Antiviral Activity of Nano Carbon Fullerene Lipidosome against Influenza Virus In Vitro

The activity of nano carbon fullerene lipidosome(NCFL) against influenza virus H1N1 in vitro was studied by observing the cytotoxicities and its activity rendered by different intensities of lighting with various periods of time.Rimantadine hydrochloride was used as the positive control drug.By using microcultural technique,the morphological changes of cells were observed and by using the gentian violet staining,antiviral activity of the NCFL against influenza virus was assayed.The results showed that:(1) The maximal concentration of the NCFL was 7 μg/mL and the 50% toxic concentration(TC50) was 13.54 μg/mL respectively;(2) NCFL had a significant activity of directly killing the influenza virus,while the activities in antiadsorption and antireplication were not obvious;(3) There was a dose-activity relationship between the dosages of NCFL and the direct killing effect against the influenza virus,and the periods of lighting-time could influence the activity partly.It was concluded that NCFL had a significant activity of directly killing the influenza virus.     

An extract from the earthworm Eisenia fetida non-specifically inhibits the activity of influenza and adenoviruses

OBJECTIVE:To test the in vitro antiviral activity of a crudetissueextract(CTE)fromtheearthwormEisenia fetida,determine any effective components in the CTE,andelucidatepossiblemechanismsofaction.METHODS:A CTE was made by homogenizing earthworms,followed by treatment with ammonium sulfate,then thermal denaturation.Inhibition of virus-induced cytopathic effect(CPE) was used to assess antiviral activity.Chromatographic analysis was used to identify effective components in the CTE.RESULTS:The CTE inhibited viral CPE at non-cytotoxic concentrations.Chromatography indicated that antiviral components corresponded to three active peaks indicative of proteases,nucleases and lysozymes.For adenoviruses,reduction in viral activity occurred for 100 μg/mL CTE.The reduction in adenoviral activity for four fractions was 100%,91.8%,86.9%,and 94.7%.For influenza viruses,reduction in viral activity of 100%,86.6%,69.1% and 88.3% was observed for 37 μg/mL CTE.In addition,three active fractions mixture had stronger antiviral activity(98.7% and 96.7%) than three fractions alone.Gel electrophoresis results indicated that nucleases from E.fetida could degrade the genome of influenza viruses and adenoviruses.CONCLUSION:The earthworm CTE displayed non-specific antiviral properties,possibly mediated by a combination of proteases,nucleases and lysozymes.Nucleases likely participate in the antiviral process,and degrade the genome of the virus thereby preventing further replication.     

An O-glycoside of Sialic Acid Derivative that inhibits Both Hemagglutinin and Sialidase Activities of Influenza Viruses

The compound Neu5Ac3αF-DSPE (4), in which the C-3 position was modified with an axial fluorine atom, inhibited the catalytic hydrolysis of influenza virus sialidase and the binding activity of hemagglutinin. The inhibitory activities to sialidases were independent of virus isolates examined.With the positive results obtained for inhibition of hemagglutination and hemolysis induced by A/Aichi/2/68 virus,the inhibitory effect of Neu5Ac3αFDSPE (4) against MDCK cells was examined, and it was found that 4 inhibits the viral infection with IC50 value of 5.6 μM based on the cytopathic effects. The experimental results indicate that compound 4 not only inhibits the attachment of virus to the cell surface receptor but also disturbs the release of the progeny viruses from infected cells by inhibiting both hemagglutinin and sialidase of the influenza viruses.The study suggested that the compound is a new class of bifunctional drug candidates for the future chemotherapy of influenza.     

