扶正化瘀方含药血清对肝星状细胞activinA/smad信号通路活化的影响

目的　探讨扶正化瘀方含药血清对肝星状细胞（HSC）激活素A（activinA）/smad信号通路活化的影响。方法　20只SD大鼠按照随机数字表法分为对照组及扶正化瘀（Fzhy）-低、中、高剂量含药血清组，每组5只，分别以蒸馏水，0.75、1.5、3.0 g/kg扶正化瘀溶液（扶正化瘀胶囊药物粉末与蒸馏水配制）灌胃1次/d，连续3 d，取血制备空白血清（对照组）和含药血清。以大鼠HSC-T6细胞为研究对象，分别用5%、10%、20%体积分数的各组含药血清培养细胞。CCK-8检测细胞存活率，选取细胞存活率在50%左右的体积分数重新分为对照组及Fzhy-低、中、高剂量组。流式细胞术检测细胞周期及凋亡率、线粒体膜电位变化和活性氧（ROS）水平；实时荧光定量聚合酶链反应检测细胞中activinA、smad3、samd7、核因子（NF）-κB的mRNA水平；蛋白质印迹法检测细胞中activinⅡA受体（ActRⅡA）、smad3、NF-κB p65、胱天蛋白酶（caspase）-3的蛋白水平。结果　各扶正化瘀含药血清组的细胞存活率均低于对照组（P＜0.05），选择体积分数为10%的含药血清进行后续实验。对照组和各扶正化瘀方含药血清组细胞周期和凋亡率差异均无统计学意义。对照组及Fzhy-低、中、高剂量组细胞内ROS水平及线粒体膜电位水平降低比例逐次升高（P＜0.05）。与对照组比较，Fzhy-中、高剂量组smad3 mRNA表达水平降低，smad7 mRNA升高，Fzhy-低、中、高剂量组NF-κB、activinA mRNA，smad3、NF-κB p65、ActRⅡA蛋白表达水平降低，Fzhy-低剂量组、中剂量组caspase-3蛋白表达水平升高（P＜0.05）；与Fzhy-低剂量组相比，Fzhy-中、高剂量组smad3 mRNA表达水平降低，Fzhy-中剂量组activinA及smad7 mRNA升高（P＜0.05）。结论　扶正化瘀方含药血清可通过影响HSC-T6细胞增殖、参与细胞的氧化应激以及调节细胞activinA/smad信号转导通路来实现抗纤维化作用。     

扶正解毒抗癌方治疗晚期非小细胞肺癌的临床效果

目的:探讨扶正解毒抗癌方对晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的疗效及对患者化疗后骨髓抑制的影响。方法:选取2016年1月至2019年3月滁州市第一人民医院收治的NSCLC患者100例,按照随机数字表法分为对照组和研究组,每组50例。对照组患者采用常规化疗,研究组患者在对照组的基础上加用扶正解毒抗癌方,两组患者连续治疗2个化疗周期。观察两组患者的临床疗效,分别于治疗前后测定白细胞计数、血小板计数及血红蛋白水平,比较两组患者治疗后骨髓抑制发生情况和卡氏(Karnofsky,KPS)评分的差异。结果:研究组患者的总有效率为62.00%(31/50),明显高于对照组的36.00%(18/50),差异有统计学意义(P<0.05)。治疗后,两组患者白细胞计数、血小板计数及血红蛋白水平均明显低于治疗前,但研究组患者上述指标水平明显高于对照组,差异均有统计学意义(P<0.05)。研究组患者骨髓抑制发生率为38.00%(19/50),明显低于对照组的68.00%(34/50),差异有统计学意义(P<0.05)。治疗后,研究组患者KPS评分提高率和稳定率分别为34.00%(17/50)、46.00%(23/50),明显优于对照组的14.00%(7/50)、16.00%(8/50),差异均有统计学意义(P<0.05)。结论:扶正解毒抗癌方联合常规化疗治疗晚期NSCLC的疗效显著,能够减轻患者化疗后骨髓抑制情况,提高患者生活质量。     

