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siRNA-Inhibition of TIGAR Hypersensitizes Human Papillomavirus-Transformed Cells to Apoptosis Induced by Chemotherapy Drugs that Cause Oxidative Stress sirna抑制TIGAR超敏化人乳头瘤病毒转化细胞对化疗药物引起氧化应激诱导的凋亡
Pub Date : 2021-05-31 DOI: 10.35248/1948-5964.21.13.223
Laçin Yapindi, B. Hernandez, R. Harrod
The high-risk subtype Human Papillomaviruses (hrHPVs), including HPV16, HPV18, HPV31, HPV33, and HPV45, infect and oncogenically transform epithelial cells and cause squamous cell carcinomas and adenocarcinomas associated with the development of cervical cancer and subsets of vulvar, vaginal, penile, and anogenital cancers, as well as head-and-neck oropharyngeal carcinomas which often have poor clinical prognoses. Many cancers have been shown to contain elevated levels of the TP53-Induced Glycolysis and Apoptosis Regulator (TIGAR)-a glycolytic enzyme and antioxidant effector which frequently correlates with an aggressive tumor phenotype and serves as a determinant of therapy-resistance. We therefore tested whether siRNA-inhibition of TIGAR protein expression could sensitize HPV18-transformed HeLa cells to genotoxic chemotherapy agents (i.e., cisplatin, etoposide, doxorubicin, and 4-hydroxycyclophosphamide) that induce oxidative stress and DNA-damage. Here we demonstrate that the siRNA-knockdown of TIGAR hypersensitized HeLa cells to low, otherwise sub-inhibitory concentrations of these drugs and markedly induced cellular apoptosis, as compared to a scrambled RNA (scrRNA) oligonucleotide negative control or a non-transformed immortalized human fibroblast cell-line, HFL1. Importantly, these findings suggest that therapeutically inhibiting TIGAR could hypersensitize hrHPV+ cervical tumor cells to low-dosage concentrations of chemotherapy drugs that induce oxidative DNA-damage, which could potentially lead to more favorable clinical outcomes by reducing the adverse side-effects of these anticancer medications and making them more tolerable for patients. Our studies have further shown that siRNA-inhibition of TIGAR sensitizes HPV18+ HeLa cells to apoptosis induced by 4-hydroxycyclophosphamide-a DNA-alkylating agent these cells were reported to have resistance to, alluding to another possible benefit of targeting TIGAR in combinatorial treatment strategies against virus-induced cancers.
高危亚型人乳头瘤病毒(hrhpv),包括HPV16、HPV18、HPV31、HPV33和HPV45,感染上皮细胞并致癌转化,引起与宫颈癌发展相关的鳞状细胞癌和腺癌,以及外阴、阴道、阴茎和肛门生殖器癌亚群,以及头颈口咽癌,临床预后往往较差。许多癌症已被证明含有高水平的tp53诱导的糖酵解和凋亡调节因子(TIGAR),这是一种糖酵解酶和抗氧化效应物,经常与侵袭性肿瘤表型相关,并作为治疗耐药性的决定因素。因此,我们测试了sirna抑制TIGAR蛋白表达是否可以使hpv18转化的HeLa细胞对诱导氧化应激和dna损伤的遗传毒性化疗药物(即顺铂、依托泊苷、阿霉素和4-羟基环磷酰胺)敏感。在这里,我们证明,与混乱RNA (scrRNA)寡核苷酸阴性对照或未转化的永生化人成纤维细胞系HFL1相比,TIGAR敲低sirna使HeLa细胞对这些药物的低浓度(否则为亚抑制浓度)高度敏感,并显著诱导细胞凋亡。重要的是,这些发现表明,在治疗上抑制TIGAR可能会使hrHPV+宫颈肿瘤细胞对低剂量浓度化疗药物过敏,从而诱导dna氧化损伤,这可能会通过减少这些抗癌药物的不良副作用并使患者更耐受这些药物而获得更有利的临床结果。我们的研究进一步表明,sirna抑制TIGAR使HPV18+ HeLa细胞对4-羟基环磷酰胺(一种dna烷基化剂)诱导的凋亡敏感,这些细胞被报道具有耐药性,这暗示了在针对病毒诱导的癌症的联合治疗策略中靶向TIGAR的另一个可能的益处。
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引用次数: 0
Coroless, effective ingredient isolated from traditional Chinese medicine for the prevention and treatment of Covid-19 从中药中分离出的预防和治疗新型冠状病毒肺炎的无色有效成分
Pub Date : 2021-02-20 DOI: 10.37421/1948-5964.21.13.217
Nie Leng, Liang Youdong, Yang Xinwei
We believe that a good medicine for the prevention and treatment of Covid-19 requires three points: 1. Effectively inhibit the cells from being infected by Covid-19; 2. Quickly repair lung damage; 3. Non-toxic and harmless. Fortunately, from the treasure depot of Chinese traditional medicine, we found Kyllinga brevifolia Rottb, which is sufficient for three points and can be used for prevention, treatment and rehabilitation. Not only that, we have isolated the active ingredient of the Kyllinga brevifolia Rottb, named it Coroless, and verified that Coroless inhibits the Covid-19 from infecting cells through in vitro cell experiments.
