丁元庆从三阳论治痤疮经验

丁元庆认为痤疮属于阳证而好发于三阳经循行之处,三阳气机郁闭,郁而化热化火,火热内结是痤疮发病的病机关键,热结、瘀阻、滞毒于三阳经脉,外蒸而结于肌肤是痤疮的病理基础。临证从三阳论治,针对郁热、火热、湿热、毒热病机,太阳痤疮用麻黄连翘赤小豆汤、翘荷汤、泻心汤、黄连解毒汤化裁;阳明痤疮用茵陈蒿汤、葛根芩连汤、桃核承气汤或茵陈蒿汤合桃核承气汤、三物白散或小陷胸汤、清胃散合升麻鳖甲汤化裁;少阳痤疮用柴胡陷胸汤、柴胡桂枝汤、桂枝茯苓丸合三物白散化裁。     

全胸腔镜与开胸三尖瓣手术后患者炎症反应及心肌损伤情况的比较

目的 全胸腔镜三尖瓣手术是一种微创治疗方式,本次研究重点比较患者接受全胸腔镜及开胸三尖瓣手术后炎症反应和心肌损伤标记物的变化趋势。 方法 回顾性分析2018年1月至2019年4月前来我院手术的88例三尖瓣疾病患者[男53例,年龄（49 ± 19）岁,左室射血分数（63 ± 6）%],其中56例患者接受全胸腔镜手术（三尖瓣成形术50例）,32例患者接受传统开胸手术,全部患者均接受单纯单瓣膜手术。我们将白细胞和C反应蛋白水平作为全身炎症反应的指标,将乳酸脱氢酶、肌酸激酶、肌酸激酶同工酶、天冬氨酸转氨酶和肌钙蛋白水平等作为心肌损伤的标志物。 结果 两组患者入院时各项指标无统计学差异,全身炎症反应于术后第2日达到高峰期,随后逐渐减少。腔镜组患者的炎症指标及心肌损伤标志物水平显著低于开胸组。 结论 与传统开胸三尖瓣手术相比,全胸腔镜三尖瓣手术能明显减少术后心肌损伤和全身炎症反应。     

靶向AKT1和COX-2的RNAi抑制U251胶质瘤细胞侵袭的体外实验

目的应用RNA干扰(RNA interference,RNAi)技术敲低恶性胶质瘤细胞系U251中AKT1和COX-2表达后,在体外对细胞侵袭转移的抑制作用,初步探讨其可能的作用机制。方法构建重组腺病毒载体rAd5-A-C同时搭载靶向AKT1和COX-2的shRNA干扰序列,将其转染至恶性胶质瘤细胞系U251细胞。Realtime PCR检测RNAi后目的基因mRNA的表达水平,Western Blot检测目的基因及MMP-2、MMP-9、TIMP-2的表达情况。酶联免疫吸附试验检测转染前后分泌到细胞外的MMP-2和MMP-9的浓度变化。划痕实验和Transwell实验评价肿瘤细胞转染前后的细胞侵袭能力的变化。结果rAd5-A-C转染组AKT1和COX-2的mRNA和蛋白表达均明显抑制;MMP-2、MMP-9表达下调,TIMP-2表达则上调。ELISA检测胞外MMP-2、MMP-9浓度明显减低;划痕实验显示细胞转移运动能力明显减弱,Transwell体外侵袭实验结果显示穿过细胞数明显减低(P0.001)。结论重组腺病毒介导的RNAi技术可以序列特异性地抑制U251细胞AKT1和COX-2的表达,在体外对U251细胞侵袭转移产生明显抑制作用,可能成为恶性胶质瘤治疗的新策略。     

反义miR-21通过RECK抑制胶质瘤细胞侵袭性生长的体内外研究

目的 探讨胶质瘤细胞LN229中RECK作为miR-21的调控靶点,在胶质瘤侵袭性生长中的作用.方法 将反义miR-21(AS-miR-21)寡核苷酸转染至人脑胶质瘤细胞LN229中.Real-time PCR检测LN229细胞中miR-21的表达量.荧光素酶实验检测miR-21对RECK的调控关系.Transwell实验评价LN229细胞侵袭能力的变化,应用Western blot检测细胞内MMP2/9和RECK蛋白水平的变化,ELISA实验检测培养基中活性MMP2/9的表达量,动物实验评价体内条件下肿瘤侵袭性的变化.结果 Real-time PCR显示转染组中miR-21的表达量与对照组相比下调60%.荧光素酶实验证明RECK是miR-21的靶点.Transwell实验证实胶质瘤细胞侵袭能力下降,Western blot和ELISA实验证实MMP2/9表达降低,动物实验及免疫荧光反映肿瘤侵袭性生长受抑制.结论 反义miR-21通过上调RECK的表达而抑制恶性胶质瘤细胞的侵袭性生长.

