Prostate cancer (PCa) is the most prevalent malignant tumor in the male genitourinary system. Carbon monoxide (CO) gas therapy and chemodynamic therapy (CDT) have emerged as alternative treatments for cancer. However, the therapeutic efficacy of both approaches is constrained by the insufficient endogenous hydrogen peroxide (H2O2) supply and the overexpression of protective glutathione (GSH) in the tumor microenvironment (TME). Herein, we developed a biomimetic cascaded nanoreactor (DCM@MCuH@MnCO) for synergistically combining CO-mediated gas therapy with CDT driven by depletion of GSH and self-supply of H2O2 in the tumor. DCM@MCuH@MnCO was constructed by integrating a copper-catechol coordinated network (CuH) and H2O2-sensitive manganese carbonyl (MnCO) within the mesoporous carbon hollow spheres (MCHS), and further modifying the surface with tumor cell membranes (DCM) to enhance its tumor-targeting ability. The CuH in the nanoreactor can disassemble, triggered by continuous consumption of GSH, releasing pro-oxidative catechol ligands to generate H2O2 for TME remodeling. Subsequently, MnCO decomposes on demand to release CO in situ while copper ions (Cu+) convert H2O2 into reactive oxygen species (ROS) via a Fenton-like reaction. These events cooperatively cause mitochondrial damage and induce a lethal ROS storm within cells, ultimately effectively killing tumor cells both in vitro and in vivo. This study demonstrates significant prospects for CO therapy and CDT, offering a novel and promising strategy for efficient PCa treatment.

Background: Pneumonia is a common acute respiratory infection that contributes to significant mortality and morbidity worldwide. The disruption of the airway microbiome in respiratory infection has been extensively reported. However, whether the changes in respiratory tract microbial communities during pneumonia were related to disease severity remains elusive. Herein, we aimed to investigate the correlation between the changes in airway microbiome and immune response in pneumonia patients. Methods: We performed metagenomic and metatranscriptomic sequencing on immunocompetent (ICO) and immunocompromised host (ICH) with pneumonia using bronchoalveolar lavage fluid (BALF), blood, sputum, and swab samples. Results: Compared to ICO patients with pneumonia, ICH patients had higher Pneumonia Severity Index (PSI) score. BALF metagenomic and metatranscriptomic sequencing showed higher microbial diversity in ICH patients, while ICH patients exhibited lower microbial diversity in sputum samples. Additionally, pneumonia patients with different PSI scores exhibited different microbial communities. Relative abundance of Human Gammaherpesvirus 4 (EBV) was positively correlated with PSI score. For ICH patients, BALF metatranscriptomic sequencing found 183 up-regulated genes and 85 down-regulated genes in EBV-detected group compared with EBV not-detected group, while there was no significant difference in ICO patients, indicating that EBV might be reactivated in ICH patients, while EBV might be latent in ICO patients. In ICH patients, we observed significant down-regulation of immune related genes and interferon stimulated genes in EBV-detected group compared to the not-detected group, including CSF1R, CXCR6, IL10, IL16, and TNFRSF25. Co-occurrence network analysis found positive correlations between EBV and Citrobacter freundii or Campylobacter concisus, indicating that synergistic effects on exacerbating the severity of pneumonia might exist between EBV and these two microbes. Conclusion: EBV might be considered as a microbial signature for disease severity, which could regulate immune-related signaling pathways. Notably, we unravel that EBV presence might inhibit the immune response of hosts, reduce anti-inflammatory responses, and increase the possibilities of infections caused by other pathogens, exacerbating the pneumonia severity.

