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COVID-19 pandemic caused by SARS-CoV-2 infection severely threatens global health and economic development. No effective antiviral drug is currently available to treat COVID-19 and any other human coronavirus infections. We report herein that a macrolide antibiotic, carrimycin, potently inhibited the cytopathic effects (CPE) and reduced the levels of viral protein and RNA in multiple cell types infected by human coronavirus 229E, OC43, and SARS-CoV-2. Time-of-addition and pseudotype virus infection studies indicated that carrimycin inhibited one or multiple post-entry replication events of human coronavirus infection. In support of this notion, metabolic labelling studies showed that carrimycin significantly inhibited the synthesis of viral RNA. Our studies thus strongly suggest that carrimycin is an antiviral agent against a broad-spectrum of human coronaviruses and its therapeutic efficacy to COVID-19 is currently under clinical investigation.

Transforming growth factor beta (TGF‐β) plays an important role in the viral liver disease progression via controlling viral propagation and mediating inflammation‐associated responses. However, the antiviral activities and mechanisms of TGF‐β isoforms, including TGF‐β1, TGF‐β2 and TGF‐β3, remain unclear. Here, we demonstrated that all of the three TGF‐β isoforms were increased in Huh7.5 cells infected by hepatitis C virus (HCV), but in turn, the elevated TGF‐β isoforms could inhibit HCV propagation with different potency in infectious HCV cell culture system. TGF‐β isoforms suppressed HCV propagation through interrupting several different stages in the whole HCV life cycle, including virus entry and intracellular replication, in TGF‐β/SMAD signalling pathway–dependent and TGF‐β/SMAD signalling pathway–independent manners. TGF‐β isoforms showed additional anti‐HCV activities when combined with each other. However, the elevated TGF‐β1 and TGF‐β2, not TGF‐β3, could also induce liver fibrosis with a high expression of type I collagen alpha‐1 and α‐smooth muscle actin in LX‐2 cells. Our results showed a new insight into TGF‐β isoforms in the HCV‐related liver disease progression.

Interferon lambda-2 (IL28A) has a wide antiviral effect with fewer side-effects. Autophagy is a host mechanism to maintain intracellular homeostasis and defends invasion of pathogenic microorganisms. HCV NS5A can disable host defense systems to support HCV replication. Thus, molecular mechanism of interaction among interferon lambda, autophagy, and HCV was concerned and explored in this study. We report that HCV NS5A activated an incomplete autophagy by promoting the autophagic ubiquitylation-like enzymes ATG3, ATG5, ATG7, ATG10, and autophagosome maker LC3B, but blocked autophagy flux; IL28A bound to NS5A at NS5A-ISDR region, and degraded HCV-NS5A by promoting autolysosome formations in HepG2 cells. A software prediction of IL28A protein conformation indicated a potential structure of IL28A homotetramer; the first α-helix of IL28A locates in the interfaces among the four IL28A chains to maintain IL28A homotetrameric conformation. Co-IP and cell immunofluorescence experiments with sequential deletion mutants demonstrate that IL28A preferred a homotetramer conformation to a monomer in the cells; the IL28A homotetramer is positively correlated with autolysosomal degradation of HCV NS5A and the other HCV proteins. Summarily, the first α-helix of IL28A protein is the key domain for maintaining IL28A homotetramer which is required for promoting formation of autolysosomes and degradation of HCV proteins in vitro.

Bicyclol is a synthetic drug for hepatoprotection in clinic since 2004. Preliminary clinical observations suggest that bicyclol might be active against hepatitis C virus (HCV) with unknown mechanism. Here, we showed that bicyclol significantly inhibited HCV replication in vitro and in hepatitis C patients. Using bicyclol as a probe, we identified glycolipid transfer protein (GLTP) to be a novel restrictive factor for HCV replication. The GLTP preferentially bound host vesicle-associated membrane protein-associated protein-A (VAP-A) in competition with the HCV NS5A, causing an interruption of the complex formation between VAP-A and HCV NS5A. As the formation of VAP-A/NS5A complex is essential for viral RNA replication, up-regulation of GLTP by bicyclol reduced the level of VAP-A/NS5A complex and thus inhibited HCV replication. Bicyclol also exhibited an inhibition on HCV variants resistant to direct-acting antiviral agents (DAAs) with an efficacy identical to that on wild type HCV. In combination with bicyclol, DAAs inhibited HCV replication in a synergistic fashion. GLTP appears to be a newly discovered host restrictive factor for HCV replication, Up-regulation of GLTP causes spontaneous restriction of HCV replication., Graphical abstract Treatment of the HCV-infected Huh7.5 cells with bicyclol decreased the level of miR-449b and thus induced the expression of glycolipid transfer protein (GLTP). The up-regulated GLTP preferentially bound host cofactor vesicle-associated membrane protein-associated protein-A (VAP-A) in competition with the HCV NS5A, causing a decrease of the HCV replicative complex level of VAP-A/NS5A, and thus inhibited HCV replication.fx1