Effect of All-trans Retinoic Acid on Airway Inflammation in Asthmatic Rats and Its Mechanism

The inhibitive effects of all-trans retinoic acid (ARTA) on airway inflammation in asthmatic rats and its mechanism on the basis of the regulation of nuclear factor kappaB (NF-kB) were explored. Thirty-two SD rats were randomly divided into 4 groups: control group, asthma group,dexamethasone treatment group and retinotic acid treatment group. The total and differential cell counts in the collected bronchoalveolar lavage fluid (BALF) were measured. The pathological changes in lung tissues were estimated by scoring. The expression of NF-kB inhibitor (IkBa), NF-kB,intercellular adhering molecule-1 (ICAM-1) in lung tissue was detected by immunohistochemical method. The results showed that in the two treatment groups, the total cell counts and proportion of inflammatory cells in BALF were significantly reduced, but there was no significant difference in differential cell counts in BALF between them. The pathological changes in lung tissues in the treatment groups were significantly attenuated as compared with asthma group. Except the epithelial injury in retinotic acid treatment group was milder than in dexamethasone treatment group, the remaining lesions showed no significant difference between them. In the two treatment groups, the expression of IkBa was increased, while the expression of NF-kB and ICAM-1 decreased with the difference between the two groups being not significant. It was concluded that the similar anti-inflammatory effects and mechanism of ATRA on airway in asthmatic rats to those of dexamethasone were contributed to the increase of cytoplasmic IkBa content and suppression of NF-kB activationand expression.     

Qiangzhi Decoction(羌跖汤) Protects Mice from Influenza A Pneumonia through Inhibition of Inflammatory Cytokine Storm

Objective:To investigate the preventive effects of Qiangzhi Decoction(羌跖汤,QZD) on influenza A pneumonia through inhibition of inflammatory cytokine storm in vivo and in vitro.Methods:One hundred ICR mice were randomly divided into the virus control,the Tamiflu control and the QZD high-,medium-,and low-dose groups.Mice were infected intranasally with influenza virus(H1N1) at 10 median lethal dose(LD_(50)).QZD and Tamiflu were administered intragastrically twice daily from day 0 to day 7 after infection.The virus control group was treated with distilled water alone under the same condition.The number of surviving mice was recorded daily for 14 days after viral infection.The histological damage and viral replication and the expression of inflammatory cytokines were monitored.Additionally,the suppression capacity on the secretion of regulated on activation normal T cells expressed and secreted(RANTES) and tumor necrosis factor- α(TNF- α) in epithelial and macrophage cell-lines were evaluated.Results:Compared with the virus control group,the survival rate of the QZD groups significantly improved in a dose-dependent manner(P0.05),the viral titers in lung tissue was inhibited(P0.05),and the production of inflammatory cytokines interferon-γ(IFN- γ),interleukin-6(IL-6),TNF- α,and intercellular adhesion molecule-1(ICAM-1) were suppressed(P0.05).Meanwhile,the secretion of RANTETS and TNF-α by epithelial and macrophage cell-lines was inhibited with the treatment of QZD respectively in vitro(P0.05).Conclusions:The preventive effects of QZD on influenza virus infection might be due to its unique cytokine inhibition mechanism.QZD may have significant therapeutic potential in combination with antiviral drugs.     

Effects of High Erucic Acid Rapeseed Oil on Fatty Acid Oxidation in Rat Liver

The effects of high erucic acid rapeseed oil (HER) on fatty acid oxidation in rat liver compared with low erucic acid rapeseed oil (LER) were studied. Weanling male SD rats were fed diets containing 20% HER or LER for 1 week or 4 weeks, or 5% HER diet for 4 weeks. The hepatic oxidation capacity of butyric acid or palmitic acid was determined by titrating the propanone produced by their oxidation. The results showed that feeding HER to rats led to an increase in the weight of liver and a decrease in the hepatic oxidation capacity ofpalmitic acid. Hepatic oxidation of butyric acid was not influenced by the intake of HER. The inhibitory action of HER on the oxidation of long-chain fatty acids probably resulted from the incorporation of erucic acid into mitochondrial membranes, interfering the fatty acyl-CoA transfering system on the membranes, but not from the fi-oxidation enzyme system itl mitochondria being directly inhibited.     