Prostate cancer (PCa) presents a formidable therapeutic challenge owing to the limited efficacy of existing treatments. In this study, a series of comparative experiments demonstrated that nanodiamonds (NDs) with different functional groups exhibit inhibitory effects on the growth, replication, and migration of prostate cancer cells without directly killing the cells. Among them, aminated nanodiamonds (aNDs) showed the most pronounced inhibitory activity. Subsequently, a combination of proteomics analysis and machine learning was employed to elucidate the molecular mechanisms by which these aNDs exerted their inhibitory effects on PCa progression. Data-independent acquisition proteomics identified the key proteins affected by the aNDs, and the Kyoto Encyclopedia of Genes and Genomes analysis revealed ribosome pathway enrichment. Bioinformatics analysis focused on ten crucial genes, thus incorporating a novel prognostic model that categorized patients based on gene expression and emphasized the significance of mitochondrial ribosomal protein L22 (MRPL22). In vivo experiments confirmed the antitumor effects and biocompatibility of the aNDs. In addition, an analysis of the response of MRPL22 to anticancer drugs in public databases revealed its relevance to a range of drugs and compounds. In summary, this study integrated diverse interdisciplinary methodologies, thereby offering insights into the mechanism of the inhibition of PCa by the aNDs and the regulatory role of MRPL22 in the ribosomal pathway.

Background Pneumonia is a common acute respiratory infection that contributes to significant mortality and morbidity worldwide. The disruption of the airway microbiome in respiratory infection has been extensively reported. However, whether the changes in respiratory tract microbial communities during pneumonia were related to disease severity remains elusive. Herein, we aimed to investigate the correlation between the changes in airway microbiome and immune response in pneumonia patients. Methods We performed metagenomic and metatranscriptomic sequencing on immunocompetent (ICO) and immunocompromised host (ICH) with pneumonia using bronchoalveolar lavage fluid (BALF), blood, sputum, and swab samples. Results Compared to ICO patients with pneumonia, ICH patients had higher Pneumonia Severity Index (PSI) score. BALF metagenomic and metatranscriptomic sequencing showed higher microbial diversity in ICH patients, while ICH patients exhibited lower microbial diversity in sputum samples. Additionally, pneumonia patients with different PSI scores exhibited different microbial communities. Relative abundance of Human Gammaherpesvirus 4 (EBV) was positively correlated with PSI score. For ICH patients, BALF metatranscriptomic sequencing found 183 up-regulated genes and 85 down-regulated genes in EBV-detected group compared with EBV not-detected group, while there was no significant difference in ICO patients, indicating that EBV might be reactivated in ICH patients, while EBV might be latent in ICO patients. In ICH patients, we observed significant down-regulation of immune related genes and interferon stimulated genes in EBV-detected group compared to the not-detected group, including CSF1R, CXCR6, IL10, IL16, and TNFRSF25. Co-occurrence network analysis found positive correlations between EBV and Citrobacter freundii or Campylobacter concisus, indicating that synergistic effects on exacerbating the severity of pneumonia might exist between EBV and these two microbes. Conclusion EBV might be considered as a microbial signature for disease severity, which could regulate immune-related signaling pathways. Notably, we unravel that EBV presence might inhibit the immune response of hosts, reduce anti-inflammatory responses, and increase the possibilities of infections caused by other pathogens, exacerbating the pneumonia severity.

Purpose: Current treatment approaches for Prostate cancer (PCa) often come with debilitating side effects and limited therapeutic outcomes. There is urgent need for an alternative effective and safe treatment for PCa. Methods: We developed a nanoplatform to target prostate cancer cells based on graphdiyne (GDY) and a copper -based metal -organic framework (GDY-CuMOF), that carries the chemotherapy drug doxorubicin (DOX) for cancer treatment. Moreover, to provide GDYCuMOF@DOX with homotypic targeting capability, we coated the PCa cell membrane (DU145 cell membrane, DCM) onto the surface of GDY-CuMOF@DOX, thus obtaining a biomimetic nanoplatform (DCM@GDY-CuMOF@DOX). The nanoplatform was characterized by using transmission electron microscope, atomic force microscope, X-ray diffraction, etc. Drug release behavior, antitumor effects in vivo and in vitro, and biosafety of the nanoplatform were evaluated. Results: We found that GDY-CuMOF exhibited a remarkable capability to load DOX mainly through pi-conjugation and pore adsorption, and it responsively released DOX and generated Cu + in the presence of glutathione (GSH). In vivo experiments demonstrated that this nanoplatform exhibits remarkable cell -killing efficiency by generating lethal reactive oxygen species (ROS) and mediating cuproptosis. In addition, DCM@GDY-CuMOF@DOX effectively suppresses tumor growth in vivo without causing any apparent side effects. Conclusion: The constructed DCM@GDY-CuMOF@DOX nanoplatform integrates tumor targeting, drug -responsive release and combination with cuproptosis and chemodynamic therapy, offering insights for further biomedical research on efficient PCa treatment.

