Objective To explore the impact of sinomenine on bleomycin A5-induced pulmonary fibrosis (PF) in rats and the underlying mechanism. Methods MRC-5 cells were cultured and treated with sinomenine to determine its optimal concentration and time through the MTT assay. Subsequently, MRC-5 cells were incubated with 80 μmol/L sinomenine for 48 hours or transfected with miR-21 mimic/a disintegrin-like and metalloproteinase with thrombospondin type 1 motif (ADAMTS-1) siRNA prior to sinomenine treatment. The expression of miR-21, ADAMTS-1, collagen type 1 (Col1) and collagen type 3 (Col3) was detected by quantitative real-time PCR (qRT-PCR) and/or Western blot analysis. Thirty SD rats were randomly divided into control group, sinomenine group and sinomenine combined with miR-21 agomir group, with 10 animals in each group. Bleomycin A5 were intratracheally administered to establish the PF model. Then, rats in control group, sinomenine group and sinomenine +miR-21 agomir group were treated with 9 g/L sodium chloride solution, sinomenine and sinomenine+miR-21 agomir, respectively. On day 28, all rats were sacrificed. HE and Masson staining was performed in pulmonary tissue. The expression of ADAMTS-1, Col1 and Col3 in pulmonary tissue were detected by qRT-PCR and/or Western blot analysis. ELISA was used to measure serum procollagen type 1 carboxyterminal propeptide (P1CP) and procollagen type 3 aminoterminal propeptide (P3NP) levels. Results Administration of sinomenine decreased miR-21 levels, up-regulated ADAMTS-1 expression, and promoted Col1 and Col3 degradation in MRC-5 cells. Importantly, interfering with the miR-21/ADAMTS-1 signaling pathway partially reversed the promotive effect of sinomenine on Col1 and Col3 degradation. Treatment of SD rats with sinomenine reduced alveolitis and PF scores, decreased serum P1CP and P3NP levels, up-regulated pulmonary ADAMTS-1 expression, and down-regulated Col1 and Col3 expression. However, these effects were reversed by miR-21 agomir. Conclusion Sinomenine promotes Col1 and Col3 degradation and inhibits PF in rats by miR-21/ADAMTS-1 pathway.

目的:探讨乌苯美司对D组慢性阻塞性肺疾病急性加重期(AECOPD)患者辅助性T细胞 17(Th17)/调节性T细胞(Treg)免疫失衡的影响.方法:选取2019 年10 月—2021 年3 月怀化市第一人民医院收治的D组AECOPD患者80 例,采用随机数字表法分为对照组和干预组,各40 例.对照组予以常规对症和支气管扩张治疗,干预组在对照组基础上加用乌苯美司胶囊治疗,共12 周.评估两组患者的住院时间和治疗前后的肺功能指标、免疫功能指标、细胞因子水平,观察不良反应情况,随访并比较12 个月内急性加重次数和再住院次数.结果:干预组患者住院时间短于对照组(P<0.05).治疗前,两组患者肺功能指标、免疫功能指标及细胞因子水平相比无显著差异(P>0.05).治疗后,两组患者FEV1、FEV1%、FEV1/FVC 均高于治疗前,且干预组均高于对照组(P<0.05);两组患者 Th17 细胞、Th17/Treg、白细胞介素(IL)-17A水平均低于治疗前,Treg细胞、IL-10 水平均高于治疗前,且干预组改善程度优于对照组,差异均有统计学意义(P<0.05).随访12 个月干预组患者急性加重次数和再住院次数明显少于对照组(P<0.05).而两组患者不良反应发生率相比无显著差异(P>0.05).结论:乌苯美司能有效改善D组AECOPD患者Th17/Treg免疫失衡,降低IL-17A表达,增高IL-10 水平,减轻炎症反应,缩短住院时间,提升肺功能,减少急性加重和再住院次数,且安全性良好.

