The Efficacy and Safety of Triazavirin for COVID-19: A Trial Protocol

根据2019年12月的报道,新冠病毒肺炎(coronavirus disease 2019, COVID-19)是由一种新型冠状病毒引起的肺炎。到目前为止,还没有发现能够治疗这种病毒的有效药物。本研究是一项在黑龙江省10个研究中心正在进行的多中心双盲随机对照试验(randomized controlled trial, RCT),其目的是研究与安慰剂相比,特力阿扎维林(triazavirin, TZV)治疗COVID-19患者的疗效和安全性。共计划招募240名COVID-19患者参加这项试验。咽拭子病毒核酸检测为阳性的受试者被随机(1∶1)分为两组:使用标准治疗加TZV或标准治疗加安慰剂,进行为期7 d的治疗和为期21 d的随访。主要结局是受试者临床改善的时间。次要结局包括临床改善率、退热时间、肺内炎症明显吸收的平均时间和人数比例、病毒核酸转阴率、病死率以及重症和危重症患者的转化率。整个试验过程将对不良事件,严重不良事件,肝功能、肾功能以及合并用药进行监测和记录。本试验的结果可为临床医生治疗COVID-19提供循证医学的证据和建议。

CPAL, as a New Mediator of Cardiomyocyte Metabolic Alterations and Pyroptosis, Regulates Myocardial Infarction Injury in Mice

Myocardial infarction (MI), the most serious of the ischemic heart diseases, is accompanied by myocardial metabolic disorders and the loss of cardiomyocytes. Increasing evidence has shown that long noncoding RNAs (lncRNAs) are involved in various pathological conditions such as cancer and cardiovascular diseases (CVDs), and are emerging as a novel biomarker for these disorders. This study aims to investigate the regulatory role and mechanisms of lncRNAs in myocardial remodeling in the setting of MI. We find that post-infarcted hearts exhibit a reduction of adenosine triphosphate (ATP) and an alteration of the glucose and lipid metabolism genes cluster of differentiation 36 (CD36), hexokinase 1 (HK1), and clucose transporter 4 (GLUT4), accompanied by cardiomyocyte pyroptosis. We then identify a previously unknown conserved lncRNA, AK009126 (cardiomyocyte pyroptosis-associated lncRNA, CPAL), which is remarkably upregulated in the myocardial border zone of MI mice. Importantly, the adeno-associated virus 9 (AAV9)-mediated silencing of endogenous CPAL by its short hairpin RNA (shRNA) partially abrogates myocardial metabolic alterations and cardiomyocyte pyroptosis during MI in mice. Mechanistically, CPAL is shown to bind directly to nuclear factor kappa B (NFκB) and to act as an activator of NFκB to induce NFκB phosphorylation in cardiomyocytes. We also find that CPAL upregulates caspase-1 expression at the transcriptional level and consequently promotes the release of interleukin (IL)-18 and IL-1β from cardiomyocytes. Collectively, our findings reveal the conserved lncRNA CPAL as a new regulator of cardiac metabolic abnormalities and cardiomyocyte pyroptosis in the setting of MI and suggest CPAL as a new therapeutic target to protect cardiomyocytes against ischemic injury in infarcted hearts.

Blue LED promotes the chemosensitivity of human hepatoma to Sorafenib by inducing DNA damage

Background:Phototherapies based on sunlight,infrared,ultraviolet,visible,and laser-based treatments present advantages like high curative effects,small invasion,and negligible adverse reactions in cancer treatment.We aimed to explore the potential therapeutic effects of blue light emitting diode(LED)in human hepatoma cells and decipher the underlying cellular and molecular mechanisms.Methods:Wound healing and transwell assays were employed to probe the inhibition of the invasion and migration of hepatocellular carcinoma cells in the presence of blue LED.The sphere-forming test was used to evaluate the effect of LED blue light irradiation on cancer stem cell properties.Immunofluorescence and western blotting were used to detect the changes inγ-H2AX.The Cell Counting Kit-8 assay,5-ethynyl-2′-deoxyuridine staining,and colony formation assay were used to detect the combined effect of blue LED and sorafenib on cell proliferation inhibition.Results:We demonstrated that the irradiation of blue LED light in hepatoma cells could lead to cell proliferation reduction along with the increase of cell apoptosis.Simultaneously,blue LED irradiation also markedly suppressed the migration and invasion ability of human hepatoma cells.Sphere formation analysis further revealed the decreased cancer stemness of hepatoma cells upon blue LED irradiation.Mechanistically,blue LED irradiation significantly promoted the expression of the phosphorylation of the core histone protein H2AX(γ-H2AX),a sensitive molecular marker of DNA damage.In addition,we found that the combined treatment of blue LED irradiation and sorafenib increased cancer cell sensitivity to sorafenib.Conclusion:Collectively,we demonstrated that blue LED irradiation exhibited anti-tumor effects on liver cancer cells by inducing DNA damage and could enhance chemosensitivity of cancer cells,which represents a potential approach for human hepatoma treatment.

