β-sitosterol (BS), a major bioactive constituent present in plants, has shown potent anti-cancer activity against many human cancer cells, but its activity in pancreatic cancer (PC) cells has rarely been reported. Gemcitabine (GEM) is one of the first-line drugs for PC therapy, however, the treatment effect is not sustained due to prolonged drug resistance. In this study, we firstly studied the anti-PC activity and the mechanism of BS alone and in combination with GEM in vitro and in vivo . BS effectively inhibited the growth of PC cell lines by inhibiting proliferation, inducing G0/G1 phase arrest and apoptosis, suppressed the NF- kB activity, and increased expression of the protein Bax but decreased expression of the protein Bcl-2. Moreover, BS inhibited migration and invasion and downregulated epithelial–mesenchymal transition (EMT) markers and AKT/GSK-3β signaling pathways. Furthermore, the combination of BS and GEM exhibited a significant synergistic effect in MIAPaCa-2 and BXPC-3 cells. More importantly, the combined treatment with BS and GEM lead to significant growth inhibition of PC xenografts. Overall, our data revealed a promising treatment option for PC by the combination therapy of BS and GEM.

目的:建立扶正通络颗粒(黄芪、当归、枸杞、丹参等)的质量标准.方法:TLC法对当归、丹参进行定性鉴别,HPLC法对阿魏酸和丹参酮ⅡA进行定量测定.结果:TLC斑点分离清晰,阴性无干扰.阿魏酸和丹参酮ⅡA分别在0.15 ～2.4 μg(r=1.000)和0.05 ～0.4 μg(r =0.999 9)范围内与峰面积呈良好的线性关系.平均回收率分别为98.06％和98.56％,RSD分别为2.25％和1.58％(n=9).两者平均含量分别为0.189 6和0.061 9 mg·g-1,RSD分别为4.78％和1.52％(n=6).结论:该方法可靠准确,专属性好,可用于扶正通络颗粒的质量控制.

Rosmarinic acid (RA), isolated from herbal balm mint plants, has demonstrated potent anti-tumor properties against liver cancer. However, the precise underlying mechanisms remain unclear. This study aimed to investigate the molecular mechanisms of RA in HepG2 cells. RA anti-tumor activity was assessed using 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays, and Hoechst 33258 staining. Apoptosis and the cell cycle distribution were evaluated by flow cytometry. A proteomics approach was used to identify differentially expressed proteins following RA treatment in HepG2 cells, and quantitative reverse transcription-quantitative polymerase chain reaction was used to validate the results. Bioinformatics analysis was also implemented to further understand the identified proteins, and western blotting was used to analyze the associated proteins. Our results suggested that RA treatment significantly inhibits the viability of HepG2 cells. The MTT and LDH assays indicated dose-dependent decreases in cell proliferation following RA treatment. Hoechst 33258 staining and flow cytometry analysis showed that RA exhibits an apoptosis-inducing effect and induces cell cycle arrest in G1. The proteomics analysis successfully identified 16 differentially expressed proteins. Bioinformatics analysis indicated that the identified proteins participated in several biological processes and exhibited various molecular functions, mainly related to inactivation of the glycolytic pathway. Further western blotting analysis showed that RA could downregulate the expression of glucose transporter-1 and hexokinase-2, leading to the suppression of glucose consumption and generation of lactate and ATP. Taken together, our study found that RA exhibits significant cytotoxic effects by inhibiting cell proliferation and inducing apoptosis and cell cycle arrest, possibly by blocking the glycolytic pathway in human HepG2 cells.

当前我国医学本科生普遍缺乏创新思维及创新实践能力,急需转变培养模式.我们尝试应用适合医学本科生的长效科研创新能力培养模式,通过兴趣激发、创新意识培养、科研创新实践和参加学术活动等方法与手段,提高医学本科生创新能力,以适应未来医疗卫生事业发展需求.