Bicyclol, a synthetic hepatoprotective and anti-inflammatory agent approved in China, was widely used to treat various hepatitis accompanied by elevated serum aminotransferases. However, the pharmacological effects and mechanisms of bicyclol on advanced liver diseases, such as fibrosis/cirrhosis and hepatocellular carcinoma (HCC), remain to be explored. Here, we revealed that bicyclol prevents from formatting severe fibrosis, slows the progression of moderate liver fibrosis, accelerates the regression of moderate liver fibrosis, decreases the malignancy of HCC in rat models induced by diethylnitrosamine (DEN), and also blocks steatohepatitis to HCC in mice induced by western diet plus carbon tetrachloride and DEN. The detailed pharmacological mechanism showed that bicyclol alleviates chronic progressive liver diseases by inhibiting the levels of IL-6 and subsequent phosphorylated STAT3. Conclusion: Bicyclol plays significant protective roles in multiply stages of fibrosis/cirrhosis-HCC and nonalcoholic fatty liver disease-related HCC via inhibiting IL-6/STAT3 signaling pathway. Therefore, bicyclol might be a promising therapeutic strategy for treating advanced liver diseases.

Background In China, an abundance of cardiovascular risk factors has contributed to the increasing prevalence of cardiovascular diseases (CVD), which caused almost 4 million deaths per year. However, comprehensive evidence on the geographical profiles of cardiovascular disease risk in China is lacking, as findings in prior studies have been limited to relatively small sample sizes, had incomplete regional coverage, or focused on a narrow risk factor spectrum. Purpose To compare the population CVD risk among different regions across China, and to describe the geographical distributions of CVD risk factors and their clusters throughout the nation. Methods In a nationwide population-based screening project covering 252 counties of China, standardized measurements were conducted to collect information on 12 major CVD risk factors. Individuals of high CVD risk were identified as those with previous CVD, or with a predicted 10-year risk of CVD greater than 10% according to the WHO risk prediction charts. We applied factor analysis to generate “clusters” that characterized the clustering of these risk factors, then explored their relationship with the local ambient temperature and per capital GDP. Results Among 983476 participants included, 9.2% were of high CVD risk, with a range of 1.6% to 23.6% across counties. Among the seven regions in China, the rate was relatively high in the Northeast (11.8%) and North China (10.4%), while low in the South China (7.2%) and Northwest (7.8%). We identified 6 clusters underlying CVD risk factors, including Obesity factor, Blood pressure factor, Staple food factor, Non-staple food factor, Smoking and alcohol factor, and Metabolic and physical activity factor (Figure). We found high risk regions were facing different leading challenges, like obesity and blood pressure for the North China, while unhealthy non-staple food for the Northeast. The South China, as the region with the lowest CVD risk, still had the highest prevalence of unhealthy staple food. Lower annual average ambient temperature was associated with higher risk in Blood pressure factor, Obesity factor and Non-staple food factor, but lower risk in Staple food factor and Metabolic and physical activity factor (p Conclusions The geographical profile of CVD risk in China is complex - population risk levels varied substantially across regions, which were contributed by different risk factors. China needs geographically targeted intervention strategies considering environmental and socio-economic factors to control CVD risk and reduce the burden related to CVD. Geographical disparity of risk clusters Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): The National Key Research and Development Program from the Ministry of Science and Technology of China; the CAMS Innovation Fund for Medical Science

Background Heart failure (HF) is a global public health problem, which causes high mortality. Purpose To characterize the pattern of 1-year mortality, including the rate and timing of death, and to identify clinical and non-clinical factors predicting 1-year all-cause mortality after hospitalization for HF. Methods We prospectively enrolled patients who were hospitalized primarily for HF and discharged alive from 52 hospitals across 20 provinces in China between August 2016 and May 2018, and followed them up by face-to-face interview or telephone. Patients' fatal status were ascertained according to investigators' report and national death registration. Cognitive function was measured by Mini-Cog score and health status was measured by Kansas City Cardiomyopathy Questionnaire (KCCQ). We fitted a Cox proportional hazards model accounting for hospital as random effects to identify patient characteristics associated with 1-year all-cause mortality. Results Among 4899 patients, 4882 (97.7%) completed 1-year follow-up. The median (IQR) age was 67 (57, 75) years and 37.5% were women. The main etiology of HF was ischemic heart disease (53.1%), 66.0% of patients had history of HF before the index hospitalization. Among 813 deaths within one year after discharge, 13.4% and 58.4% occurred within one months and six months, respectively. Systolic blood pressure Conclusion In China, one in six patients hospitalized for HF die within 1-year, with the majority occurring during the first 6 months after hospital discharge. Both clinical and non-clinical factors predict 1-year mortality. Funding Acknowledgement Type of funding source: Other. Main funding source(s): The National Key Research and Development Program from the Ministry of Science and Technology of China; the CAMS Innovation Fund for Medical Science; the 111 Project from the Ministry of Education of China