Efficacy and Safety of Triazavirin Therapy for Coronavirus Disease 2019:A Pilot Randomized Controlled Trial

目前,尚无有效疗法可治愈由严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)引起的新型冠状病毒肺炎(coronavirus disease 2019,COVID-19)。为了评估抗病毒药物特力阿扎维林治疗COVID-19的有效性和安全性,本研究针对感染COVID-19的成年住院患者开展了一项随机双盲对照试验。本研究从10个分中心招募COVID-19患者,并将患者按1:1的比例随机分为两组。试验组的患者每天服用250 mg的特力阿扎维林药物3~4次,对照组患者则服用安慰剂,为期共7 d。主要结局指标为临床改善时间,临床改善时间的定义为随机分组28d内患者的体温、呼吸频率、血氧饱和度、咳嗽频率和肺CT(计算机断层扫描,computed tomography)所显示的肺部感染吸收情况全部恢复正常的时间。次要结局指标包括主要结局指标的5个组成指标,及肺部感染吸收的平均时间和吸收比例,以及用咽拭子采样法连续两次SARS-CoV-2核酸检测阴性的转阴率。与此同时,记录合并的治疗药物、不良事件和严重不良事件。由于需要进行住院治疗的新增感染病例的减少,本研究在招募52名患者后便停止招募。将52名受试者随机分为服用特力阿扎维林药物的试验组(n=26)和服用安慰剂的对照组(n=26)。结果显示,两组临床改善时间并无明显差异[中位数,7 d vs.12 d;风险比(RR)为2.0;95%置信区间(CI)为0.7~5.6;p=0.2]。服用特力阿扎维林的试验组中有10名患者发生临床改善,服用安慰剂的对照组中有6名患者出现临床改善(38.5%vs.23.1%,RR为2.1;95%CI为0.6~7.0;p=0.2)。除了肺部感染的吸收情况外(试验组50.0%,对照组26.1%),其余主要结局所观察指标均在28 d内恢复正常。此外,试验组的患者在呼吸系统、心脏、肾、肝或凝血功能等方面的合并治疗较对照组少。尽管特力阿扎维林药物对COVID-19患者的疗效尚未达到统计学上的显著性水平,但本研究结果表明,由于特力阿扎维林药物具有抗病毒作用,将其用于COVID-19的治疗也可能具有一定的疗效。对此,需要进一步的研究进行验证。

Altered expression profile of long non-coding RNAs during heart aging in mice

Objective:Long noncoding RNAs(lncRNAs)play an important role in regulating the occurrence and development of cardiovascular diseases.However,the role of lncRNAs in heart aging remains poorly understood.The objective of this study was to identify differentially expressed lncRNAs in the heart of aging mice and elucidate the relevant regulatory pathways of cardiac aging.Materials and methods:Echocardiography was used to detect the cardiac function of 18-months(aged)and 3-months(young)old C57BL/6 mice.Microarray analysis was performed to unravel the expression profiles of lncRNAs and mRNAs,and qRT-PCR to verify the highly dysregulated lncRNAs.Results:Our results demonstrated that the heart function in aged mice was impaired relative to young ones.Microarray results showed that 155 lncRNAs were upregulated and 37 were downregulated,and 170 mRNAs were significantly upregulated and 44 were remarkably downregulated in aging hearts.Gene ontology analysis indicated that differentially expressed genes are mainly related to immune function,cell proliferation,copper ion response,and cellular cation homeostasis.KEGG pathway analysis showed that the differentially expressed mRNAs are related to cytokine-cytokine receptor interaction,inflammatory mediator regulation of TRP channels,and the NF-kappa B signaling pathway.Conclusion:These results imply that the differentially expressed lncRNAs may regulate the development of heart aging.This study provides a new perspective on the potential effects and mechanisms of lncRNAs in heart aging.

Electrocardiogram abnormalities and higher body mass index as clinically applicable factors for predicting poor outcome in patients with coronavirus disease 2019

Background:Patients with coronavirus disease 2019(COVID-19)have high resource utilization.Identifying the causes of severe COVID-19 is helpful for early intervention to reduce the consumption of medical resources.Methods:We included 103 patients with COVID-19 in this single-center observational study.To evaluate the incidence,predictors,and effects of COVID-19,we analyzed demographic information,laboratory results,comorbidities,and vital signs as factors for association with severe COVID-19.Results:The incidence of severe COVID-19 was 16.5%and the percent poor outcome(including mortality,entering in ICU or transferred to a superior hospital)was 6.8%.The majority of severe COVID-19 patients had abnormal electrocardiogram(ECG)(82.35%),hypertension(76.47%)and other cardiac diseases(58.82%).Multivariate logistic regression was used to determine the predictors of severe illness.Abnormal body mass index(BMI)and ECG(P<0.05)were independent predictors of severe COVID-19.ECG abnormality was associated with increased odds of poor outcome(area under the receiver operating characteristic curves[AUC],0.793;P=0.010)and severe COVID-19(AUC,0.807;P<0.0001).Overweight was also associated with increased odds of poor outcome(AUC,0.728;P=0.045)and severe illness COVID-19(AUC,0.816;P<0.0001).Conclusion:Overweight and electrophysiological disorders on admission are important predictors of prognosis of patients with COVID-19.