To screen target gene cluster by bioinformatics analysis and verify them by in vitro experiment and clinicopathological correlation analysis. We try to find a new biomarker with prognostic value for prostate cancer (PCa). 42 candidate marker genes were constructed by protein protein interaction (PPI) network and enriched by KEGG pathway to find out the gene cluster we are interested in. Prognostic model was established to preliminarily analyze the prognostic value of this gene cluster in PCa, and Cox risk regression was used for comparative analysis. Immunohistochemistry was used to detect the expression of each gene in clinical tissue microarray. Finally, we analyzed the correlation between each gene and their clinicopathological features of PCa combined with TCGA clinical data. Based on the analysis of PPI and KEGG, we found the target gene cluster (FCGR3A, HAVCR2, CCR7 and CD28). Prognostic model analysis showed that this gene cluster had the ability to predict biochemical recurrence, and the survival rate and ROC analysis showed favorable prediction effect. Univariate Cox regression analysis showed that the risk scores of Gleason score (GS), T stage, N stage and PSA were significantly different (P<0.05), and the risk ratio of high expression was 2.30 times that of low expression (P=0.004). However, it was not statistically significant in multivariate Cox regression analysis (P>0.05). The results of tissue microarray showed that FCGR3A and HAVCR2 were highly expressed in PCa (P<0.01), while the expression of CCR7 and CD28 had no significant difference (P>0.05). Kaplan-Meier analysis showed that there was significant difference in BCR free survival of FCGR3A and HAVCR2 (FCGR3A, P=0.010; HAVCR2, P=0.018), while the expression of CCR7 and CD28 had no significant difference on the survival and prognosis of PCa patients (P>0.05). TCGA clinical data analysis found that the expression of FCGR3A had a unique correlation with the clinicopathological features of PCa, which was closely related to the tumor stage. The expression of FCGR3A is related to BCR free survival of PCa patients. Therefore, FCGR3A is a new biomarker with potential prognostic value of PCa.

目的 探讨双黄连雾化吸入(SHL Inh)对小鼠H9 N2亚型禽流感病毒感染的疗效.方法 将45只BALB/c小鼠随机分为正常对照组、病毒对照组、奥司他韦组(阳性药对照)、双黄连腹腔注射(SHL Ip)组和SHL Inh组,每组9只.正常对照组经鼻滴入磷酸盐缓冲液(PBS),其余4组均经鼻接种等量H9N2禽流感病毒.奥司他韦组、SHL Ip组和SHL Inh组在接种病毒后分别给予相应药物治疗,正常对照组和病毒对照组给予等量蒸馏水,连续给药5d.每日记录小鼠体质量,接种后第5天处死,取肺称重,取部分肺组织制作HE病理切片,其余肺组织匀浆后测定病毒滴度、炎症因子表达水平.结果 病毒对照组和SHL Ip组小鼠肺指数均高于正常对照组,差异有统计学意义(P＜0.01).SHL Inh组肺指数低于病毒对照组,高于SHL Ip组,差异有统计学意义(P＜0.01).病毒对照组肺病毒滴度为(4.06±0.41) lgTCID50/g,奥司他韦组、SHL Inh组和SHL Ip组病毒滴度依次为(3.29±0.27)、(3.44±0.46)、(3.33±0.43) lgTCID50/g,均低于病毒对照组病毒滴度,差异有统计学意义(P＜0.05或P＜0.01).SHL Inh并未改善小鼠体质量下降,也未降低肺炎症因子水平.SHL Ip组与SHL Inh组在体质量下降、肺炎症因子水平、肺病毒滴度和肺病理炎症方面的差异无统计学意义(P＞0.05).结论 SHL Inh具有抗H9N2亚型禽流感病毒的疗效,且雾化给药与注射给药在降低病毒滴度方面的疗效相当.

