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.     

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     

“Real life” polycyclic aromatic hydrocarbon (PAH) mixtures modulate hCG,hPL and hPLGF levels and disrupt the physiological ratio of MMP-2 to MMP-9 and VEGF expression in human placenta cell lines

Using JEG-3 and BeWo cells, we examined the effect of “real life” mixtures of polycyclic aromatic hydrocarbons (PAHs), at doses reported in maternal blood (Mix I) and in placental tissue (Mix II), on human chorionic gonadotropin (hCG), placental lactogen (hPL) and placental growth factor (hPLGF) secretion, protein expression and immunolocalization. Additionally, the action of PAH mixtures on basal and hormone-stimulated matrix metalloproteinase-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) protein expression was evaluated. Under basal conditions, the PAH mixtures increased hCG and decreased hPLGF levels in both cell lines, while hPL expression was stimulated in JEG-3 and inhibited in BeWo. There was no effect on the MMP-2/MMP-9 ratio or VEGF expression. In hormone-stimulated cells, PAH mixtures changed the MMP-2/MMP-9 ratio in JEG-3 cells in favor of MMP-9, while in BeWo MMP-2 was favored. The effect on VEGF expression was cell specific and dependent on the mixture. In hCG-treated cells, only Mix II inhibited VEGF expression in JEG-3 cells. Neither PAH mixtures affected this protein in BeWo cells. In hPL-treated cells, Mix I had a stimulatory effect in JEG-3 cells, while Mix II exerted an inhibitory effect in BeWo cells. In hPLGF-treated cells, Mix II decreased in JEG-3 cells, but in BeWo cells, both mixtures increased VEGF expression. Considering that the evaluated protein hormones play crucial roles in angiogenesis and neovascularization in the placenta, “real life” PAH mixtures by disrupting protein hormones levels, the MMP-2/MMP-9 ratio and VEGF expression can lead to insufficiency and many pregnancy-related disorders.     

Identifying the most likely contributors to a Y-STR mixture using the discrete Laplace method

In some crime cases, the male part of the DNA in a stain can only be analysed using Y chromosomal markers, e.g. Y-STRs. This may be the case in e.g. rape cases, where the male components can only be detected as Y-STR profiles, because the fraction of male DNA is much smaller than that of female DNA, which can mask the male results when autosomal STRs are investigated. Sometimes, mixtures of Y-STRs are observed, e.g. in rape cases with multiple offenders. In such cases, Y-STR mixture analysis is required, e.g. by mixture deconvolution, to deduce the most likely DNA profiles from the contributors.We demonstrate how the discrete Laplace method can be used to separate a two person Y-STR mixture, where the Y-STR profiles of the true contributors are not present in the reference dataset, which is often the case for Y-STR profiles in real case work. We also briefly discuss how to calculate the weight of the evidence using the likelihood ratio principle when a suspect's Y-STR profile fits into a two person mixture. We used three datasets with between 7 and 21 Y-STR loci: Denmark (n = 181), Somalia (n = 201) and Germany (n = 3443). The Danish dataset with 21 loci was truncated to 15 and 10 loci to examine the effect of the number of loci. For each of these datasets, an out of sample simulation study was performed: A total of 550 mixtures were composed by randomly sampling two haplotypes, h₁ and h₂, from the dataset.We then used the discrete Laplace method on the remaining data (excluding h₁ and h₂) to rank the contributor pairs by the product of the contributors’ estimated haplotype frequencies. Successful separation of mixtures (defined by the observation that the true contributor pair was among the 10 most likely contributor pairs) was found in 42–52% of the cases for 21 loci, 69–75% for 15 loci and 92–99% for 10 loci or less depending on the dataset and how the discrete Laplace model was chosen. Y-STR mixtures with many loci are difficult to separate, but even haplotypes with 21 Y-STR loci can be separated.     

