目的 观察交泰丸水煎液对失眠小鼠昼夜节律紊乱的改善作用,并探讨其作用机制.方法 30只雄性ICR小鼠随机分为交泰丸组、模型组和对照组.交泰丸组按450 mg/kg腹腔注射对氯苯丙氨酸(PCPA),每天1次、连续2 d,建立小鼠失眠模型;然后灌胃交泰丸水煎液0.4 mL/次,连续7 d.模型组参照上述方法建立小鼠失眠模型后,灌胃交泰丸水煎液等量蒸馏水,连续7 d;对照组腹腔注射PCPA等量生理盐水每天1次、连续2 d后,灌胃交泰丸水煎液等量蒸馏水,连续7 d.然后采用旷场实验、跑台实验评价小鼠活跃度及运动能力(反映昼夜节律紊乱情况),RT-qPCR法检测小鼠脑组织时钟基因CLOCK、PER1、BMAL1及食欲素前体(HCRT),ELISA法检测小鼠外周血清DNA甲基转移酶1(DNMT1)和组蛋白去乙酰化酶(HDAC)活性,Western blotting法检测小鼠脑组织中转录调控轴SIRT1和FOXO1蛋白.结果 与对照组相比较,模型组小鼠被电击次数增加、夜间总活动距离降低,脑组织中CLOCK、BMAL1、HCRT mRNA相对表达量下降,血清DNMT1水平上升、HDAC水平下降,脑组织SIRT1蛋白水平下降、FOXO1蛋白水平上升(P均<0.05).与模型组相比较,交泰丸组小鼠在夜间总活动距离增加、被电击次数减少(P均<0.05),脑组织中CLOCK、HCRT mRNA相对表达量升高(P均<0.05),血清DNMT1活性下降、SIRT1蛋白水平升高、FOXO1蛋白水平降低(P均<0.05).结论 交泰丸水煎液能使失眠小鼠昼夜节奏紊乱改善,其作用可能是通过SIRT1/FOXO1轴上调CLOCK、BMAL1等时钟基因以及HCRT的转录水平,并抑制DNMT1活性而实现的.

Background:With the continuous advancement of clinical application and experimental research of JTP, the application prospect of JTP in nervous system diseases and metabolic diseases is becoming increasingly clear. Jiaotai Pill (JTP) is a traditional Chinese medicine formula for insomnia, consisting of Coptidis rhizoma and Cinnamomi cortex, which dates back to Han Shi Yi Tong in the Ming Dynasty of China.Objective:Based on the brain-gut axis theory, this paper aims to explore the potential mechanism of JTP in the intervention of insomnia by using intestinal microbiome and metabolomics technology, taking the animal model of insomnia as the research object, so as to provide experimental basis for its further application and research.Methods:The insomnia mouse model was induced by intraperitoneal injection of para-chlorophenylalanine (PCPA). The clinical equivalent dose of JTP was administered by gavage for one week. The efficacy of JTP was evaluated by behavioral tests, serum biochemical detection, and brain histomorphological observation. The contents of cecum were analyzed by microbiomics and metabolomics.Results:The results show that insomnia caused by PCPA led to daytime dysfunction, higher HPA axis hormone levels, and morphologically impaired hippocampus. JTP reversed these anomalies. Omics research indicates that JTP significantly reduced gut α diversity; at the phylum level, JTP reduced the relative abundance of Firmicutes, Deferribacterota, Cyanobacteria, and Actinobacteriota and increased the relative abundance of Verrucomicrobiota, Proteobacteria, and Desulfobacterota. At the genus level, JTP reduced the relative abundance of Muribaculaceae, Lachnospiraceae_NK4A136_group, Alistipes, Colidextribacter, Muribaculum, and Mucispirillum and increased the relative abundance of Bacteroides and Akkermansia. JTP also reversed the activation of the linoleic acid metabolism pathway induced by insomnia. The combined analysis of omics suggests that JTP may play a role by regulating the inflammatory state of the body. Further gene expression analysis of brain tissue confirmed this.Conclusions:We hypothesize that JTP may achieve insomnia relief by eliminating inflammation-causing bacteria in the gut and reducing inflammation levels through the brain-gut axis, pointing to potential targets and pathways for future research on JTP.