A series of novel biaryl amide derivatives were synthesized and evaluated for anti-HCV virus activity. Some significant SARs were uncovered. The intensive structural modifications led to fifteen novel compounds with more potent inhibitory activity compared to the hit compounds IMB 26 and IMB1f. Among them, compound 80 was the most active, with EC50 values almost equivalent to the clinical drug telaprevir (EC50 = 15 nM). Furthermore, it also had a good safety and in vitro and oral pharmacokinetic (oral bioavailability in rats: 34%) profile, suggesting a highly drug-like nature. Compound 80represents a more promising scaffold for anti-HCV virus activity for further study., Graphical abstract Image 1

Background Patients with cardiovascular diseases (CVD) are at high risk for recurrent major cardiovascular events. Effective public health strategies to lower blood pressure (BP) are necessary to reduce risk of cardiovascular disease. However, substantial uncertainty remains about the optimal target level to lower BP in patients with cardiovascular disease. Purpose To assess the effects on the incidence of major cardiovascular events in patients with CVD during the scheduled treatment period of greater reduction in blood pressure with a systolic BP (SBP) target Methods This study is a multicenter, open-label, randomized controlled trial comparing two strategies for lowering SBP: lowering SBP to the standard target of Results Regarding the results, we hypothesize that comparing with SBP target of Conclusion This study can provide reliable evaluation on whether more intensive SBP target of

Background Cardiovascular diseases (CVD) is the leading cause of death in China. Secondary prevention medications can improve the prognosis of CVD, yet little is known about the current use, variation and associated factors of these therapies in China. Purpose The aim of this study was to describe the current use of secondary prevention medications among patients with established CVD in the community setting in China, assess variations across population subgroups, and identify the individual characteristics associated with these therapies. Methods We studied 2.6 million participants aged 35–75 years from all 31 provinces in the China Patient-Centered Evaluative Assessment of Cardiac Events (PEACE) Million Persons Project, a government-funded public health program conducted from 2014 to 2018. Participants self-reported their history of ischemic heart disease (IHD) or ischemic stroke (IS) and medication use in an interview. Among participants with IHD and/or IS, we assessed the reported use of secondary prevention medications (anti-platelet drugs and statins) in the overall population and in 1,530,408 population subgroups, defined by all possible combinations of 16 factors (age, sex, urbanity, geographic region, ethnicity, occupation, annual household income, education, marital status, medical insurance, current smoker, current drinker, history of hypertension, history of diabetes, body mass index and years since diagnosis). Multivariable mixed models with a logit link function and community-specific random intercepts were fitted to assess the associations of demographic, socioeconomic and health behavior factors with the reported use of secondary prevention mediations. Results Among 2,613,035 screened participants, 2.9% (74,830) had history of IHD and/or IS (1.2% for IHD, 2.4% for IS). Overall, the reported use rate either anti-platelet drugs or statins was 21.9% (18.3% anti-platelet drugs, 11.0% statins, and 7.4% both). Among the 1,530,408 population subgroups, the use of secondary prevention medications varied substantially (3.4% to 52.0%). Multivariable analyses found that that younger people, women, those living in rural areas, current smokers, current drinkers, people without hypertension or diabetes, and those with established CVD for more than 2 years were less likely to take anti-platelet drugs or statins (Figure). Forest plot of multivariable mixed model Conclusions The current use of secondary prevention drugs is suboptimal and varies substantially across population subgroups in China. Our study identifies target populations for interventions to improve secondary prevention of CVD. Acknowledgement/Funding This study was supported by the Ministry of Finance of China and National Health Commission of China

