Biochemistry course is the basic course of Chinese medicine colleges and universities, which has the characteristics of both foundation and practice. How to carry out teaching reform, transform boring theoretical knowledge into students’ inner knowledge and ability, and combine teaching content with ideological and political education to guide students’ active learning has been a problem that teachers have been thinking deeply and actively exploring. Starting from the integration of scientific research cases into theory, practice teaching and research topics,this paper analyzes the way of scientific research to help teaching, reflects on the existing problems and improvement strategies of scientific research into teaching, combines the cases with the characteristics of traditional Chinese medicine and biochemistry courses, so as to make a certain exploration of the reform of scientific research to help biochemistry teaching and promote teaching and learning.

目的 观察益气通络方对糖尿病周围神经病变大鼠体重、空腹血糖(FBG)、坐骨神经传导速度、高迁移率族蛋白(HMG)B1蛋白和mRNA表达的影响,探讨益气通络方改善糖尿病周围神经病变的作用机制.方法 采用腹腔注射链脲佐菌素(STZ)复制糖尿病大鼠模型.造模成功大鼠随机分为模型对照组、益气通络方高、低剂量组、西药对照组,另设正常对照组,造模成功后分别给予益气通络方、二甲双胍灌胃给药,干预12 w后,观察大鼠一般状态,检测体重、FBG、坐骨神经传导速度,坐骨神经组织中HMGB1蛋白及mRNA表达.结果 与模型对照组比较,经益气通络方干预后,大鼠体重、FBG、坐骨神经传导速度明显改善;坐骨神经组织中HMGB1蛋白和mRNA表达量均显著下调(P<0.05),其中,益气通络方高剂量组对HMGB1蛋白和mRNA表达的改善更为显著.结论 益气通络方能够下调HMGB1蛋白和mRNA表达,改善坐骨神经传导速度,防治DPN大鼠糖尿病周围神经病变损伤,其机制可能与对HMGB1的调节有关.

目的 探讨蛹虫草活性成分虫草素和虫草酸的抗炎症作用机制.方法 以蛹虫草为研究材料,以蛹虫草的活性成分虫草素和虫草酸为研究对象,基于网络药理学的原理,构建蛹虫草活性成分虫草素和虫草酸抗炎症作用蛋白质互作网络.并依据该网络,以脂多糖诱导的RAW264.7细胞为模型,在确立虫草素和虫草酸合适的干预浓度和时间后,对在互作网络中筛选到的8个关键节点蛋白质的基因表达情况进行分析.结果 蛹虫草活性成分虫草素和虫草酸的抗炎症作用相关靶点86个,构建虫草素和虫草酸的抗炎症作用靶点的蛋白质相互作用网络,虫草素/虫草酸抗炎症作用靶点通路主要富集在26条通路中,对互作网络中筛选到的8个关键节点蛋白质的基因表达情况分析发现,虫草素和虫草酸干预24 h后8个关键靶点蛋白均明显下调.结论 蛹虫草活性成分虫草素和虫草酸对抗炎症作用的多条信号通路有调控介导作用,通过调控多种关键靶蛋白表达情况参与抗炎症作用机制,为进一步揭示蛹虫草的抗炎症作用分子机制提供理论依据.

知识网络构建对于中医药院校医学生非常重要,可以帮助学生对分散的、碎片化式的知识点进行整合,从整体上把握知识的主线和主要内容以及知识点间的相互联系.而思维导图具有发散性思维的特征,能够将复杂的东西简单化,抽象的东西具体化,有助于教学.同时学会思维导图的方法可以培养学生良好的学习习惯,提高创新性思维和综合分析问题解决问题的能力.因此在生物化学理论教学和实验教学中尝试利用思维导图训练,促进师生间的沟通交流,提高学生的学习兴趣,加强学生对生物化学内容的把握、进行章节复习以及培养学生核心素养的提高等方面都有一定的作用.