目的 观察大鼠交泰丸含药血清预处理对人脑胶质细胞(HEB)氧化损伤的改善作用并探讨其分子机制.方法 SD大鼠随机分为两部分,分别通过肉桂及黄连水提物灌胃、同体积蒸馏水灌胃制备交泰丸含药血清及空白血清,将DMEM培养基与血清配制成10％完全培养基.将HEB分为对照组、模型组及交泰丸组,对照组及模型组加入空白血清培养基、交泰丸组加入含药血清培养基.培养24 h后,交泰丸组及模型组使用过氧化氢处理建立HEB氧化损伤模型,对照组使用等体积PBS处理.采用β-半乳糖苷酶染色法观察各组细胞氧化损伤比例,CCK-8法观察细胞活力,流式细胞术观察细胞凋亡率,活性氧荧光染色法观察细胞氧化应激水平,qRT-PCR法检测细胞衰老相关基因沉默信息调节因子1(SIRT1)、趋化因子联接因子1(CXCL1)、叉头框转录因子O亚族1(FOXO1)、P16、白细胞介素1α(IL-1α)、胰岛素样生长因子结合蛋白3(IGFBP3)、IL-6 mRNA表达.结果 细胞氧化损伤比例模型组>交泰丸组>对照组,细胞活力对照组>交泰丸组>模型组,细胞凋亡率模型组>交泰丸组>对照组(P均<0.05).对照组细胞核膜边界清晰,ROS荧光强度低;模型组细胞核膜发生部分裂解,ROS荧光强度增强;交泰丸组细胞膜及核膜结构较模型组更加完整,ROS荧光强度降低.与对照组比较,模型组SIRT1、FOXO1 mRNA表达下降,CXCL1 mRNA表达上升;与模型组比较,交泰丸组SIRT1、FOXO1 mRNA表达下降(P均<0.05),三组其他基因比较差异均无统计学意义.结论 大鼠交泰丸含药血清预处理可抑制过氧化氢诱导的HEB氧化损伤,其机制可能与增强细胞活力、抗细胞凋亡、减少细胞氧化应激水平以及抗炎症有关.

目的 探讨黄芪发酵产物减轻PM2.5诱导炎症反应肺损伤的影响.方法 60只Wistar大鼠随机分成空白对照组、模型对照组、富马酸福莫特罗片对照组、黄芪发酵产物的低剂量组、中剂量组和高剂量组,每组10只,根据文献采用气管滴注方法复制PM2.5肺损伤的动物模型,各组进行相应的处理4周,最后一次滴注24～36 h内处死,检测血清和右肺上叶组织匀浆中TNF-α、IL-6、IL-1的含量,进行右肺中叶的病理切片观察,使用胞核蛋白提取试剂盒提取左肺组织中的蛋白,将TLR4 、Src、p-Src、NFκB、ICAM-1的条带通过β-actin进行归一化,并用图像分析软件进行半定量分析.结果 黄芪发酵产物高剂量组与模型组比较,血清中的TNF-α、IL-1和右肺上叶组织匀浆中的TNF-α、IL-6、IL-1及左肺组织匀浆中的TLR4、Src、NFκB、ICAM-1均有统计学意义(P＜0.01),中剂量组与模型组比较,有些指标也有统计学意义,如Src、NFκB、ICAM-1等.结论 黄芪发酵产物在一定程度上能够拮抗PM2.5诱导的肺脏中炎性因子水平增加,通过抑制TLR4-Src-NFκB-ICAM-1通路拮抗PM2.5诱发的炎性反应.