Arsenic Induced Inhibition of δ-aminolevulinate Dehydratase Activity in Rat Blood and its Response To Meso 2,3-dimercaptosuccinic Acid and Monoisoamyl DMSA

Objective The objective of this study was to investigate arsenic induced changes in blood δ-aminolevulinic acid dehydratase (ALAD) after in vitro and in vivo exposure to this element and its response to co-administration of meso 2,3-dimercaptosuccinic acid (DMSA) and monoisoamyl DMSA (MiADMSA) either individually or in combination.Methods Rat whole blood was exposed to varying concentrations (0.1, 0.2 and 0.5mmol/L) of arsenic (Ⅲ) or arsenic (Ⅴ),to assess their effects on blood ALAD activity. Varying concentrations of MiADMSA and DMSA (0.1,0.5 and 1.0mmol/L) were also tried in combination to determine its ability to mask the effect of arsenic induced (0.5mmol/L) inhibition of blood ALAD in vitro. In vitro and in vivo experiments were also conducted to determine the effects of DMSA and MiADMSA either individually or in combination with arsenic,on blood ALAD activity and blood arsenic concentlation.Results In vitro experiments showed significant inhibition of the enzyme activity when 0.1-0.5 mmol/L of arsenic (Ⅲ and Ⅴ) was used.Treatment with MiADMSA increased ALAD activity when blood was incubated at the concentration of 0.1mmol/L arsenic (Ⅲ) and 0.1mmol/L MiADMSA. No effect of 0.1mmol/L MiADMSA on ALAD activity was noticed when the arsenic concentration was increased to 0.2 and 0.5 mmol/L. Similarly, MiADMSA at a lower concentration (0.1mmol/L) was partially effective in the turnover of ALAD activity against 0.5 mmol/L arsenic (Ⅲ),but at two higher concentrations (0.5 and 1.0mmol/L) a complete restoration of ALAD activity was observed. DMSA at all the three concentrations (0.1,0.5 and 1.0mmol/L) was effective in restoring ALAD activity to the normal value Conclusions The results thus suggest that arsenic has a distinct effect on ALAD activity.Another important toxicological finding of the present study,based on in vivo experiments further suggests that combined administration of DMSA and MiADMSA could be more beneficial for reducing blood ALAD inhibition and blood arsenic concentration than the individual treatment.     

Combined antiviral activity of interferon-α and RNA interference directed against hepatitis C without affecting vector delivery and gene silencing

The current standard interferon-alpha(IFN-α)-based therapy for chronic hepatitis C virus(HCV) infection is only effective in approximately half of the patients,prompting the need for alternative treatments.RNA interference(RNAi)represents novel approach to combat HCV by sequence-specific targeting of viral or host factors involved in infection.Monotherapy of RNAi,however,may lead to therapeutic resistance by mutational escape of the virus.Here,we proposed that combining lentiviral vector-mediated RNAi and IFN-a could be more effective and avoid therapeutic resistance.In this study,we found that IFN-α treatment did not interfere with RNAi-mediated gene silencing.RNAi and IFN-α act independently on HCV replication showing combined antiviral activity when used simultaneously or sequentially.Transduction ofmouse hepatocytes in vivo and in vitro was not effected by IFN-α treatment.In conclusion,RNAi and IFN-a can be effectively combined without crossinterference and may represent a promising combinational strategy for the treatment of hepatitis C.     

Combined antiviral activity of interferon-α and RNA interference directed against hepatitis C without affecting vector delivery and gene silencing

The current standard interferon-alpha(IFN-α)-based therapy for chronic hepatitis C virus(HCV) infection is only effective in approximately half of the patients, prompting the need for alternative treatments. RNA interference(RNAi) represents novel approach to combat HCV by sequence-specific targeting of viral or host factors involved in infection. Monotherapy of RNAi, however, may lead to therapeutic resistance by mutational escape of the virus. Here, we proposed that combining lentiviral vector-mediated RNAi and IFN-α could be more effective and avoid therapeutic resistance. In this study, we found that IFN-α treatment did not interfere with RNAi-mediated gene silencing. RNAi and IFN-α act independently on HCV replication showing combined antiviral activity when used simultaneously or sequentially. Transduction ofmouse hepatocytes in vivo and in vitro was not effected by IFN-α treatment. In conclusion, RNAi and IFN-α can be effectively combined without crossinterference and may represent a promising combinational strategy for the treatment of hepatitis c.     