Chronic obstructive pulmonary disease (COPD) is a worldwide chronic inflammatory lung disease, and influenza A virus (IAV) infection is a common cause of acute exacerbations of COPD (AECOPD). Therefore, targeting viral infections represents a promising strategy to prevent the occurrence and development of inflammatory flare ups in AECOPD. Jianpiyifei II (JPYFII) is a traditional herbal medicine used in China to treat patients with COPD, and its clinical indications are not well understood. However, investigation of the anti-inflammatory effects and underlying mechanism using an animal model of smoking have been reported in a previous study by our group. In addition, some included herbs, such as Radix astragali and Radix aupleuri, were reported to exhibit antiviral effects. Therefore, the aim of the present study was to investigate whether JPYFII formulation relieved acute inflammation by clearing the IAV in a mouse model that was exposed to cigarette smoke experimentally. JPYFII formulation treatment during smoke exposure and IAV infection significantly reduced the number of cells observed in bronchoalveolar lavage fluid (BALF), expression of proinflammatory cytokines, chemokines, superoxide production, and viral load in IAV-infected and smoke-exposed mice. However, JPYFII formulation treatment during smoke exposure alone did not reduce the number of cells in BALF or the expression of Il-6, Tnf-a, and Il-1β. The results demonstrated that JPYFII formulation exerted an antiviral effect and reduced the exacerbation of lung inflammation in cigarette smoke (CS)-exposed mice infected with IAV. Our results suggested that JPYFII formulation could potentially be used to treat patients with AECOPD associated with IAV infection.

Ethnopharmacological relevance: Influenza virus infection is widely believed to cause mild symptoms, but can lead to high mortality and severe disease complicated by secondary bacterial pneumonia. Traditional Chinese medicine (TCM) has been proposed as a promising agent to treat respiratory viral infections. A herbal formula Lianhuaqingwen capsule (LHQW) comprising two prescriptions: Maxing Shigan decoction and Yinqiao San, has been used clinically to treat respiratory infection with immune regulatory effects. However, little is known about the capacity of LHQW against influenza-induced secondary bacterial pneumonia. Aim of study: This study aimed to evaluate the efficacy and underlying mechanism of LHQW on influenza A virus A/PR/8/34 (PR8) secondary methicillin-resistant Staphy-lococcus aureus (MRSA) infection. Methods: The anti-adhesion activity of LHQW against PR8-induced MRSA infection was assessed in human lung epithelial (A549) cells and the effect of LHQW on the expression of intracellular adhesion molecule 1 (ICAM-1) was detected. Also, the mRNA expression levels of inflammatory cytokines upon lipopolysaccharide (LPS) stimulation in PR8-infected A549 cells were determined. The body weight change, survivals, viral titers, colonies and the pathological parameters after LHQW treatment in severe pneumonia model have all been systematically determined. Results: LHQW significantly reduced the adhesion of MRSA to PR8-infected A549 cells in a dose-dependent manner by suppressing the up-regulation of bacterial receptors. LHQW also markedly declined the over expression of IL-6, IL-8, and TNF-alpha induced by LPS stimulated-A549 cells following influenza virus infection. Furthermore, the abnormal changes of lung index in dual-infection mice were relieved after administered with LHQW in preventive and therapeutic mode, but with no significantly difference (P > 0.05). LHQW could not effectively improve survival rate or prolong the survival time of mice (P > 0.05). LHQW (1000 mg/kg/d) administered prophylactically significantly decreased the lung viral titers (P < 0.05), slightly downregulated IL 6 but TNF-alpha, IL-1 beta levels and improved lung pathological inflammation including neutrophil infiltration, necrosis, which is consistent with the expression of inflammatory factors. Conclusions: LHQW inhibited influenza-induced bacterial adhesion by down-regulating the adhesion molecules with the improvement trend on severe pneumonia, indicating that it can be used as an adjuvant medication in severe viral-bacterial pneumonia therapy rather than as a single medication.

