Identification of Flt3 internal tandem duplications downstream targets by high-throughput immunoblotting protein array system

The receptor tyrosine kinase Flt3 plays an important role in proliferation and survival of hematopoietic cells. Flt3 is the most frequently mutated gene (20-30%) in cases of acutemyeloid leukemia (AML). The majority of Flt3 mutations are internal tandem duplications (ITD) in the juxtamembrane domain of Flt3 receptor. This mutation results in the constitutive activation of STAT5 and Ras/mitogen-activated protein kinase pathways, leading to the aberrant growth of AML cells. In this study, to better understand the mechanisms of Flt3-ITD to the downstream pathways, a high-throughput immunoblotting protein array system was employed. As a result, c-Jun and c-Raf were markedly induced, suggesting that these factors are functional downstream targets of Flt3-ITD.     

High glucose induces apoptosis in AC16 human cardiomyocytes via macrophage migration inhibitory factor and c‐Jun N‐terminal kinase

1. It is known that high glucose can induce cardiomyocyte apoptosis and that macrophage migration inhibitory factor (MIF) may be involved in the development of diabetes. However, the relationship between high glucose and MIF in diabetic cardiomyopathy remains unclear. 2. In the present study, AC16 human cardiomyocytes were cultured in the presence of 25 mmol/L glucose for 20, 30 and 60 min before being subjected to western blot analyses to determine MIF expression and c‐Jun N‐terminal kinase (JNK) activation. In addition, AC16 cells were pretreated with 2.5 μmol/L SP600125 (a JNK inhibitor), 40 μmol/L (s,r )‐3‐(4‐hydroxyphenyl)‐4,5‐dihydro‐5‐isoxazole acetic acid methyl ester (ISO‐1; an MIF antagonist) or 0.1% dimethylsulphoxide (DMSO; vehicle) for 1 h prior to exposure to 25 mmol/L glucose and culture for 72 h, followed by annexin V–fluorescein isothiocyanate/propidium iodide staining and flow cytometry analysis. Caspase 3 activity and phosphorylation of JNK were also analysed by western blotting. 3. The high concentration of glucose increased expression of endogenous MIF and JNK phosphorylation in AC16 cardiomyocytes. Pretreatment of cells with SP600125 and ISO‐1 reduced glucose‐induced apoptosis and caspase 3 activity. Furthermore, JNK phosphorylation was attenuated by inhibition of endogenous MIF. 4. In conclusion, myocardial cell apoptosis induced by high glucose involves the overexpression of MIF and activation of the JNK signalling pathway. The identification of a high glucose–MIF–JNK pathway will help determine potential new targets in the treatment of diabetic cardiomyopathy.     

香砂六君丸治疗糖尿病胃轻瘫随机平行对照研究

[目的]观察香砂六君丸治疗糖尿病胃轻瘫疗效。[方法]使用随机平行对照方法,将100例住院患者按随机数字表法分为两组。对照组50例多潘立酮,10mg/次,3次/d,口服。治疗组50例香砂六君丸,4.5g/次,3次/d,口服。连续治疗7d为1疗程。观测临床症状、中医证候积分、不良反应。连续治疗4疗程,判定疗效。[结果]治疗组痊愈10例,显效14例,有效23例,无效3例,总有效率94.00%。对照组痊愈5例,显效12例,有效20例,无效13例,总有效率74.00%。治疗组疗效优于对照组(P0.05)。中医证候积分两组均有改善(P0.05),治疗组改善优于对照组(P0.05)。[结论]香砂六君丸治疗糖尿病胃轻瘫效果显著,值得推广。     

杨兆钢老中医临床经验录

杨兆钢教授提出芒针"疏弹趋动,技巧术行"独特的针刺治疗理念,应用芒针,运用中医理论,在临床治愈了很多疑难杂症,现简要阐述马凡氏综合征、癃闭、子宫肌瘤的中医辨证、中医治法、针刺取穴、临床疗效以及体会。     

罗格列酮对OLETF大鼠肺组织TGF-β1／JNK信号转导途径的影响

目的 探讨罗格列酮对自发性2型糖尿病OLETF大鼠肺组织转化生长因子-1β（TGF-β1）／c—Jun氨基末端激酶（JNK）信号转导途径的影响。方法将16只30周龄成模OLETF大鼠分为OLETF组和罗格列酮治疗组（OLETF／L纽）,设LETO大鼠8只为对照组。免疫组织化学分析肺组织TGF—p1、磷酸化（P）-JNK、I型胶原表达。结果与LETO组大鼠相比,OLETF组大鼠肺组织TGF—β1、P—JNK表达（7．4438±0,6397vs1．1844±0．2066、5．7025±0．5530VS1．5875±0．1727）及Ⅰ型胶原的阳性面积比、平均光密度（0．1705±0．0110VS0．0550±0．0048、0．0597±0．0066VS0．0292±0．0019）均显著增加（P〈0．05）。与OLETF组大鼠相比,OLETF／L组大鼠肺组织TGF-β1、PJNK表达（1．8398±0．3591VS7．4438±0．6397、1．7500±0．1690vs5．7025±0．5530）及Ⅰ型胶原的阳性面积比、平均光密度（0．0856±0．0068VS0．1705±0．0110、0．0351±0．0059VS0．0597±0．0066）均显著减少（P〈0．05）。结论罗格列酮可能通过调控TGF-β1／JNK信号转导途径改善糖尿病肺纤维化。     