Aim To investigate the efficacy and safety of ticagrelor combined with a lower dose of aspirin than that recommended by guidelines. Methods Hospitalized patients received ticagrelor (90 mg twice daily) plus aspirin (50–100 mg/day) for up to 12 months. The rates of major adverse cardiovascular events (MACEs), bleeding events and ticagrelor adherence were compared among the groups treated with 50 mg and 75–100 mg aspirin. Results MACE risk was not significantly different between the two groups (OR=0.829, 95% CI: 0.279–2.461, P=0.736). However, 75–100 mg aspirin was associated with a greater risk of bleeding events (OR=1.524, 95% CI: 1.082–2.146, P=0.016), particularly mild and moderate bleeding events (OR=1.480, 95% CI: 1.047–2.092, P=0.026). Moreover, lower-dose aspirin was associated with a lower rate of ticagrelor withdrawal (OR=1.850, 95% CI: 1.025–3.339, P=0.041), mainly because of the decrease in ticagrelor withdrawal due to bleeding (OR=4.565, 95% CI: 1.081–19.270, P=0.039). MACEs and bleeding events within 1 year Endpoint 75–100 mg (n=744) 50 mg (n=188) Unadjusted (95% CI) P value Adjusted# (95% CI) P value MACEs 18 (2.4) 4 (2.1) 0.876 (0.296–2.588) * 0.876 0.829 (0.279–2.461)* 0.736 Bleeding 311 (41.8) 60 (31.9) 1.532 (1.091–2.152)** 0.014 1.524 (1.082–2.146)** 0.016 Severe bleeding 14 (1.9) 2 (1.1) 1.784 (0.402–7.916)** 0.447 1.807 (0.405–8.069)** 0.438 Mild bleeding 297 (39.9) 58 (30.8) 1.489 (1.057–2.098)** 0.023 1.480 (1.047–2.092)** 0.026 #Adjusted for age, sex, LDL-C, hs-CRP, diabetes mellitus, hypertension, hyperlipidaemia and MI history. *Hazard ratio (high vs low); **Odds ratio (high vs low). Conclusion Among patients who took ticagrelor (90 mg twice daily), 50 mg aspirin daily is associated with a lower rate of bleeding events and ticagrelor withdrawal but does not increase the MACE risk compared with 75–100 mg aspirin daily. Acknowledgement/Funding The study was supported by AstraZeneca's fund for the TIFU study (ESR-15-11199).

Background Hospital readmission after heart failure (HF) is common, costly, and potentially preventable. Current patterns of 30-day readmission rates after HF discharge and patient characteristics associated with these readmissions in China are unknown. Purpose To calculate the rates and timing of all-cause readmissions within 30-day after HF hospitalization and to identify patient characteristics associated with readmissions. Methods We included consecutive patients who were hospitalized primarily for HF and discharged alive from 52 geographically diverse hospitals in China followed for 30 days after discharge. Patients were invited for face-to-face interviews to collect information about any readmission. If a patient was unwilling or unable to attend, information was obtained by telephone contact with the patient, his or her relatives, or physicians. We fit mixed models with a logit link function accounting for patient clustering by hospital to identify patient factors associated with 30-day readmission. To estimate the variability in hospital-level, we calculated the median odds ratio (OR) from the fully adjusted hierarchical model, which represents the average likelihood of a statistically identical patient readmitted at one random hospital versus another. Results Among 4649 eligible patients, 4542 (97.7%) had 30-day follow-up. the median (IQR) age was 67 (57, 75) years; 62.5% were men. The length of index hospital stay was 9 (7, 13) days. Readmissions for any cause within 30 days of discharge occurred in 562 patients (12.4%), of which 30.1% occurred within 7 days of discharge and 58.9% 15 days. Half of readmission diagnosis within 30 days were HF (283, 50.4%). Prior HF (OR 1.3, 95% CI 1.09–1.55), valvular heart disease (OR 1.64, 95% CI 1.26–2.1), chronic obstructive pulmonary disease (OR 1.35, 95% CI 1.05–1.73), diabetes (OR 1.44, 95% CI 1.19–1.74) and longer length of index hospitalization stay (HR 1.02, 95% CI 1.01–1.03 per day) were associated with higher risks of 30-day readmission. The C-statistic was 0.615. The median odds ratio was 1.32 (1.05–1.48). Conclusion In China, one in eight patients hospitalized for HF are readmitted within 30-days, with the majority occurring within 2 weeks of discharge. Both patient characteristics and variability across hospitals accounted for the variability in readmissions, suggesting an important opportunity for improving the transitions of care in HF.