目的 香烟暴露对流感病毒感染小鼠气道灌洗液免疫细胞的影响.方法 香烟烟雾暴露4d,第5天给予小鼠3个滴度(6、12、24 PFU)的流感病毒H1N1(A/PR/8/34)[假熏烟假攻毒组、假熏烟攻毒组(6PFU)、假熏烟攻毒组(12PFU)、假熏烟攻毒组(24PFU)、熏烟联合攻毒组(6PFU)、熏烟联合攻毒组(12PFU)、熏烟联合攻毒组(24PFU)].分别观察实验后第3、5、7天气道灌洗液免疫细胞及相关改变.观察内容为:支气管灌洗液中的细胞总数及分类计数;化学发光法检测灌洗液细胞活性氧簇(ROS)的产生;以及BCA法检测小鼠灌洗液蛋白浓度.结果 细胞计数结果表明H1N1病毒感染导致气道灌洗液细胞计数及分类计数呈滴度依赖性升高,并在攻毒后3 d(24PFU)、7 d(12PFU)达到应答高峰,并伴随灌洗液细胞ROS的产生速率在相应时点升高.香烟暴露的H1N1病毒感染小鼠气道的灌洗液细胞计数和分类计数在攻毒后5d(24PFU)达到应答高峰,并伴有灌洗液细胞ROS产生速率的增加.在攻毒后3、5、7d,H1N1病毒感染小鼠气道灌洗液蛋白含量均明显升高,且熏烟联合攻毒组与假熏烟联合攻毒组小鼠灌洗液蛋白含量在各时点无明显差异.结论 香烟暴露延迟了气道的免疫细胞的急性募集以及杀伤力的高峰,但没有改变气道炎症介质分泌的总体水平.

Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, China; State Key Laboratory of Respiratory Disease, Guangzhou Medical University, Guangzhou, Guangdong 510006, China; Department of Infection Control, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, China; State Key Laboratory of Quality Research in Chinese Medicine, Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology,

At the end of December 2019, Wuhan reported a number of cases of unidentified pneumonia whose pathogen was later identified as a novel coronavirus, by DNA sequencing.As of 8 a.m. on May 6, 2020, novel coronavirus infections have been reported in 214 countries and regions worldwide, posing a huge threat to global public health.China has taken a series of prompt measures of prevention and control while providing medical services, which effectively contained the epidemic domestically.Rapid and accurate laboratory testing plays a crucial role in the etiological diagnosis of COVID-19.In order to improve the understanding of laboratory testing methods for COVID-19 and summarize the cumulative knowledge on the subject, the Guangdong Provincial Department of Science and Technology organized experts in the field of clinical virology diagnosis and treatment (including virology, clinical laboratory, clinical medicine, and preventive medicine), summarized the research progress and experience of novel coronavirus laboratory testing technologies, both locally and abroad, and developed a consensus-based report for novel coronavirus testing.This report covers the aspects of etiology, nucleic acid testing, serological screening, biosafety, and containment in relation to the novel coronavirus and suggests some possible problems that may be encountered in actual testing. EtiologyThe novel coronavirus belongs to the subfamily Sarbecovirus β coronavirus.It is a positive-sense, single-stranded RNA virus and often a pleomorphic enveloped virus.The viral particle is round or ovular with a diameter of 60-140 nm.The novel coronavirus genome encodes four major types of structural proteins, including spike protein (S), nucleocapsid protein (N), membrane protein (M), and envelope protein (E).Its gene sequence is similar to that of the SARS-CoV gene sequence, both of which most likely originated from bats.The homology of the two S proteins in the primary amino acid sequence is 87.2%,