目的:探讨青藤碱(SIN)对氧化应激损伤、肺纤维化的保护作用及与Kelch样ECH相关蛋白1(Keap1)/核因子E2相关因子2(Nrf2)信号通路的关系.方法:通过过氧化氢(H2O2)诱导MRC-5细胞建立氧化应激损伤模型,并予SIN处理,采用CCK-8法检测细胞活力,生化试剂盒检测丙二醛(MDA)含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性,Western blot检测Keap1、Nrf2蛋白表达.将30只SD大鼠随机分为对照组、博莱霉素A5(BLM)组和BLM+SIN组,每组包含10只动物,BLM组、BLM+SIN组气管内注射BLM建立大鼠肺纤维化模型,对照组气管内注射等体积9 g/L氯化钠溶液,造模后次日,BLM+SIN组以SIN灌胃,其余两组给予9 g/L氯化钠溶液灌胃,第28天处死大鼠,分离肺组织,通过HE和Masson染色观察肺组织病理学改变,测定肺组织MDA含量及SOD、GSH-Px、CAT活性,Western blot检测Keap1、Nrf2蛋白表达.结果:相对于H2O2 组,SIN干预后MRC-5细胞活力增加,MDA含量降低,SOD、GSH-Px和CAT活性升高,下调Keap1表达,促进Nrf2核移位.与BLM组相比,SIN给药降低大鼠肺泡炎、肺纤维化病变和评分及肺组织MDA含量,增加肺组织SOD、GSH-Px和CAT活性,下调肺组织Keap1表达,升高胞核Nrf2水平.结论:SIN可能通过激活Keap1/Nrf2信号通路抑制氧化应激损伤,减轻大鼠肺纤维化.

Objective To observe the role of tumor necrosis factor-α (TNF-α) and platelet-derived growth factor-B (PDGF-B) in kiwi fruit essence-mediated protection of radiation-induced lung injury (RILI) in rats. Methods 96 male healthy Sprague-Dawley rats were divided into normal control group, model group, and kiwi fruit essence treatment group(60 and 240 mg/kg) by the random number table method, with 24 animals in each group. The whole lungs underwent 6 MV X-ray irradiation (18 Gy) to induce RILI animal models in rats of the latter three groups. On the next day after irradiation, rats in the latter two groups were intragastrically administrated with 60 or 240 mg/kg kiwi fruit essence, once a day. The rats in the normal control and model groups were treated with 9 g/L sodium chloride solution. Eight rats in the latter three groups were randomly sacrificed on days 14, 28, and 56, while normal control rats were sacrificed on day 56 as the overall control. Blood samples were collected and separated. Serum concentrations of TNF-α and PDGF-B were detected using ELISA. The lung tissues were isolated for HE and Masson staining to evaluate alveolitis and pulmonary fibrosis (PF). The hydroxyproline (HYP) content in lung tissues was detected. The mRNA and protein expression of pulmonary TNF-α and PDGF-B were determined by quantitative real-time PCR and immunohistochemistry. Results Compared with the model group, treatment with 60 and 240 mg/kg kiwi fruit essence group significantly reduced alveolitis on days 14 and 28 as well as PF lesions on days 28 and 56. Compared with the normal control group, HYP content in the lung tissue of the model group increased on day 28 and day 56, while TNF-α and PDGF-B levels in the serum and lung tissues increased at each time point. Compared with the model group during the same period, 60 and 240 mg/kg kiwi fruit essence element treatment group reported the diminished levels of serum and pulmonary TNF-α on day 14 and day 28. Consistently, the lung tissue HYP content and serum and pulmonary PDGF-B levels on day 28 and day 56 were reduced. In addition, the above indicators in the 240 mg/kg kiwi fruit essence treatment group were lower than those for the 60 mg/kg kiwi fruit essence treatment group. Conclusion Kiwi fruit essence can alleviate RILI in rats, which is related to the down-regulation of TNF-α expression at the early stage and decreased PDGF-B level at the middle and late stages.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的探讨生长阻滞特异性转录本5（growth arrest-specific transcript 5,GAS5）对肺纤维化的影响及分子机制。方法采用生物信息学、荧光素酶报告基因分析GAS5、miR-21、ADAMTS-1间的竞争性内源RNA（ceRNA）关系。SD大鼠分为对照组、LV-GAS5组和LV-GAS5+miR-21 agomir组,经气管内注入博莱霉素A5建立肺纤维化模型后,分别给予尾静脉注射0.2 mL的PBS、LV-GAS5、LV-GAS5+miR-21 agomir。d 28,处死大鼠,收集血液,采用ELISA分析血清PICP和PⅢNP水平,提取肺组织,HE染色和Masson染色观察病理改变,并用qRT-PCR检测GAS5、miR-21、ADAMTS-1表达,Western blot检测ADAMTS-1、ColⅠ、ColⅢ表达。结果GAS5、miR-21、ADAMTS-1之间存在ceRNA调控模式。与对照组比较,GAS5过表达减轻肺组织病理改变,降低血清PICP、PⅢNP水平,上调肺组织GAS5、ADAMTS-1表达,降低肺组织miR-21、ColⅠ、ColⅢ水平（P &lt;0.01）。此外,miR-21agomir处理逆转了GAS5过表达对肺纤维化大鼠肺组织病理改变、胶原沉积和ADAMTS-1表达的影响（P &lt;0.01）。结论GAS5通过内源性吸附miR-21,上调ADAMTS-1表达,促进ColⅠ、ColⅢ降解,减轻肺纤维化病变。</span>