祛寒逐风合剂联合西医常规疗法治疗膝骨关节炎风寒痹阻证临床研究

目的观察祛寒逐风合剂联合西医常规疗法对膝骨关节炎风寒痹阻证的临床疗效,以及对关节症状及相关实验室指标的影响。方法采用随机数字表法将94例膝骨关节炎风寒痹阻证患者分为观察组和对照组各47例。对照组采用常规药物疗法+康复训练,观察组在对照组基础上予祛寒逐风合剂,每次50m L,每日3次,口服。2组均连续治疗2周。比较2组临床疗效,观察2组治疗前后美国西大略湖和麦克马斯特大学骨关节炎指数(WOMAC)评分、视觉模拟评分法(VAS)评分、压痛指数、中医症状评分,白细胞介素(IL)-1β、IL-6、血管内皮生长因子(VEGF)、肿瘤坏死因子(TNF)-α、骨钙素、抗酒石酸盐酸性磷酸酶异构体(TRACP)-5b、骨特异性碱性磷酸酶(BALP)、纤维蛋白原、红细胞沉降率和红细胞聚集指数,对2组进行安全性评价。结果观察组总有效率为89.36%(42/47),对照组为74.47%(35/47),观察组明显优于对照组(P<0.05)。与本组治疗前比较,2组治疗后WOMAC评分、VAS评分、压痛指数、中医症状评分明显下降(P<0.05);2组治疗后比较,观察组WOMAC评分、VAS评分、压痛指数、中医症状评分明显低于对照组(P<0.05)。与本组治疗前比较,2组治疗后IL-1β、IL-6、TNF-α、VEGF、TRACP-5b水平明显下降,骨钙素、BALP水平明显升高(P<0.05);2组治疗后比较,观察组IL-1β、IL-6、TNF-α、VEGF、TRACP-5b水平明显低于对照组,骨钙素、BALP水平明显高于对照组(P<0.05)。与本组治疗前比较,观察组治疗后纤维蛋白原、红细胞沉降率、红细胞聚集指数明显下降(P<0.05);2组治疗后比较,观察组血液流变学各项指标明显低于对照组(P<0.05)。2组均未发生不良反应。结论祛寒逐风合剂联合西医常规疗法治疗膝骨关节炎风寒痹阻证患者疗效较好,可明显改善关节症状及相关实验室指标。     

Microhaplotypes in forensic genetics

Microhaplotype loci (microhaps, MHs) are a novel type of molecular marker of less than 300 nucleotides, defined by two or more closely linked SNPs associated in multiple allelic combinations. The value of these markers is enhanced by massively parallel sequencing (MPS), which allows the sequencing of both parental haplotypes at each of the many multiplexed loci. This review describes the features of these multi-SNP markers and documents their value in forensic genetics, focusing on individualization, biogeographic ancestry inference, and mixture deconvolution. Foreseeable applications also include missing person identification, relationship testing, and medical diagnostic applications. The technique is not restricted to humans.     

扶正抑癌汤联合腹腔镜辅助小切口根治手术治疗胃癌临床研究

目的:探讨扶正抑癌汤联合腹腔镜辅助小切口根治手术治疗胃癌的效果及对患者免疫功能的影响。方法:选择80例胃癌患者,按随机数字表法分为观察组和对照组各40例。对照组行腹腔镜辅助小切口根治手术治疗,观察组予以扶正抑癌汤联合腹腔镜辅助小切口根治术手术治疗,比较2组患者临床疗效及免疫功能。结果:治疗前,2组中医症状评分比较,差异无统计学意义(P>0.05)。治疗后,2组中医症状评分较治疗前下降(P<0.05),且观察组中医症状评分低于对照组(P<0.05)。治疗前,2组癌胚抗原(CEA)、糖类抗原199 (CA199)、糖类抗原125 (CA125)水平比较,差异无统计学意义(P>0.05)。治疗后,2组CEA、CA199、CA125水平较治疗前下降(P<0.05),且观察组CEA、CA199、CA125水平低于对照组(P<0.05)。治疗前,2组CD4^+、CD8^+、CD4^+/CD8^+水平比较,差异无统计学意义(P>0.05)。治疗后,2组CD4^+、CD8^+、CD4^+/CD8^+水平较治疗前升高(P<0.05),且观察组CD4^+、CD8^+、CD4^+/CD8^+水平高于对照组(P<0.05)。结论:扶正抑癌汤联合腹腔镜辅助小切口根治手术治疗胃癌效果显著,可有效降低相关肿瘤标志物水平,并改善患者免疫功能。     