青蒿琥酯对甲型流感病毒肺炎的治疗作用研究

  目的  探讨青蒿琥酯（artesunate, ART）对甲型流感病毒肺炎的治疗作用。  方法  将36只小鼠随机分为6组:正常对照组（C组）、溶剂对照组（M组,10%DMSO）、阳性药物组（P组,奥司他韦1.25 mg/kg）、ART高剂量组（ART-G组,ART 120 mg/kg）、ART中剂量组（ART-Z组,ART 60 mg/kg）和ART低剂量组（ART-D组,ART 30 mg/kg）,每组6只。除C组不进行病毒干预和腹腔注射外,其余5组小鼠鼻腔滴入感染甲型流感病毒（influenza A virus, IAV）,12 h后按分组剂量进行每日一次腹腔注射;治疗过程中观察小鼠体征、体质量和存活情况;治疗7 d后,取小鼠肺组织,计算肺指数,HE染色观察小鼠肺组织病理变化,RT-qPCR和Western blot分别检测肺组织中Toll 样受体 4（Toll-like receptor 4, TLR4）、核因子κB（nuclear factor kappa-B, NF-κB）（p65）、肿瘤坏死因子α（tumor necrosis factor α, TNF-α)、白细胞介素-6（interleukin-6, IL-6）和白细胞介素-1β（interleukin-1, IL-1β）mRNA和蛋白表达水平。  结果  与C组比较,M组小鼠体征变差、体质量和存活率降低,肺指数增加,肺组织出现严重病理变化,肺组织中TLR4、NF-κB (p65)、TNF-α、IL-6和IL-1β mRNA和蛋白表达水平升高（P<0.05）;与M组比较,ART各组小鼠体征明显改善、体质量和存活率升高,肺指数降低,肺组织病理变化得到改善,肺组织中TLR4、NF-κB (p65)、TNF-α、IL-6和IL-1β mRNA和蛋白表达水平降低（P<0.05）,且上述指标变化以ART-G组最显著。  结论  ART对甲型流感病毒肺炎具有治疗作用,其机制与抑制TLR4/NF-κB (p65)信号通路活化和抗炎相关。     

Metabolomic fingerprinting of porcine lung tissue during pre-clinical prolonged ex vivo lung perfusion using in vivo SPME coupled with LC-HRMS

Normothermic ex vivo lung perfusion (NEVLP) has emerged as a modernized organ preservation technique that allows for detailed assessment of donor lung function prior to transplantation. The main goal of this study was to identify potential biomarkers of lung function and/or injury during a prolonged (19 h) NEVLP procedure using in vivo solid-phase microextraction (SPME) technology followed by liquid chromatography-high resolution mass spectrometry (LC-HRMS). The use of minimally invasive in vivo SPME fibers for repeated sampling of biological tissue permits the monitoring and evaluation of biochemical changes and alterations in the metabolomic profile of the lung. These in vivo SPME fibers were directly introduced into the lung and were also used to extract metabolites (on-site SPME) from fresh perfusate samples collected alongside lung samplings. A subsequent goal of the study was to assess the feasibility of SPME as an in vivo method in metabolomics studies, in comparison to the traditional in-lab metabolomics workflow. Several upregulated biochemical pathways involved in pro- and anti-inflammatory responses, as well as lipid metabolism, were observed during extended lung perfusion, especially between the 11th and 12th hours of the procedure, in both lung and perfusate samples. However, several unstable and/or short-lived metabolites, such as neuroprostanes, have been extracted from lung tissue in vivo using SPME fibers. On-site monitoring of the metabolomic profiles of both lung tissues through in vivo SPME and perfusate samples on site throughout the prolonged NEVLP procedure can be effectively performed using in vivo SPME technology.     

