免疫学是中医药院校研究生必修的一门西医基础课程,为培养学生综合学习的能力,学校对2019级中医、中药和中西医结合等专业研究生的免疫学课程教学模式进行改革,充分利用网络背景下多种多样新的教学方法,如慕课、PBL等,结合传统的授课方式重新组织教学资源、实施混合式学习,以达到提高教学效率的目的 .结果 显示,混合式教学可实现"以学生为中心"的教育理念,激发学生的学习兴趣,提升学生自主学习的能力.

微生物与免疫学是药学专业的一门专业基础课,在培养现代化药学人才方面有着重要意义,因此推动微生物学与免疫学教学是培养专业的药学人才的重要任务.文章围绕微生物学与免疫学的学科特点和学生专业特点从教学计划设置、教学方法和手段、考评方式等几个方面探讨如何提高课程教学质量,调动学生学习的积极性和主动性,达到教学目的.我们认为,在制定合理的教学计划,围绕专业特色开展教学的前提下,结合教学内容灵活运用多种教学方法和手段,加强随堂测试,多种考评方式相结合,可能是保证教学效果,达到教学目标的关键和保障.

病原生物学是一门重要的基础课程,病毒学是该课程中要求重点掌握的章节.相比其他章节,病毒学部分的知识点琐碎而抽象,对学生的学习造成很大的困难.文章从病毒学课程的特点出发,突出以学生为主体,充分利用网络资源与网络工具,使用微课、慕课、翻转课堂、PBL等教学方法对病毒学的教学模式进行改革.通过这种网络背景下病毒学教学模式改革的尝试,改善病毒学的教学质量与教学效率,拉近教学与科研的距离.

中医药事业的发展急需应用型创新型人才的培养.为提高中医药学生的科研创新能力,开展教学科研相结合的中医药抗病毒技术科研培训研究,同时设立未参加培训的对照纽.研究结果显示:试验组学生与对照组相比,科研创新思维、技能、知识、能力都得到了显著地提高.本研究为创新型人才培养模式的探索提供了新的思路.

目的 探讨缺氧对舌苔形成相关细胞凋亡的影响.方法 将CoCl2作用于撤血清舌鳞癌Tca-8113细胞,建立化学性缺氧模型.MTT法检测细胞增殖活性,荧光染色检测细胞凋亡,琼脂糖电泳观察DNA Ladder,流式细胞术检测细胞周期分布和凋亡,Western blotting检测Akt、NF-κB p50、NF-κB p65、Bax、Bcl-2、HSP 70的表达水平.结果 50～400μmol/L CoCl2作用细胞8～24h能明显抑制撤血清舌鳞癌细胞增殖(P＜0.05),并具有量效和时效关系.Hoechst荧光染色显示CoCL2可引起撤血清舌鳞癌细胞凋亡.流式细胞术显示100,200,400μmol/L CoC12 12h缺氧-12h复氧可促进撤血清舌鳞癌细胞凋亡(P＜0.05),且凋亡率随CoCl2的浓度增高.Western blotting表明12h缺氧-12h复氧可上调Bax、HSP 70、NF-κB p50,下调NF-κB P65表达(100μ,mol/L CoCl2),促进舌鳞癌细胞凋亡.结论 缺氧是促进舌苔形成相关细胞凋亡的重要因素,其发生机制可能受NF-κB、Bax、HSP 70蛋白的调控.

对于大一、二年级学生而言，因为没有相关基础知识储备，在学习免疫学这门课时往往感觉比较抽象。这要求在教学过程中尽量把抽象的知识具体化。文章讨论了在免疫学教学过程中的实践，在形式上，运用动画教学模式把抽象理论变成具体的过程；在内容上建立学生理解免疫知识的分子平台：蛋白质结构变化而改变其功能从而通过信号传导调节细胞和组织器官功能，使学生掌握理解免疫学知识的内涵与外延。结果显示上述方法有效提高了学生对难点问题求索的兴趣与课堂理解能力。

