以宁夏医科大学2020 级临床医学专业2 个班的本科生为研究对象,分为对照组和实验组.对照组采用传统教学模式,实验组采用思维导图教学模式(课前学生利用思维导图预习,课堂上教师利用思维导图讲解,课后学生用思维导图复习总结),教学后对两组学生的主观评价(调查问卷)及客观评价(期末考试成绩)进行统计学分析.实践表明,思维导图教学模式能收到良好的教学效果,值得在生物化学教学中推广和实施.

"生物化学"是医学各专业的一门重要基础课程,具有涵盖内容庞杂、概念复杂抽象的特点,因此,一些学生对知识点的理解和记忆难度较大,容易产生畏难情绪.思维导图能够梳理知识体系,方便学生对知识点进行归纳,但是由于性质所限,思维导图并不能详细阐述知识点.Rubric作为形成性成绩评价工具,可以指导学生以思维导图为课堂笔记,对知识点进行自主整理,并进行互相评阅,应用客观数据及时反馈思维导图的应用效果.Rubric在学生思维导图笔记中的应用增加了学生之间的相互交流,促进了学生自主学习的能力,提高了教学效果.

目的 观察枸杞多糖(LBP)对高糖诱导的人视网膜色素上皮细胞(ARPE-19)中NOD样受体热蛋白结构域相关蛋白 3(NLRP3)炎症小体/细胞焦亡信号通路的影响.方法 将体外培养的ARPE-19 细胞,随机分为 5 组,即正常组(5.5 mmol·L-1 葡萄糖)、高糖组(55.5 mmol·L-1 葡萄糖)、LBP低浓度组(55.5 mmol·L-1 葡萄糖+100 μg·mL-1 LBP)、LBP中浓度组(55.5 mmol·L-1 葡萄糖+200 μg·mL-1 LBP)和LBP高浓度组(55.5 mmol·L-1葡萄糖+400 μg·mL-1 LBP).采用免疫荧光观察各组细胞中NLRP3 蛋白的表达,Western blot检测炎症小体NLRP3、半胱氨酸蛋白酶-1(Caspase-1)、凋亡相关斑点样蛋白(ASC)和其下游细胞焦亡蛋白消皮素D(GSD-MD)的相对表达量,ELISA检测细胞上清液中白细胞介素(interleukm,IL)-1β和IL-18 的表达水平.结果 免疫荧光结果显示,NLRP3 蛋白主要在视网膜色素上皮细胞胞质中表达,且与正常组相比,高糖组NLRP3 蛋白的表达水平升高(P＜0.05);而与高糖组相比,LBP中、高浓度组NLRP3 蛋白的表达水平降低(P＜0.01).West-ern blot结果显示,与正常组相比,高糖组细胞中NLRP3、Caspase-1、ASC、GSDMD蛋白的相对表达量升高(P均＜0.05);与高糖组相比,LBP中、高浓度组中NLRP3、Caspase-1、ASC、GSDMD蛋白的相对表达量降低(P均＜0.05).ELISA结果显示,与正常组相比,高糖组细胞中IL-1β和IL-18 表达水平均升高(P均＜0.05);与高糖组相比,LBP中、高浓度组中IL-1β、IL-18 表达水平均降低(P均＜0.05).结论 LBP可通过抑制NLRP3 炎症小体/细胞焦亡信号通路的激活,从而对高糖环境下的人视网膜色素上皮细胞产生保护作用.

目的 研究枸杞多糖(LBP)对高糖干预的人晶状体上皮(HLEB3)细胞中沉默信息调节因子 1(SIRT1)及沉默信息调节因子 6(SIRT6)表达水平的影响.方法 将对数生长期的HLEB3 细胞随机分为对照组(CON组,5.5 mmol·L-1)、高糖组(HG组,55.5 mmol·L-1)、HG+12.5 mg·L-1 LBP组、HG+25 mg·L-1 LBP组、HG+50 mg·L-1 LBP组,培养 24h后,分别用相应培养基干预 48 h,用显微镜观察各组细胞的形态及密度变化;采用CCK-8 试剂盒检测各组细胞的存活率;采用免疫荧光法对各组细胞中SIRT1 和SIRT6 蛋白进行定位检测;采用Western blot法测定各组细胞中SIRT1与SIRT6蛋白的表达水平.结果 光镜下观察发现,CON组HLEB3 细胞主要呈梭形贴壁生长,HG组细胞形态呈不规则状,细胞密度降低;与HG组相比,HG+12.5 mg·L-1 LBP组、HG+25 mg·L-1 LBP组、HG+50 mg·L-1 LBP组细胞密度依次增加.CCK-8 检测结果显示,与CON组相比,HG组HLEB3 细胞活性降低(P＜0.05);与HG组相比,HG+50 mg·L-1 LBP组HLEB3 细胞活性增加(P＜0.05).免疫荧光结果显示,HLEB3 细胞中SIRT1 蛋白主要在细胞核中表达,细胞质中有少量表达;SIRT6 蛋白表达定位在细胞核.Western blot结果显示,与CON组相比,HG组HLEB3 细胞中SIRT1 和SIRT6 蛋白表达均下调(P均＜0.05);与HG组相比,HG+50 mg·L-1 LBP组HLEB3 细胞中SIRT1 和SIRT6 蛋白表达均上调(P均＜0.01).结论 LBP可以增加高糖干预的HLEB3 细胞的增殖活性,并上调SIRT1 和SIRT6 蛋白的表达.

