Optimization of separation methods and culture system of chicken embryonic stem cells in vitro

背景:胚胎干细胞是从动物早期胚胎的内细胞团或原始生殖细胞分离出来的具有发育全能性的一种未分化的无限增殖细胞系。而鸡胚胎干细胞则是从X期鸡胚的胚盘分离而来。目的:优化鸡胚胎干细胞分离方法和离体培养体系。方法:采用滤纸纸环-发环的方法从X期鸡胚分离胚盘细胞,并采用STO细胞作为饲养层和大鼠肝细胞(brl)条件培养基(CM)+细胞因子作为离体培养体系对分离的胚盘细胞进行培养。结果与结论:滤纸纸环-发环法获得的完整胚盘率为75%~85%,克隆形成率约为50%。brl-CM+饲养层培养体系,鸡胚胎干细胞可传至7代,而brl-CM+饲养层+细胞因子培养体系,鸡胚胎干细胞可传至25代。分离到的鸡胚胎干细胞,经碱性磷酸酶染色、SSEA-1染色鉴定,表明鸡胚胎干细胞处于未分化状态。提示,实验不仅优化了鸡胚胎的分离方法,获得完整且杂质少的胚盘,而且进一步优化了鸡胚胎干细胞体外培养体系。BACKGROUND:Embryonic stem cells are undifferentiated permanent cell line derived from inner cell mass cells and primordial germ cells of animal's early embryos.Chicken embryonic stem cells are derived from the blastodermal of a X-stage embryo.OBJECTIVE:To optim the separation method and in vitro cultural system of chicken embryonic stem cells.METHODS:The X-stage chicken embryos were isolated by using a small square of ?lter paper with a hole punched in the center,and the blastodermal cells were isolated by using the hair loop.STO cells were used to make feeder layer;at the same time,BRL-CM and cytokine were also used for chicken embryonic stem cells in vitro cultural system.RESULTS AND CONCLUSION:The filter paper loop and the hair loop could obtain complete the blastoderm,and the successful percentage was 75%-85%.The colony formation rate was about 50%.After culture in the BRL-CM + feeder layer + cytokine culture system,the passage of CES cells is the seventh generation;BRL-CM + feeder layer + cytokines,cultured chicken embryonic stem cells could passage to the 25th generation.Isolated chicken embryonic stem cells were in an undifferentiated state detected by alkaline phosphatase staining and SSEA-1 staining.The findings indicate that this experiment not only optimized the isolation method of chicken embryonic stem cells to obtain complete and pure embryos,but also further improved the in vitro culture system of chicken embryonic stem cells.国家973项目(2009CB941600)资助;国家自然科学基金项目(31072101)资助---

Mechanism of geniposide in improving free fatty acid metabolism in rats with non-alcoholic fatty liver disease