组成型光形态建成蛋白1介导p53及Jun激酶泛素化通路的研究进展

组成型光形态建成蛋白1(COP1)于多种类型肿瘤中过度表达,在促进细胞增殖、转化和肿瘤进展等多种生物学过程中起重要的作用。COP1与p53呈负调节并参与致癌基因Jun激酶的信号通路。目前COP1表达在人类肿瘤发生、发展中的具体作用仍存在许多争论,通过解析COP1与p53和Jun激酶的相互关联,进一步探讨与肿瘤的侵袭及转移关系。     

The Ethanol Extract from Lonicera japonica Thunb. Regresses Nonalcoholic Steatohepatitis in a Methionine- and Choline-Deficient Diet-Fed Animal Model

Nonalcoholic steatohepatitis (NASH) is characterized as fat accumulation in the hepatic tissue associated with various degrees of inflammation and progressive fibrosis. The potent anti-inflammatory and ethnopharmacological properties of Lonicera japonica Thunb. (Caprifoliaceae) make it an excellent source of novel medicinal targets for the treatment of NASH. The aim of the study was to investigate the effects of L. japonica ethanol extract (LJEE) on NASH in mice. C57BL/6J mice were fed with methionine-choline-deficient diet (MCDD) for eight weeks to promote the development of NASH. After development of the model, the mice were administered LJEE once daily via oral gavage at doses of 100, 200, or 300 mg/kg for another four weeks. Simultaneous treatments with LJEE (300 mg/kg/day) resulted in pronounced improvements in liver steatosis, ballooning degeneration, and inflammation. LJEE prevented MCDD-induced plasma level increases in aspartate aminotransferase and alanine aminotransferase. LJEE significantly reduced hepatic malondialdehyde level and ameliorated hepatic inflammation and fibrosis in MCDD-fed mice, which were associated with down-regulation of cytochrome P450 2E1 suppression of multiple proinflammatory and profibrotic genes. LJEE can prevent hepatic steatosis by reducing hepatic peroxisome acyl-CoA:diacylglycerol acyltransferase 2 expression, as well as by inducing proliferator-activated receptor α expression. In addition, the LJEE treatments caused significant reduction in the phosphorylated form of Jun N-terminal kinase along with an increase in the phosphorylated level of extra cellular signal-regulated kinase 1/2. Our study demonstrated the protective role of LJEE in ameliorating nutritional steatohepatitis.     

杨道文教授治疗小儿过敏性鼻炎-哮喘综合征经验

杨道文教授结合小儿生理特点及小儿CARAS的临床特征,认为主要病机是风邪致病、正气亏虚和气机失调,其中强调风邪致病需注意外风与内风并重,正气亏虚主要表现在脾肺两脏。从治风、补虚、调气三方面组方化裁论治。治风时以辛散治外风,以镇敛治内风;补虚时注重肺脾两脏,且补脾之中兼顾运脾;调气时强调肺胃同降,以降为主,降中寓宣。附典型案例1则,以资验证。     

基于网络药理学探究兰艾双香方治疗细菌感染的机制研究*

目的 旨在通过网络药理学的研究方法寻找兰艾双香方在治疗细菌感染方面存在的潜在的分子机制及相关分子通路,为进一步研究兰艾双香方提供理论依据。方法 通过中药数据分析平台（TCMSP）筛选兰艾双香方的活性成分及对应靶点,对兰艾双香方进行复方成分分析,利用Cytoscape软件构建兰艾双香方治疗细菌感染的“单味药-活性成分-作用靶点”网络,再通过Gene Cards,OMIM和DigSee 3个数据库整合细菌感染的相关靶点,将靶点数据导入jvenn平台做出韦恩图,导入String平台得到PPI,导入Metascape中对兰艾双香方治疗细菌感染的作用靶点进行GO功能富集分析和KEGG通路富集分析,根据KEGG富集得到的通路绘制相关气泡图,将KEGG分析得到的通路导入Cytoscape软件构建“靶点通路”网络。结果 ①筛选得到兰艾双香方共110个药效成分和1 788个作用靶点,关键靶点包括过氧化物酶体增殖物激活受体、血管内皮生长因子A、肿瘤坏死因子和表皮生长因子受体等;②细菌感染相关靶点1 442个;③蛋白互作核心网络共97个蛋白,分析得到4个蛋白模块;④获得2 401个GO生物过程,KEGG通路富集筛选得到324条相关信号通路,通路类型包括癌症的路径、糖尿病并发症中的AGE-RAGE信号通路、乙型肝炎、肿瘤坏死因子信号通路、小细胞肺癌、利什曼病、HIF-1信号通路、军团杆菌病等,这些通路均与细菌感染的发生发展相关;⑤结果证实,兰艾双香方可能通过调节细胞代谢、增殖与凋亡、脂代谢等多种途径达到治疗细菌感染的目的,该目的是多成分、多靶点和多通路相互作用的结果。结论 兰艾双香方在癌症、糖尿病和乙型肝炎疾病进展过程中并发细菌感染的治疗可能有更好的疗效。