目的:研究CpG ODN与小鼠肝癌细胞(H22)裂解物(TCL)联用对小鼠原位移植性肝癌的抑制作用及机制初探.方法:制备H22细胞裂解物和建立BALB/c小鼠原位移植性肝癌模型.将模型鼠随机分为四组PBS、TCL、CpG ODN和CpG ODN+TCL,观察各组小鼠肿瘤生长情况及小鼠的生存期.采用细胞毒性淋巴细胞(CTL)杀伤实验观察各组小鼠淋巴细胞对肝癌细胞的杀伤情况.用ELISA方法检测CpG ODN协助TCL诱导Th1型细胞因子分泌水平.结果:与其他组相比CpG ODN+TCL明显抑制了肿瘤生长、延长了荷瘤鼠的生存期(P<0.05).体内外实验显示,CpG ODN能协同TCL刺激小鼠产生Th1型细胞因子和诱导CTL反应.结论:以CpG ODN为佐剂的TCL具有抑制小鼠原位移植性肝癌的作用,且这种作用可能由于CpG ODN协同TCL诱导小鼠产生的Th1型细胞因子和CTL反应实现的.

目的:观察单宁酸对人肾癌细胞OS-RC-2增殖及凋亡的影响,并初步探讨其作用机制.方法:培养人肾癌细胞OS-RC-2,将其分为空白对照组、不同浓度TA组(20、40、80和100μmol/L),观察OS-RC-2的细胞形态,CCK8法检测细胞增殖,流式细胞术检测细胞凋亡,DCF法测定细胞内ROS,化学显示法检测抗氧化酶活性,ELISA法细胞上清中ICAM-1蛋白表达,Western blot法检测Bcl-2、Bax及Caspase-3蛋白的表达.结果:TA可有效抑制OS-RC-2肾癌细胞增殖,并促进其凋亡,呈显著剂量效应关系.TA可诱导OS-RC-2肾癌细胞内ROS生成增加,增加细胞中MDA含量,降低GSH-Px及SOD的活性,降低OS-RC-2细胞ICAM-1蛋白的分泌,下调细胞核中Bcl-2蛋白表达及上调Bax及Caspase-3凋亡蛋白的表达.结论:TA可能通过干预肾癌细胞内ROS及氧化酶的活性、抑制ICAM-1蛋白分泌及靶向线粒体凋亡等机制参与抑制肾癌细胞生长及促进其凋亡.

Objective To explore the protective mechanism of Danggui Sini Decoction on diabetic peripheral neuropathy (DPN) by observing the changes of NF-κb signaling pathway.Methods Forty-eight DPN rats were randomly divided into blank group, high-dose Danggui Sini Decoction group, low-dose Danggui Sini Decoction group and metformin group with 12 rats in each group after intraperitoneal injection of streptozotocin (STZ).To observe the effects of Danggui Sini Decoction on blood glucose, sciatic nerve conduction velocity, NF-κb protein and mRNA in diabetic rats.Results Compared with the model group, the Danggui Sini Decoction group could significantly reduce the blood glucose content, increase the conduction velocity of sciatic nerve, and inhibit the development of DPN by decreasing the expression levels of NF-κb protein and mRNA.Conclusion Danggui Sini Decoction has protective effect on sciatic nerve function in DPN rats.The mechanism may be related to the anti-inflammatory effect of Danggui Sini Decoction on decreasing NF-κb signal.

目的:观察ICAM-1和8-OHdG在肾癌组织中的表达情况,并分析两者的表达与肾癌患者临床病理参数之间的关系,探讨其在肾癌发生和进展中的作用.方法:采用免疫组化的方法检测46例肾癌组织和18例远离癌组织的正常肾组织中ICAM-1和8-OHdG的表达,通过阳性染色强度及阳性细胞百分比进行综合评分,观察不同组织者中两者的表达差异,同时分别统计分析ICAM-1和8-OHdG的表达与肾癌患者年龄、性别、肿瘤大小、肿瘤分期以及是否有淋巴结转移或远处转移等临床病理参数之间的关系.结果:ICAM-1在正常肾组织中的表达多为弱阳性或阴性表达,而在肾癌组织中ICAM-1阳性表达增加(占比82.6％),且与肿瘤的分化程度、淋巴结转移和远处转移具有显著相关性(P<0.05或P<0.01).绝大多数正常肾组织中8-OHdG呈阴性表达,而肾癌组织中8-OHdG阳性表达增加(占比63％),与肿瘤大小、淋巴结转移和远处转移以及无进展生存期(PFS)之间具有显著相关性(P<0.05或P<0.01).结论:ICAM-1和8-OHdG在肾癌组织中高表达,二者在肾癌的发生和进展中可能发挥着重要的作用.

