensic science. The driving factors behind PCR evolution for DNA profiling are discussed along with a critical comparison of cycling conditions used in commercial PCR kits. Newer PCR methods that are currently used in forensic practice and beyond are examined, and possible future directions of PCR for DNA profiling are evaluated.
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  • 文章类型: Case Reports
    单侧急性特发性黄斑病变(UAIM)是一种罕见的疾病,在流感样疾病后可能导致年轻人的单侧视力丧失。偶尔,它与手足口病(HFMD)有关,通常未被诊断。在这里,我们报告了一例UAIM与HFMD合并急性肠道病毒感染相关的病例。根据使用多模态诊断成像的临床发现,包括眼底图像,光学相干层析成像,荧光血管造影术,以及泛肠道病毒RNA聚合酶链反应的血清学测试,我们详细介绍了UAIM合并HFMD相关脉络膜炎的临床过程,并推测其发病机制。这项研究可以提醒每位医生UAIM在手足口病中可能导致的视力丧失,将患者转诊到眼科检查对消除潜在的视力损害很重要。
    Unilateral acute idiopathic maculopathy (UAIM) is a rare disease that may cause unilateral vision loss in young adults after a flu-like illness. Occasionally, it is associated with hand-foot-mouth disease (HFMD) and is often underdiagnosed. Herein, we report a case with characteristics of UAIM associated with HFMD with acute enteroviral infection. On the basis of the clinical findings using multimodality diagnostic imaging, including fundus image, optical coherence tomography, fluorescence angiography, and serological test for pan-enterovirus RNA polymerase chain reaction, we detailed the clinical course and postulated the pathogenesis of UAIM with choroiditis associated with HFMD. This study could remind every doctor of the potential visual loss caused by UAIM in HFMD, and referring patients to the ophthalmologic survey is important to eliminate potential visual impairment.
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  • 文章类型: Systematic Review
    卵巢结核是一种罕见的非特异性临床表现,诊断困难,和具体的医疗管理。结核病(TB)的卵巢受累可能以两种形式发生,即,骨膜炎和卵巢炎。结核病的体质症状,如厌食症,减肥,盗汗,据报道,高达45%的患者出现夜间体温升高。误诊和延迟诊断很常见。直接的组织病理学证明是最好的诊断方式。细针抽吸细胞学(FNAC)是选择的研究,聚合酶链反应(PCR)测定法提高了其灵敏度。对于孤立的卵巢结核和广泛的疾病,建议使用6个月的标准短期抗结核药物。建议治疗12个月。手术是为药物治疗失败和脓肿形成而保留的。关于泌尿生殖道结核的研究很多,但仍缺乏定义诊断研究和管理指南的详细研究。本文旨在介绍和分享有关卵巢结核的英文文献,以更好地了解病因和诊断方法,并为其管理提供指导。
    Ovarian tuberculosis is a rare entity with non-specific clinical manifestations, difficult diagnosis, and specific medical management. Ovarian involvement in tuberculosis (TB) may occur in two forms, namely, perioophoritis and oophoritis. The constitutional symptoms of tuberculosis such as anorexia, weight loss, night sweats, and evening rise in temperature have been reported in up to 45% of patients. Misdiagnosis and delayed diagnosis are common. A direct histopathological demonstration is the best diagnostic modality. Fine needle aspiration cytology (FNAC) is the study of choice and polymerase chain reaction (PCR) assay increases its sensitivity. The standard short-course antituberculous for 6 months is recommended for isolated ovarian tuberculosis and for widespread disease, 12 months of therapy is recommended. Surgery is reserved for failure of medical therapy and abscess formation. There are many studies on genito-urinary tuberculosis but a detailed study defining diagnostic studies and management guidelines is still lacking. This article aims to present and share a review of the English-language literature on ovarian tuberculosis to gain a better understanding of etiopathogenesis and diagnostic methods and to provide guidelines for its management.
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