观察栀子苷改善非酒精性脂肪性肝病的效果,从游离脂肪酸探讨栀子苷调节非酒精性脂肪性肝病的作用机制。健康雄性Wistar大鼠40只,随机分为4组:正常组、模型组、栀子苷组和血脂康组,每组大鼠10只,正常组大鼠给予正常大鼠饲料喂养,其余3组大鼠均采用高脂饲料喂养以诱导非酒精性脂肪性肝病,造模时间为8周,从第5周起至第8周末,栀子苷组和血脂康组分别灌服相应的药物。记录大鼠体重、肝湿重、脂肪质量;用相应方法检测肝组织TG,FFA,FAS,AMPK,ACCase及Malonyl-Co A含量,血清CHO,LDL-C的含量,血清AST,ALT的活性;观察肝脏组织肉眼及病理变化(HE染色法)。结果显示,与正常组相比,模型组大鼠的体重;肝湿重;脂肪重;血清CHO,LDL-C,ALT,AST;肝组织TG,FFA,FAS,ACCase及Malonyl-Co A含量皆显著升高(P<0.01);肝组织AMPK活性显著降低(P<0.01),肝组织肉眼外观及病理切片脂肪变性明显,并出现炎症损伤;与模型组相比,栀子苷组大鼠体重、脂肪质量、肝组织FFA含量、血清ALT,AST活性均显著降低(P<0.01),肝湿重,肝组织TG,FAS,ACCase及Malonyl-Co A含量明显降低(P<0.05),肝组织AMPK活性明显增多(P<0.05),肝脏肉眼外观及病理学表现均有所改善;与模型组相比,血脂康组大鼠的肝湿重、脂肪质量、肝组织TG,FFA和血清LDL-C水平明显降低(P<0.05);与血脂康组相比,栀子苷组大鼠的体重、脂肪质量、肝组织FFA含量均显著降低(P<0.01),其他方面无明显差异。结果表明,栀子苷具有显著的改善高脂饮食诱导的大鼠非酒精性脂肪性肝病的药理效应;其改善大鼠非酒精性脂肪性肝病的游离脂肪酸代谢是通过调节"AMPK-ACCase-Malonyl-Co A-FFA"轴来实现的。To observe the effect of geniposide on non-alcoholic fatty liver disease( NAFLD),and discuss the mechanism of geniposide for NAFLD from the aspect of free fatty acid,forty healthy Wistar male rats were randomly divided into normal group,model group,geniposide and Xuezhikang group. The rats in normal group were fed with normal diets,and the rats in other 3 groups were given with high-fat diet for 8 weeks to induce the NAFLD models. From the week 5 to end of week 8,the rats in geniposide and Xuezhikang group were intervened with corresponding medicines. The body weight,liver wet weight,and fat weight of the rats were recorded.Visual and pathological changes in hepatic tissues were observed with HE staining. The contents of TG,FFA,FAS,AMPK,ACCase and Malonyl-Co A in hepatic tissue,contents of CHO and LDL-C in serum and activities of AST and ALT in serum were detected by using corresponding methods. The results showed that the body weight,liver wet weight,and fat weight of the rats,CHO,LDL-C,ALT and AST levels in serum,TG,FFA,FAS,ACCase and Malonyl-Co A levels in hepatic tissues of the rats in model group were significantly higher than those in normal group( P < 0. 01),while AMPK activity was significantly lower than that of the normal group( P < 0. 01),with obvious visual and pathological steatosis in hepatic tissues,and inflammatory injury occurred in model group. Compared with the model group,body weight of the rat,fat weight,levels of FFA in hepatic tissues,ALT and AST activities in serum,liver wet weight,TG,FAS,ACCase and Malonyl-Co A levels were significantly decreased in geniposide group( P < 0. 01),while the AMPK activity in hepatic tissues was significantly increased( P < 0. 05),with improvement in visual and pathological performance.Compared with the model group,liver wet weight,fat weight,TG and FFA levels in hepatic tissues,and LDL-C level in serum were significantly decreased in Xuezhikang group( P < 0. 05). Compared with Xuezhikang group,the body weight of rat,fat weight and FFA level in hepatic tissues were significantly lower in geniposide group( P < 0. 01),but with no significant difference in other aspects. These findings indicated that geniposide was highly effective in improving the pharmacological effect of NAFLD induced by highfat diet,and the mechanism was achieved through AMPK-ACCase-Malonyl-Co A-FFA axis.国家自然科学基金项目(81274155;81503529);; 福建省自然科学基金项目(2014J01374);; 福建省高等学校新世纪人才计划项目;; 厦门市科技计划项目(3502Z20134020);; 福建省中医药科研项目(WZPW201308

