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。     

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.     

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。     

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.     

敲低miR-19a、miR-19b抑制人脑胶质瘤细胞生长的体外研究

目的 探究敲低microRNA(miR)-19a和miR-19b表达对人脑胶质瘤细胞系SNB19细胞生物学特征的影响.方法 常规培养SNB19细胞,脂质体介导miR-19a、miR-19b抑制物转染SNB19细胞,同时设未转染组(对照组)、无义序列转染组和miR-19a+miR-19b抑制物转染组.应用RT-PCR检测转染后细胞miR-19a、miR-19b的表达,MTT、流式细胞术分别检测转染后细胞增殖能力和细胞周期的变化,Transwell实验检测转染后细胞的侵袭能力.结果 与对照组和无义序列转染组比较,抑制物转染组细胞miR-19a和miR-19b的表达、增殖和侵袭能力均降低,差异有统计学意义(P＜0.05),而且miR-19a+miR-19b抑制物转染组细胞miR-19a和miR-19b的表达、转染后2、3、4、5 d细胞的增殖能力、侵袭能力均低于miR-19a、miR-19b抑制物转染组,差异有统计学意义(P＜0.05);抑制物转染组较对照组和无义序列转染组细胞周期延迟,且miR-19a+miR-19b抑制物转染组较miR-19a、miR-19b抑制物单独转染组细胞延迟更为明显.结论 miR-19a和miR-19b可能是癌微RNA,有望作为人脑胶质瘤基因治疗的侯选靶点.

Abstract：

objective To investigate the effects of knocking down of miR-19a and miR-19b on the biological characteristics of SNB19 glioblastoma cells. Methods Oligonucleotides inhibitor of miR-19a and miR-19b (miR-19a inhibitor or miR-19b inhibitor) mediated by lipofectamine2000 were transfected to SNB19 cells to knock down miR-19a and miR-19b; control group (without transfection),group D (performing transfection with nonsense sequence) and group E (performing transfection with both miR-19a inhibitor and miR-19b inhibitor) were established. Real time PCR was conducted to detect the expressions ofmiR-19a and miR-19b in these groups after the transfection. The cell proliferation rate and cell cycle kinetics were detected by 3-(4, 5-Dime- -thylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively; the cell invasive ability was evaluated by Transwell assay.Results As compared with those in control group and group D, the expressions of miR-19a and miR-19b, proliferation activity and invasive ability of cells in the miR-19a/19b inhibitor transfected cells (group A/B) were significantly reduced (P＜0.05). The expressions of miR-19a and miR-19b and the proliferation activity and invasive ability of cells 2, 3, 4 and 5 d after the transfection in group E were significantly reduced as compared with those in group A/B (P＜0.05). Delayed cell cycle in group A/B and group E was noted as compared with that in control group and group D; and group E enjoyed more obviously delayed eell cycle than group A/B (P＜0.05). Conclusion MiR-19a and miR-19b might be oncomiRs, and may be candidate target miRNAs for gene therapy of glioma.     

反义miRNA-221/222上调p27kip1抑制胶质瘤细胞生长的体外研究

目的 探讨敲低miRNA-221/222表达上调p27kip1抑制U251人脑胶质瘤细胞株生长的效果及机制.方法 脂质体共转染反义miRNA-221/222下调U251人脑胶质瘤细胞株miRNA-221、miRNA-222的表达.使用Northern blot方法 鉴定转染后U251细胞miRNA-221、miRNA-222表达水平下调;MTT法评价反义miRNA-221/222抑制U251细胞生长效果;流式细胞术检测转染后U251细胞周期分布;Western blot分析p27kip1蛋白的表达变化.结果 Northern blot显示反义miRNA-221/222共转染组使miRNA-221、miRNA-222的表达水平明显下降.反义miRNA-221转染组仅使miRNA-221的表达水平明显下降,反义miRNA-222转染组仅使miRNA-222的表达水平明显下降.转染无意序列组及对照组的miRNA-221、miRNA-222表达水平没有改变.MTT结果 显示反义miRNA-221/222共转染组肿瘤细胞生长速度小于对照组、转染无意序列组、转染反义miRNA-221组、转染反义miRNA-222组.流式细胞术检测可见反义miRNA-221/222共转染组细胞周期存在G0/G1期阻滞且明显高于其他各组.Western blot显示反义miRNA-221/222共转染组的p27kip1蛋白表达明显上调.结论 反义miRNA-221/222通过上调p27kip1蛋白表达来抑制胶质瘤细胞U251的生长.     