目的 探讨PCPA致失眠大鼠的时钟基因表达变化.方法 雄性SD大鼠一次性腹腔注射PCPA复制失眠动物模型,采用旷场实验评价失眠大鼠的行为学变化,采用ELISA法检测外周血清中HPA轴激素水平,采用RT-qPCR技术检测大鼠脑部、肝脏核心时钟基因(Clock、Baml1、Per1、Per2、Cry1和Cry2)的转录水平.结果 与对照组比较,模型组大鼠行为学得分显著升高(P<0.01);脑部Clock、Bmal1、Cry1表达下调(P<0.05),Cry2的表达显著上升(P<0.05);肝脏Per1表达上升(P<0.01).结论 PCPA诱导的大鼠失眠可能与脑部Clock、Bmal1、Cry1和Cry2基因和肝脏Per1基因的表达改变有关.

目的:探讨黄芪发酵产物拮抗PM2.5肺损伤的干预效果,为黄芪研究及其发酵产物的开发利用提供参考依据.方法:通过电感耦合等离子体发射光谱仪确定PM2.5重金属的主要成分.将36只Wistar大鼠随机分成空白对照组,染毒组,黄芪组,黄芪发酵产物低、中、高剂量组,根据文献采用气管滴注方法复制PM2.5肺损伤的动物模型,各组相应处理4周,最后1次滴注后,进行微循环动态可视化观察研究,并进行血气分析检测.录制直径在30～50 μm的肺细静脉白细胞黏附动态视频,计算200 μm长的肺细静脉内粘附白细胞数量.结果:本研究PM2.5的重金属成分以铁(Fe)、锌(Zn)、铬(Cr)为主.与空白对照组比较,染毒组肺组织白细胞聚集严重(P＜0.01),PCO2显著升高(P＜0.05),PO2和SO2显著降低(P＜0.05).与染毒组比较,黄芪组及黄芪发酵产物中、高剂量组黏附于肺细静脉血管壁的白细胞数量显著减少(P＜0.05),PCO2显著降低(P＜0.05),P02和S02显著升高(P＜0.05).结论:黄芪发酵产物能使PM2.5损伤的肺组织炎性程度降低,肺功能得到改善,对临床上呼吸系统疾病防治及其黄芪科学研究具有一定的启示作用.

专业课的课程思政是当前高校思想政治工作的新理念新模式.为了扎实推进高等教育"立德树人"根本任务,我们对中医药院校的生物化学课程思政进行了探索.本文从学校层面的课程思政理念输入、教师层面的思政育人能力培养和具体的课程思政设计工作三方面探讨将课程思政融入生化课堂的一系列方法和举措,从而实现在培养学生专业技能的同时进行价值引领的教学目标.

目的:探究小檗碱联合松果菊苷抗人脑胶质细胞衰老的作用及其分子机制.方法:采用依托泊苷诱导人脑胶质细胞衰老,使用小檗碱与松果菊苷联合干预,采用β-半乳糖苷酶染色法色观察衰老阳性细胞比例,采用CCK-8法检测细胞活力,采用流式细胞术检测细胞周期,采用Western blot法检测细胞中衰老相关蛋白FoxO1、CAT、SOD2、Bcl-2的表达水平.结果:小檗碱联合松果菊苷可显著减少依托泊苷诱导的衰老阳性细胞比例,显著增强细胞活力,显著降低G2/M期细胞比例,显著提高FoxO1、CAT、SOD2、Bcl-2的蛋白水平.结论:小檗碱联合松果菊苷可显著抑制依托泊苷诱导的人脑胶质细胞衰老,可能与增强细胞活力、推进细胞周期进程、降低胞内氧化应激水平、抗凋亡等途径有关.

