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Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress

Biomedical and Environmental Sciences 2018年第12期31卷 867-875页

作者： LUO Xiao Nuan yao hai lan SONG Juan SONG Qin Qin SHI Bing Tian XIA Dong HAN Jun State Key Laboratory of Infectious Disease Prevention and Control Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention Molecular Immunology Laboratory Capital Institute of Pediatrics

Objective Autophagy is a highly conserved intracellular degradation pathway. Many picornaviruses induce autophagy to benefit viral replication, but an understanding of how autophagy occurs remains incomplete. In this study, we explored whether coxsackievirus B3(CVB3) infection induced autophagy through endoplasmic reticulum(ER) stress. Methods In CVB3-infected HeLa cells, the specific molecules of ER stress and autophagy were detected using Western blotting, reverse transcription polymerase chain reaction(RT-PCR), and confocal microscopy. Then PKR-like ER protein kinase(PERK) inhibitor, inositol-requiring protein-1(IRE1) inhibitor, or activating transcription factor-6(ATF6) inhibitor worked on CVB3-infected cells, their effect on autophagy was assessed by Western blotting for detecting microtubule-associated protein light chain 3(LC3). Results CVB3 infection induced ER stress, and ER stress sensors PERK/eIF2α, IRE1/XBP1, and ATF6 were activated. CVB3 infection increased the accumulation of green fluorescent protein(GFP)-LC3 punctuation and induced the conversion from LC3-Ⅰ to phosphatidylethanolamine-conjugated LC3-1(LC3-Ⅱ). CVB3 infection still decreased the expression of mammalian target of rapamycin(mTOR) and p-mTOR. Inhibition of PERK, IRE1, or ATF6 significantly decreased the ratio of LC3-Ⅱ to LC3-Ⅰ in CVB3-infected HeLa cells. Conclusion CVB3 infection induced autophagy through ER stress in HeL a cells, and PERK, IRE1, and ATF6 a pathways participated in the regulation of autophagy. Our data suggested that ER stress may inhibit mTOR signaling pathway to induce autophagy during CVB3 infection.

关键词： Coxsackievirus B3(CVB3) Autophagy Endoplasmic reticulum(ER) stress Unfolded protein response(UPR)

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The Viral Load of Epstein-Barr Virus in Blood of Children after Hematopoietic Stem Cell Transplantation

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Biomedical and Environmental Sciences 2022年第9期35卷 804-810页

作者： WANG Wen Jun FENG Shun Qiao HE Feng DU hai Jun FENG Miao WANG Rui Fang MEI Guo Yong LIU Mi LIU Rong yao hai lan HAN Jun State Key Laboratory of Infectious Disease Prevention and Control National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Department of Hematology Children’s Hospital of Capital Institute of PediatricsBeijing 100020China Department of Biochemistry&Immunology Capital Institute of PediatricsBeijing 100020China

Objective To detect the Epstein-Barr virus(EBV)viral load of children after hematopoietic stem cell transplantation(HSCT)using chip digital PCR(cdPCR).Methods The sensitivity of cdPCR was determined using EBV plasmids and the EBV B95-8 *** specificity of EBV cdPCR was evaluated using the EBV B95-8 strain and other herpesviruses(herpes simplex virus 1,herpes simplex virus 2,varicella zoster virus,human cytomegalovirus,human herpesvirus 6,and human herpesvirus 7).From May 2019 to September 2020,64 serum samples of children following HSCT were *** infection and the viral load of serum samples were detected by *** epidemiological characteristics of EBV infections were analyzed in HSCT *** The limit of detection of EBV cdPCR was 110 copies/mL,and the limit of detection of EBV quantitative PCR was 327 copies/mL for the pUC57-BALF5 *** result of EBV cdPCR was up to 121 copies/mL in the EBV B95-8 strain,and both were more sensitive than that of quantitative *** cdPCR,the incidence of EBV infection was 18.75%in 64 children after *** minimum EBV viral load was 140 copies/mL,and the maximum viral load was 3,209 copies/mL using *** average hospital stay of children with EBV infection(184±91 days)was longer than that of children without EBV infection(125±79 days),P=*** EBV cdPCR had good sensitivity and *** incidence of EBV infection was 18.75%in 64 children after HSCT from May 2019 to September *** cdPCR could therefore be a novel method to detect EBV viral load in children after HSCT.