基于网络药理学挖掘溃疡性结肠炎“扶正”治则的潜在机制

目的:通过数据库检索与网络药理学方法探讨溃疡性结肠炎"扶正"治则的潜在机制。方法:从中医E百数据库检索溃疡性结肠炎相关方剂,检索方式为"主治肠澼/久痢/痢疾",筛选所检索到的处方并进行药物种类、用药频次统计分析,选择溃疡性结肠炎治疗的常用扶正药物。将选择后的扶正药物输入中药系统药理学分析平台检索所有化学成分和作用靶点,构建化合物-靶点相互作用网络图;通过Digsee数据库筛选溃疡性结肠炎相关靶点,构建疾病-靶点相互作用网络图;筛选两个网络图的核心靶点,利用DAVID工具对核心靶点进行富集分析。结果:数据库共检索到溃疡性结肠炎相关方剂731首,根据纳入、排除标准筛选后获得处方304首,中药220味,经用药频次统计后,以甘草、人参、白术为溃疡性结肠炎扶正治则的代表药物。基于药代动力学ADME参数对上述药物的化学成分进行筛选和靶标预测,共得到136个蛋白靶点;将化合物-靶点、疾病-靶点的蛋白质—蛋白质相互作用网络图合并,以"节点连接度""节点介度"和"节点紧密度"为评价标准,筛选共有靶点129个;对入选靶标进行京都基因与基因组百科全书(KEGG)富集分析,以P值筛选出前20条通路在溃疡性结肠炎方面具有作用。结论:扶正药物治疗溃疡性结肠炎作用的发挥是多靶点、多通路的整体效应,挖掘了溃疡性结肠炎扶正治则的潜在机制,为进一步实验研究提供了理论基础。     

Qingjie Fuzheng Granules regulates cancer cell proliferation,apoptosis and tumor angiogenesis in colorectal cancer xenograft mice via Sonic Hedgehog pathway

BackgroundSonic Hedgehog (SHh) signaling pathway plays a critical role in cell proliferation, apoptosis, and tumor angiogenesis in various types of malignancies including colorectal cancer (CRC). Qingjie Fuzheng Granules (QFG) is a traditional Chinese medicinal formula, which has been clinically used in various cancer treatments, including CRC. In this study, we explored the potential molecular mechanisms of QFG treatment effects on CRC via the SHh pathway.MethodsA CRC HCT-116 xenograft mouse model was utilized for all experiments. Mice were treated with intra-gastric administration of 1 g/kg of QFG or saline 6 days a week for 28 days (4 weeks). Body weight, length and shortest diameter of the tumor were measured every 3 days. At the end of the treatment, the tumor weight was measured. TUNEL staining assays were used to detect tumor apoptosis. Western blot and immunohistochemistry (IHC) assays were used to detect the expression of relative proteins.ResultsIn our results, QFG inhibited the increase of tumor volume and weight, and exhibited no impact on mouse body weight. Furthermore, QFG significantly decreased the expression of SHh, Smo and Gli proteins, indicating the action of SHh signaling. Consequently, the expression of pro-proliferative survivin, Ki-67, Cyclin-D1 and CDK4 were decreased and expression of anti-proliferative p21 was increased. The pro-apoptotic Bax/Bcl-2 ratio, cle-caspase-3 and TUNEL-positive cell percentage in tumor tissues were increased. Meanwhile, the pro-angiogenic VEGF-A and VEGFR-2 expression was down-regulated.ConclusionsQFG inhibited CRC cell proliferation and promoted CRC cell apoptosis and tumor angiogenesis in vivo through the suppression of SHh pathway, suggesting that QFG could be a potential therapeutic drug for CRC.     