我们认为,做好防控新冠肺炎的药,需要三点。有效抑制细胞被新冠病毒感染;2. 快速修复肺部损伤;3.无毒无害。幸运的是,我们从中药宝库中找到了足足三穴,可以预防、治疗、康复的短叶针。不仅如此,我们还分离出了短叶藻(Kyllinga brevifolia Rottb)的活性成分,将其命名为Coroless,并通过体外细胞实验验证了Coroless对新冠病毒感染细胞的抑制作用。
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引用次数: 0
An Exploratory Study on COVID-19 and the Rights of Children Based on Keyword Network Analysis 基于关键词网络分析的COVID-19与儿童权利探索性研究
Pub Date : 2021-02-12 DOI: 10.21203/RS.3.RS-156498/V1
Seoyeon Lee
COVID-19 has become a worldwide health crisis. Around March 2020, the entire country was shut down, including schools. This resulted in significant changes in the lives of children. In this study, the researcher conducted a keyword network analysis utilizing Ucinet ver 6.716 and NetDraw ver 2.173, after gathering the data using Textom in order to examine the current status of the rights of children in a COVID-19 pandemic. The findings of this study were that the degree centrality was higher with poverty, educational institutions, parents, teachers, income support, child care, child-rearing, caring, online classes, and child welfare, etc. Therefore, it can be said that there is an urgent need for the implementation of the respect of the rights of children all over the world in this COVID-19 pandemic.
COVID-19已成为一场全球性的健康危机。2020年3月左右,包括学校在内的整个国家都关闭了。这使儿童的生活发生了重大变化。在本研究中,研究者在使用texttom收集数据后,利用Ucinet版本6.716和NetDraw版本2.173进行了关键词网络分析,以研究COVID-19大流行中儿童权利的现状。研究发现,贫困程度、教育机构、家长、教师、收入支持、儿童保育、育儿、关爱、网络课程、儿童福利等因素的中心性较高。因此,可以说,在2019冠状病毒病大流行期间,迫切需要在世界各地落实对儿童权利的尊重。
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引用次数: 1
Comorbidities of COVID-19 Patients with Low Cycle Threshold (Ct) Value of Nucleocapsid (N) Gene: An Application to Cluster-Based Logistic Model. 核衣壳(N)基因低周期阈值(Ct)患者的合共病:基于聚类的Logistic模型的应用
Pub Date : 2021-01-19 DOI: 10.21203/RS.3.RS-147576/V2
Sujan Rudra, Shuva Das, Md. Ehsanul Hoque, A. Kalam, Mohammad Arifur Rahman, Swagata Nandy Shizuka, T. Rahman
Background: Coronavirus disease 2019 (COVID-19) is a health crisis throughout the world. The widely used Real-time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCR) method is most capable of describing the patient’s condition. Comorbidities can make patients more critical.Methods: In this study, we shed light on the low cycle threshold (Ct) value of the N gene in the rRT-PCR test of the COVID-19 patients who had comorbidities, cure rate, and the needfulness of ICU (Intensive Care Unit) management. We had conducted the research in the Molecular Biology Laboratory of Chittagong Medical College between May and August 2020, then took the telephone interview with 300 positive patients who fulfilled the study criteria. We applied cluster-based logistic regression to analyze the data.Results: Low Ct value of the N gene found 1.324 times more in Type 2 DM patients and 1.871 times higher in hypertensive patients, and hospitalized patients are 2.480 times more vulnerable to shift in ICU.Conclusions: While infection with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) frequently causes severe diseases, suspected cases with comorbid conditions should go through the rRT-PCR as early as possible.