Objective: This study was designed to investigate the effect of Hedysarum polybotrys polysaccharides 3 (HPS-3) on splenic T-lymphocytes differentiation. Methods: The isolated spleen lymphocytes were incubated with HPS-3 at serial concentrations for 72 h and the cell viability was detected using MIT. Concanavaline (ConA) was used as a positive control. The production of Th1 (IFN-gamma, TNF-alpha and IL-2) and Th2 (IL-4) cytokines in HPS-3 treated lymphocytes were detected using ELISA. Distribution of T cell subsets was detected using flow cytometry. Cellular ultrastructure of lymphocytes was investigated using the TEM scanning. Results The proliferation of lymphocytes was enhanced by HPS-3 in a dose-dependent manner, with the highest proliferation ability at 100 mg/mL. HPS-3 treament significantly upregulated the production of Th1 cytokines (IFN-gamma, TNF-alpha, and IL-2) and reduced IL-4. Moreover, the percentages of CD3+ T cells and CD3(+)CD8(+) T cells subsets were upregulated by HPS-3, enven higher than those of ConA treated cells. At last, we found the lymphocytes treated by HPS-3 had more organelles, especially more mitochondria, suggesting HPS-3 promoted lymphocytes activity. Conclusion: We determined HPS-3 administation promoted the proliferation of lymphocytes and might alter the balance of Th1/Th2 ratio, showing the immunomodulatory effect of HPS-3.

目的 探讨含红芪和含黄芪的益气养血汤抗免疫衰老的可能作用机制并比较两者的作用效果.方法 30只青龄昆明种小鼠随机分成红芪益气养血汤组、黄芪益气养血汤组、空白组,每组10只,红芪益气养血汤组及黄芪益气养血汤组分别给予相应生药6g/ (kg· d),灌胃量为0.4 ml/ (20g·d),空白组灌胃相同体积生理盐水,连续灌胃14天,末次灌胃2h后眼球摘除取血,制备益气养血汤含药血清和空白血清.调整SAMP8小鼠脾脏T淋巴细胞浓度为所需浓度,分为红芪益气养血汤组(含药血清)、黄芪益气养血汤组(含药血清)、空白血清组(空白血清)、青龄鼠组(完全培养液)、SAMP8组(完全培养液).培养72h后,检测各组细胞增殖能力,T淋巴细胞表面分子包括CD8+、CD4+、CD4 +/CD8+、CD28+ CD152+表达情况,以及细胞培养上清液中白细胞介素2 (IL-2)和γ干扰素(IFN-γ)水平和细胞中PI-3K mRNA表达、β-半乳糖苷酶染色细胞阳性率.结果 与青龄鼠组比较,SAMP8组小鼠脾脏T淋巴细胞增殖能力、细胞表面CD8+分子表达、培养上清液IL-2和IFN-γ水平、细胞中PI3K mRNA表达明显降低,CD4+、CD4 +/CD8+、CD28+ CD152+双阳性表达及β-半乳糖苷酶衰老细胞阳性率均明显升高(P＜0.05).与空白血清组比较,红芪益气养血汤组和黄芪益气养血汤组均能提高T淋巴细胞的增殖能力、培养上清液中IL-2和INF-γ水平和细胞中PI3K mRNA表达,减少β-半乳糖苷酶染色细胞阳性率(P＜0.05),并且在改善细胞增殖能力、提高IL-2水平及PI3K mRNA表达方面红芪益气养血汤组优于黄芪益气养血汤组(P＜0.05).结论 含红芪和含黄芪的益气养血汤含药血清均能增强SAMP8小鼠脾脏T淋巴细胞的体外增殖能力,可能通过促进IL-2及IFN-γ分泌、上调PI3K mRNA表达量、降低β-半乳糖苷酶表达,从而改善T淋巴细胞的免疫衰老状态,并且含红芪的益气养血汤作用效果优于含黄芪的益气养血汤.