目的 探讨紫外线照射对人视网膜色素上皮细胞(ARPE-19细胞)活性和沉默信息调节因子6(SIRT6)表达的影响.方法 将对数生长期的ARPE-19细胞按照照射剂量随机分为对照组(0 mJ·cm-2)及7.5 mJ·cm-2、15.0 mJ·cm-2、30.0 mJ·cm-2、60.0 mJ·cm-2、120.0 mJ·cm-2、240.0 mJ·cm-2照射剂量组进行干预培养.照射后继续常规培养24 h,采用倒置荧光显微镜观察细胞形态并拍照,CCK-8法检测细胞存活率,Western blot检测各组细胞中SIRT6的表达情况,采用免疫荧光染色对对照组、30.0 mJ·cm-2照射剂量组和240.0 mJ·cm-2照射剂量组ARPE-19细胞中SIRT6进行定位检测.结果 对照组ARPE-19细胞呈梭形贴壁生长,边界清楚,连接紧密,细胞状态良好;30.0 mJ·cm-2照射剂量组ARPE-19细胞形态不规整,细胞数量略有减少,细胞间连接疏松.CCK-8法检测结果显示:与对照组相比,ARPE-19细胞在不同剂量紫外线照射后,细胞存活率显著降低且呈剂量依赖性(均为P<0.05).与对照组相比,除较低照射剂量组(7.5 mJ·cm-2、15.0 mJ·cm-2)外,其余各组SIRT6蛋白相对表达量均降低,且随着照射剂量的增加呈剂量依赖性逐渐降低(均为P<0.05).经过30.0 mJ·cm-2紫外线照射后,ARPE-19细胞随着再培养时间的延长,细胞中SIRT6蛋白的相对表达量逐渐下降(均为P<0.05).对照组、30.0 mJ·cm-2照射剂量组和240.0 mJ·cm-2照射剂量组SIRT6主要表达于ARPE-19细胞核内;与对照组相比,30.0 mJ·cm-2照射剂量组和240.0 mJ·cm-2照射剂量组ARPE-19细胞中SIRT6的荧光强度显著降低,且240.0 mJ·cm-2照射剂量组降低更明显.结论 紫外线照射能够降低ARPE-19细胞的增殖活性,同时也能下调细胞中SIRT6的表达.

随着"互联网+"教育的推进,医学生物化学与分子生物学实验课改革势在必行,在线学习平台是基于此背景开发的混合式数字化教学平台.本研究将在线学习平台应用于生物化学与分子生物学实验教学中,构建实验课翻转课堂,通过签到、资料推送、预习作业及实验报告等4个课堂活动研究了辅助实验教学模式,得到了较好的教学效果.

注重学生自主学习能力的培养是当前教育教学改革的一个重要方向.在临床医学专业试点班进行平时成绩考核方式的革新,以总结作业、随堂测试和课堂讨论等形式的多元组合作为学生平时成绩的综合测评,有别于传统的考勤、课后总结作业或思考题为主的评定方式,也的确能够调动学生的学习兴趣、提升学习效果、培养学生自主学习的能力.