Background Leukocyte telomere length, as an emerging marker of biological age, has been shown to associate with hypertension. However, it has not been studied whether telomere attrition rate in patients with hypertension is related to the heterogeneity of blood pressure (BP) response to antihypertensive therapy. Purpose Our aim is to investigate the relationship between telomere attrition rate and BP lowering in a longitudinal Chinese hypertensive cohort. We also aim to explore the potential association between telomere attrition rate and the differences in antihypertensive treatment response. Methods A community-based, prospective study was conducted at BenXi county, Liaoning province, in the northern China. A total of 3,671 hypertensive patients were recruited from 2013 t 2015 and of whom 1,382 provided blood samples at baseline. After a median follow-up period of 2.2 (range 1.5–2.4) years, the blood samples were collected from 1,197 patients again in 2016, and 185 patients were not reached to obtain blood sample because of immigration. In addition, 89 blood samples were excluded due to insufficient quality. Finally, 1,108 patients who are available for blood samples both at baseline and at follow-up, were included in the analysis for telomeres change. Annual telomere attrition rate was calculated as (follow-up telomere length-baseline telomere length)/follow-up year, and then categorized into two groups: the shorten (annual telomere attrition rate 0). Multivariable linear regression model was used to examine the association of annual telomere attrition rate with blood pressure lowering and antihypertensive treatment. Cox Proportional Hazards model was used to examine the association between annual telomere attrition rate and cardiovascular disease risk. Results In multivariable linear regression models, the telomere lengthening was significantly associated with decrease in systolic blood pressure (SBP) (β: −4.13; p=0.006) and pulse pressure (PP) (β: −3.22; p=0.007) during the follow-up, but not associated with diastolic blood pressure (DBP) change. And the associations were observed age- and gender-specific difference. The lengthen was significantly associated with ΔSBP and ΔPP in women and younger patients (age ≤60 years old). Furthermore, the associations were observed in patients who treated with calcium channel blocker (CCB) and angiotensin receptor blocker (ARB), but not in diuretics. Then we found no significant association between annual telomere attrition rate and incident cardiovascular events during the follow-up. Conclusion(s) Our data showed that the increasing of leukocyte telomere length is associated with the decreasing of SBP and PP, particularly for the patients who received CCB and ARB therapy. These data showed that annual telomere attrition rate could be a marker of treatment response and will help in clinical management. Acknowledgement/Funding the National Science and Technology Pillar Program during the Twelfth Five-year Plan Period (No. 2011BAI11B04)

With the development of more effective direct-acting antivirals (DAAs), dual- or triple-therapy regimens represent the major strategy used to cure chronic hepatitis C virus (HCV) infection. Thus, shorter treatment duration regimens with low burden, few adverse effects and good patient adherence are urgently needed. This study theoretically demonstrates a proof-of-concept approach for shortening therapy duration by examining HCV-infected Huh7.5 cells after treatment with a high or low fixed dose of three DAAs (simeprevir + daclatasvir + sofosbuvir) for 6–15 days. The results demonstrated that HCV-infected Huh7.5 cells achieved an ultrarapid virologic response with undetectable HCV RNA and protein and were cured after treatment with the triple-therapy regimen for 15 days. When the treatment duration was shortened, virologic relapse might occur after treatment with a low fixed dose of the three DAAs for 9 days and did occur after treatment with a low fixed dose for 6 days, although HCV was below detectable levels at the end of treatment. However, virologic relapse could be avoided with treatment of a high fixed dose of the three DAAs for 9 or 6 days. Although a virologic breakthrough occurred after an intermittent treatment regimen at the low fixed dose, the high fixed dose cured HCV-positive Huh7.5 cells with intermittent treatment. In conclusion, HCV is persistently present below detectable levels in HCV-infected Huh7.5 cells for a long time after treatment, and a shortened therapy duration is associated with an increased risk of virologic relapse, but virologic relapse or breakthrough might be avoided by treatment with a combination of more highly effective DAAs.

Central nervous system (CNS) metastases are very common in patients with non-small-cell lung cancer (NSCLC). We aimed to explore the clinical impact of osimertinib, a third-generation epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI), on CNS metastases in patients with advanced NSCLC in real-world setting. Patients with advanced NSCLC who received osimertinib after progression of early-generation EGFR-TKIs and CNS metastases on baseline brain scan were retrospectively collected. Primary outcomes were disease control rate (DCR) and progression-free survival (PFS), and secondary objectives were objective response rate (ORR), time to tumor response, median best percentage change from baseline in CNS target lesion (TL) size and safety. Between Apr 1, 2017, and Dec 30, 2017, 22 patients met selection criteria, 15 with ≥ 1 measurable CNS lesion (RECIST 1.1) were included in CNS evaluable for response (cEFR) set. Among the 22 patients, ORR and DCR were 40.9% and 86.4%, respectively, with median PFS of 8.5 months (95% CI 4.1, 13.0). Median intracranial PFS was not reached. Of 15 patients in cEFR set, CNS DCR was 80.0% with complete response reported in 3 patients (20.0%). Median best percentage change from baseline in CNS TL size was − 40% (range − 100 to + 60%) and median time to CNS tumor response was 1.3 months. CNS ORR was 53.3%. The safety profile was acceptable and no new unexpected findings were found. This real-world analysis further confirmed that osimertinib indeed demonstrated clinically meaningful efficacy against CNS metastases in Chinese patients with advanced NSCLC.