目的 从心脏和肾脏DNA甲基化和组蛋白乙酰化修饰探讨交泰丸干预失眠动物模型的机制.方法 48只雄性SD大鼠随机分为正常组、模型组和交泰丸组,每组16只.实验第1天,模型组和交泰丸组按300 mg/kg一次性腹腔注射氯苯丙氨酸(PCPA)建立失眠模型,正常组一次性腹腔注射等体积生理盐水.实验第3天,交泰丸组灌胃交泰丸水提液5 g/kg,1次/d,连续3 d,正常组和模型组灌胃等体积生理盐水.ELISA法检测心脏和肾脏表观遗传修饰主要酶DNA甲基转移酶1(DNMT1)、组蛋白去乙酰化酶(HDAC)活性及乙酰化组蛋白H3(Acetyl-Histone H3)水平,比色法检测心脏和肾脏DNA甲基化水平.结果 PCPA致失眠大鼠心脏DNMT1、HDAC活性、DNA甲基化水平显著上升,肾脏DNMT1活性、HDAC活性、DNA甲基化水平显著下降,心脏Acetyl-Histone H3水平显著下降,肾脏Acetyl-Histone H3水平显著上升.交泰丸干预可显著降低失眠大鼠心脏DNMT1、HDAC活性和DNA甲基化水平,显著上调肾脏DNMT1活性和DNA甲基化水平,显著下调肾脏Acetyl-Histone H3水平.结论 交泰丸干预可调节失眠大鼠心脏和肾脏的DNA甲基化和组蛋白乙酰化修饰水平,从表观遗传学角度看,心肾不交可能与心脏的DNA甲基化、组蛋白去乙酰化以及肾脏的DNA去甲基化、组蛋白乙酰化的两极发展有关.

实验动物学是一门医学基础学科,一方面为医学生奠定医学基本理论及实践方法的必要基础,另一方面也与较多的社会问题密切相关,容易引起学生对于现实世界的困惑.在课程思政建设的背景下,利用实验动物学课堂教学主渠道,将思想政治教育与课程内容相结合,充分发挥二者的协同育人作用,让学生在获得专业知识的同时,更能从日常生活中发现问题、解决问题,从而获得价值观念及能力层面的双重提升,成为具有科学精神和人文情怀的高素质新型人才.

随着中医药国际化进程的加快,双语教学成为教学改革的重点.医学生物学是一门基础性学科,也是一门极具挑战的生命科学,由于医学生物学的进展动态多是从英文文献数据库中获得,双语教学势在必行.思维导图作为一种思维工具,可将难记忆的英文或其他专业术语联系在一起,提供一个正确而快速的学习方法,近来受到广泛关注.本文结合中医院校的专业特点及培养目标、学情分析等情况,探讨思维导图在医学生物学课程双语教学过程中的应用及效果,解决双语教学过程中遇到的问题,有效提高学生的学习兴趣和能力,为其他医学专业基础课的双语学习奠定基础.

目的:探讨中药单体丹皮酚对人类风湿关节炎滑膜成纤维细胞体外增殖与凋亡的影响.方法:以人类风湿关节炎滑膜成纤维细胞株MH7A作为研究材料,采用MTT法检测丹皮酚对MH7A细胞体外增殖的影响,采用流式细胞术检测丹皮酚对MH7A细胞凋亡的影响,采用qRT-PCR检测丹皮酚处理后对Ki-67、Caspase-3的mRNA水平,采用Western blot检测丹皮酚处理后Ki-67、Caspase-3的蛋白水平.结果:丹皮酚对MH7A细胞体外增殖有显著抑制作用,且抑制率与剂量和作用时长呈正相关.各剂量丹皮酚处理72 h后MH7A的凋亡水平显著上升,且Ki-67的表达水平显著降低,Caspase-3的表达水平显著升高.结论:丹皮酚可通过降低Ki-67表达,抑制MH7A增殖;通过促进Caspase-3表达,诱导MH7A凋亡.

随着中医药事业的蓬勃发展和中医药人才国际化交流的日益推进,双语教学成为了现代化中医药人才培养基础教育中的重要组成部分.为提高双语教学效果,北京中医药大学在近年来医学生物学双语课程建设中,应用了以问题为基础的教学方法,科学规划和完善了问题设计,并围绕核心问题综合运用多种教学手段,切实加强了双语授课效果,达到了既夯实生命科学基础知识,又有所拓展创新的教学目标,使课程更好地为现代化、外向型中医药人才的培养做出贡献.