背景:2019冠状病毒病(COVID-19)是一场全球性的健康危机。实时逆转录聚合酶链式反应(rRT-PCR)是最能描述患者病情的方法。合并症会使病人更加危急。方法:本研究探讨新冠肺炎合并合并症患者的rRT-PCR检测中N基因的低周期阈值(Ct)、治愈率及重症监护病房管理的必要性。我们于2020年5月至8月在吉大港医学院分子生物学实验室进行了研究,并对300名符合研究标准的阳性患者进行了电话访谈。我们采用基于聚类的逻辑回归对数据进行分析。结果:N基因低Ct值在2型糖尿病患者中多出1.324倍,在高血压患者中多出1.871倍,住院患者易转ICU患者多出2.480倍。结论:严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)感染往往会导致严重的疾病,但有合并症的疑似病例应尽早进行rRT-PCR检测。
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引用次数: 3
A Questionnaire-Based Survey to Assess the Timing of Intubation in COVID-19 Pneumonia. 评估COVID-19肺炎插管时机的问卷调查
Pub Date : 2021-01-11 DOI: 10.21203/RS.3.RS-141346/V1
Samir Samal, S. Mishra, E. Patra, Rajesh Kasimahanti
BackgroundMany COVID19 pneumonia patients progress to Acute respiratory distress syndrome and end up in Intensive Care Units. Given that it is a novel viral infection, the progress of the disease, its management and associated outcomes are yet to be studied in detail. ARDS associated with COVID 19 is the same as before or different and the timing of intubation in such patients is a topic up for debate. This survey aimed to assess the opinion regarding management of COVID 19 ARDS and the timing of intubation in those patients.Methods292 clinicians including anesthesiologists, intensivists and others involved in managing COVID 19 ARDS patients at various centres were surveyed with web-based questionnaire cross sectionally within the time period of 10th June 2020 to 31st August 2020 after taking prior consent. Their responses were recorded and analyzed with statistical software IBM SPSS version 25.0.Results Among 292 included participants, 172 were intensivist, 84 were anesthesiologists and rest were others. Most of the intensivists (51.2%) had seen more than 100 COVID 19 severe ARDS patients. Around 82% of clinicians were agreed that COVID 19 ARDS was different from another form of ARDS. 67.1% of participants were agreed with patient induced self-inflicted injury could have happened in this disease. Likewise, around 91.8% of doctors involved in managing patients were believed that HFNC could be helpful if there were falling of saturation. 37% of participants were not agreed with early intubation, which may increase the risk of mortality and nosocomial infections.Conclusions and RelevanceThere was confusion in most doctors with intubation timing even if there was an indication for intubation. These confusions may be due to non-availability of specific recommendation regarding intubation in COVID 19 severe ARDS patients. However, most of the literature recommended for early intubation in these patients when indicated.
许多covid - 19肺炎患者进展为急性呼吸窘迫综合征,最终进入重症监护病房。鉴于这是一种新型病毒感染,该疾病的进展、治疗和相关结果还有待详细研究。与COVID - 19相关的ARDS与以前相同或不同,这些患者的插管时间是一个值得讨论的话题。本调查旨在评估对COVID - 19急性呼吸窘迫综合征(ARDS)患者的管理和插管时机的看法。方法在征得事先同意后,于2020年6月10日至2020年8月31日对各中心292名临床医生(包括麻醉师、重症监护医师和其他参与管理COVID - 19 ARDS患者的临床医生)进行横断面调查。采用IBM SPSS 25.0统计软件对患者的反应进行记录和分析。结果292名参与调查人员中,重症医师172名,麻醉师84名,其余为其他医师。大多数重症医师(51.2%)见过100例以上重症ARDS患者。约82%的临床医生认为COVID - 19 ARDS与另一种形式的ARDS不同。67.1%的参与者认同本病可能发生患者自伤。同样,参与患者管理的医生中,约有91.8%的人认为,如果血液饱和度下降,HFNC可能会有所帮助。37%的参与者不同意早期插管,这可能会增加死亡和医院感染的风险。结论与相关性即使有插管指征,大多数医生对插管时机也存在混淆。这些混淆可能是由于没有关于COVID - 19严重急性呼吸窘迫综合征患者插管的具体建议。然而,大多数文献建议在有指征时对这些患者进行早期插管。
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引用次数: 1
Abundant transmission of Corona Virus Nsp2 RNA Topoisomerse I120F Mutants with Concurrence D614G Spike Protein Mutation in Australia 冠状病毒Nsp2 RNA拓扑异构体I120F突变体并发D614G刺突蛋白突变在澳大利亚的大量传播