养正消积胶囊辅助GP方案治疗晚期非小细胞肺癌临床研究

目的观察养正消积胶囊辅助GP方案治疗晚期非小细胞肺癌(NSCLC)的临床疗效。方法采用随机数字表法将89例晚期NSCLC患者分为对照组44例及观察组45例。2组均采用GP方案;观察组并予养正消积胶囊,每次4粒,每日3次,口服。2组均21d为1个周期,连续治疗2个周期,随访1年。观察2组治疗前后Ki67、Bax、Bcl-2蛋白表达及肿瘤标志物[血管内皮生长因子(VEGF)、骨桥蛋白(OPN)、癌胚抗原(CEA)、糖类抗原199(CA199)]、中医症状评分、生活质量评分,比较2组客观缓解率(ORR)、疾病控制率(DCR)、无进展生存时间(PFS),监测不良反应。结果观察组ORR、DCR明显优于对照组(P<0.05)。与本组治疗前比较,观察组治疗后、随访1年VEGF、OPN、CEA、CA199水平明显降低(P<0.05);对照组治疗后上述指标明显降低(P<0.05),随访1年CEA水平明显降低(P<0.05)。与本组治疗前比较,2组治疗后Ki67、Bcl-2蛋白表达明显降低,Bax蛋白表达明显升高(P<0.05);2组治疗后比较,观察组Ki67、Bcl-2蛋白表达低于对照组(P<0.05),Bax蛋白表达高于对照组(P<0.05)。与本组治疗前比较,2组治疗后中医症状评分(神疲乏力、胃纳少馨、腰膝无力、夜尿频多、头晕目眩)、生活质量评分(躯体功能、认知功能、社会功能、情绪功能)明显改善(P<0.05);2组治疗后比较,观察组上述评分改善明显优于对照组(P<0.05)。观察组PFS明显长于对照组(P<0.05),不良反应明显低于对照组(P<0.05)。结论养正消积胶囊辅助GP方案治疗晚期NSCLC可有效降低患者肿瘤标志物水平,改善生存质量,减少吉西他滨和顺铂的不良反应,提高疗效。福建省卫生厅中医药科研专项课题(wst201210

Study on the dose ratio of Chlorogenic acid-Geniposide to treat rats with experimental fatty liver based on uniform design

目的:运用均匀设计法优化绿原酸-栀子苷组合治疗实验性脂肪肝大鼠的最佳剂量配比,探讨中药有效成分配伍配比规律。方法:运用数学模型均匀设计法对绿原酸; -栀子苷组合中的2种中药有效成分进行分组设计,分为6种组合对高脂饮食诱导的实验性脂肪肝大鼠模型进行治疗,观察各组大鼠肝脏的病理变化,并以肝组织甘; 油三酯(TG)、血清低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、总胆固醇(CHO)、谷丙转氨酶(ALT)、谷草转氨酶(A; ST)测定作为筛选指标,优选二者的最优剂量配比,并根据所得重新分组验证。结果:与正常组比较,模型组脂肪肝大鼠肝组织TG、血清LDL-C、CHO含; 量明显升高(P<0.01),血清HDL-C含量明显降低(P<0.01),血清ALT、AST活性明显升高(P<0.01),肝组织存在明显脂肪变性。; 经多元逐步回归分析,绿原酸-栀子苷组合治疗脂肪肝的最佳剂量配比是绿原酸90mg+栀子苷90mg。结论:应用均匀设计与药效学分析的方法可有效优化中; 药有效成分组合的最佳剂量配比。Objective: To explore the active ingredients and dose ratio of medicine,; combined with the optimal dose ratio of Chlorogenic acid-Geniposide to; treat rats with experimental fatty liver. Methods: The active; ingredients and optimized prescription were screened out, adopting; uniform design to divide Chlorogenic acid-Geniposide into six; combinations, by using experiment on high fat diet-induced fatty liver; rat models, and the pathological results were observed. The serum LDL-C,; HDL-C, CHO, ALT, AST and liver TG were taken as the screening-indexes.; Results: Compared with the normal groups, the serum LDL-C, CHO, ALT, AST; and liver TG in model groups were significantly higher (P<0.01), the; serum HDL-C in model group was significantly decreased (P<0.01), and the; pathology of liver tissue got obvious steatosis. According to stepwise; regression analysis of uniform design, the best ratio of Chlorogenic; acid-Geniposide to anti-NAFLD were Chlorogenic acid 90mg with Geniposide; 90mg. Conclusion: Uniform design is the appropriate way to confirm the; optimized prescription for active ingredient of Chinese medicine.国家自然科学基金面上项目; 福建省自然科学基金面上项目; 福建省高等学校新世纪人才计划项

基于神经网络的光子器件逆设计研究进展

光与纳米结构的相互作用一直是纳米光子学的重要研究内容之一,核心部件的纳米结构对光子器件的功能和性能具有决定性作用。纳米光子器件的设计存在两种思路:一是从物理原理出发的直观设计;二是根据所需光学响应探索最优结构的逆设计。近年来,逆设计在纳米器件中取得了一系列重要进展,尤其是最近将深度学习方法引入进来,开启了高性能纳米光子器件智能高效设计的新篇章。文章围绕纳米光子器件智能逆设计方法,分析归纳了这一新兴研究方向的产生背景、重要进展和典型应用,对智能逆设计面临的挑战及未来发展方向进行了展望