Objective: To investigate the effect of Jiaotai Pill on serum HPA axis and epigenetic modification in PCPA-induced insomnia rats. Methods: The model of insomnia was induced by intraperitoneal injection of PCPA in rats. The rats were administrated with Jiaotai Pill(Cinnamon:Berberine=10:1) for 3 days. Levels of ACTH, CORT, and CRF, enzyme activities of epigenetic modification, such as DNMT1 and HDAC, as well as serum level of Acetyl-Histone H3 were measured by means of ELISA mothod. Results: Serum hormones of HPA-axis were increased in PCPA-induced insomnia rats, but there was no significant difference between the model rats and the normal rats; Jiaotai Pill could sig-nificantly reduce the hormone levels. Activity of serum DNMT1 was significantly higher in the insomnia rats than that in the normal rats; which could be obviously reduce by the intervention of Jiaotai Pill. Serum levels of Acetyl-Histone H3 was increased in insomnia rats, however, there was no significant difference between the insomnia rats and the normal rats. Jiaotai Pill could significantly reduced the level of Acetyl-Histone H3. Conclusion: Jiaotai Pill can regulate the hormone levels of HPA -axis when treating insomnia of the pattern of abnormal physiological coordiantion between heart-yang and kidney-yin, which is achieved by restoring homeostasis and by epigenetic modifications, such as DNA demethylation and histone acetylation.

Objective It is to investigate the inhibitory effects of Chinese medicine small molecule compound TanshinoneⅡA on proliferation and COX-2 expression in breast cancer cells. Methods Three kinds of breast cancer cell lines T47D,MCF-7 and MDA-MB-231 were used as experimental models in vitro. The proliferation depending on the different dosage and duration of Tanshinone IIA in the 3 kinds of breast cancer cells was detected by MTT assays. Westernblot was applied to detect the protein level of COX-2 in the 3 kinds of breast cancer cells after treated with Tanshinone ⅡA for 48 hours. Plasmids pCOX2-Luc,pNF-κB -Luc and pAP1-Luc were transfected into breast cancer cells respectively and the dual luciferase reporter gene assay system was used to detect the transcription level of COX-2 and the activity of NF-κB and AP-1. Results The results of MTT assays showed that Tanshinone ⅡA inhibited the proliferation of breast cancer cell lines T47D,MCF-7 and MDA-MB-231,and the inhibiting rate was positively correlated with the dosage and duration of TanshinoneⅡA. Westernblot revealed that the protein level of COX-2 in the 3 kinds of breast cancer cells were markedly decreased after treated with Tanshinone ⅡA for 48 hours. Luciferase assay results showed that the treatment of TanshinoneⅡA for 48 hours apparently reduced the transcription level of COX-2 in the cells T47D and MCF-7(P＜0.05),and obviously inhibited the activity of AP-1 and NF-κB in the 3 kinds of breast cancer cells. Conclusion Tanshinone ⅡA may inhibit the expression of COX-2 through depressing the activity of its transcription factors AP-1 and NF-κB,thereby suppress the proliferation of breast cancer cells.

生理教学评价改革是生理教学改革中的重要部分.回顾了形成性评价的原理及主要方法学,分析了我校中医学专业生理学教学评价中存在的问题,提出了构建科学、客观、有效的评价体系的几点建议与原则,以期真正实现以评促教的教学改革目标.