Pub Date : 2021-01-09 DOI: 10.35248/1948-5964.13.S16.001
A. Chakraborty
We previously predicted Nsp2 Corona virus protein as RNA topoisomerase through amino acid homology among Vibrio haemolytica DNA topoisomerase IA/IV as well as DNA primase, DNA gyrase and bi-subunit Trypanosoma brucei DNA topoisomerase IB. Many DNA topoisomerase I/III have RNA topoisomerase activity and such ubiquitous enzymes are conserved and involved in the regulation of replication and transcription. We have checked here mutational profile of Nsp2 RNA topoisomerase analyzing >10000 orf1a 4405 amino acid length Corona virus polyprotein. Mutant proteins were selected by BLAST search having 99.84% sequence similarity and 181-818aa portion Nsp2 protein (protein id. QIU82057) was analyzed using CLUSTAL Omega software. We found 26 different mutations where most changes were selected at Isoleucine and Alanine into Valine or Leucine into Phenylanaline pinpointing conserved nature of the Corona virus RNA topoisomerase. Major nonsense very abundant mutations were found at I120F (Isoleucine to Phenylalanine). Other important mutations were R27C, I198V, T85I, L410F, I559V and P583S. The I120F mutation was abundant in Australian isolates and its spread was seen in the Bangladesh and other countries like USA. We suggest that abundant I120F mutation of Nsp2 Topoisomerase may increase transmission of Corona virus by stabilizing RNA structure for efficient virus pakaging. Interestingly, such mutations were found in association of D614G mutation of Spike protein, known to >70% increase infectivity. On the contrary, all P583S Nsp2 mutants analyzed had no concurrence D614G spike protein mutation. Many silent mutations (5-7) were detected by genome wide analysis but no N501Y Spike protein mutation. This is first report that predicts a link of greater Corona virus transmission with Nsp2 protein I120F and spike protein D614G mutations.
我们之前通过溶血弧菌DNA拓扑异构酶IA/IV、DNA引物酶、DNA回转酶和双亚基布氏锥虫DNA拓扑异构酶IB的氨基酸同源性预测Nsp2冠状病毒蛋白为RNA拓扑异构酶,许多DNA拓扑异构酶I/III具有RNA拓扑异构酶活性,这些酶是普遍存在的,并参与复制和转录的调控。我们在这里检查了Nsp2 RNA拓扑异构酶的突变谱,分析了>10000 orf1a 4405氨基酸长度的冠状病毒多蛋白。BLAST搜索得到序列相似度为99.84%、Nsp2蛋白181-818aa部分(蛋白id)的突变蛋白。采用CLUSTAL Omega软件对QIU82057进行分析。我们发现了26个不同的突变,其中异亮氨酸和丙氨酸被选择为缬氨酸或亮氨酸被选择为苯analine,从而确定了冠状病毒RNA拓扑异构酶的保守性。在I120F(异亮氨酸到苯丙氨酸)上发现了大量的无义突变。其他重要的突变有R27C、I198V、T85I、L410F、I559V和P583S。I120F突变在澳大利亚分离株中大量存在,在孟加拉国和美国等其他国家也有传播。我们认为,Nsp2拓扑异构酶丰富的I120F突变可能通过稳定RNA结构以提高病毒包装效率来增加冠状病毒的传播。有趣的是,这些突变被发现与穗蛋白D614G突变相关,已知该突变可使感染性增加70%以上。相反,所有的P583S Nsp2突变体都没有并发的D614G刺突蛋白突变。全基因组分析检测到许多沉默突变(5-7),但未发现N501Y刺突蛋白突变。这是第一份预测冠状病毒传播与Nsp2蛋白I120F和刺突蛋白D614G突变有关的报告。
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引用次数: 0
Remdesivir and Lopinavir/Ritonavir as Potential Drugs to Treat Corona Virus Disease 2019 瑞德西韦和洛匹那韦/利托那韦作为治疗冠状病毒病的潜在药物2019
Pub Date : 2021-01-04 DOI: 10.35248/1948-5964.20.S6.002
Reda Bzikha, Bouhmou Ayoub, Bzikha Ilham, S. Bouchnafati
Drugs that are efficacious against SARS-CoV-2 have yet to be established. Remdesivir and Lopinavir/ritonavir have garnered considerable attention for their potential to treat coronavirus disease 2019 (COVID-19). Remdesivir not only in vivo but also in vitro testing shows the inhibition of human coronavirus replication, including SARS-CoV aswell as Lopinavir/ritonavir that shows promising antiviral drug against SARS-CoV-2.