Silencing the expression of Cbl-b enhances the immune activation of T lymphocytes against RM-1 prostate cancer cells in vitro

BackgroundThe ubiquitin ligase Cbl-b potently modulates T lymphocyte immune responses and is critical in modulating tumor-induced immunosuppression. The influence of Cbl-b in modulating T lymphocyte activity against prostate cancer remains ill defined. We have determined the effects of silencing Cbl-b expression in T lymphocytes and their subsequent cytotoxic activity against prostate cancer cells.MethodsT lymphocytes were isolated from the spleens of C57BL/6 mice. Lipofectamine-directed transfection of T lymphocytes with specific small interfering RNA (siRNA) silenced Cbl-b expression, which was confirmed by Western immunoblotting. The siRNA species were chosen that promoted the greatest transfection efficiency and dampened Cbl-b expression in T lymphocytes. The expression of CD69, CD25, and CD71 by the transfected T lymphocytes was determined by flow cytometry. T lymphocyte proliferation was assessed by CCK-8 assay. Enzyme-linked immunosorbent assay (ELISA) was used to measure the secretion of interleukin (IL)-2, interferon (IFN)-γ, and tumor necrosis factor (TNF)-β. The objective was to compare the cytotoxic activity of transfected T lymphocytes and nontransfected (i.e., negative control) T lymphocytes against the murine prostate cancer cell line target RM-1 in vitro.ResultsWe selected a specific siRNA that decreased T lymphocyte Cbl-b expression to 15%. The siRNA-transfected T lymphocytes showed higher proliferation; higher CD69, CD25, and CD71 expression (p < 0.001); and higher IL-2, IFN-γ, and TNF-β secretion (p < 0.05), compared to the nontransfected cells. Transfected T lymphocytes were also more potent at killing RM-1 prostate cancer cells, compared to the negative control in vitro.ConclusionSilencing Cbl-b significantly enhanced T lymphocyte function and T lymphocyte cytotoxicity activity against a model prostate cancer cell line in vitro. This study suggests a potentially novel immunotherapeutic strategy against prostate cancer.     

清透邪热方对甲型H1N1流感病毒感染Ana-1细胞TLR-7、MyD88及NF-κB mRNA及蛋白表达的影响

目的 研究清透邪热方体外抗甲型H1N1流感病毒效应机制。方法 将清透邪热方制备成0.388、0.194、0.097、0.0485、0.02425mg/mL不同浓度,用H1N1甲型流感病毒感染小鼠Ana-1巨噬细胞,给予不同浓度清透邪热方及达菲进行干预,检测肿瘤坏死因子α（TNF-α）、干扰素诱导蛋白-10（IP-10）和白细胞介素-6（IL-6）表达,Toll样受体7（TLR7）及其下游髓样分化因子88(MyD88)、核因子-κB（NF-κB）mRNA和蛋白表达。结果 清透邪热方能够抑制甲型H1N1流感病毒感染Ana-1细胞TNF-α、IP-10、IL-6的表达;其中清透邪热方0.02425mg/mL组降低H1N1病毒感染组TNF-α、IL-6显著（P＜0.01）;清透邪热方0.097mg/mL组降低IP-10显著（P＜0.01）。清透邪热方能够抑制H1N1流感病毒感染后Ana-1细胞后TLR-7、MyD88及NF-κB mRNA及蛋白表达升高;其中清透邪热方0.0485mg/mL组抑制H1N1病毒感染组TLR-7 mRNA、MyD88 mRNA表达显著（P＜0.01）;清透邪热方0.02425mg/mL组抑制H1N1病毒感染组NF-κB mRNA表达显著（P＜0.01）;清透邪热方0.0485mg/mL组抑制H1N1病毒感染组TLR-7蛋白表达升高显著（P＜0.01）,清透邪热方0.388mg/mL组抑制H1N1病毒感染组MyD88蛋白、NF-κB蛋白表达显著（P＜0.01）。结论 清透邪热方通过抑制甲型H1N1流感病毒感染Ana-1细胞TLR-7、MyD88及NF-κB mRNA及蛋白表达升高,减少TNF-α、IP-10和IL-6表达,起到抗病毒作用。      