关键词： Chip digital PCR Epstein-Barr virus Hematopoietic stem cell transplantation Quantitative PCR

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Exploration of IRES Elements within the ORF of the Coxsackievirus B3 Genome

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Biomedical and Environmental Sciences 2022年第4期35卷 322-333页

作者： SONG Qin Qin LUO Xiao Nuan SHI Bing Tian LIU Mi SONG Juan XIA Dong XIA Zhi Qiang WANG Wen Jun yao hai lan HAN Jun State Key Laboratory of Infectious Disease Prevention and Control National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Molecular Immunology Laboratory Capital Institute of PediatricsBeijing 100020China

Objective This study aimed to identify internal ribosome entry sites(IRESs)in the open reading frame(ORF)of the Coxsackievirus B3(CVB3)*** The sequences of P1,P2,or P3 of the CVB3 genome or the truncated sequences from each antithymocyte globulin(ATG)to the end of the P1,P2,or P3 gene were inserted into the *** transfection,possible IRES-dependent green fluorescent protein(GFP)-fused proteins were detected by anti-GFP western *** sequences of possible IRESs were inserted into specific Fluc/Rluc bicistronic vectors,in which the potential IRESs were determined according to the Fluc/Rluc activity *** of Fluc and Rluc mRNA of the bicistronic vector was detected by *** After transfection of full length or truncated sequences of the P1,P2,or P3 plasmids,six GFPfused protein bands in P1,six bands in P2 and nine bands in P3 were detected through western *** IRESs in VP2(1461–1646 nt)and VP1(2784–2983 nt)of P1;one IRES in 2C(4119–4564 nt)of P2;and two IRESs in 3C(5634–5834 nt)and 3D(6870–7087 nt)of P3 were identified according to Fluc/Rluc activity *** cryptic promoter was also excluded by *** Five IRESs are present in the CVB3 coding region.

关键词： CVB3 ORF P1 P2 P3 IRESs

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Laparoscopic liver resection utilizing the ventral avascular area of the inferior vena cava:A retrospective cohort study

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World Journal of Gastroenterology 2025年第1期31卷 42-57页

作者： Kun Huang Zhu Chen Heng Xiao hai-Yang Hu Xing-Yu Chen Cheng-You Du Xiang lan Department of Hepatobiliary Surgery The First Affiliated Hospital of Chongqing Medical UniversityChongqing 400000China Department of General Surgery Mianyang Hospital of Traditional Chinese MedicineMianyang 621000Sichuan ProvinceChina

BACKGROUND Laparoscopic liver resection(LLR)can be challenging due to the difficulty of establishing a retrohepatic tunnel under *** the third hepatic hilum before parenchymal transection often leads to significant liver mobilization,tumor compression,and bleeding from the short hepatic veins(SHVs).This study introduces a novel technique utilizing the ventral avascular area of the inferior vena cava(IVC),allowing SHVs to be addressed after parenchymal transection,thereby reducing surgical complexity and improving outcomes in in situ *** To introduce and evaluate a novel LLR technique using the ventral avascular area of the IVC and compare its short-term outcomes with conventional *** The clinical cohort data of patients with pathologically confirmed hepatocellular carcinoma or intrahepatic cholangiocarcinoma who underwent conventional LLR and novel LLR between July 2021 and July 2023 at the First Affiliated Hospital of Chongqing Medical University were retrospectively *** novel LLR,we initially separated the caudate lobe from the IVC using dissecting forceps along the ventral avascular area of the ***,we transected the parenchyma of the left and right caudate lobes from the caudal side to the cephalic side using the avascular area as a ***,we addressed the SHVs and finally dissected the root of the right hepatic vein or left hepatic *** short-term postoperative outcomes and oncological results of the two approaches were evaluated and *** A total of 256 patients were included,with 150(58.59%)undergoing conventional LLR and 106(41.41%)undergoing novel *** novel technique resulted in significantly larger tumor resections(6.47±2.96 cm vs 4.01±2.33 cm,P<0.001),shorter operative times(199.57±60.37 minutes vs 262.33±83.90 minutes,P<0.001),less intraoperative blood loss(206.92±37.09 mL vs 363.34±131.27 mL,P<0.001),and greater resection volume(345.11±31.40 mL vs 264.38±31.98 mL,P<0.001)comp

关键词： Laparoscopic liver resection Inferior vena cava Retrohepatic tunnel Short hepatic veins Complex liver tumors Intraoperative bleeding control