ETHNOPHARMACOLOGICAL RELEVANCE:Lycium barbarum polysaccharide (LBP) extracted from the Lycium barbarum L. has been widely used to improve diabetes and its relative complications. However, the mechanisms have not fully understood. A recent study has demonstrated that LBP upregulates suituin 1 (SIRT1). OBJECTIVE:This study was to define the role of Sirt1 and its downstream signaling pathways in diabetic cataract using in vitro and in vivo models. MATERIALS AND METHODS:Human lens epithelial cell line SRA01/04 cells were cultured under high glucose (HG) medium with treatment of LBP or vehicle. Cell viability, apoptosis, protein and/or mRNA levels of Sirt1, BAX, Bcl-2, active-caspase-3, FOXO1, p27 and acetylated p53 were measured. SIRT1 upregulated- and knocked-down cells were generated and tested in high glucose culture. Diabetes mellitus was induced in rats by streptozotocin injection. Body weight, blood glucose levels, lens transparency and retinal function were assessed and SIRT1, as well as the aforementioned biomarkers were measured using Western blotting and qPCR in the animal lens samples. RESULTS:The results showed that HG decreased cell viability and LBP prevented the decrease. The reduced viability in HG cultured SRA01/04 cells was associated with increased levels of BAX, active caspase 3, FOXO1, p27, and p53 and decreased levels of SIRT1 and Bcl-2. Further experiments using sirt1 gene modulated cells showed that upregulation of Sirt1 improved viability, increase cell division as reflected by an increased proportion of S phase in the cell cycle, reduced the number of apoptotic cell death and suppressed p53 acetylation and caspase 3 activation. Opposite results were observed in SIRT1 knock-down cells. Treating diabetic animals with LBP reduced body weight loss and blood glucose content in diabetic animals. Similarly, LBP hindered the development of cataract in lenses and improved retinal function. The beneficial effect of LBP on diabetic cataract was associated with the supression of p53, caspase 3, FOXO1, BAX, p27 and elevation of SIRT1 and Bcl-2, which were consistent with the in vitro findings. CONCLUSION:Our findings showed that diabetes caused cataract is associated with suppression of SIRT1 and Bcl-2 and activation of other cell death related genes. LBP prevented diabetic cataract in animals by upregulating Sirt1 and Bcl-2 and suppressing cell death related genes.

2020春节后的疫情特殊时期,很多高校的本科生并未返校,而是通过网络完成学业.钉钉平台具有强大的后台服务系统成为这段时期的主力军并被广大中小学生及大学生所认可,但钉钉直播缺乏课后及学习自主性活动,因此我们利用"钉钉直播+在线学习平台"的双重模式完成了分子生物学教学任务,包括课前预习、上课讲授、课后作业、期中考试、实验课程和期末考试等所有内容,大大提高了学生在家上课的学习效率.文章基于"钉钉直播+在线学习平台"实施的一学年度分子生物学的在线教学过程实践,为其他高校的分子生物学教学及同类型课程的在线教学提供参考.

生物化学是医学生必修的基础课程.随着现代医学的飞速发展,急需医学生具备全面的自主学习能力,而数字化教学资源库就是为拓展学生的自主学习提供的有效的教育平台.在生物化学教学中,将教师课堂授课和学生自主学习与数字化教学资源进行有效结合,对于拓宽教学渠道、丰富教学手段、提高教学质量具有重要作用.

目的 观察高浓度葡萄糖对人晶状体上皮细胞(SRA01/04)活性及沉默信号调节蛋白6(SIRT6)表达的影响.方法 将SRA01/04细胞在不同葡萄糖浓度(5.5 mmol·L-1、15.5 mmol·L-1、25.5 mmol·L-1、35.5 mmol·L-1、45.5 mmol·L-1、55.5 mmol·L-1、75.5 mmol·L-1)培养基中培养48 h后,用光镜观察SRA01/04细胞形态;CCK-8试剂盒检测各组细胞的活性;Western blot检测各组细胞SIRT6蛋白的表达水平.然后将SRA01/04细胞用55.5 mmol·L-1葡萄糖干预,培养不同时间(0 h、6 h、12 h、24 h、48 h、72 h)后,Western blot检测不同时间SRA01/04细胞SIRT6蛋白表达水平.免疫荧光技术检测对照组(5.5 mmol·L-1葡萄糖)和高糖组(55.5 mmol·L-1葡萄糖)细胞的SIRT6蛋白表达情况.结果 随着葡萄糖浓度增加,SRA01/04细胞的数量逐渐降低.CCK-8检测结果显示,与5.5 mmol·L-1葡萄糖组相比,随着葡萄糖浓度增加,SRA01/04细胞的活性逐渐降低,差异均有统计学意义(均为P<0.05).Western blot检测结果显示,SRA01/04细胞的SIRT6蛋白表达与葡萄糖呈浓度和时间依赖性,45.5 mmol·L-1、55.5 mmol·L-1、75.5 mmol·L-1葡萄糖组与5.5 mmol·L-1葡萄糖组比较,以及24 h组、48 h组和72 h组与0 h组比较,差异均有统计学意义(均为P<0.05).细胞免疫荧光结果显示,SRA01/04细胞中的SIRT6蛋白主要在细胞核表达,高糖组SRA01/04细胞中SIRT6蛋白表达较对照组显著降低(P<0.05),且高糖组细胞核出现明显的核固缩和核碎裂现象.结论 高浓度葡萄糖可以降低S RA01/04细胞的增殖活性,降低S I RT6蛋白的表达水平.