OBJECTIVE:HSV-2 infection has increased significantly in recent years, which is closely associated with cervical cancer and HIV infection. The lack of success in vaccine development and the emergence of drug resistance to commonly used drugs emphasize the urgent need for alternative antivirals against HSV-2 infection. Arbidol (ARB) has been demonstrated to be a broad spectrum antiviral drug that exhibits immunomodulatory properties that affect the HSV-2 life cycle. This study investigated the efficacy and mechanism of ARB against HSV-2 in vivo and in vitro to further explore the clinical application of ARB.METHODS:The efficacy of ARB on HSV-2 infection in vitro was examined by CPE and MTT assays. A vaginitis model was established to monitor changes in histopathology and inflammatory cytokine (IL-2, IL-4, TNF-α and TGF-β) expression by H&E staining and ELISA, respectively, and the efficacy of ARB was evaluated accordingly. Furthermore, flow cytometry was used to determine the ratio of CD4+/CD8+ T cells in the peripheral blood of the vaginitis animals. Considering the balance of efficacy and pharmacokinetics, ARB ointment was strictly prepared to observe formulation efficacy differences compared to the oral dosing form.RESULTS:The results showed that, in vitro, the TC50 and IC50 of ARB were 32.32 μg/mL and 4.77 μg/mL (SI = 6.82), respectively, indicating that ARB presents effective activity against HSV-2 in a dose-dependent manner. The results of the time-course assay suggested that 25 μg/mL ARB affected the late stage of HSV-2 replication. However, ARB did not inhibit viral attachment or cell penetration. The in vivo results showed that ARB ointment can improve the survival rate, prolong the survival time and reduce the reproductive tract injury in mice infected with HSV-2, regulate cytokine expression; and balance the CD4+ and CD8+ T lymphocyte ratio in the peripheral blood to participate in the regulation of immune response.CONCLUSION:ARB showed anti-HSV-2 activity in vitro in a dose-dependent manner and played a role in inhibiting the late replication cycle of the virus. The vaginitis model was successfully established, according to immunomodulation outcomes, responded better to ARB in ointment form than in oral form.

Outbreaks of avian influenza virus continue to pose threats to human health. Animal models such as the mouse, ferret, and macaque are used to understand the pathogenesis of avian influenza virus infection in humans. We previously reported that the tree shrew (Tupaia belangeri, family Tupaiidae), which is regarded as a "low-level primate", has α2,3- and α2,6-linked sialic acid receptor distributions similar to those of humans and is potentially a useful mammalian model for studying mild human influenza (H1N1) virus infection. In this study, we used the tree shrew experimental model to investigate the pathogenesis of avian influenza A (H9N2) virus infection and the effect of the E627K mutation in the PB2 gene, an adaptation to mammalian hosts. Evidence of disease, virus titers in the upper and lower respiratory tract, histopathology and induction of proinflammatory cytokines are described. We also established ex vivo culture models of tree shrew respiratory tissues to study the tropism and replication of the H9N2 virus. Our results demonstrated that the tree shrew is a viable new in vivo experimental model for avian influenza research that provides results comparable to those observed in ferrets. The disease spectrum and pathogenesis in tree shrews correlate well with what is observed in humans.

目的 评价阿比多尔与连花清瘟胶囊抑制中东呼吸综合征冠状病毒(Middle East respiratory syndrome coronavirus,MERS-CoV)复制的体内外药效.方法 以MTT方法检测阿比多尔和连花清瘟胶囊在非洲绿猴肾细胞的半数有毒浓度(TC50).建立MERS-CoV感染非洲绿猴肾细胞模型,药物分别处理感染细胞48 h后收获上清,通过空斑试验检测培养上清病毒滴度;建立MERS-CoV感染表达hCD26小鼠模型,将小鼠随机分成3组,分别为病毒对照组、阿比多尔组[90 mg/(kg·d)]和连花清瘟胶囊组[1300 mg/(kg·d)],每组6只,药物处理后于感染后第3、5天,采用空斑试验检测鼠肺病毒滴度,分析阿比多尔组、连花清瘟胶囊组与病毒对照组的病毒滴度差异.结果 阿比多尔和连花清瘟胶囊在非洲绿猴肾细胞的TC50分别为36.78μg/mL和4023.27μg/mL,其体外抑制MERS-CoV复制的最低浓度分别为4μg/mL和900μg/mL,治疗指数分别为2.19和0.47;两种药物处理MERS-CoV感染小鼠后病毒滴度与病毒对照组差异无统计学意义(P>0.05).结论 阿比多尔具有体外抑制MERS-CoV复制的作用.