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基于MSP430的微功耗无线压力变送器研究与设计

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仪表技术与传感器 2011年第2期 19-21,25页

作者：侯培国邢耀双姚海兰李铮燕山大学电气工程学院河北秦皇岛066004

基于压阻式压力传感器的测压原理,以MSP430F449单片机为核心,设计了一种微功耗无线压力变送器,用来实现对井口压力的快速准确测量。系统设计中,为了降低功耗,外部电路均采用微功耗设计方式,并通过精确电源管理,关闭不用的模块,使设备处... 详细信息

基于压阻式压力传感器的测压原理,以MSP430F449单片机为核心,设计了一种微功耗无线压力变送器,用来实现对井口压力的快速准确测量。系统设计中,为了降低功耗,外部电路均采用微功耗设计方式,并通过精确电源管理,关闭不用的模块,使设备处于低功耗状态。同时,采用微功耗编程技术,进一步延长了电池的使用寿命。设计中,选用TR3001芯片作为无线发射接收模块,实现了测量数据的无线传输。通过合理的研究与设计,提高了压力数据的采集精度,解决了系统微功耗问题。

关键词：压力传感器微功耗 MSP430单片机无线通讯

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短双链RNA对CVB3病毒细胞内清除作用的可行性研究及其机制探讨

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中华实验和临床病毒学杂志 2008年第4期22卷 260-262页

作者：肖宗慧韩继生姚海兰刘哲伟首都儿科研究所分子免疫室北京100020

目的通过观察短双链RNA（Short interfering RNA，siRNA）体外转染HeLa细胞评价siRNA抗CVB3病毒感染的可行性和抑制作用机制。方法通过细胞病变作用保护实验，选择合适的siRNA剂量。Western Blot、***等方法在体外检测siRNA抗CVB3病毒... 详细信息

目的通过观察短双链RNA（Short interfering RNA，siRNA）体外转染HeLa细胞评价siRNA抗CVB3病毒感染的可行性和抑制作用机制。方法通过细胞病变作用保护实验，选择合适的siRNA剂量。Western Blot、***等方法在体外检测siRNA抗CVB3病毒复制作用及作用途径。结果siRNA．3753通过脂质体转染试剂能高效转入HeLa细胞，转染效率可达98．77％，在细胞内可稳定长达48h。siRNA-3753的浓度为0．6gmol／L对病毒抑制率高同时对细胞不会产生毒性作用，该浓度作为本次研究的实验剂量。siRNA-3753抗CVB3病毒感染作用是通过siRNA直接降解病毒基因得到的，而不是通过激活PKR或干扰素等其他未知因素起效的。结论siRNA可以高效、较长时间的转染入HeLa细胞内，在低剂量时即产生较好的抗病毒感染作用，通过直接降解病毒基因组的途径特异性抑制CVB3病毒的复制及感染。

关键词： RNA 柯萨奇病毒感染脂质体转染试剂抗病毒感染作用细胞

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甘薯自然开花自交结实特异材料的遗传分析

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华北农学报 2011年第B12期26卷 24-27页

作者：刘兰服马志民姚海兰张松树河北省农林科学院粮油作物研究所河北省作物遗传改良重点实验室河北石家庄050035

利用甘薯自然开花并自交结实的特异材料,进行自交、杂交、正反交等方法,创制分离群体,研究后代的遗传变异规律。从后代中培育自然开花自交结实品系,可打破甘薯自交杂交遗传障碍,进行育种材料的基因纯合累加,提供甘薯育种自交亲本及简化... 详细信息

利用甘薯自然开花并自交结实的特异材料,进行自交、杂交、正反交等方法,创制分离群体,研究后代的遗传变异规律。从后代中培育自然开花自交结实品系,可打破甘薯自交杂交遗传障碍,进行育种材料的基因纯合累加,提供甘薯育种自交亲本及简化制种新技术。

关键词：甘薯自然开花自交结实遗传分析

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基于MSP430和模糊PID油田注水流量控制系统

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仪表技术与传感器 2011年第4期 103-106页

作者：侯培国姚海兰邢耀双周志爽河北省测试计量技术及仪器重点实验室(燕山大学) 河北秦皇岛066004

介绍了一种基于MSP430单片机的低功耗油田注水流量控制系统,该系统以阀门为控制对象,利用MSP430的Tim er A模式,产生PWM输出控制信号,实现电机的PWM控制,从而通过控制注水阀门的开度,调节注水流量。采用模糊自调整PID控制,实现油田注水... 详细信息