Autophagy-related 10 (ATG10) is essential for autophagy since it promotes ATG5-ATG12 complex formation. Our previous study found that there are two isoforms of the ATG10 protein, ATG10 (a longer one) and ATG10S, which have identical sequences except an absence of a 36-amino acid fragment (peptide B) in ATG10S, yet exhibit distinct effects on HCV genome replication. Here, we report the existence of two amino acids, cysteine at residue 44 and 135 (Cys44 and Cys135, respectively), in ATG10 being related to differential effects of ATG10 on HCV replication and autophagy flux. Through a series of ATG10 mutation experiments and protein modeling prediction, we found that Cys44 was involved in the dual role of the two isoforms of ATG10 protein on HCV replication and autophagy flux, and that Cys135 plays similar roles as Cys44, but the disulfide bond of Cys44-Cys135 was not verified in the ATG10 protein. Further analyses by full HCV virion infection confirmed the roles of -SH of Cys44 and Cys135 on HCV replication. ATG10 with deleted or mutated Cys44 and/or Cys135 could activate expression of innate immunity-related genes, including il28a, irf-3, irf-7, and promote complete autophagy by driving autophagosomes to interact with lysosomes via IL28A-mediation. Subcellular localization assay and chromatin immunoprecipitation assay showed that ATG10 with the sulfydryl deletion or substitution of Cys44 and Cys135 could translocate into the nucleus and bind to promoter of IL28A gene; the results indicated that ATG10 with Cys44 and/or Cys135 absence might act as transcriptional factors to trigger the expression of anti-HCV immunological genes, too. In conclusion, our findings provide important information for understanding the differential roles on HCV replication and autophagy flux between ATG10 and ATG10S, and how the structure-function relationship of ATG10 transformed by a single -SH group loss on Cys44 and Cys135 in ATG10 protein, which may be a new target against HCV replication.

The hepatitis C virus (HCV) infection is associated with various extrahepatic manifestations, which are correlated with poor outcomes, and thus increase the morbidity and mortality of chronic hepatitis C (CHC). Therefore, understanding the internal linkages between systemic manifestations and HCV infection is helpful for treatment of CHC. Yet, the mechanism by which the virus evokes the systemic diseases remains to be elucidated. In the present study, using gene set enrichment analysis (GSEA) and signaling pathway impact analysis (SPIA), a comprehensive analysis of micro-array data of mRNAs was conducted in HCV-infected and -uninfected Huh7.5 cells, and signaling pathways (which are significantly activated or inhibited) and certain molecules (which are commonly important in those signaling pathways) were selected. Forty signaling pathways were selected using GSEA, and eight signaling pathways were selected with SPIA. These pathways are associated with cancer, metabolism, environmental information processing and organismal systems, which provide important information for further clarifying the intrinsic associations between syndromes of HCV infection, of which seven pathways were not previously reported, including basal transcription factors, pathogenic Escherichia coli infection, shigellosis, gastric acid secretion, dorso-ventral axis formation, amoebiasis and cholinergic synapse. Ten genes, SOS1, RAF1, IFNA2, IFNG, MTHFR, IGF1, CALM3, UBE2B, TP53 and BMP7 whose expression may be the key internal driving molecules, were selected using the online tool Anni 2.1. Furthermore, the present study demonstrated the internal linkages between systemic manifestations and HCV infection, and presented the potential molecules that are key to those linkages.

Autophagy and immune response are two defense systems that human-body uses against viral infection. Previous studies documented that some viral mechanisms circumvented host immunity mechanisms and hijacked autophagy for its replication and survival. Here, we focus on interactions between autophagy mechanism and innate-immune-response in HCV-subgenomic replicon cells to find a mechanism linking the two pathways. We report distinct effects of two autophagy-related protein ATG10s on HCV-subgenomic replication. ATG10, a canonical long isoform in autophagy process, can facilitate HCV-subgenomic replicon amplification by promoting autophagosome formation and by combining with and detaining autophagosomes in cellular periphery, causing impaired autophagy flux. ATG10S, a non-canonical short isoform of ATG10 proteins, can activate expression of IL28A/B and immunity genes related to viral ds-RNA including ddx-58, tlr-3, tlr-7, irf-3 and irf-7, and promote autophagolysosome formation by directly combining and driving autophagosomes to perinuclear region where lysosomes gather, leading to lysosomal degradation of HCV-subgenomic replicon in HepG2 cells. ATG10S also can suppress infectious HCV virion replication in Huh7.5 cells. Another finding is that IL28A protein directly conjugates ATG10S and helps autophagosome docking to lysosomes. ATG10S might be a new host factor against HCV replication, and as a target for screening chemicals with new anti-virus mechanisms.

