福建省常见尸食性蝇类的COⅠ及16S rDNA序列鉴定

目的利用线粒体细胞色素c氧化酶亚基Ⅰ(cytochrome c oxidase subunitⅠ,COⅠ)以及16S核糖体脱氧核糖核酸(ribosomal deoxyribonucleic acid,rDNA)基因片段序列对福建省常见尸食性蝇类种属进行分子鉴定,探讨两种遗传标记的鉴别效力。方法收集福建9个地区案件现场捕获的常见尸食性蝇类样本22只,经形态学鉴定后提取DNA,扩增COⅠ及16S rDNA基因片段,测序后上传GeneBank数据库,使用BLAST、MEGA 10.0软件进行序列比对、同源性分析以及种内和种间遗传距离分析,并采用非加权组平均法(unweighted pair-group method with arithmetic means,UPGMA)分别构建福建省常见尸食性蝇类COⅠ及16S rDNA基因片段序列的系统发育树。结果经形态学鉴定,共收集到3科5属6种常见尸食性蝇类。基因序列分析结果显示,16S rDNA基因片段序列的种间遗传距离均数为1.8%~8.9%,种内遗传距离均数为0.0%~2.4%;COⅠ基因片段序列的种间遗传距离均数为7.2%~13.6%,种内遗传距离均数为0.0%~6.3%。结论COⅠ和16S rDNA基因片段序列均能够对不同蝇种的种属进行鉴别,16S rDNA对于福建省常见丽蝇科尸食性蝇类的种属鉴别价值更高。

微信平台及绿色通道在急性ST段抬高型心肌梗死患者转诊无缝对接中的作用

目的探讨应用微信平台及绿色通道对急性ST段抬高型心肌梗死(ST-segment elevation myocardial infarction,STEMI)患者转诊无缝对接效率的影响。方法徐州医科大学附属连云港医院于2016年7月1日启用微信平台结合绿色通道的模式对急性胸痛患者进行院前急救和转运,建立由120、急诊科、心内科以及不具备经皮冠状动脉介入(percutaneous coronary intervention,PCI)治疗能力的连云港市区及四县内各医院急诊科参与的微信群,利用微信平台传输病史和心电图资料,并开放绿色通道尽快行急诊PCI治疗,其中急性STEMI并符合条件的患者绕行急诊。本文采用回顾性分析我院2016年1月1日至2017年3月31日收治的急性STE-MI并行急诊PCI治疗患者的临床资料,将应用微信平台及绿色通道之前170例(2016年1月1日至2016年6月31日)患者设为A组(对照组),将应用微信平台及绿色通道之后339例(2016年7月1日至2017年3月31日)患者设为B组(观察组),对比分析两组首次医疗接触-球囊扩张(FMC-to-B)时间、入门-球囊扩张(D-to-B)时间、PCI治疗成功率、平均住院时间、人均住院费用、院内病死率等指标。结果两组PCI治疗成功率、住院时间及院内病死率比较,差异均无统计学意义(P>0.05);而首次医疗接触-球囊扩张时间和入门-球囊扩张时间均较微信平台及绿色通道应用前显著减少,住院费用减少,差异均有统计学意义(P<0.05)。结论微信平台和绿色通道的应用有助于急性STEMI患者的无缝转诊对接,明显缩短首次医疗接触-球囊扩张时间和入门-球囊扩张时间,提高救治效率。

PI3K／AKT／GSK-3β信号通路在转录后水平调控ICAM-1的表达

目的探讨磷脂酰肌醇-3-激酶（PBK）／丝氨酸苏氨酸蛋白激酶（AKT）／糖原合成酶激酶-3β（GSK-3β）信号通路调控脂多糖（LPS）诱导的人脐静脉内皮细胞（HUVEC）表达细胞间黏附分子-1（ICAM-1）及其机制。方法①LPS（10mg／L）刺激HUVEC0、6、12、24h或者0、4、8、12h或者0、30、60、120min后，Western blot检测ICAM-1、PI3K、P—PI3K、AKT、P—AKT、GSK-3β和P—GSK-3β的表达，实时荧光定量PCR（qPCR）检测ICAM-1 mRNA的表达。②将P13K、AKT和GSK-3B的siRNA分别转染HUVEC，转染浓度梯度为40、80和100nmol／L，对照组转染control siRNA，Western blot检测P13K、AKT和GSK-3①总蛋白的表达。③将PI3K、AKT和GSK-3β的siRNA分别转染HUVEC，对照组转染control siRNA，LPS刺激细胞30min、1h、8h和12h，Westernblot检测P—AKT、P—GSK-3B和ICAM-1的表达，qPCR检测ICAM-1mRNA的表达。④转染方法同上，LPS刺激HUVEC8h后加入放线菌素D（ActD）5ms／L抑制转录，于0、1、2、5和4h收集细胞，qPCR检测ICAM—lmRNA的表达并计算mRNA的半衰期。结果①LPS刺激HUVEC后ICAM-1的表达12h达高峰，与其他时间点有显著差异（P〈0．05或P〈0．01）；LPS刺激HUVEC后ICAM-1mRNA的表达8h达高峰，与其他时间点有显著差异（P〈0．05或P〈0．01）；LPS刺激HUVEC后PI3K、AKT和GSK-3B蛋白的磷酸化水平分别在刺激30min、1h和1h后达高峰。②PI3K、AKT和GSK-3B转染组的最适转染浓度为100nmol／L。③PI3K转染组的P—AKT和P—GSK-3β表达显著下降（P〈0．01），AKT转染组P—GSK-3β表达显著下降（P〈0．01）；PBK和AKT转染组的ICAM-1的蛋白和mRNA表达均显著升高（P〈0．05或P〈0．01），相反GSK-3β转染组ICAM-1的蛋白和mRNA表达均显著下降（P〈0．01）。④PI3K、AKT的沉默表达显著延长ICAM-1的mRNA半衰期（P〈0．01），相反GSK-3B的沉默表达显著缩短ICAM-1的mRNA半衰期（P〈0．01）。结论LPS通过激活PI3K／AKT／GSK-3β信号通路负向调控HUVEC表达ICAM-1。PBK、AKT和GSK-3B在转录后水平调控ICAM-1mRNA的稳定性。