目的:比较含红芪和含黄芪的益气养血汤对快速老化小鼠P8(SAMP8)小鼠免疫功能的影响,并探讨其可能的作用机制.方法:取昆明种青龄小鼠10只为青龄鼠组,SAMP8小鼠40只随机分为4组,分别为胸腺肽组、生理盐水组、益气养血红芪组和益气养血黄芪组.分别以生理盐水,胸腺肽每日4.6 mg· kg-1,含红芪、黄芪的益气养血汤每日6 g·kg-1剂量ig小鼠,每日1次,连续ig15次,末次ig后2h眼球摘除取血及脾脏.酶联免疫吸附测定(ELISA)检测其血清白细胞介素-2(IL-2),γ干扰素(IFN-γ)含量,流式细胞术检测脾脏淋巴细胞表面CD3,CD4,CD8,CD28及CD152分子的表达情况,实时荧光定量聚合酶链式反应(Real-time PCR)检测p38 MAPK mRNA的表达情况,蛋白质免疫印迹(Western blot)及免疫组化法检测其p38MAPK蛋白表达.结果:与青龄小鼠比较,生理盐水组SAMP8小鼠IL-2,IFN-γ含量降低,CD8,CD28分子表达降低,CD4分子表达及CD28CD152分子双阳性表达增高,p38 MAPK mRNA及蛋白表达降低(P＜0.05).与生理盐水组比较,胸腺肽组及中药组均能提高SAMP8小鼠血清中IL-2,IFN-γ含量,胸腺肽组含量高于中药组,红芪组IL-2含量高于黄芪组;胸腺肽组及中药组均能提高CD28分子表达,降低CD28CD152双阳性分子表达,胸腺肽组及黄芪组双阳性表达率低于红芪组;中药组能提高SAMP8小鼠脾淋巴细胞表面CD8分子表达,且黄芪组CD8表达高于红芪组;胸腺肽组及黄芪组能降低CD4分子表达;胸腺肽组及中药组均能提高p38 MAPK mRNA表达,且胸腺肽组mRNA表达高于中药组,黄芪组高于红芪组;胸腺肽组及中药组均能提高p38MAPK蛋白含量及蛋白阳性率,且胸腺肽组优于中药组(P＜0.05).结论:含红芪、黄芪的益气养血汤均能改善SAMP8小鼠的免疫衰老现象,增加p38MAPK的表达.

目的 探索广东异地引种甘肃道地药材红芪的可行性.方法 取同一批红芪种子分别在广东广州和甘肃陇西两地种植栽培.通过土壤营养物质含量测定,比较两地土壤状态对红芪生长的影响.HPLC比较两地种植红芪的指纹图谱,测定其中指标成分毛蕊异黄酮、芒柄花素含量.苯酚-硫酸法测红芪含糖量.结果 广州和陇西的土壤样本中营养物质含量略有区别.广州红芪样本中芒柄花素含量远高于陇西种植的红芪.指纹图谱比对结果显示,环境改变没有使红芪品质发生明显改变.结论 控制好土壤环境可保证广东异地引种红芪品质不发生改变.

成体干细胞应用于器官移植比胚胎干细胞更具优势,因为病人可以使用自己的成体干细胞,制造替代器官,同时也能够减少器官买卖,保证公平,防止道德滑坡,并且有着较高的社会认可度.为了更好地维护人类的健康利益,需对两者的结合进行科学的管理和伦理规范.这样,才会使再生医学的研究在器官移植领域取得进步.

In the disease phenomenon of various links in the form of interdependence,logical relation of causality occupies a very important position.logical relation of causality is a fundamental problem in the philosophy,the Characteristic of interrelated between things.Use of logical relation of causality is the issue and intuitive in medical,it's empirical in theory and practice combined with each other.Based on the understanding and grasp of causation,it can be help the physician in epistemological basis to better grasp the essence of disease specific medical practice,to achieve a rapid and accurate screening of the disease before the appearance of clinical complex disease,to play a guiding role.

It was aimed to establish an HPLC method for simultaneous determination of three kinds of components in Shenju Zhike drop pill i.e.cinnamic acid,aurantriamarin and harpagoside.Samples were analyzed by gradient elution on Hypersil-C18 column,MeOH-0.2% acetic acid was the mobile phase,the detection wavelength was 280 nm and the temperature of column was 25 ℃.The results showed that the linear ranges of cinnamic acid,aurantriamarin and harpagoside were 0.008 14～0.203 50,0.009 68～0.242 00,0.006 84～0.102 60 mg·mL-1,respectively.The average recovery rate was 98%～100%,RSD<2.19%.Therefore,the simple,rapid and accurate method could be used for the quality control of Shenju Zhike drop pill.