Background ROS1 rearrangement, a novel specific molecular subtype of non-small cell lung cancer (NSCLC), has become a validated therapeutic target. However, the efficacy of crizotinib and platinum-based chemotherapy as first-line treatment between different ROS1 fusion variants in advanced Chinese NSCLC patients has not yet been fully elucidated. This study explored which treatment regimen would be better in such patients. Methods Retrospective real-world analyses of clinical and treatment outcome with ROS1-positive advanced NSCLC patients from multi-center study were explored in China, from Jan 9th 2011 to Jan 1st 2019. The study evaluated first-line regimens including crizotinib treatment and platinum-based chemotherapy. The primary endpoint was progression-free survival (PFS), and second endpoints were the objective response rate (ORR) and overall survival (OS). Kaplan-Meier survival analysis was carried out by using SPSS 16.0. Results A total of 102 patients, 75 (73.5%) females and 27 (26.5%) males were enrolled. The median age of patients was 52 years and 99 (97.1%) were identified with adenocarcinoma type. 57 cases were administered crizotinib and 45 received platinum-based chemotherapy. For the first-line treatment, the median PFS was significantly longer for crizotinib treatment compared with platinum-based chemotherapy (median,15.0 vs. 9.7months, p = 0.006), respectively. 58 cases were confirmed ROS1 fusion variants by next-generation sequence (NGS) including CD74 fusion (n = 32) and non-CD74 fusion(n = 26). Patients receiving crizotinib with the CD74 fusion had a significant longer PFS than those by platinum-based chemotherapy (median,18.3 vs. 8.8months, p = 0.002); in contrast, a statistical significance was not observed on PFS in the non-CD74 fusion subgroup (12.2 vs. 9.0 months, p = 0.544). The ORR of crizotinib treatment was higher than that of platinum-based chemotherapy (80.7% vs. 55.6%, p = 0.006). OS was not reached owing to immature data. Conclusions Crizotinib as first-line treatment tends to have more benefit than platinum-based chemotherapy for advanced Chinese NSCLC patients with CD74 ROS1 fusion variant. Legal entity responsible for the study The authors. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

Host heat shock cognate 70 (Hsc70) protein is packaged into hepatitis C viral (HCV) particles as a structural component of the virus in the assembly process. It helps HCV RNA release into the cytoplasm in the next infection cycle. The goal of this study is to investigate whether chemically down-regulating host Hsc70 expression could be a novel strategy to interrupt HCV replication. Compounds were screened with an Hsc70 messenger RNA (mRNA) assay. IMB-DM122 was found to be an effective and safe inhibitor for Hsc70 mRNA/protein expression in human hepatocytes. IMB-DM122 inhibited HCV replication through destabilization of Hsc70 mRNA, and the half-life of host Hsc70 mRNA was reduced by 78% after the compound treatment. The Hsc70 mRNA 3′ untranslated region sequence is the element responsible for the effect of IMB-DM122 on Hsc70 mRNA. The compound appears to be highly efficient in inhibiting Hsc70-related HCV replication. Treatment of the HCV-infected hepatocytes with IMB-DM122 reduced the virion encapsidation of Hsc70, and therefore disrupted HCV replication and the infection cycle. IMB-DM122 showed considerable good safety in vitro as well as in vivo with no indication of harmful effect on liver and kidney functions. Conclusion: Hsc70 might be a new drug target and mechanism to inhibit HCV proliferation. (HEPATOLOGY 2010;)

The cluster of differentiation 36 (CD36) is a membrane protein related to lipid metabolism. We show that HCV infection in vitro increased CD36 expression in either surface or soluble form. HCV attachment was facilitated through a direct interaction between CD36 and HCV E1 protein, causing enhanced entry and replication. The HCV co-receptor effect of CD36 was independent of that of SR-BI. CD36 monoclonal antibodies neutralized the effect of CD36 and reduced HCV replication. CD36 inhibitor sulfo-N-succinimidyl oleate (SSO), which directly bound CD36 but not SR-BI, significantly interrupted HCV entry, and therefore inhibited HCV replication. SSO’s antiviral effect was seen only in HCV but not in other viruses. SSO in combination with known anti-HCV drugs showed additional inhibition against HCV. SSO was considerably safe in mice. Conclusively, CD36 interacts with HCV E1 and might be a co-receptor specific for HCV entry; thus, CD36 could be a potential drug target against HCV.

