目的 探讨赖氨酰氧化酶样蛋白2(LOXL2)在结直肠癌中的表达水平及临床预后意义,并结合免疫浸润初步探究其机制.方法 应用生物信息学和免疫组织化学染色方法评估LOXL2在结直肠癌中的表达情况及其预后潜能,并分析LOXL2与结直肠癌免疫浸润水平的关系.结果 与正常结直肠组织比较,LOXL2在结直肠癌中表达上调,且LOXL2高表达与结直肠癌患者较差的总生存期相关(P＜0.05).LOXL2与结直肠癌中巨噬细胞的浸润水平及其细胞标志物关系密切.结论 LOXL2与结直肠癌临床预后及免疫浸润水平相关.

目的:应用网络药理学方法,从双黄连方剂中预测具有抗肿瘤活性的组分中药,并通过离体细胞实验与动物实验对其抗CT26结肠癌活性进行初步验证.方法:基于中药系统药理学数据库(Traditional Chinese Medicine Systems Pharmacology Database,TCMSP),《中药活性成分分析手册》,药品生物信息学和化学信息学数据库(DrugBank),信号通路数据库(Reactome)及人类蛋白质参考数据库(Human protein reference database,HPRD),构建蛋白-蛋白相互作用网络(proteinprotein interactions network,PPI)和化合物靶标网络,挖掘化合物的药理作用,发现与PPI网络节点距离较短的化合物,确定组成中药的配伍.以CT26结肠癌细胞为实验材料,结合离体和在体实验初步验证组分中药的抗肿瘤作用.结果:从双黄连组分中预测并发现“黄芩苷、连翘酯苷A和绿原酸”3个化合物配伍具有潜在抗肿瘤活性.验证结果表明,细胞实验中,与空白组比较,双黄连组分中药能显著抑制CT26细胞的增殖和迁移(P＜0.01);动物实验中,与荷瘤模型组比较,双黄连组分中药治疗组小鼠皮下肿瘤体积增长缓慢、肿瘤重量显著降低(P＜0.01).结论:结合数据库挖掘和化合物靶点网络构建、分析等方法从双黄连方剂中发现具有抗肿瘤活性的组分中药“黄芩苷、连翘酯苷A和绿原酸”.经验证,该组分中药具有较好的抗CT26结肠癌活性.

Objective To establish a new-type virtual screening predictive model of Chinese medicinal compounds with anti-fibrosis effects, and to verify the predictive performance of the model.Methods The dimension reduction and characteristic optimization of molecular fingerprints were implemented by using random forest (RF) algorithm and gradient boosting decision tree (GBDT) algorithm.A hybrid model of characteristic optimization-machine learning was established, and optimized characteristics were input into logistic regression (LR) and machine learning algorithm of artificial neural network (ANN) for training.Precision, recall rate and F1 value were used for reviewing the performances of various model combinations.The virtual screening predictive model of Chinese medicinal compounds with anti-fibrosis effect was determined according to results of model performance reviewing.The predictive results of anti-fibrosis activity of Chinese medicinal compounds were compared between the virtual screening predictive model and molecular docking model for further verifying the predictive efficiency of the virtual screening predictive model.Results The precision of RF model was 0.76, recall rate was 0.75 and F1 value was 0.74 (AUC=0.818).The precision that of GBDT model was 0.76, recall rate was 0.74 and F1 value was 0.72 (AUC=0.829).The precision of ANN model was 0.75, racall rate was 0.75 and F1 value was 0.75 (AUC=0.802) , and that of model of RF+LR was 0.77, recall rate was 0.76 and F1 value was 0.75 (AUC=0.840).The precision of model of RF+LR was 0.74, recall rate was 0.84 and F1 value was 0.79 (AUC=0.850) , and that of model of GBDT+LR was 0.80, recall rate was 0.80 and F1 value was 0.79 (AUC=0.872).The precision of model of GBDT+ANN was 0.73, recall rate was 0.91 and F1 value was 0.81 (AUC=0.837).The results of molecular docking activities of Chinese medicinal compounds including curcumin, glycyrrhizic acid, hydro-xysafflor yellow A, emodine and gypenoside were accordance with the predictive results of the virtual screening predictive model.Conclusion The model based on RF+LR is better than the models established based on other methods.The virtual screening predictive model has good performance in prediction of Chinese medicinal compounds through comparing with molecular docking model.The method has feature of highthroughput screening and can make up the shortage of compound screening efficiency in molecular docking.It provides a new way for virtual screening prediction of Chinese medicinal compounds with anti-fibrosis effects.