目的 通过运用行为阶段模型(TTM)对医学生临床技能学课程学习动机进行评估,探索虚拟仿真教学方法的积极作用.方法 选取该校处于本科第3学年2学期临床医学专业学生240名,在均衡原则下随机分配入虚拟仿真教学组及传统教学组,每组120名.进入分组前和学期末,学生完善T T M评估量表;同时搜集学期末临床技能诊断部分的实践考核成绩进行统计学分析.结果 学期末,虚拟仿真教学组学习动机处于第四、五阶段学习阶段学生所占比例优于传统教学组,差异有统计学意义(P<0.05).虚拟仿真教学组平均成绩[(80.13±3.32)分]高于传统教学组[(74.26±4.25)分],差异有统计学意义(t=6.42,P<0.05).结论 TTM直观显示了虚拟仿真教学方法在提升医学生学习动机方面的优势,值得在教学评价实践中推广.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的观察生长阻滞特异性转录本5（growth arrest-specific transcript 5,GAS5）对肺成纤维细胞Ⅰ型胶原（ColⅠ）和Ⅲ型胶原（ColⅢ）表达的影响,并探讨其机制。方法培养人胚肺成纤维细胞MRC-5,用GAS5慢病毒过表达载体（LV-GAS5）处理,荧光实时定量PCR（qRT-PCR）检测GAS5、miR-21水平,qRT-PCR和Western blot测定I型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-1（ADAMTS-1）、ColⅠ、ColⅢ表达。生物信息学、双荧光素酶报告基因分析GAS5、ADAMTS-1 3′-非翻译区与miR-21的靶向结合情况。以miR-21模拟物（mimic）或ADAMTS-1 siRNA转染MRC-5细胞48 h,再用LV-GAS5处理细胞相同时间,qRT-PCR和Western blot检测ADAMTS-1、ColⅠ、ColⅢ表达。结果过表达GAS5降低miR-21水平,抑制ColⅠ、ColⅢ蛋白表达,增加ADAMTS-1 mR NA与蛋白水平（P &lt;0.01）。GAS5、miR-21、ADAMTS-1之间存在竞争性内源RNA （ceR NA）作用模式。miR-21 mimic或ADAMTS-1 siR NA预处理几乎完全逆转过表达GAS5对ColⅠ、ColⅢ降解的促进作用（P &lt;0.01）。结论 GAS5作为ceR NA竞争性结合miR-21,上调ADAMTS-1表达,从而促进MRC-5细胞ColⅠ、ColⅢ降解。</span>