Ovarian cancer is the most lethal gynecologic malignancy due to the lack of symptoms until advanced stages, and new diagnosis and treatment strategy is in urgent need. In this study, we found higher expression of miR-19a-3p in ovarian cancer tissues compared with that in the adjacent normal tissues. By chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assay (EMSA) analysis, we showed that nuclear factor-kappaB (NF-kappa B) binds to the promoter of miR-19a-3p, leading to reduced expression in ovarian cancer cells. Further study indicated that miR-19a-3p inhibits the expression of insulin-like growth factor binding protein-3 (IGFBP-3), resulting in enhanced growth and migration of ovarian cancer cells in vitro and tumor growth in vivo. These results showed that miR-19a-3p enhances the oncogenesis of ovarian cancer through inhibition of IGFBP-3 expression, and which can be inhibited by NF-kappa B, suggesting an NF-kappa B/miR-19a-3p/IGFBP-3 pathway in the oncogenesis of ovarian cancer, which expands our understanding of ovarian cancer and they may contribute to the development of new diagnosis and treatment of ovarian cancer.

以生物化学课程为重点的前提下,站在学生的视角将临床与基础进行有机的整合,更新授课切入点、重新组织课程,提前(早期)适度引入临床课程及实际临床工作中与生化课程内容密切相关的知识、理念、思维等,继而融合成模式与系统,从而达到激发学生学习兴趣、提高学习效果的目的.

目的 观察枸杞多糖(LBP)对高糖诱导的人晶状体上皮细胞(SRA0 1/04)凋亡及SIRT1表达的影响.方法 将SRA (OVOA)细胞随机分为正常对照组、高糖组及高糖+LBP低、中、高3个剂量组.各条件下培养48 h后采用CCK-8法检测各组细胞的活性,流式细胞技术(FCM)检测细胞的凋亡率,蛋白免疫印迹法检测各组细胞中SIRT1蛋白的相对表达量.结果 CCK-8结果显示,与正常对照组相比,高糖组细胞活性降低;而与高糖组相比,LBP高剂量组细胞活性升高,差异有统计学意义(P＜0.05).流式细胞术显示,与正常对照组相比,高糖组细胞凋亡率显著上升;与高糖组相比,LBP组细胞凋亡率显著下降,差异有统计学意义(P＜0.05).蛋白印迹法结果显示,与正常对照组相比,高糖组细胞中的SIRT1蛋白表达降低;与高糖组相比,LBP组细胞中的SIRT1蛋白表达升高,差异有统计学意义(P＜0.05).结论 LBP可能通过调节SIRT1的表达对高糖环境下人晶状体上皮细胞凋亡产生抑制作用.

目的 探讨基于移动学习App的混合式教学模式在《生物化学》课程教学中的应用.方法 选取宁夏医科大学2017级护理专业82名学生为研究对象,护理1班采用混合式教学模式,护理2班采用传统的讲授教学模式,比较两组学生考核成绩和教学满意度的差别.结果 在考试成绩上,混合式教学组学生成绩与对照组相比,其差异无统计学意义(72.26±3.89比71.22±4.12,P=0.265).教学满意度评价结果显示,在提高学习积极性,激发学生课堂学习氛围,提升学习效率和学习能力,促进语言表达能力,增加沟通交往能力方面,混合式教学组评分明显优于普通教学模式组,两组差异具有统计学意义(P＜0.005).结论 本研究发现,基于移动学习App的混合式教学模式在《生物化学》课程教学中效果较好,学生满意度较高,在取得更多教学经验后,可以在今后的《生物化学》课程教学中推广.

通过分析实验教学现存问题和不足,以学生自主性学习为导向,提出在传统实验教学模式基础上增设自主学习、自主实验总结、自主实验设计等实验教学环节,提高实验教学质量,并就实践效果进行探讨,旨在为生物化学与分子生物学实验教学改革提供参考.