OBJECTIVE:To study the synergistic effect on hepatoma cell(SMMC-7721) and the reduction killing effect on normal liver cells(LO-2) treated with sodium cantharidinate (SCA) in combination with fluorouracil(5-FU) or cisplatin(DDP) as well as the related mechanism.METHODS:MTT assay was used to select the best ratio of SCA with 5-FU or SCA with DDP which had less toxicity on LO-2 cell line and had synergistic effect on SMMC-7721 cell line; Flow cytometry assay was used to analyze the apoptosis-induction of the different ratio of drugs on both cell lines; Hoechst-33258 fluorescent staining assay was used to observe the nuclear morphological changes of cells; Immunoblotting assay was used to analyze the Ras/Raf/ERK1/2 signaling pathway and the apoptosis related signaling pathway in both cell lines.RESULTS:MTI assay indicated that the proliferation inhibition of SCA,5-FU and DDP on SMMC-7721 cell line was in a time-and dose-dependent manner respectively. Among them, SCA had a more significant inhibition on SMMC-7721 cell line than on LO-2 after 12 h or 24 h treatment (P <0. 01). Moreover, after a treatment of 48 h,the ratio of 2. 5 µg/mL SCA and 2 µg/mL DDP showed a more significant inhibition on SMMC-7721 cell line than on LO-2 cell line,which was then be considered as the optimal concentration ratio for the following experiment. Co-treatment of SCA (2. 5 µg/mL) with DDP (2 µg/mL) induced a more significant apoptosis on SMMC-7721 cell line compared with single treatment with SCA (2. 5 µg/mL) or DDP (2 µg/mL) respectively (P < 0. 01). After a 48 h treatment of the optimal ratio of drugs, the significant morphological apoptotic characteristics were observed both under inverted microscope and by Hoechst-33258 fluorescent staining assay in both cell lines. The results of Western blot assay showed that this ratio of drugs could significantly increase the protein expression of Bax,P53 and P21 and decreased the expression of BCL-2, Casepase-3, p-Erk, p-Ras and p-c-Raf in SMMC-7721 cells. Meanwhile,the effect on the proteins mentioned above was lesser in LO-2 cells.CONCLUSION:These results indicates that 2. 5 µg/mL SCA + 2 µg/mL DDP showed a higher inhibition on the hepatic carcinoma cells and a relatively lower cytotoxicity on normal liver cells. The major anti-cancer mechanism is related with the inhibition on Erk signaling pathway and the induction of apoptosis through the mitochondrial pathway.

Morroniside is a water-soluble compound extracted from the fruit of Cornus officinalis and is used to protect lung activity against aging. In the present study, the manner in which morroniside regulates normal lung and cancer cells was examined. The human embryonic lung fibroblast (HELF) cell line and lung cancer A549 cell line, and their responses to morroniside treatment, were examined. Results showed that morroniside reverses the apoptotic effect of H2O2 on HELF cell growth, protecting cell proliferation and normal cell morphology and inhibiting apoptosis. However, these effects were not present in A549 cells. Western blotting showed that morroniside also markedly downregulated retinoblastoma protein in HELF cells. These results suggest that morroniside treatment exhibits different effects on apoptosis in HELF and A549 cells, making it a viable compound for decreasing the side effects of anticancer medicines in normal cells.

探讨加味苇茎汤含药血清及成分芹菜素对肺腺癌A549细胞增殖、细胞周期及凋亡的影响,选用肺癌A549细胞体外培养,在不同浓度含药血清及芹菜素作用下,用MTT法检测细胞生长抑制率、流式细胞术检测细胞周期及凋亡率、hoechst33342染色检测细胞凋亡形态.结果显示,加味苇茎汤含药血清及芹菜素对A549的生长抑制率:24 h分别为31％、34％,48 h分别为45％、53％;流式细胞术检测细胞周期发现A549细胞经加味苇茎汤含药血清处理后主要阻滞在G2期,而芹菜素能明显引起细胞凋亡.荧光显微镜下观察到较典型的细胞凋亡形态甚至细胞脱落,视野内细胞核数明显减少.该研究结果显示,加味苇茎汤含药血清及芹菜素能明显抑制A549细胞的增殖,引起细胞周期阻滞、诱导其凋亡.