通过分析甲基CpG结合蛋白2(methyl CpG binding protein 2,MeCP2)对山姜素作用下鼠巨噬细胞(RAW246.7)合成IL-6和TNF-α的影响及作用机制,揭示山姜素与MeCP2联合应用对于炎症性疾病的干预潜力.用重组质粒pEGFP-C1-MeCP2以及空白质粒pGL-basic分别转染RAW246.7,后将细胞随机分为对照组、山姜素组(终浓度1 mg/mL)以及山姜素+GW9662(0.1 mmol/L)组.孵育72 h后,ELISA检测培养液中IL-6和TNF-α表达水平,Western blotting分别检测细胞核内过氧化物酶体增殖物激活受体(peroxisome proliferators-activated receptor,PPAR)、MeCP2以及胞质中NLRP3、pro-Caspase-1和各聚集状态凋亡相关微粒蛋白(apoptosis-associated speck-like protein containing CARD,ASC)含量,并采用免疫荧光染色法检测ASC斑点形成情况.结果 显示,与空白质粒转染比较,重组质粒转染RAW246.7在山姜素作用下合成IL-6和TNF-α的水平更低;而在对照组以及GW9662组,转染重组质粒对炎性因子合成无明显影响.山姜素对MeCP2表达无促进作用,且重组质粒转染对PPAR表达亦无影响.另外,山姜素可明显抑制RAW246.7胞质NLRP3合成,但对pro-Caspase-1含量以及ASC聚集状态无显著影响;而过表达MeCP2可遏制pro-Caspase-1合成并促进ASC解聚.由此,MeCP2可通过抑制炎症小体通路以促进山姜素对鼠巨噬细胞炎性因子合成的抑制,提示山姜素与MeCP2联合有望成为炎症性疾病防治的新策略.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的:探讨混合性评价模式在诊断学体格检查实践教学中的实施效果。方法按数字表法随机抽取我校2015级本科临床医学专业两个教学班共242名学生为研究对象,将其分为研究组（119人）和对照组（123人）,研究组实施混合性评价模式,对照组采用传统评价模式,结束教学后,根据诊断学体格检查实践考核成绩、出勤率、问卷调查、评价模式满意度调查及学生访谈进行教学效果的评价。结果研究组诊断学体格检查实践考核成绩明显高于对照组（P&lt;0.01）,同时,研究组在出勤率、问卷调查、评价模式满意度调查和学生访谈上对诊断学体格检查实践教学效果的评价优于对照组（P&lt;0.01）。结论混合性评价模式能明显提高诊断学体格检查实践教学效果,值得在以后的教学中推广使用。</span>

目的 探讨情景模拟结合混合式教学法在诊断学教学中对提高医学生问诊能力的作用.方法 按随机数字表法选取2018年3—6月湖南医药学院2015级本科临床医学专业两个教学班232名学生为研究对象,将其分为实验组(112人,实施情景模拟结合混合式教学法)和对照组(120人,采用传统教学法),教学结束后,比较两组学生诊断学问诊考核成绩,并通过问卷调查、教学满意度和学生访谈进行教学效果的评价.结果 实验组诊断学问诊考核成绩(31.59±4.52)分明显高于对照组(27.08±5.03)分(t=7.163,P<0.01),并且,实验组在问卷调查、教学满意度和学生访谈等方面对诊断学问诊教学效果的评价优于对照组(P<0.01).结论 情景模拟结合混合式教学法可显著提高医学生问诊能力,值得在教学中推广使用.

目的 通过运用行为阶段模型(TTM)对不同层次本科医学生临床课程学习动机进行分析,揭示不同层次本科医学生在学习动机方面的区别并探讨学习动机对学习效果的影响.方法 随机从南华大学衡阳医学院和湖南医药学院各筛选60名学生,根据学生实际情况,制定针对学习动机分析的TTM评估量表.在接受临床课程学习半学期后,请学生完善调查量表;再搜集临床课程的期末理论及迷你临床演练评估(Mini-CEX)测试成绩进行统计学分析.结果 2所高校医学生临床课程学习动机行为阶段分布比较,差异有统计学意义(P＜0.05),且学习动机所处行为阶段对学习效果有一定影响,尤其是Mini-CEX成绩.结论 根据实际情况制定的TTM可全面细致地分析和评估医学生学习动机,值得在以后的教学工作中不断改进和推广.

目的 探讨microRNA-27a-3p(miR-27a-3p)对肺成纤维细胞I型胶原(ColⅠ)和III型胶原(ColⅢ)合成的影响及分子机制.方法 培养人胚肺成纤维细胞MRC-5,转染miR-27a-3p模拟物/抑制物,实时荧光定量PCR(qPCR)检测miR-27a-3p水平;qPCR和Western blot测定ColⅠ、ColⅢ、Wnt3a表达;Western blot分析细胞核内 β-catenin水平.生物信息学预测miR-27a-3p与Wnt3a 3′-非翻译区(3′-UTR)靶向结合情况,并用双荧光素酶报告基因检测miR-27a-3p模拟物对Wnt3a 3′-UTR野生型和突变型荧光素酶活性的影响.给予Wnt3a/β-catenin信号通路抑制剂Dkk1预处理MRC-5细胞6 h,再用miR-27a-3p抑制物转染细胞48 h,qPCR和Western blot检测ColⅠ、ColⅢ表达.结果 miR-27a-3p模拟物明显增加miR-27a-3p水平,降低ColⅠ、ColⅢ、Wnt3a和 β-catenin表达,其抑制物的作用则相反,与对照组比较,差异均有显著性(P<0.01).Wnt3a 3′-UTR存在1个miR-27a-3p的结合位点,miR-27a-3p过表达减少野生型Wnt3a 3′-UTR荧光素酶活性(P<0.01),但对其突变型荧光素酶活性无影响(P>0.05).Dkk1预处理几乎完全逆转miR-27a-3p抑制物对ColⅠ、ColⅢ合成的诱导作用(P<0.05).结论 miR-27a-3p抑制肺成纤维细胞ColⅠ、ColⅢ生物合成,其机制与拮抗Wnt3a/β-catenin信号通路有关.