目的 观察当归四逆汤对糖尿病大鼠坐骨神经传导速度的影响及糖基化终末产物(AGEs)和AGEs受体(RAGE)的调节作用.方法 采用链脲佐菌素(STZ)复制糖尿病大鼠模型,模型鼠随机分为中药组、西药组、模型组,另设健康鼠为正常组.分别干预8 w后,检测糖尿病大鼠坐骨神经传导速度、坐骨神经病理形态、AGEs含量、RAGE mRNA表达水平.结果 与模型组相比,中药能够保护坐骨神经结构,显著降低AGEs含量、下调RAGE mRNA水平(P<0.05).结论 当归四逆汤能够通过下调AGEs/RAGE含量提高坐骨神经传导速度、保护坐骨神经结构,进而抑制大鼠糖尿病周围神经病变(DPN)的发生发展.

目的 探讨寡居脱氧核苷酸(ODN)协同扶正清解方对肝癌细胞侵袭转移的抑制作用.方法 将对数生长期的肝癌细胞HEPG2分为3组培养,Medium组、ODN组及ODN+扶正清解方组,分别在细胞培养液、含ODN的细胞培养液、含ODN和扶正清解方的细胞培养液中培养12、24、36、48、60、72、84 h.然后通过MTT法测定各组肝癌细胞的增殖情况、划痕实验检测肝癌细胞迁移能力、Transwell小室检测肝癌细胞侵袭转移能力.结果 MTT实验表明ODN+扶正清解方组的HepG2细胞的生长在第24h开始受到抑制,其细胞增值能力明显低于Medium组及单独ODN组(P<0.05);迁移实验显示ODN+扶正清解方组的肝癌细胞与对照细胞相比,更少的细胞发生了迁移(P<0.05);Transwel实验显示,ODN+扶正清解方组HepG2细胞加入Transwel上室以后24 h,穿过Matrigel胶的细胞数目显著低于对照组细胞(P<0.05).结论 ODN协同扶正清解方通过抑制肝癌细胞的增殖、迁移,降低了肝癌细胞的侵袭转移能力.

目的:通过研究HMGB1/TLR4信号通路变化,探索通络消渴方改善2型糖尿病大鼠心脏损伤的可能机制.方法:采用链脲佐菌素(STZ)腹腔注射方法建立2型糖尿病大鼠模型,成模后将大鼠随机分为模型组、二甲双胍组及通络消渴方高、低剂量组,并设正常对照组.造模成功后即开始灌胃给药,中药高剂量组(7.36 g·kg-1·d-1)、中药低剂量组(1.84 g·kg-1·d-1)和二甲双胍组(140 mg·kg-1·d-1),空白组和模型组灌胃同体积0.9％ NaCl溶液,每日1次.8周后处死大鼠,取样检测各组大鼠空腹血糖、心肌病理及HMGB1和TLR4蛋白表达.结果:与正常对照组比较,模型组大鼠血糖显著升高、体质量明显下降;与模型组比较,通络消渴方高、低剂量组均可抑制大鼠血糖升高,且中药高剂量组可明显抑制体质量下降,差异具有显著性(P＜0.05).病理结果显示,通络消渴方高、低剂量组和二甲双胍组大鼠心肌病理改变较糖尿病模型组明显减轻;免疫组化结果显示,通络消渴方高剂量组、低剂量组和二甲双胍组均可降低大鼠HMGB1和TLR4蛋白表达,与模型组比较差异有统计学意义(P＜0.05).结论:通络消渴方可改善糖尿病大鼠心脏功能,其机制可能与降低HMGB1/TLR4信号蛋白表达,从而发挥抗炎作用有关.

目的 观察当归四逆汤对糖尿病周围神经病变(DPN)大鼠的防治作用及对乙二醛酶Ⅰ(GLO1)的影响.方法 采用链脲佐菌素(STZ)复制糖尿病大鼠模型,模型鼠随机分为模型组、西药组、中药组,另设健康鼠为正常组.观察当归四逆汤对大鼠DPN的防治作用,干预8 w后检测糖尿病大鼠坐骨神经传导速度;GLO1活性、mRNA及蛋白表达水平.结果 与模型组相比,中药组大鼠坐骨神经传导速度、结构明显有改善,GLO1的活性增强、蛋白及mRNA表达均上调(P<00.5).结论 当归四逆汤能够提高坐骨神经GLO1活性、上调蛋白及mRNA表达,对大鼠DPN具有防治作用,其机制与对GLO1的调节有关.