SB203580减轻体外循环肺组织炎症介质产生的实验研究

目的 研究体外循环肺组织中P38丝裂原活化蛋白激酶(P38 MAPK)的变化对肺组织炎症反应的作用及其机制.方法 54只SD大鼠随机分为3组:全麻开胸组(S组)、体外循环组(CPB组)、体外循环+SB203580组(SB组).不同时间段处死动物,留取标本,Western blotting检测肺组织中P38 MAPK、磷酸化P38 MAPK.EMSA检测核因子(NF)-κB的DNA结合活性变化,ELIASA分别检测TNF-α和IL-1β产量.结果 CPB组磷酸化P38MAPK较S组增加.NF-κB活性水平也较S组明显增加,肺组织中TNF-α和IL-1β产量增加.SB203580减轻了肺组织中磷酸化P38MAPK活性水平,减少了肺组织中炎症闪子的产生.结论 (1)P38MAPK通过影响NF-κB的激活而参与体外循环术后肺组织炎症因子的产生;(2)SB203580通过阻断P38MAPK的激活而减轻体外循环术后肺炎症因子的产生.

Abstract：

Objective To examine the changes in P38MAPK during and after cardiopulmonary bypass (CPB) and the effect of SB203580, a specific P38MAPK inhibitor, on CPB-induced pulmonary inflammatory response. Metholds Fifty-four SD rats were randomized into 3 groups (each=18), namely sham CPB group, CPB group, and SB203580 group in which rats underwent CPB with SB203580 pretreatment. The lungs were excised immediately after the rats were sacrificed at scheduled time points and p38, nuclear factor-κB (NF-ΚB), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected. Results The activities of P38 MAPK and NF-ΚB were significantly increased in CPB group as compared with those in sham CPB group. CPB resulted in increased TNF-α and IL-1β production in the lung tissues. Administration of SB203580 prevented up-regulation of lung phosphorylated P38 MAPK, and decreased proinflammatory cytokine productions in the lung tissues. Conclusion P38 MAPK is activated in the lung tissue during and after CPB to affect the activation of NF-κB in the lung; SB203580 selectively inhibits P38 MAPK activation to reduce proinflammatory cytokine production after CPB.     

肺气虚模型大鼠介导TNF-α-MAPK信号途径对肾AQP2表达的影响

目的:通过观测肺气虚大鼠血浆中肿瘤坏死因子(TNF-α)含量,肾组织水通道蛋白2(AQP2),丝裂原激活蛋白激酶(p38MAPK)和核因子-κB(NF-κBp65)表达的变化,探讨中医藏象中肺与肾在水液代谢之间关系的现代科学内涵。方法:将SPF级健康雄性Wistar大鼠20只随机分为正常组和模型组,采用ELISA法检测血浆TNF-α含量、Western法检测肾组织AQP2、免疫组化法检测肾组织p38MAPK和NF-κB p65表达变化。结果:与正常组比较,模型组血浆中TNF-α含量升高,肾组织AQP2、p38MAPK和NF-κB p65表达上调(P<0.01)。结论:肺气虚状态下肾组织AQP2表达变化可能是通过TNF-α-MAPK信号转导途径实现的。     

中医药调控肺癌相关信号通路研究进展

MAPK信号通路是细胞内信号传导的主要途径,利用MAPK信号通路可对肺癌进行有效治疗,今后还需研究MAPK信号通路的相关蛋白在肺癌中的作用及潜在的分子机制,发现新的药物作用靶点,为肺癌的治疗提供新途径.中医药在干预Wnt/β - catenin信号通路的研究方面较为薄弱,主要以COX -2、β -catenin、GSK...