Diabetic retinopathy (DR) has become the most frequent cause of impaired visual acuity and blindness in working-age population in developed countries. Here we use diabetic rats to clarify the role of Lycium barbarum polysaccharides (LBP) on DR. We treated diabetic rats with LBP (400 mg/kg/d or 200 mg/kg/d) orally for 20 weeks. Electroretinogram (ERGs) and Laser Doppler blood flow were measured to assess the retinal function, routine histology and ultrastructural studies were performed to evaluate the morphological alterations, and immunohistochemistry, western blotting, and RT-PCR were conducted to detect the protein and mRNA levels of pro- and antiangiogenic factors. The results showed that diabetes suppressed the amplitudes of a-wave, b-wave, and oscillatory potential in ERG, reduced retinal blood flow, decreased the thickness of the retina, and increased the thickness of basement membrane of the retinal capillary. Furthermore, diabetes increased the mRNA and protein expressions of proangiogenic GFAP and VEGF and suppressed the levels of antiangiogenic PEDG. Treatment with LBP either completely or partially reversed the alterations caused by diabetes. It is concluded that the LBP protects retinal function and morphology in diabetic rats, probably through reinstallation of the balance between proangiogenic and antiangiogenic factors, which reduces neovascularization. LBP could be used as a therapeutic drug for DR.

目的 探讨三七总皂苷对糖尿病大鼠视网膜病变血管絮张素Ⅱ(AngⅡ)和内皮素-1(ET-1)的影响.方法 采用链脲佐菌素(65 mg/kg)腹腔注射的方法建立大鼠模型,选取50只SD大鼠,将其随机分为5组:正常对照组,糖尿病模型组,三七总皂苷高剂量组(150 mg/kg),三七总皂苷低剂量组(50 mg/kg)以及导升明组(167 mg/kg).灌胃给药20周后处死大鼠,取大鼠血清检测AngⅡ和ET-1的含量,摘取眼球,剥离大鼠眼球的视网膜组织,采用免疫组织化学法及荧光定量PCR法检测大鼠视网膜组织中ET-1蛋白及mRNA的表达水平.结果 与正常对照组相比,糖尿病大鼠血清中AngⅡ和ET-1的含量明显增加(P＜0.05),三七总皂苷组中的大鼠血清AngⅡ和ET-1的含量[(336.9±85.9) ng/L,(59.8±8.4) ng/L)]明显低于糖尿病模型组[(557.9±86.2)ng/L,(89.8±7.2) ng/L,P＜0.05)].同时,糖尿病大鼠视网膜组织中ET-1蛋白及mRNA的表达增加,而三七总皂苷组和导升明组大鼠视网膜组织中ET-1的表达明显低于糖尿病模型组大鼠(P＜0.05).结论 三七总皂苷可能通过降低大鼠血清中AngⅡ和ET-1以及糖尿病大鼠视网膜组织中ET-1蛋白与mRNA的表达水平,从而延缓糖尿病大鼠视网膜病变的发生和发展.

2012年9月,宁夏医科大学启动卓越医生教育培养计划,并以此为契机,对"卓越医生班"生物化学与医学遗传学两门专业课进行了整合,针对学科特点,在教学设置、教学方法 及考核方式等方面进行了一系列改革,收到了良好的教学效果.

Objective To observe the effect of shRNA interference lentivirus vector targeting rat Sirt1 gene on the expression of Sirt1 in retinal ganglion cell (RGC). Methods Four short hairpin (sh) RNA interference sequences targeting rat Sirt1 gene were designed. The target sequences of Oligo DNA were synthesized and annealed to double strand DNA, which was subsequently connected with pGLV3 lentivirus vector to build the lentiviral vector. The positive clones were identified by polymerase chain reaction (PCR) and DNA sequencing. The lentiviral vector construct and lentiviral packaging plasmids were co-transfected into 293T cells, then the titer of lentivirus were determined. The RGC were divided into 6 groups including blank group, negative control group and si-Sirt1-1, si-Sirt1-2, si-Sirt1-3, si-Sirt1-4 groups. Real-time PCR and Western blotting were used to detect the expression of Sirt1 mRNA and protein in the RGC cells. Results PCR and DNA sequencing analysis confirmed that the shRNA sequence was successfully inserted into the lentivirus vector. The concentrated titer of virus suspension was 8×108 TU/ml after the recombinant lentiviral vector successfully transfected and harvested in 293T cells. Comparing with NC group, the expression of Sirt1 mRNA and protein were significantly decreased in the si-Sirt1-1, si-Sirt1-2, si-Sirt1-3 and si-Sirt1-4 groups (F=27.682, 1185.206; P=0.000, 0.000). The si-Sirt1-2 group had the strongest effect in reducing the expression of Sirt1 mRNA and protein. Conclusion The 4 lentiviral vectors harboring RNAi targeting rat Sirt1 gene can effectively down regulate the expression of Sirt1 mRNA and protein in RGC cells.