Shiqi herbal tea reduces the susceptibility to H1N1 influenza virus in stressed mice

Objective:It is well known that stress plays a critical role in immune response and susceptibility to diseases.Among many stressors,restraint stress has been shown to suppress immune function and increase susceptibility to infections.In this study,we employed a restraint-mouse model to investigate the effect and preliminary mechanism of ShiQi herbal tea(SQHT),a traditional Chinese medicine,on H1N1 virus infection.Methods:Mice were exposed to restraint stress and infected with H1N1 influenza virus by intranasal inoculation.SQHT(936 and 1872 mg/kg/d)was orally administrated to mice for 7 days from the first day of restraint stress.The survival rate of mice in each group was monitored daily for 21 days.Histopathological changes,inflammatory cells infiltration,and virus titer in lungs were examined.For the study of mechanisms,we investigated whether SQHT could promote interferon-β(IFN-β)generation and interferon stimulated genes(ISG)expression.Results:Our results suggested that SQHT(936 mg/kg/d)significantly reduced H1N1-induced mortality,the level of complement C5a and lung tissue inflammation,and viral replication in restraint-stressed mice.Further results revealed that in restraint-stressed mice model,SQHT(936 mg/kg/d)administration markedly improved IFN-βgeneration,and increased MX1 and IFITM3 gene expression.Conclusion:Our study demonstrates that SQHT reduces restraint stress-induced susceptibility to H1N1 infection via improving IFN-βantiviral pathway,which provides a certain basis for the clinical use of SQHT to treat H1N1 infection.

Mangiferin ameliorates hyperglycemia by inhibiting oxidation and α-glucosidase activity

目的：芒果苷（MF）是从知母根提取出来的-种多元酚.本研究旨在探讨芒果苷对胰岛素抵抗和链脲霉素（STZ）引起的糖尿病动物模型的高血糖的影响.方法：通过多次皮下注射氢化可的松琥珀酸钠（HCSS）（70mg/kg）或单次静脉内注射STZ（130mg/kg）建立糖尿病模型.同时连续口服10天不同剂量（50,100和200mg/kg）的MF.腹腔注射胰岛素（0.5U/kg）后,收集不同时间的血糖数值来检测胰岛素抵抗.同时测定肾脏氧自由基吸收能力（ORAC）和超氧化物歧化酶（SOD）活性.建立体外实验,研究MF对α-葡萄糖苷酶的抑制能力.结果：口服芒果苷能够显著抑制由注射HCSS引起的胰岛素抵抗.STZ诱导的糖尿病症状也有所改善,包括空腹血糖,糖化血红蛋白,血浆甘油三酯,肝糖原,肾脏SOD和ORAC水平.体外实验证明芒果苷具有较强的α-葡萄糖苷酶抑制活性.结论：结果表明芒果苷能改善胰岛素抵抗和STZ诱导的糖代谢紊乱.芒果苷通过提高抗氧化能力,促进肝糖原合成,抑制α-葡萄糖苷酶活性发挥保护作用.