Objective To investigate the expression of AF-6mRNA protein and its effect on the invasion of hepatocellular carcinoma.Methods Using real-time quantitative PCR detection method, the expression levels of AF-6mRNA in 28 pairs of cancer cells, ANLT and cell lines were detected, and the inlfuence of AF-6mRNA on the invasion was analyzed.Results 26 cases of AF-6mRNA showed low expression in hepatocellular carcinoma, the incidence rate was 92.9%, the rod was signiifcantly lower than the normal cell line AF-6mRNA expression in hepatocellular carcinoma cell line (P<0.05), was obviously higher than that of invasion and metastasis of AF-6mRNA cells expressing ability in cell lines with low metastasis (P<0.05). ConclusionLow expression of AF-6mRNA in hepatocellular carcinoma may be related to high invasive ability.

ObjectiveTo explore the application value of miR-30c in reducing the invasion and metastasis of hepatocellular carcinoma.Methods To detect the expression of miR-30c in hepatocellular carcinoma and hepatocellular carcinoma cell lines,and to analyze the role of the invasion of hepatocellular carcinoma cells.Results The expression of miR-30c was significantly lower than that of no vascular metastasis group,and the Snail expression was controlled by transfection of miR-30c.Conclusion miR-30c expression up regulation,can inhibit the expression of snail,reduce the invasion.

Objective To study the Effect of inhibitory ODN on mice with ulcerative colitis.Methods Ulcerative colitis(UC)was induced by DSS in mice.The mice with UC were randomly divided into three groups.Mice with UC were injected with follows by i.p:PBS,A151,A151control respectively.Disease active index(DAI)was observed every day.At 7days after last injection the mice were sacrificed and their colons were taken out to undergo histological examination.Results The weight gain of mice with UC was induced by A151.Moreover,the DAI score and histological score in the group of A151+UC were significantly lower than those in other UC groups(P0.05).Conclusion Inhibitory ODN might have the potency to therapy UC.

生物化学是医学教学的基础课程,是生命科学领域中的重要学科,对于后续医学课程的学习和工作都是非常必要,同时对学生来说又是一门非常抽象难于理解的学科。本文就如何提高医学院校学生学习生物化学的兴趣做了详细阐述,以期对教师在讲授医学课程和学生学习过程中能有所帮助。

教学实验室的管理体制改革是提高实验教学水平,保证人才培养质量的重要手段。本文阐述了实验室工作体制、实验教学创新、健全实验室管理制度、建立人才激励机制等方面的实验改革内容。

Objective To study the effect of activated TLR9 in intestinal epithelium on ulcerative colitis induced by DSS in mice.Method Mice were randomly divided into six groups:PBS group,CpG ODN group,control ODN group,DSS+PBS group,DSS+CpG ODN group and DSS+control ODN group.The mice in PBS group,CpG ODN group and control ODN group were fed normally and were injected intraperitoneally with PBS,CpG ODN,and control ODN,respectively.The mice in DSS+PBS group,DSS+CpG ODN group and DSS+control ODN group were fed with DSS in water for drinking and were injected intraperitoneally with PBS,CpG ODN,and control ODN,respectively.Disease active index(DAI)was recorded every day.After ten days the mice were killed and their colons were taken out to undergo histological examination.Results The loss of weight in DSS+CpG ODN group was more than those in other groups(P0.05).Moreover,the DAI score and histological score in the group of DSS+CpG ODN were significantly higher than those in other groups(P0.05).Conclusion The CpG ODN could aggravate the ulcerative colitis induced by DSS in mice.

Objective To study if inhibitory ODN could decrease the levels of IL-8 and TNF-αsecreted by lamina propria mononuclear cells(LPMCs) from the patients with ulcerative colitis(UC).Methods LPMC were isolated from intestinal mucosal biopsy specimens from 10 patients with UC,and cultured with or without CpG ODN,inhibitory ODN(A151) and dexamethasone(DEX).The levels of IL-8 and TNF-α were tested by enzyme linked immunosorbent assay(ELISA) and the expression of IL-8 and TNF-αmRNA was measured by reversal transcription-polymerase chain reaction(RT-PCR).Results A151 resulted in down-regulation of the expression of IL-8 and TNF-α mRNA,and strikingly decreased the levels of IL-8and TNF-α,and the inhibitory effects were greater than those induced by DEX(P0.05).Conclusion The application of inhibitory ODN may serve as a novel molecular approach for the treatment of patients with UC.