降黄合剂Ⅱ号对肝内胆汁淤积大鼠肝损伤及肝组织中NTCP和BSEP表达的影响

目的探讨降黄合剂Ⅱ号对复合因素诱导的阴黄证肝内胆汁淤积模型大鼠肝损伤及钠-牛黄胆酸盐共转运多肽(NTCP)、胆盐输出泵(BSEP)表达的影响。方法将60只SD雄性大鼠随机分为正常组、模型组、降黄合剂Ⅱ号高剂量组、降黄合剂Ⅱ号中剂量组、降黄合剂Ⅱ号低剂量组,每组12只。正常组大鼠予常规饲养;其余各组大鼠造模阶段施以寒湿条件、大黄煎剂灌胃及高糖高脂饮食,造模3周,第22天时模型组及降黄合剂Ⅱ号各组给予0.5%的ANIT 20 mg/100 g灌胃1次,正常组予以相同体积生理盐水灌胃,建立阴黄证肝内胆汁淤积大鼠模型。降黄合剂Ⅱ号各组于造模第8天开始每天下午给相应剂量的降黄合剂Ⅱ号灌胃,一直到造模后14 d,共用药28 d,正常组和模型组均予相同剂量生理盐水灌胃。观察各组大鼠一般状态,检测干预结束后各组大鼠血清碱性磷酸酶(ALP)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBil)水平,镜下观察各组大鼠肝组织病理变化,采用Western blot法检测各组大鼠肝组织中BSEP、NTCP蛋白表达水平。结果正常组大鼠饮食量、活动量、被毛、体质量、排便排尿情况均正常;模型组大鼠形体均较消瘦,饮食量减少,大便增多,尿液明显变黄、增多,被毛枯黄、粗糙且易脱毛,精神状态不佳;降黄合剂Ⅱ号各组大鼠干预后情况均较模型组明显好转,以大剂量组最为明显。模型组大鼠血清ALP、ALT、AST、TBil水平均明显高于正常组(P均<0.05);降黄合剂Ⅱ号大剂量组和降黄合剂Ⅱ号中剂量组上述指标水平均明显低于模型组(P均<0.05),且降黄合剂Ⅱ号大剂量组各指标水平均明显低于降黄合剂Ⅱ号小剂量组(P均<0.05),其中血清ALP、ALT、TBil水平明显低于降黄合剂Ⅱ号中剂量组(P均<0.05)。HE染色显示降黄合剂Ⅱ号大、中剂量组肝组织较模型组中炎细胞浸润减少,毛细胆管淤堵减轻,肝细胞肿胀缓解,细胞核清晰,形态较完整,可见少量炎性细胞。模型组大鼠肝组织中NTCP、BSEP相对表达量均明显低于正常组(P均<0.05);降黄合剂Ⅱ号大剂量组和降黄合剂Ⅱ号中剂量组大鼠肝组织中NTCP、BSEP相对表达量均明显高于模型组(P均<0.05),降黄合剂Ⅱ号小剂量组与模型组比较差异均无统计学意义(P均>0.05)。结论降黄合剂Ⅱ号可改善阴黄证肝内胆汁淤积模型大鼠肝损伤,机制可能与上调NTCP和BSEP表达有关。     

Distinguishing mode of action of compounds inducing craniofacial malformations in zebrafish embryos to support dose-response modeling in combined exposures

Knowledge on mode-of-action (MOA) is required to understand toxicological effects of compounds, notably in the context of risk assessment of mixtures. Such information is generally scarce, and often complicated by the existence of multiple MOAs per compound. Here, MOAs related to developmental craniofacial malformations were derived from literature, and assembled in a MOA network. A selection of gene expression markers was based on these MOAs. Next, these markers were verified by qPCR in zebrafish embryos, after exposure to reference compounds. These were: triazoles for inhibition of retinoic acid (RA) metabolism, AM580 and CD3254 for selective activation of respectively RA-receptor (RAR) and retinoid-X-receptor (RXR), dithiocarbamates for inhibition of lysyl oxidase, TCDD for activation of the aryl-hydrocarbon-receptor (AhR), VPA for inhibition of histone deacetylase (HDAC), and PFOS for activation of peroxisome proliferator-activated receptor-alpha (PPARα). Next, marker gene profiles for these reference compounds were used to map the profiles of test compounds to known MOAs. In this way, 2,4-dinitrophenol matched with the TCDD and RAR profiles, boric acid with RAR, endosulfan with PFOS, fenpropimorph with dithiocarbamates, PCB126 with AhR, and RA with triazoles and RAR profiles. Prochloraz showed no match. Activities of these compounds in ToxCast assays, and in silico analysis of binding affinity to the respective targets showed limited concordance with the marker gene expression profiles, but still confirmed the complex MOA profiles of reference and test compounds. Ultimately, this approach could be used to support modeling of mixture effects based on upfront knowledge of (dis)similarity of MOAs.     

止咳平喘十二味合剂联合信必可都保治疗痰热壅肺型哮喘-慢阻肺重叠综合征疗效观察

<正>哮喘-慢阻肺重叠综合征(ACOS)是一类具有慢性气道疾病症状同时具备哮喘和慢性阻塞性肺病临床特征的疾病。目前尚缺乏满意的治疗方案,以吸入糖皮质激素联合长效支气管扩张剂为基础的治疗方案虽能在一定程度上改善临床症状,但长期应用存在依从性差、不良反应多等缺点。ACOS归属中医"哮病""喘症""肺胀"等范畴,其证型构成复杂多样,其