Influenza pandemic is a constant major threat to public health caused by influenza A viruses (IAVs). IAVs are subcategorized by the surface proteins hemagglutinin (HA) and neuraminidase (NA), in which they are both essential targets for drug discovery. While it is of great concern that NA inhibitor oseltamivir resistant strains are frequently identified from human or avian influenza virus, structural and functional characterization of influenza HA has raised hopes for new antiviral therapies. In this study, we explored a structure-activity relationship (SAR) of pinanamine-based antivirals and discovered a potent inhibitor M090 against amantadine-resistant viruses, including the 2009 H1N1 pandemic strains, and oseltamivir-resistant viruses. Mechanism of action studies, particularly hemolysis inhibition, indicated that M090 targets influenza HA and it occupied a highly conserved pocket of the HA2 domain and inhibited virus-mediated membrane fusion by "locking" the bending state of HA2 during the conformational rearrangement process. This work provides new binding sites within the HA protein and indicates that this pocket may be a promising target for broad-spectrum anti-influenza A drug design and development.

Influenza virus infections are the main contagious respiratory disease with high levels of morbidity and mortality worldwide. Antiviral drugs are indispensable for the prophylaxis and treatment of influenza and other respiratory viral infections. In this study, the Arbidol hydrochloride (ARB), which has been licensed in Russia and China, is used to investigate its anti-viral and anti-inflammatory efficacy in vitro and in vivo. The antiviral results in vitro showed that ARB had a better inhibition on Influenza virus A/PR/8/34 (H1N1), A/Guangdong/GIRD07/09 (H1N1), A/Aichi/2/68 (H3N2), A/HK/Y280/97 (H9N2) with IC50 ranging from 4.4 to 12.1μM. The further mechanisms study demonstrated that ARB is able to inhibit hemagglutinin-mediated hemolysis at concentration of 3.91-15.63μg/mL. The anti-inflammatory efficacy in vitro indicated that IL-6, IP-10, MCP-1, RANTES and TNF-α levels were diminished by ARB at concentrations of 22.6 and 18.8μM. The in vivo results in mice model displayed that the survival rates of mice administered 25mg/mL and 45mg/mL ARB were 40% and 50% respectively. And also, ARB can inhibit the decrease of body weight at 45mg/mL and inhibit the increase of mice lung index at 25mg/mL and 45mg/mL comparing to virus group. In ferret model, the ARB-treated ferrets showed a fever that peaked at 2 dpi and gradually decreased beginning at 3 dpi while relatively high temperatures were observed until 4 dpi in the virus group. The ARB-treated group scored 0-1 in the activity level at 2 dpi and 3 dpi at all time points. The transcription levels of cytokines in the respiratory tract of ferrets were detected at 3 dpi. Several proinflammatory cytokines induced by influenza (IL-10, TNF-α, IL-8 and IL-6) were down-regulated by post-treatment with ARB. The histopathological results of ferret lung displayed that ARB can alleviate the influenza virus induced lung lesions. Our results clarified the activity of ARB in both suppressing virus propagation and modulating the expression of inflammatory cytokines in vitro and in vivo, it can be as an effective drug to treat the influenza virus infection.

Influenza A viruses with the presence of mutations in M2 still circulate and threaten to avian species and human in China. A novel M2 inhibitor pinanamine was previously identified as an antiviral agent by an in vitro assay. In this study, we monitored the activity of pinanamine against influenza A/FM1/47 (H1N1) virus infection in cell culture and mice. Pinanamine showed more potent antiviral effect than ribavirin, and was as effective as oseltamivir carboxylate and amantadine in Madin-Darby canine kidney (MDCK) cells. Pinanamine at dose of 50 mg/kg/d administrated once a day for 6 d starting 24 h prior to virus exposure promoted survival rate of infected mice to 100% (p<0.001) and produced significant reduction (p<0.001) in lung virus yields and lung index. Even lower the dose of 3.1 mg/kg/d, pinanamine was 60% protective (p<0.05), which was equivalent to treatment with amantadine at 50 mg/kg/d. Our finding highlights the potential of pinanamine as a promising lead compound for influenza A virus.

China-made Peramivir, an anti-influenza neuraminidase inhibitor drug, is manufactured and widely used in China. Although effective if initiated within 48 h of the onset of symptoms, yet we observed that this drug shows an inconclusive efficacy if treatment is delayed in clinical. Thus we evaluated the efficacy of delayed treatment of China-made Peramivir in a mouse model.