目的 探讨基于物理诊断网络考评系统的混合式教学法在诊断学心电图教学中的应用效果.方法 将湖南医药学院医学院2015级本科临床医学专业两个教学班232名学生随机分为对照组(120人)和研究组(112人),对照组和研究组分别采用传统教学法、基于物理诊断网络考评系统的混合式教学法进行心电图教学.教学结束后,比较两组学生心电图考核成绩,并通过问卷调查、教学满意度调查和学生访谈评价教学效果.结果 研究组学生心电图考核成绩高于对照组(P<0.05);研究组在问卷调查、教学满意度调查和学生访谈等方面对心电图教学效果的评价均优于对照组(均P<0.05).结论 基于物理诊断网络考评系统的混合式教学法能显著提高诊断学心电图教学效果,值得推广应用.

目的 全科医学在我国是一门较为新兴的发展中的学科.随着医疗卫生体制改革,各地政府及医学院校更加重视全科医学的人才培养,湖南医药学院也将《全科医学概论》列为医学教育的必修课程.全科医学概论是以人为中心,面向社区和家庭,整合临床医学、预防医学、康复医学以及人文社会学科相关内容于一体的综合性医学专业学科.本文对以病例为中心、问题为基础、社区为导向的教学法(case-centered、problem-based、community-o-riented teaching mode,CPC)联合传统讲授式教学法(lecture-based learning,LBL)在全科医学概论课程中的应用效果进行初步探索,为以后的教学提供借鉴与参考.方法 按数字表法随机选取2015级本科临床医学专业2个教学班248名学生为研究对象,将其分为对照组(120人)和实验组(128人),对照组采用LBL教学法,实验组应用CPC联合LBL教学法,课程结束后根据全科医学概论理论考试成绩、见习考核成绩、问卷调查和教学满意度进行教学效果的评价.结果 实验组学生全科医学概论理论考试成绩和见习考核成绩均明显高于对照组(均P＜0.01),同时,实验组在问卷调查和教学满意度上对全科医学概论教学效果的评价优于对照组(均P＜0.01).结论 CPC联合LBL教学法是一种科学有效的适合医学院校全科医学概论的教学方法,其能提升医学生对全科医学概论的认知,培养医学生的综合能力、知识运用能力,提高医学生的岗位胜任力,值得在今后的教学中推广.

Objective To observe the effect of miR-27a-3p on bleomycin A5-induced pulmonary fibrosis (PF) in rats and explore the underlying mechanism. Methods Forty-five male SD rats were randomly divided into control group, miR-27a-3p agomir group and miR-27a-3p antagomir group. Each group contained 15 animals. All rats were injected intratracheally with bleomycin A5 to establish PF models. On the first day after bleomycin A5 administration, the rats in the control group, miR-27a-3p agomir group and miR-27a-3p antagomir group were injected at the caudal vein with physiological saline, agomir and antagomir, respectively. Injection was given one time each three days, totally nine times. On day 28, blood samples were collected and then underwent enzyme linked immunosorbent assay for procollagen type 1 carboxyterminal propeptide (P1CP) and procollagen type 3 aminoterminal propeptide (P3NP) concentrations. Subsequently, all rats were sacrificed to remove pulmonary tissue. Both HE and Masson staining were performed to evaluate the pathological changes of PF. The expression of miR-27a-3p, collagen type 1 (Col1), and collagen type 3 (Col3) were detected using fluorescence real time quantitative PCR. Western blotting was used to examine Col1, Col3, Wnt3a and β-catenin levels. Results The miR-27a-3p agomir markedly increased miR-27a-3p expression in the pulmonary tissue, whereas its antagomir decreased it, showing higher transfection efficacy. The pulmonary inflammation and fibrosis degree was alleviated in the miR-27a-3p agomir group while aggravated in the miR-27a-3p antagomir group. In comparison with control group, serum P1CP and P3NP levels decreased in the miR-27a-3p agomir group but increased in the miR-27a-3p antagomir group. Treatment with miR-27a-3p agomir down-regulated the expression of Col1, Col3, Wnt3a and β-catenin in the pulmonary tissue, while miR-27a-3p antagomir up-regulated their expression. Conclusion The miR-27a-3p inhibits the Wnt3a/β-catenin signaling pathway, leading to the down-regulation of Col1 and Col3 expression and the subsequent alleviation of PF.