ObjectiveTo investigate the mechanism of Zhou's prescription on inducing apoptosis related to the formation of tongue coating. MethodsThe two kinds of human tongue squamous cell carcinoma were cultured in RPMI-1640 with serum to simulate the damnification and rehabilitate of the lingual dorsum epidermis in vitro.The effective part which was extracted from Zhou's prescription by chemical methods,role in the cells.After the two kinds of human squamous cell carcinoma were incubated in RPMI-1640 with Zhou' s prescription,MTT assay evaluated the activities of cell proliferation,and flow cytometric analysis was used to analyze the cell cycle distribution and apoptosis.Detected the level of the PGE2 in the cell culture supernatant with enzyme-linked immunosorbent assay(ELISA). ResultsIn comparison with the control,MTT assay,of nhich the result showed the Zhou's prescription could induce inhibitory effects on the growth in SAS and TCA-8113 cells.The flow cytometric analysis showed that the Zhou's prescriptionc could induce the apoptosis of human squamous cell carcinoma.Besides the result of The ELISA assay result showed that with the increasing of the concentration of the component,the Zhou's prescription could decrease the PGE2 level in the cell culture supernatant. ConclusionThe results suggested that the inhibitory effects of Zhou's prescription on apoptosis which related to the formation of tongue coating resulted from the inhibition of PGE2 releasing.

Objective: To investigate the mechanism of Qianjin Weijing prescription(QJWJ) on promoting apoptosis of the human tongue squamous cell carcinoma SAS and TCA-8113 with serum withdrawal.Method: The SAS and TCA-8113 were treated with serum withdrawal to imitate the condition caused by the chemical therapy of anti-angiogenesis.The assistant effects on anti-tumor of three parts extracted from QJWJ were investigated by SAS and TCA-8113 with serum withdrawal in vitro.MTT assay was used to evaluate the activities of cell proliferation,and flow cytometric analysis was conducted to analyze the cell cycle distribution and apoptosis.The level of the prostaglandin E 2(PGE 2) in the cell culture supernatant was detected by enzyme-linked immunosorbent assay(ELISA).Result: In comparison with the control,the results showed the 07 part of QJWJ could distinctly promote the inhibitory effects on the activities of cell proliferation in SAS and TCA-8113 cells.The maximum growth inhibition rate 34.36%,and the apoptosis of human squamous cell carcinoma(P < 0.05) by flow cytometric analysis.The level of the PGE 2 in the cell culture supernatant was inhibited by 07 part of the QJWJ by ELISA assay(P < 0.05).Conclusion: The results suggested that the promoted effects of QJWJ on apoptosis of the hunman tongue squamous cell carcinoma with serum withdrawal was mediated by the inhibition of PGE 2 releasing.The research provided a new way to explore the scientific interests of QJWJ in adjuvant treatment of malignant cancers.

中医学与免疫学具有类似的思想内涵,免疫学联系中医药学经典理论与现代生命科学成果的重要桥梁.本文论述了中医院校研究生《免疫学》课程教学中开展启发式教学的必要性、可行性和实施体会,对培养高素质、复合型中医药人才具有借鉴意义.

Objective To study the effects and mechanisms of Cornus water extraction in human embryonic lung fibroblast cells(MRC-5) induced aging by D-galactose.Methods To observe growth conditions by MTT method about the water extraction of Cornus on human embryonic lung fibroblast cells(MRC-5) induced aging by D-galactose;To detect anti-aging pathway of P16,P21 in Western blot about Cornus water extraction on MRC-5 cells induced aging by D-galactose.In experiment of Telomere PNA Kit / FITC for Flow Cytometry,study telomere length influences of Cornus water extraction on MRC-5 cell.Results The results of MTT method showed the water extraction of Cornus can promote reproduce of human embryonic lung fibroblast cells(MRC-5) induced aging by D-galactose;Western blot method and Telomere PNA Kit/FITC for Flow Cytometry results reveal,compared with the control group,Cornus water extraction could make P16 and P21 expression reduced of MRC-5cell induced aging by D-galactose;and delay the telomere shortening of MRC-5 cell.Conclusion Cornus water extraction have anti-aging function.