BMSCs作为HSV-tk载体联合CX43增强旁效应治疗胶质瘤的实验研究

目的 以骨髓间充质干细胞(BMSCs)为自杀基因HSV-tk载体,联合连接蛋白CX43介导的旁效应,对以BMSCs肿瘤追踪特性为基础的胶质瘤进行治疗研究.方法 以CX43质粒转染C6胶质瘤细胞,以腺病毒介导HSV-tk转染BMSCs;建立C6胶质瘤模型,实验动物分为4组:C6对照组、CX43-C6组、tk-BMSCs/C6/GCV治疗组和tk-BMSCs/CX43-C6/GCV治疗组.观察动物生存期,MRI监测肿瘤体积;脑切片行PCNA、原位凋亡检测肿瘤中央与边缘部位细胞增殖与凋亡情况.结果 CX43组、tk/GCV组生存期长于对照组,二者联合治疗组生存期最长,MRI示部分肿瘤消失;肿瘤内部与边缘细胞增殖活性下降,凋亡明显.结论 tk-BMSCs/GCV体系可有效杀伤侵袭瘤细胞在内的胶质瘤灶,连接蛋白CX43可增强此治疗效果.     

肾燥不藏机理与证治举隅

闭藏是肾的主要生理功能。闭藏之职是由肾中阴阳相互协调共同完成的，其中阴主静，是肾主闭藏的重要生理基础；阴主滋润，阴虚则燥，导致闭藏失职，称为肾燥不藏，由此可导致诸多的病证。因此，滋肾润燥，是治疗肾燥不藏的基本方法。     

反义miR-21抑制异种移植U251人脑胶质瘤生长的体内研究

目的 探讨敲低miR-21表达抑制裸鼠荷皮下U251人腩胶质瘤生长的疗效和机制. 方法 原位注射miR-21反义寡聚核苷酸(AS.miR-21)治疗裸鼠皮下荷U251人脑胶质瘤.定时测量肿瘤大小评估原位注射AS-miR-21治疗效果:使用Real time PCR和原位杂交方法鉴定治疗后miR-21表达水平下调;采用HE染色和免疫组织化学染色(PCNA、PTEN、MMPs、和Septins)评价治疗后肿瘤牛物学性状改变;TUNEL法检测肿瘤细胞凋亡. 结果 肿瘤生长曲线显示AS-miR-21治疗组肿瘤生长速度及体积小于空白对照与无义序列治疗组(F=52.458,P=0.000);Real-time PCR法:AS-miR-21治疗组miR-21表达下调为对照组的0.018±0.009:原位杂交显示AS-miR-21治疗组miR-21表达水平较对照与无义序列治疗组下调:组织病理学检测表明AS-miR-21治疗后肿瘤恶性度降低;TUNEL法检测可见AS-miR-21治疗组细胞凋亡数显著高于对照与无义序列治疗组(F=141.021,P=0.000).结论 以miR-21作为靶点抑制异种移植U251人脑胶质瘤生长,miR-21可能通过上调抑癌基因PTEN抑制胶质瘤的生长,可以作为人脑胶质瘤基因治疗的侯选靶点.     

肝火的形成致病机制及证治规律探微

肝火主要成于情志不畅，肝郁化火。肝火即成，动而不居，上熏下灼，内燔外蒸，伤脏腑，损气血，灼津液，扰神明，为害甚广，其证候特点则可分为肝火本证与肝火证两方面。清泻肝火为其治疗大法，处方用药有治肝火与治肝火所犯脏腑两大要点。