Objective To observe the effects of microRNA-7 (miR-7) on intracerebral hemorrhage-induced brain injury in rats and explore the underlying mechanism. Methods Seventy-five SD rats were used to establish intracerebral hemorrhage model through injection of VII collagenase into the pallidum. These rats were then divided into control, miR-7 agomir, agomir control, miR-7 antagomir and antagomir control groups, containing 15 animals in each group. On day 2 after modeling, rats were injected with 10 μL of physiological saline, miR-7 agomir, agomir control, miR-7 antagomir and antagomir control via the lateral ventricle, respectively. On day 7 after modeling, the neurological function score was evaluated, and then all rats were killed to obtain brain tissue. HE staining was conducted to observe the pathological changes of cerebral tissue. Brain water content was examined by the dried and wet mass. The expressions of miR-7, glial fibrillary acidic protein (GFAP) and epidermal growth factor receptor (EGFR) in the cerebral tissue around hemotomas were detected using real-time quantitative PCR. Western blotting was used to analyze the levels of GFAP, EGFR, signal transducers and activators of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) in the cerebral tissue around hemotomas. In addition, bioinformatics was used to predict the binding of miR-7 to EGFR. Both wild type luciferase reporter gene vector and corresponding mutant vector were constructed and then transfected into HEK293T cells along with miR-7 mimic for detecting the luciferase activity. Results In comparison with control group, miR-7 agomir markedly increased miR-7 level, alleviated the pathological impairment of cerebral tissues, reduced neurological function score and brain water content, and attenuated the expression levels of GFAP and EGFR and p-STAT3/STAT3 ratio in the cerebral tissue around hemotomas. An opposite effect was observed in response to miR-7 antagomir. However, their negative controls had no impact on the above indicators. There was a binding site between 3'-untranslated region within the EGFR and miR-7. Moreover, miR-7 mimic markedly decreased the luciferase activity of the reporter gene of wild type, but not its mutant. Conclusion The miR-7 can inhibit the EGFR/STAT3 signaling pathway to antagonize astrocyte activation, leading to the protection against brain injury after intracerebral hemorrhage in rats.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的:探讨基于微博平台的混合式教学法在诊断学教学中的实施效果。方法:按数字表法随机选取我校2015级临床医学专业两个教学班学生为研究对象,将其随机分为对照组（123人）和实验组（119人）,对照组采用传统教学法,实验组实施基于微博平台的混合式教学法,教学结束后根据诊断学成绩、期末主题演讲、问卷调查和教学满意度进行教学效果的评价。结果:实验组诊断学期末理论考试成绩及实践操作考核成绩显著高于对照组（P&lt;0.01）。同时,实验组在期末主题演讲、问卷调查和教学满意度上对诊断学教学效果的评价优于对照组（P&lt;0.01）。结论:基于微博平台的混合式教学法可提高诊断学教学效果,是一种行之有效的教学方法。</span>