介绍了一种基于MSP430单片机的低功耗油田注水流量控制系统,该系统以阀门为控制对象,利用MSP430的Tim er A模式,产生PWM输出控制信号,实现电机的PWM控制,从而通过控制注水阀门的开度,调节注水流量。采用模糊自调整PID控制,实现油田注水流量的闭环调节,同时,利用GPRS模块将数据远距离传送给PC机,实现远程控制,在低功耗设计上,除了在硬件上选用低功耗芯片外,还利用软件设计进一步降低了功耗,仿真结果表明:采用PWM和模糊自调整PID控制流量具有良好的动态、稳态性,从而证明了这种设计的合理性和优越性。

关键词： MSP430 PWM 模糊PID 低功耗

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Small extracellular vesicles derived from human induced pluripotent stem cell-differentiated neural progenitor cells mitigate retinal ganglion cell degeneration in a mouse model of optic nerve injury

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Neural Regeneration Research 2025年第2期20卷 587-597页

作者： Tong Li Hui-Min Xing hai-Dong Qian Qiao Gao Sheng-lan Xu Hua Ma Zai-Long Chi State Key Laboratory of Ophthalmology Optometry and Visual ScienceEye HospitalWenzhou Medical UniversityWenzhouZhejiang ProvinceChina National Clinical Research Center for Ocular Diseases Eye HospitalWenzhou Medical UniversityWenzhouZhejiang ProvinceChina

Several studies have found that transplantation of neural progenitor cells(NPCs)promotes the survival of injured ***,a poor integration rate and high risk of tumorigenicity after cell transplantation limits their clinical *** extracellular vesicles(sEVs)contain bioactive molecules for neuronal protection and *** studies have shown that stem/progenitor cell-derived sEVs can promote neuronal survival and recovery of neurological function in neurodegenerative eye diseases and other eye *** this study,we intravitreally transplanted sEVs derived from human induced pluripotent stem cells(hiPSCs)and hiPSCs-differentiated NPCs(hiPSC-NPC)in a mouse model of optic nerve *** results show that these intravitreally injected sEVs were ingested by retinal cells,especially those localized in the ganglion cell *** with hiPSC-NPC-derived sEVs mitigated optic nerve crush-induced retinal ganglion cell degeneration,and regulated the retinal microenvironment by inhibiting excessive activation of *** analysis further revealed that hiPSC-NPC derived sEVs transported neuroprotective and anti-inflammatory miRNA cargos to target cells,which had protective effects on RGCs after optic nerve *** findings suggest that sEVs derived from hiPSC-NPC are a promising cell-free therapeutic strategy for optic neuropathy.

关键词： exosome miRNA neural progenitor cell neurodegeneration neuroinflammation neuroprotection optic nerve crush optic neuropathy retinal ganglion cell small extracellular vesicles

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ITK蛋白调节小鼠脾淋巴细胞增殖分化的研究

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中华实验和临床病毒学杂志 2009年第4期23卷 269-271页

作者：肖宗慧何峰姚海兰韩继生刘哲伟首都儿科研究所分子免疫室北京100020

目的应用RNA干扰技术特异性抑制Itk基因的表达，观察Itk蛋白在淋巴细胞增殖和相应免疫因子合成分泌中的作用，进行Itk作为药物靶标的确认。方法设计合成针对Itk基因的siRNA，将siRNA电转染至小鼠脾淋巴细胞，WestemBlot检测Itk蛋白的表... 详细信息

目的应用RNA干扰技术特异性抑制Itk基因的表达，观察Itk蛋白在淋巴细胞增殖和相应免疫因子合成分泌中的作用，进行Itk作为药物靶标的确认。方法设计合成针对Itk基因的siRNA，将siRNA电转染至小鼠脾淋巴细胞，WestemBlot检测Itk蛋白的表达、MTS方法检测细胞增殖率、ELISA方法检测细胞因子的含量。结果实验组Itk—siRNA在细胞水平上有效抑制了Itk蛋白的表达；Itk受抑制后细胞增殖率与对照组相比明显降低，差异有统计学意义，P〈0．05；细胞因子IL-2，IL-4，IL-5，IFN-γ的分泌水平下降，P〈0．05。结论Itk表达受抑制后有效降低脾淋巴细胞的增殖率及细胞因子分泌的减少，研究验证了Itk在细胞增殖和炎性细胞因子分泌方面的重要免疫调节作用，为将其作为药物的靶基因提供实验依据。

关键词：蛋白质酪氨酸激酶 RNA 细胞培养的细胞因子类

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