Objective To study the mechanism of Wuzi Yanzong Wan (Five-Seed Progeny Pill) on improving heat stress resistance of sertoli cells in rats based on data from the whole genome microarray.Methods Sertoli cells in primary culture were randomly divided into Wuzi Yanzong Wan (WZYZW) group, normal group and model group.The cells in three groups were given WZYZW solution and equal volume of PBS respectively.After 24 hours, rats in WZYZW group and model group were placed in water bath at 43 ℃ for 15 minutes, while rats in normal group were put in water bath at 37 ℃ for 15 minutes, then total RNA of three groups were extracted respectively and data was analyzed with gene chip technology.Results Compared with the model group, there were totally 1,203 differently genes in WZYZW group based on Sertoli cell gene expression profiles, in which 697 were down regulated and 506 were up-regulated.There were totally 473 differently expressed genes in the normal group, including 168 up-regulated genes and 305 down regulated genes.Among these down-regulated gene expressions, RGD1305298, Trpt1, Dll3, Abi3 and Ppp1r12b were significant in the WZYZW group and normal group, while Dusp7 was significant among up-regulated gene expressions.The genes mentioned above were related to cell cycle, apoptosis, cell metabolism, cell adhesion and Notch signal pathway, regulation of actin cytoskeleton, MAPK signaling pathway and tight junction pathway, etc.Conclusion The mechanism of Wuzi Yanzong pill in improving heat stress resistance of sertoli cells may involve maintenance of tight junctions between sertoli cells, inhibition of sertoli cells apoptosis and promotion of cell differentiation and maturation.

Objective:To explore the effects of Astragalus glycoproteins on collagen-induced arthritis mice′s specific transcription factors, T-bet and GATA-3, expression, and to clarify the immune mechanism of Astragalus glycoprotein on the treatment in collagen-induced arthritis mice.Methods:CIA model was induced by bovine Ⅱcollagen in mice.Mice were randomly divided into a CIA model group, a hydrocortisone positive control group, a Astragalus glycoprotein low dose group, a middle dose group and a high dose group.HE was used to observe the CIA mice spleen tissues damage.IFN-γ and IL-4 level were measured with flow cytometry in CIA mice.Western blot was used to analyze T-bet and GATA-3 protein expression.Results:Compared with the model group, Astragalus glycoprotein improved spleen tissue damage in CIA mice, and inhibited the infiltration of inflammatory cells.Astragalus glycoprotein reduced IFN-γand IL-4 level (P<0.05) and T-bet and GATA-3 protein expression (P<0.05).Conclusion:The mechanisms of action of Astragalus glycoprotein on CIA mice may be related to the decrease of T-bet and GATA-3 expression and function balance of the Th1 and Th2 cells.

Qihuang Guoyi Class of BUCM aims at training top students majoring in traditional Chinese medicine in the fields of clinical practice, disease prevention and scientific research. It is an important task of our reform to nurture excellent high school students to be top creative professionals of TCM. It is on this premise that we reformed the traditional verifying physiological experiment to a multilevel experiment teaching mode which featured basic techniques, verifying and comprehensive experiment, and autonomous design all together. Students’learning interest was raised. Teaching resources were optimized, and efficiency was increased. From this teaching reform practice we come to realize that it is of great significance to train students’problem-solving and creative thinking abilities in an integrated multi-level experimental teaching system.

Objective:To investigate the effect of Paeonol on the proliferation and COX -2 transcription of SiHa cells . Methods:Human cervical squamous cell carcinoma SiHa cell line was used as a model in vitro .The proliferation of SiHa cells treated by Paeonol was detected by MTT assays .Plasmids pCOX2-Luc and pAP1-Luc were transfected into SiHa cells and the dual luciferase reporter gene assay system was used to detect the transcription level of COX -2 and the ac-tivity of AP-1 .Results:The results of MTT assays showed that Paeonol significantly inhibited the proliferation of SiHa cells and the effect was associated positively with the dose and treatment time of paeonol .Luciferase assay results showed that Paeonol apparently reduced the transcription level of SiHa COX -2 cells ( P<0 .01 ) , and obviously inhibited the activity of AP-1(P<0.01).Conclusion:Paeonol may inhibit COX-2 transcription by depressing the activity of tran-scription factor AP-1,thereby suppress the proliferation of SiHa cells .