商宪敏教授认为慢性支气管炎以肺脾肾虚、功能失调为本,痰饮、外邪为标,病性往往是虚实错杂,本虚标实.在治疗上,标本兼顾,攻补兼施;注重辨分期、辨寒热、辨诱因、辨痰性、辨脏腑治疗,注意辨证辨病相结合,临床取得了较好的疗效.

目的 以体外培养的肥大细胞为实验材料,考察注射用双黄连诱发肥大细胞脱颗粒早期的生物学效应,初步探讨中药注射剂安全性评价的质控指标.方法 肥大细胞为RBL-2H3细胞株(ATCC-6562),注射用双黄连购自哈药集团中药二厂(Z20043425);细胞活力检测采用MTT法,β-氨基己糖苷酶检测采用ELISA法;采用扫描电镜观察肥大细胞形态,用激光共聚焦显微镜检测活细胞内钙离子浓度.结果 注射用双黄连细胞毒性剂量及致类过敏剂量为≥0.08 mg/mL;肥大细胞脱颗粒反应出现在药物作用早期(≤1分钟);较低浓度注射用双黄连诱发肥大细胞内钙离子浓度峰值出现在4分钟左右.结论 注射用双黄连作用肥大细胞早期,可激活Ca2+相关信号通路、启动肥大细胞脱颗粒反应、大量释放β-氨基己糖苷酶.该结果提示:基于RBL-2H3细胞的Ca2+浓度及致敏介质检测平台有望成为中药注射剂安全性评价的质控指标之一,具有重要应用前景.

One of the histopathological features of Chronic kidney disease (CKD) is renal fibrosis which is accompanied by the deposition of extracellular matrix (ECM).MicroRNAs are short non-coding RNAs that regulate most of important processes about renal physiological functions and homeostatsis.miR-29s are involved in the pathogenesis of fibrosis by regulating ECM production and deposition,and epithelial-mesenchymal transition (EMT).In this review,we describe interactions of miR-29s with multiple pro-fibrotic and inflammatory pathways and miR-vs as a promising therapeutic reagent or target for the treatment of renal fibrosis.We also review the mechanism of microRNA-29s associated with renal fibrosis.

目的 观察补肾活血方对去卵巢大鼠子宫的雌激素受体及周期蛋白(Cyclin)B表达的影响,探讨其治疗围绝经期综合征的作用机制.方法 选择SD雌性大鼠54只,随机分为假手术组、模型组、雌二醇组、补肾活血方组、联合用药组、补肾活血方+假手术组,每组9只.以双侧卵巢切除法建立大鼠围绝经期模型.各组灌胃给药40 d后取血清和子宫称重;酶联免疫吸附(ELISA)法检测血清中雌二醇(E2)、黄体生成素(LH)、卵泡刺激素(FSH)浓度;HE染色观察子宫组织病理改变并测定子宫内膜厚度;免疫组化法观察雌激素受体(ER)α、ERβ在子宫组织中分布与表达情况;蛋白质印迹法(Western blot)法检测子宫组织中周期蛋白(Cyclin B)蛋白表达.结果 补肾活血方治疗40 d后,与模型组比较,血清LH、FSH明显降低(P＜0.05或P＜0.01),血清E2水平升高(P＜0.05);同时,子宫脏器指数、子宫内膜厚度、子宫组织中ERα、ERβ和Cyclin B蛋白表达均明显升高(P＜0.05),与雌激素组呈现相似的作用,但作用强度均低于雌激素;补肾活血方+假手术组大鼠各项指标与假手术组比较无明显差异.结论 补肾活血方具有雌激素样作用,但作用强度弱于雌激素,对正常大鼠子宫组织无显著影响.