Selenium (Se) is an essential trance element in testis. However, the potential protective effects of Se against cadmium (Cd)-induced reproductive toxicity remained to be elucidated. Male ICR mice were orally administered by gavage with Na2SeO3 (0.1, 0.2, 0.4 mg/kg BW) for 1 h prior to CdCl2 (5 mg/kg BW) alone or in combination for 15, 25 or 35 days. Cd exposure caused a significant decrease in body weight, sperm concentration and motility as well as plasma testosterone level which was accompanied by decreased antioxidant enzymatic activity of SOD and GSH-Px and by increased lipid peroxidation (as malondialdehyde, MDA). Se pretreatment compensated deficits in the sperm parameters (concentration, motility and morphology) induced by Cd. Se (0.4 mg/kg BW) treatment significantly increased serum testosterone level that was reduced by Cd (on 15th, 25th and 35th day) (P < 0.01). Se treatment ameliorated Cd-induced reduction in testicular steroidogenic acute regulatory (StAR) and 17β-hydroxysteroid dehydrogenase (17β-HSD) activities. The present study suggest that the protective potential of Se against Cd-induced reprotoxicity might be due to up-regulation StAR and testosterone synthetic enzyme activity, which could be useful for increasing testosterone synthesis for achieving optimum protection in sperm quality and spermatogenesis.

目的 研究山茱萸水煎液对老年小鼠免疫器官的保护作用.方法 运用流式细胞术测定小鼠骨髓、胸腺、脾脏细胞的细胞周期、凋亡率等.结果 衰老组小鼠与青年组小鼠相比,骨髓、胸腺、脾脏细胞Go-G1期明显增多,S期明显减少,细胞增殖明显减缓；胸腺细胞、脾细胞凋亡率明显增多.山茱萸水煎液能明显抑制老年小鼠胸腺细胞的凋亡(P＜0.05),并且明显改善老年小鼠骨髓细胞、脾细胞的增殖抑制,表现为脾细胞G0-G1期明显减少(P＜0.05),S期明显增高(P＜0.05),骨髓细胞G0-G1期明显减少(P＜0.05).结论 山茱萸水煎液具有一定的抑制老年小鼠免疫衰退的作用.

Objective To discuss the effect of active sites of Radix Isatidis on apoptosis of Hep-2,Hela,U937,Jukart and on expression of IL-10 and TNF-α in RAW264.7 cells.Methods Hep-2,Hela,U937 and Jukart were pretreated by 0.5 g/L active sites in Radix Isatidis for 24 hours,and the apoptosis were assayed by flow cytometry.U937,infected by HSV-1,was incubated with 0.5 g/L medicine for 24 hours,and the apoptosis were assayed by flow cytometry.The expression of IL-10 and TNF-α in RAW264.7 induced by LPS were detected by ELISA.Results ①The result showed that 0.5 g/L active sites of Radix Isatidis did not affect the apoptosis of Hep-2,Hela,U937 and Jukart,but the apoptosis rate of U937 infected by HSV-1 was reduced from 9.72% to 6.71%,with significant difference between anti-viral group and viral group(P0.01);the G1 phase arrest caused by medicating the Hela cell,and the S phase was shortened,with significant difference between the normal Hela cell and infected Hela cell(P0.05).②In medication group,the apoptosis rates were 19.35%,28.14% and 7.52% after LPS-induced apoptosis in RAW264.7 cells by 1 g/L,0.5 g/L and 0.25 g/L medicine.The apoptosis rates in medication group were lower than that in LPS model group significantly(P0.01).Compared with cell control group,the apoptosis rates in 1 g/L medication group was increased significantly(P0.01),and there was no significant apoptosis promoted effect in 0.5 g/L and 0.25 g/L medication groups.The apoptosis rates in 0.5 g/L and 0.25 g/L medication groups were decreased significantly(P0.01);compared with cell control group,the G1 phase and S phase were shortened,while the G2 phase was extended in 1 g/L medication group,with significant difference between the two groups(P0.01).The G1 phase was extended,but the S phase was shortened in 0.5 g/L medication group significantly(P0.01).③After stimulated RAW264.7 cells by LPS,compared with cell control group,the IL-10 and TNF-α in LPS group were increased significantly(P0.05);the IL-10 in 0.5 g/L and 0.25 g/L medication groups were decreased significantly,and the TNF-α in 1 g/L medication group was increased significantly(P0.05).Compared with LPS model group,the IL-10 in 1 g/L,0.5 g/L and 0.25 g/L medication groups were decreased significantly(P0.05).Conclusion Active sites of Radix Isatidis can inhibited apoptosis in U937 and RAW264.7 cells infected by HSV-1 and LPS.It can decrease the expression of IL-10 in RAW264.7 cells after intervened by LPS for 24 hours.