Aim To explore the effects of miR-7 on astrocyte activation and the underlying mechanisms. Methods Following isolation and culturing, astro-cytes extracted from rat cortex were treated with culture solution (control group), ciliary neurotrophic factor (CNTF, an agonist of astrocyte activation), miR-7 mimic+CNTF, miR-7 mimic control+CNTF, miR-7 inhibitor+CNTF and miR-7 inhibitor control+CNTF, respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA ex-pression of glial fibrillary acidic protein (GFAP) and epidermal growth factor receptor(EGFR). The protein expression of GFAP, EGFR, signal transducers and activators of transcription 3(STAT3) and phosphoryla-ted STAT3 (p-STAT3) was measured using Western blot. Wild type pGL3-EGFR and mutant pGL3-EGFR-m recombinant plasmids were constructed and then co-transfected with miR-7 mimic into HEK293T cells,re-spectively. The luciferase activity of reporter gene was measured. In addition,astrocytes were treated with ei-ther EGFR siRNA or S31-201 (an inhibitor of STAT3),followed by the incubation with miR-7 inhib-itor and CNTF. Both qRT-PCR and Western blot were subsequently used to detect the mRNA and protein lev-els of GFAP. Results The expression levels of GFAP and EGFR as well as p-STAT3/STAT3 ratio in CNTF group were higher than those in control group (P <0.01). When compared with CNTF group,GFAP and EGFR levels and p-STAT3/STAT3 ratio significantly decreased in miR-7 mimic+CNTF group but increased in miR-7 inhibitor+CNTF group(P<0.01). In com-parison with control group, transfection with miR-7 mimic markedly reduced the luciferase activity of wild type EGFR (P <0.01). Moreover, miR-7 inhibitor-induced up-regulation of GFAP expression was almost completely reversed by either EGFR siRNA or S31-201 pretreatment (P<0.01). Conclusion miR-7 antag-onizes the activation of astrocytes from rats by inhibi-ting the EGFR/STAT3 signaling pathway.

Objective To screen the nucleus located-methyltransferase in murine macrophages (RAW246.7 cells) after peroxisome proliferator-activated receptors (PPAR) is activated by alpinetin so as to prove the epigenetic modification effect of alpinetin. Methods RAW246.7 cells were divided into control group, alpinetin group (final concentrations including 100, 200, 500, 1000 μg/mL) and 1000 μg/mL alpinetin combined with 0.1 mmol/mL GW9662 group. Firstly, bioinformatics database String was searched for the methyltransferases which might interact with PPAR. Then co-immunoprecipitation was used to screen the specific nucleus-located methyltransferase interacting with PPAR. Finally, the expressions of the related methyltransferases were validated by fluorescent quantitative PCR. Results Co-immunoprecipitation proved that EZH2, DNMT3α and TDG were the specific methyltransferases which interacted with the activated PPAR in the nucleus when induced by a certain concentration of alpinetin, which was basically consistent with the search result of the String database. No methyltransferase was found to interact with PPAR if GW9662 was added. Furthermore, only by a high concentration of alpinetin (1 000 μg/mL), could the synthesis of TDG mRNA be promoted, yet the synthesis of DNMT3α and EZH2 were not influenced. Conclusion Alpinetin, the PPAR activator, could promote the synthesis and interaction of specific methyltransferases with PPAR in the nucleus, which indicates that methylation modification on histone or cytosine may be the interpretation for the effect of gene expression regulation caused by alpinetin.

In order to reveal the role of H3K9 deacetylation in IL-6 expression from RAW246.7 caused by alpinetin,we detected the acetylation state of H3K9 and the expression level of transcription factors (P40 and CREB) combined in promoter region or nucleus.Firstly,RAW246.7 were divided into control group,alpinetin groups (with final concentration of 50,100,200 μg/ml),200 μg/ml alpinetin+GW9662 group,200 μg/ml alpinetin+ HDAC1,and Pgene SiRNA group.After incubation,concentration of IL-6 in media was tested by ELISA,while Western blot was used to detect P40,CREB and PPAR in nucleus;content of deacetylated H3K9,P40 and CREB binding in promoter region of IL-6 were tested by chip-qPCR.Date showed that alpinetin promoted the expression of PPAR and deacetylated H3K9 in IL-6 promoter in a dose-dependent manner,meanwhile inhibited generation of IL-6,accompanied by a decline of transcription factor both binding in promoter and nucleus.Compared with 200 μg/ml alpinetin group,GW9662 could suppress the promotion of PPAR and deacetylated H3K9 binding caused by alpinetin,recover the content and binding capacity of transcription factors in nucleus,and improve synthesis of IL-6;HDAC1 intervention had no influence on total synthesis of P40 and CREB in nucleus and promotion of PPAR,but could also repress the promotion of deacetylated H3K9 and promote the down-regulated binding content of P40 and CREB,thus increased synthesis of IL-6,which was still lower than GW9662 group.In conclusion,the PPAR agonist alpinetin could promote deacetylation state of H3K9 combined in the promoter of IL-6 via specific deacetylase,which could affect the binding capacity of corresponding transcription factors,thus regulate gene expression.However,synthesis of transcription factors are not interfered by deacetylation of H3K9.