对SARS-CoV-2有效的药物尚未确定。瑞德西韦和洛匹那韦/利托那韦因其治疗2019冠状病毒病(COVID-19)的潜力而引起了相当大的关注。瑞德西韦不仅在体内,而且在体外测试中都显示出对人类冠状病毒复制的抑制作用,包括SARS-CoV以及洛匹那韦/利托那韦,后者显示出对SARS-CoV-2有希望的抗病毒药物。
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引用次数: 2
Some Retroviral Diseases of Humans and Animals: A Prospectus 一些人类和动物的逆转录病毒疾病:简介
Pub Date : 2021-01-01 DOI: 10.35248/1948-5964.21.S21.002
Eltayb Abuelzein, M. Elamin, O. Ibrahim
This prospectus, gives an account on five, highly epidemiologically significant, virus members of the family Retroviridae and the ailments they cause. These are the human (HIV-1 and HIV-2) and three animal retroviral diseases of veterinary importance viz. the Enzootic Bovine Leucosis Virus (EBLV); the Caprine Arthritis-Encephalitis Virus (CAEV) and the Equine Infectious Anemia Virus (EIAV). This is expected to give a dimension that can aid field veterinarians, especially in developing countries, to help in differential clinical diagnosis, between diseases that may have similar clinical picture to retrovirus infections in animals. This article also gives enlightenment on the history, classification, properties, ecology and the interesting peculiarities of the family Retroviridae.
本说明书介绍了逆转录病毒科的五种具有高度流行病学意义的病毒成员及其引起的疾病。这些是具有兽医重要性的人类(HIV-1和HIV-2)和三种动物逆转录病毒疾病,即地方性牛白血病病毒(EBLV);山羊关节炎-脑炎病毒(CAEV)和马传染性贫血病毒(EIAV)。预计这将提供一个维度,可以帮助现场兽医,特别是发展中国家的兽医,帮助区分临床诊断可能与动物中逆转录病毒感染具有相似临床表现的疾病。本文还介绍了逆转录病毒科的历史、分类、性质、生态学和有趣的特点。
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引用次数: 0
Side Effects of Antiretroviral Therapy 抗逆转录病毒治疗的副作用
Pub Date : 2021-01-01 DOI: 10.35248/1948-5964.21.13.E229
Laura Sherry
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引用次数: 0
Measles Virus as a Vector for the HPV Vaccine Development 麻疹病毒作为HPV疫苗研制的载体
Pub Date : 2021-01-01 DOI: 10.35248/1948-5964.21.13.211
N. Penta, Gaurav K Gupta, R. Glueck
The Human Papilloma Virus (HPV) vaccine prevents infection with certain species of human papillomavirus associated with the development of cervical cancer, genital warts and some less common cancers. But, the development of vaccine that prevents HPV infection represents an important opportunity to prevent cervical cancer whilst a therapeutic immunization would be valuable in treating premalignant and malignant disease. Attenuated MV strains are highly efficient and safe vaccines, typically preventing recipients from Measles for their entire life. To benefit from these extraordinary vaccination properties in the present study MV cDNA plasmids have been constructed to produce precisely initiated and terminated MV anti-genome related to the Edmonston B vaccine strains. These transgenes were expressed at different levels, according to their genomic position and were stably maintained over many viral generations. The ability to construct new recombinant and chimeric MVs opens the prospect to develop new vaccines based on MV. In an effort to generate an inexpensive candidate HPV vaccine with improved prophylactic potential, a MV vector based on Berna-commercial Measles vaccine strain (Edmonston- Zagreb) inducing against HPV was developed.
人乳头瘤病毒(HPV)疫苗可预防感染与子宫颈癌、生殖器疣和一些不常见癌症有关的某些种类的人乳头瘤病毒。但是,预防HPV感染的疫苗的开发代表了预防宫颈癌的重要机会,而治疗性免疫在治疗癌前和恶性疾病方面将是有价值的。减毒MV毒株是高效和安全的疫苗,通常可使受者终生免受麻疹感染。为了利用这些特殊的疫苗接种特性,本研究构建了MV cDNA质粒,以产生与Edmonston B疫苗株相关的精确启动和终止的MV抗基因组。这些转基因根据其基因组位置在不同水平上表达,并在许多病毒世代中稳定保持。构建新的重组和嵌合MV的能力为开发基于MV的新疫苗开辟了前景。为了产生一种具有更好预防潜力的廉价HPV候选疫苗,我们开发了一种基于Berna-commercial麻疹疫苗株(Edmonston- Zagreb)诱导HPV的MV载体。
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引用次数: 0
期刊
Journal of Antivirals & Antiretrovirals
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