Human APOBEC3G (hA3G) is a cytidine deaminase that restricts replication of certain viruses. We have previously reported that hA3G was a host restriction factor against hepatitis C virus (HCV) replication, and hA3G stabilizers showed a significant inhibitory activity against HCV. However, the molecular mechanism of hA3G against HCV remains unknown. We show in this study that hA3G’s C-terminal directly binds HCV non-structural protein NS3 at its C-terminus, which is responsible for NS3’s helicase and NTPase activity. Binding of hA3G to the C-terminus of NS3 reduced helicase activity, and therefore inhibited HCV replication. The anti-HCV mechanism of hA3G appeared to be independent of its deamination activity. Although early stage HCV infection resulted in an increase in host hA3G as an intracellular response against HCV replication, hA3G was gradually diminished after a long-term incubation, suggesting an unknown mechanism(s) that protects HCV NS3 from inactivation by hA3G. The process represents, at least partially, a cellular defensive mechanism against HCV and the action is mediated through a direct interaction between host hA3G and HCV NS3. We believe that understanding of the antiviral mechanism of hA3G against HCV might open an interesting avenue to explore hA3G stabilizers as a new class of anti-HCV agents.

Applications of bone marrow-derived mesenchymal stem cells (BM-MSCs) have been documented for diseases occur in the sports system, the central nervous system, the cardiovascular system etc. However, poor viability of donor stem cells after transplantation limits their therapeutic efficiency. Although the autophagy theory has been reported, the underlying mechanisms are still poorly understood. Isolation and culture methods of mesenchymal stem cells are currently concentrate on four ways. Overall, BM-MSCs have both important research significance and clinical application value in cell replacement therapy, gene therapy and reconstruction of tissues as well as organs especially for myocardial infarction (MI). In this article, we review the biological characteristics of BM-MSCs and its research progress especially in MI.

BACKGROUND: Human tissue kallikrein (hTK) plays an essential role in the physiological and pathological mechanisms of blood vessels. This study aimed to determine whether angiogenesis induced by endothelial progenitor cells (EPCs) transduced with the adenovirus-mediated hTK gene could improve blood flow in rat hindlimb ischemia in vivo and to establish a promising mechanism in vitro. METHODS: EPCs transduced with adenovirus encoding hTK-162 (i.e., Ad/hTK-transduced EPCs or Ad/GFP-transduced EPCs) were administered to Wister rats with hindlimb ischemia through therapeutic neovascularization. Muscular capillary density (MCD), blood flow (BF), and the number of myofibers were measured at days 7, 14, and 21 after treatment. Expressions of integrin αvβ3 and endothelial nitric oxide synthase (eNOS) were detected on the surface of EPCs. RESULTS: MCD, BF, and the number of myofibers in rats with Ad/hTK-transduced EPCs remarkably increased at day 21 after treatment compared with rats with Ad/GFP-transduced EPCs or the control group (P

Heat-stress cognate 70 (Hsc70) is a host factor that helps hepatitis C virus (HCV) to complete its life cycle in infected hepatocytes. Using Hsc70 as a target for HCV inhibition, a series of novel N-substituted benzyl matrinic/sophoridinic acid derivatives was synthesized and evaluated for their anti-HCV activity in vitro. Among these analogues, compound 7c possessing N-p-methylbenzyl afforded an appealing ability to inhibit HCV replication with SI value over 53. Furthermore, it showed a good oral pharmacokinetic profile with area-under-curve (AUC) of 13.4 µM·h, and a considerably good safety in oral administration in mice (LD50>1000 mg/kg). As 7c suppresses HCV replication via an action mode distinctly different from that of the marketed anti-HCV drugs, it has been selected as a new mechanism anti-HCV candidate for further investigation, with an advantage of no or decreased chance to induce drug-resistant mutations.

Pluripotency makes human pluripotent stem cells (hPSCs) promising for regenerative medicine, but the teratoma formation has been considered to be a major obstacle for their clinical applications. Here, we determined that the downregulation of miR-302 suppresses the teratoma formation, hampers the self-renewal and pluripotency, and promotes hPSC differentiation. The underlying mechanism is that the high endogenous expression of miR-302 suppresses the AKT1 expression by directly targeting its 3'UTR and subsequently maintains the pluripotent factor OCT4 at high level. Our findings reveal that miR-302 regulates OCT4 by suppressing AKT1, which provides hPSCs two characteristics related to their potential for clinical applications: the benefit of pluripotency and the hindrance of teratoma formation. More importantly, we demonstrate that miR-302 upregulation cannot lead OCT4 negative human adult mesenchymal stem cells (hMSCs) to acquire the teratoma formation in vivo. Whether miR-302 upregulation can drive hMSCs to acquire a higher differentiation potential is worthy of deep investigation.