Abstract：

Objective To investigate the regulation of miR - 21 on invasion growth of human glioma cells by RECK.Method The human glioma LN229 cells were transfected with AS - miR - 21 or scrambled sequences by Lipofectamine2000.Real time PCR was conducted to detect the expression of miR-21.Luciferase experiment was performed to detect the relationship between miR-21 and RECK.The expression of RECK was evaluated by Western blot.The invasion ability was evaluated by transwell assay and subcutaneous models.Western Blot, ELISA and immunofluorescence were used to estimate the changes of MMP2/9.Results The expression of miR - 21 in LN229 cells decreased after transfection with AS-miR-21. It was proved that RECK was a direct target of miR -21 by luciferase experiment.Meanwhile, the high expression of RECK protein in AS - miR -21 group conformed its important function in this mechanism.Transwell assay demonstrated decreased invasion capability of LN229 cell lines transfected with AS- miR- 21.Western blot, ELISA, and immunofluorescence demonstrated the levels of MMP2/9 were down -regulated in AS -miR -21 group compared with control and scrambled group.Conclusions AS - miR -21 could depress the invasion of glioma cells owing to up - regulating the level of RECK which could inhibit MMP2/9 activities both in vitro and vivo.     

反义miR-30a-5p抑制脑胶质瘤生长的体内实验研究

ObjectiveTo study the inhibitory effect of knocking down miR-30a-5p on the U87 human glioma xenograft growth and its possible mechanism. MethodsNude mice bearing subcutaneous U87 human glioblastoma were established and treated with miR-30a-5p antisense oligonucleotides (AS-miR-30a-5p) subcutaneous injection. Tumor size was measured every other day until the observation period ended. Researchers executed the animals after the treatment, stripped tumor tissues and extracted RNA and protein. Real-time PCR were conducted to detect the expression of miR-30a-5p. The histopathological characteristics and proliferation and apoptosis biological characters (including SEPT7, PCNA, cyclinD1, MMP-2, apoptosis related factor P53, bcl-2 and caspase3)were evaluated by HE and immunohistochemical staining, Western blot analysis respectively, and the cell apoptosis was detected by TUNEL method.ResultsIn AS-miR-30a-5p treated group, the tumor growth was delayed and the final tumor volume was smaller than that in the control and scr-ODN treated group (F=7.167, P＜0.05), and the expression of miR-30a-5p was knocked down. The expression of PCNA、cyclinD1 were significantly down-regulated while P53、SEPT7 and caspase3 up-regulated. Apoptotic index was increased significantly. ConclusionAs-miR-30a-5p suppresses the growth of U87 human gliomas xenografts significantly. Malignant phenotype of tumors are reversed to a considerable degree. Therefore, miR-30a-5p can be a candidate for targeted therapy of human glioma.     

益气化痰方降脂及抗凝作用的实验研究

益气化痰方以治疗“中风不语，痰蒙清窍”的古代名方“涤痰汤”为基础化裁而成。研究以实验性高脂血症家兔为对象，观察了益气化痰方的降脂作用及其对凝血功能的影响，结果表明，益气化痰方能明显降低高脂血症家兔的血浆胆固醇（ＴＨ）、甘油三酯（ＴＧ），并使高密度脂蛋白——胆固醇含量增高（ＨＤＬ－ＣＨ）；延长血浆凝血酶原时间（ＰＴ）、凝血酶时间（ＴＴ）、白陶土部分凝血活酶时间（ＫＰＴＴ），显示出对凝血过程的抑制。由此提示，益气化痰方既能降低血脂，又可抗凝，认为这是其治疗心脑血管疾病，祛除痰浊，消除瘀滞，畅通血脉的作用机制之一。