Disturbed Yin-Yang balance:stress increases the susceptibility to primary and recurrent infections of herpes simplex virus type 1

Herpes simplex virus type 1(HSV-1),a neurotropic herpes virus,is able to establish a lifelong latent infection in the human host.Following primary replication in mucosal epithelial cells,the virus can enter sensory neurons innervating peripheral tissues via nerve termini.The viral genome is then transported to the nucleus where it can be maintained without producing infectious progeny,and thus latency is established in the cell.Yin-Yang balance is an essential concept in traditional Chinese medicine(TCM)theory.Yin represents stable and inhibitory factors,and Yang represents the active and aggressive factors.When the organism is exposed to stress,especially psychological stress caused by emotional stimulation,the Yin-Yang balance is disturbed and the virus can re-engage in productive replication,resulting in recurrent diseases.Therefore,a better understanding of the stress-induced susceptibility to HSV-1 primary infection and reactivation is needed and will provide helpful insights into the effective control and treatment of HSV-1.Here we reviewed the recent advances in the studies of HSV-1 susceptibility,latency and reactivation.We included mechanisms involved in primary infection and the regulation of latency and described how stress-induced changes increase the susceptibility to primary and recurrent infections.

Inhibitory effects of baicalein against herpes simplex virus type 1

Herpes simplex virus type 1(HSV-1)is a ubiquitous and widespread human pathogen,which gives rise to a range of diseases,including cold sores,corneal blindness,and encephalitis.Currently,the use of nucleoside analogs,such as acyclovir and penciclovir,in treating HSV-1 infection often presents limitation due to their side effects and low efficacy for drug-resistance strains.Therefore,new anti-herpetic drugs and strategies should be urgently developed.Here,we reported that baicalein,a naturally derived compound widely used in Asian countries,strongly inhibited HSV-1 replication in several models.Baicalein was effective against the replication of both HSV-1/F and HSV-1/Blue(an acyclovir-resistant strain)in vitro.In the ocular inoculation mice model,baicalein markedly reduced in vivo HSV-1/F replication,receded inflammatory storm and attenuated histological changes in the cornea.Consistently,baicalein was found to reduce the mortality of mice,viral loads both in nose and trigeminal ganglia in HSV-1 intranasal infection model.Moreover,an ex vivo HSV-1-EGFP infection model established in isolated murine epidermal sheets confirmed that baicalein suppressed HSV-1 replication.Further investigations unraveled that dual mechanisms,inactivating viral particles and inhibiting IκB kinase beta(IKK-β)phosphorylation,were involved in the anti-HSV-1 effect of baicalein.Collectively,our findings identified baicalein as a promising therapy candidate against the infection of HSV-1,especially acyclovir-resistant strain.

Novel insights into stress-induced susceptibility to influenza:corticosterone impacts interferon-βresponses by Mfn2-mediated ubiquitin degradation of MAVS

Although stress has been known to increase the susceptibility of pathogen infection,the underlying mechanism remains elusive.In this study,we reported that restraint stress dramatically enhanced the morbidity and mortality of mice infected with the influenza virus(H1N1)and obviously aggravated lung inflammation.Corticosterone(CORT),a main type of glucocorticoids in rodents,was secreted in the plasma of stressed mice.We further found that this stress hormone significantly boosted virus replication by restricting mitochondrial antiviral signaling(MAVS)protein-transduced IFN-βproduction without affecting its mRNA level,while the deficiency of MAVS abrogated stress/CORT-induced viral susceptibility in mice.Mechanistically,the effect of CORT was mediated by proteasome-dependent degradation of MAVS,thereby resulting in the impediment of MAVS-transduced IFN-βgeneration in vivo and in vitro.Furthermore,RNA-seq assay results indicated the involvement of Mitofusin 2(Mfn2)in this process.Gain-and loss-offunction experiments indicated that Mfn2 interacted with MAVS and recruited E3 ligase SYVN1 to promote the polyubiquitination of MAVS.Co-immunoprecipitation experiments clarified an interaction between any two regions of Mfn2(HR1),MAVS(C-terminal/TM)and SYVN1(TM).Collectively,our findings define the Mfn2-SYVN1 axis as a new signaling cascade for proteasome-dependent degradation of MAVS and a‘fine tuning’of antiviral innate immunity in response to influenza infection under stress.

急诊科医生应用床旁超声抢救不明原因休克的效果及可行性分析

目的:探讨急诊科医生应用床旁超声抢救不明原因休克的效果及可行性。方法:我院自2017-07-01开始在急诊抢救室由经过重症超声培训合格的急诊科医生应用床旁超声对急危重症患者进行救治。对不明原因休克患者的探查按照《不明原因休克急诊超声临床实践专家共识》推荐的THIRD流程进行。本文采用回顾性分析我院急诊科2016-01-01-2018-08-31收治的不明原因休克患者,其中123例由超声科医生实行床旁超声探查,称为超声医生组,159例由急诊科医生按此方案实行床旁超声探查,称为急诊医生组。对比分析两组患者诊断时间,计算急诊医生组诊断敏感性、特异性、阳性预测值、阴性预测值以及总准确率等指标。结果:急诊医生组诊断时间较超声组显著缩短,差异均有统计学意义(P<0.05);急诊医生组患者敏感性、特异性、阳性预测值、阴性预测值及总准确率较高。结论:急诊科医生应用床旁超声按照THIRD流程探查不明原因休克患者,有助于快速明确诊断,诊断效率及准确性高,提倡推广应用。