OBJECTIVE To explore in-vitro cell toxicity of anti-tumor drug disodium cantharidinate on normal human hepatocyte(LO2).METHODS MTT experiment,Hoechst-33258 fluorescent staining and electron microscope detection were applied.RESULTS Medication groups with dose of 0.625,1.25 and 2.5μg/mL respectively presented different degrees of proliferation inhibition after disodium cantharidinate acted on 24h,and inhibitory rates were 9.1%,17.1%and 23.9%respectively;Fluorescent staining and electron microscope detection indicated that cells manifested morphological changes,injured chromatin and apoptosis in 2.5μg/mL group.CONCLUSION Anti-tumor drug disodium cantharidinate has the inhibitory effect on the proliferation of cell LO2 and can evoke LO2 apoptosis under certain doses.

目的：观察槲皮素（quercetin）对人肝癌细胞SMMC-7721增殖与细胞凋亡的影响,探讨其对SMMC-7721细胞PI3K/AKT信号通路的影响。方法：采用MTT法检测槲皮素对SMMC-7721细胞生长的抑制,流式细胞术检测细胞周期变化,Western blot检测槲皮素对SMMC-7721细胞PI3K/AKT信号通路凋亡相关蛋白表达的影响。结果：槲皮素抑制SMMC-7721肝癌细胞增殖作用明显,且呈浓度和时间依赖性。顺铂和槲皮素40,80,160,320μmol.L-148 h抑制率分别为62.19%,25.47%,27.18%,36.96%,51.28%。流式细胞术结果提示,槲皮素80,160,320μmol.L-1可使SMMC-7721肝癌细胞周期阻滞于G0/G1期。Western blot凋亡相关蛋白表达检测表明,药物组AKT的表达受抑制,PTEN,Caspase-9蛋白的表达率随着药物浓度的增加而增加。结论：槲皮素能诱导SMMC-7721肝癌细胞凋亡,其机制可能是使肝癌细胞SMMC-7721周期阻滞于G0/G1期,PTEN的过表达抑制AKT活化,激活Caspase-9从而促进细胞凋亡。

Objective:To explore in-vitro cell toxicity of anti-tumor drug gallic acid(GA)on normal human hepatocyte(LO2).Methods:MTT experiment,flow cytometry,Hoechst-33258 fluorescent staining and electron microscope detection were applied.Results:0,6.25,12.5,25,50,100 and 200μmol/L medication groups presented different degrees of proliferation inhibition after GA acted on 24h,and inhibitory rates were 0,18.2%,25.5%,42.0%,42.5%,51.8% and 60.6% respectively;0,25,50 and 100μmol/L medication groups were detected by flow cytometer,whose apoptosis rates were 1.37%,4.63%,4.64% and 12.53% respectively;fluorescent staining and electron microscope detection indicated that cells manifested morphological changes,nuclear pycnosis and fragmentation in 100μmol/L group.Conclusion:Anti-tumor monomer GA also has the inhibitory effect on the proliferation of cell LO2 and can evoke LO2 apoptosis.