Objective:To observe the effects of fuzhen yiliu granule on the P53 and BCL-2genes protein expression related to apoptosis in tumor cells of H22 solid tumor mice.Methods:48mice were divided into 4 groups:model group,control group,low and high dose groups,all mice were inoculated H22 tumor strain to build models,4 groups were respectively given NS,tianxian capsule,low and high dose fuzhen yiliu granule for 18 days.P53 and BCL-2 genes protein expression were determined by flow cytometry technique.Results:The P53 genes protein expression was lower and the BCL-2 expression was higher obviously in model group,the P53 expression was elevated and BCL-2 expression was lowered in fuzhen yiliu granule groups,there was significant difference between fuzhen yiliu granule groups and model group(P<0.01).Conclusion:Fuzhen yiliu granule can promote the P53 expression and inhibit the BCL-2 expression in tumor cells,which maybe one of the mechanism that fuzhen yiliu granule can effect on tumor.

目的:观察复方中药制剂扶正抑瘤颗粒(FYK)对荷瘤(H22)小鼠T淋巴细胞凋亡相关基因Fas/FasL蛋白表达的影响。方法:将48只小鼠分为中药大、小剂量组、天仙胶囊阳性对照组和模型组,造模形成实体瘤后,分别灌胃FYK和天仙胶囊、生理盐水,18天后取血应用流式细胞仪测定Fas和FasL的蛋白表达。结果:各治疗组Fas蛋白表达较模型组下降,与模型组相比较有显著性差异(P<0.001)。各治疗组FasL蛋白表达较模型组升高,与模型组相比较有显著性差异(P<0.001)。结论:FYK可显著减轻荷瘤(H22)动物T淋巴细胞凋亡,在肿瘤的免疫逃逸中有重要意义。

目的:观察复方中药制剂扶正抑瘤颗粒（FYK）对荷瘤（H22）小鼠T淋巴细胞凋亡相关基因Fas/FasL蛋白表达的影响。方法:将48只小鼠分为中药大、小剂量组、天仙胶囊阳性对照组和模型组,造模形成实体瘤后,分别灌胃FYK和天仙胶囊、生理盐水,18天后取血应用流式细胞仪测定Fas和FasL的蛋白表达。结果:各治疗组Fas蛋白表达较模型组下降,与模型组相比较有显著性差异（P＜0.001）。各治疗组FasL蛋白表达较模型组升高,与模型组相比较有显著性差异（P＜0.001）。结论:FYK可显著减轻荷瘤（H22）动物T淋巴细胞凋亡,在肿瘤的免疫逃逸中有重要意义。

Aim An HPLC method for analyzing eleutheroside B (ELU B) and eleutheroside E (ELU E), two of the main active substances of Acanthopanax preparations were studied. Methods The samples were analyzed on a kromasil ODS column with water-acetonitrile as a gradient mobile phase. The flow rate was 0.8 mL·min-1 and detecting wavelengths were 206 nm for ELU B, 220 nm for ELU E, solid phase extraction (SPE) and internal standard-salicin were selected. Results The recoveries of Acanthopanax tablets and injection were 90.4%-96.8% and 96.4%-99.8% for ELU B, 87.7%-93.3% and 95.7%-98.5% for ELU E, respectively. The linear ranges were 4.45-22.25 μg· mL-1(r=0.999 8) and 5.11-25.55 μg·mL-1 (r=0.999 7) respectively. Conclusion This method can save the time for cleaning the chromatographic system and improve sensitivity for Acanthopanax preparations, thus providing a way to evaluate the quality of Acanthopanax preparations.

目的:探讨了应用中药制剂扶正抑瘤颗粒对肿瘤细胞增殖周期影响.方法:采用流式细胞术.结果:使用药物后,能够改变肿瘤细胞增殖周期的各期细胞分布比例,与模型组比较有显著性差异.结论:中药扶正抑瘤颗粒能够改变肿瘤细胞周期时项分布,具有明显的抑瘤作用.

OBJECTIVE:To study on chromatography fingerprint(CFP) of Huangqi herbs.METHOD:HPLC was used on a Hypersil ODS column with water-acetonitriles gradient elution. CFP was referenced to formononetin, calycosin and then the contents of astragaloside IV, formononetin and calycosin was determinted.RESULT:26 common peaks were pointed out, their similarity of 12 samples were more than 90%.CONCLUSION:The handling of samples were simple. The study method on chromatography fingerprint of Huangqi were reliable. Good repetition and their similarity accorded with the demand on "the technology guide to experiment studies on chromatography fingerprint of injection of traditional Chinese medicine (TCM)", which was issued by the nation pharmacopoeial commission. The chromatography fingerprint and determination of index composition could be used to comprehensively control the quality of Huangqi.