LncRNA ZFAS1 regulates cardiomyocyte differentiation of human embryonic stem cells

Background:Cardiomyocytes derived from human embryonic stem cells(hESCs)are regulated by complex and stringent gene networks during differentiation.Long non-coding RNAs(lncRNAs)exert critical epigenetic regulatory functions in multiple differentiation processes.However,the involvement of lncRNAs in the differentiation of hESCs into cardiomyocytes has not yet been fully elucidated.Here,we identified the key roles of ZFAS1(lncRNA zinc finger antisense 1)in the differentiation of cardiomyocytes from hESCs.Methods:A model of cardiomyocyte differentiation from stem cells was established using the monolayer differentiation method,and the number of beating hESCs-derived cardiomyocytes was calculated.Gene expression was analyzed by quantitative real-time PCR(qRTPCR).Immunofluorescence assays were performed to assess the expression of cardiac troponin T(cTnT)andα-actinin protein in cardiomyocytes.Results:qRT-PCR showed that ZFAS1 expression in the mesoderm was significantly higher than that in embryonic stem cells,cardiac progenitor cells,and cardiomyocytes.Knockdown of ZFAS1 inhibited cardiomyocyte differentiation from hESCs,which was characterized by reduced expression of the cardiac-specific markers cTnT,α-actinin,myosin heavy chain 6(MYH6),and myosin heavy chain 7(MYH7).In contrast,ZFAS1 overexpression remarkably increased the percentage of spontaneously beating cardiomyocytes.In terms of the mechanism,we found that ZFAS1 is an antisense lncRNA at the 5′end of the protein-coding gene ZNFX1.Knockdown of ZFAS1 could increase the mRNA expression level of ZNFX1.Furthermore,qRT-PCR demonstrated that the silencing of ZNFX1 led to an increase in cardiac-specific markers that predicted the promotion of cardiomyocyte differentiation.Conclusion:Altogether,these data suggest that lncRNA-ZFAS1 is required for cardiac differentiation by functionally inhibiting the expression of ZNFX1,which may provide a reference for the treatment of heart disease to a certain extent.

Regulation of cardiomyocyte fate plasticity: a key strategy for cardiac regeneration

With the high morbidity and mortality rates,cardiovascular diseases have become one of the most concerning diseases worldwide.The heart of adult mammals can hardly regenerate naturally after injury because adult cardiomyocytes have already exited the cell cycle,which subseqently triggers cardiac remodeling and heart failure.Although a series of pharmacological treatments and surgical methods have been utilized to improve heart functions,they cannot replenish the massive loss of beating cardiomyocytes after injury.Here,we summarize the latest research progress in cardiac regeneration and heart repair through altering cardiomyocyte fate plasticity,which is emerging as an effective strategy to compensate for the loss of functional cardiomyocytes and improve the impaired heart functions.First,residual cardiomyocytes in damaged hearts re-enter the cell cycle to acquire the proliferative capacity by the modifications of cell cycle-related genes or regulation of growth-related signals.Additionally,non-cardiomyocytes such as cardiac fbroblasts,were shown to be reprogrammed into cardiomyocytes and thus favor the repair of damaged hearts.Moreover,pluripotent stem cells have been shown to transform into cardiomyocytes to promote heart healing after myocardial infarction(MI).Furthermore,in vitro and in vivo studies demonstrated that environmental oxygen,energy metabolism,extracellular factors,nerves,non-coding RNAs,etc.play the key regulatory functions in cardiac regeneration.These fndings provide the theoretical basis of targeting cellular fate plasticity to induce cardiomyocyte proliferation or formation,and also provide the clues for stimulating heart repair after injury.

Cyclin L1 controls cardiomyocyte proliferation and heart repair after injury

Dear Editor,Myocardial infarction(MI)is characterized by the loss of functional cardiomyocyte(CM)in the heart,resulting in cardiac systolic dysfunction and heart failure.1,2 Increasing evidence suggested that in the heart of neonatal mice after apical resection(AR),the CM can proliferate and regenerate myocardium to repair the heart.While in the heart of adult mice after MI,the CM loses the ability to re-enter the cell cycle but undergoes hypertrophic growth.