丹芩消郁合剂结合刺络拔罐法治疗肝郁湿热型痤疮35例

目的:观察丹芩消郁合剂结合刺络拔罐法治疗肝郁湿热型痤疮的临床疗效。方法:选择肝郁湿热型中度痤疮患者70例,随机分为治疗组和对照组,每组各35例。治疗组口服丹芩消郁合剂结合刺络拔罐治疗,对照组口服丹参酮胶囊治疗。观察两组患者的临床疗效,比较两组患者治疗前后中医证候积分变化情况。结果:治疗组有效率为94.29%,对照组有效率为88.57%,两组患者有效率比较,差异无统计学意义(P0.05);治疗组愈显率为74.29%,对照组愈显率为51.43%,两组患者愈显率比较,差异具有统计学意义(P0.05)。治疗后两组患者中医证候积分较治疗前显著降低,且治疗组低于对照组,差异具有统计学意义(P0.05)。结论:丹芩消郁合剂结合刺络拔罐法治疗肝郁湿热型痤疮,可有效改善患者的临床症状。     

蛭草茶合剂对糖尿病小鼠视网膜Müller细胞的保护作用及炎症因子表达的影响

目的:观察蛭草茶合剂对糖尿病小鼠视网膜Müller细胞的保护作用及对炎症因子表达的影响。方法:健康清洁级雄性C57BL/6J小鼠75只,采用随机数字表法随机分为正常对照组、糖尿病组、低浓度蛭草茶合剂治疗组(低浓度治疗组)、中浓度蛭草茶合剂治疗组(中浓度治疗组)、高浓度蛭草茶合剂治疗组(高浓度治疗组),每组15只。糖尿病组、不同浓度治疗组小鼠按60 mg/kg剂量腹腔注射STZ诱导糖尿病动物模型。建模后4周,低浓度治疗组、中浓度治疗组、高浓度治疗组大鼠分别按30、60、120 ml/kg的浓度行蛭草茶合剂灌胃治疗。每两周测定各组小鼠体重及血糖。第8周后,采用Western blot检测各组小鼠视网膜Müller细胞中胶质纤维酸性蛋白(GFAP)表达;免疫荧光检测GFAP、谷氨酰胺合成酶(GS)表达;实时荧光定量PCR(RT-qPCR)、ELISA分别检测VEGF、TNF-α、IL-1β、IL-6 mRNA和蛋白表达。组间比较采用单因素方差分析。结果:糖尿病组小鼠体重较正常对照组降低,血糖升高;不同浓度治疗组小鼠血糖呈降低趋势。与正常对照组比较,糖尿病组小鼠视网膜Müller细胞GFAP蛋白表达升高(t=38.318,P<0.001),GS蛋白表达降低(t=29.737,P<0.001),差异有统计学意义;与糖尿病组比较,低浓度治疗组、中浓度治疗组、高浓度治疗组小鼠Müller细胞GFAP蛋白表达下降(t=13.677、19.387、16.305,P<0.05),GS蛋白表达升高(t=5.170、19.399、6.705,P<0.05),差异均有统计学意义。糖尿病组小鼠视网膜Müller细胞VEGF、TNF-α、IL-1β、IL-6 mRNA和蛋白表达均升高,不同浓度治疗组均降低。结论:蛭草茶合剂可降低糖尿病小鼠血糖,减轻糖尿病视网膜炎症反应。     

复方臭灵丹合剂质量标准研究

目的建立复方臭灵丹合剂的质量标准。方法采用TLC对桔梗和鱼腥草进行定性鉴别,依据《云南省中药材(民族药材)质量标准研究技术指导原则(试行)》对其进行分析方法验证及重现性考察。采用HPLC对合剂中的洋艾素和金丝桃苷进行定量分析,同时进行相关方法学的考察。结果桔梗和鱼腥草TLC鉴别色谱图中,供试品与对照药材相应位置上均显相同颜色的特征斑点,阴性无干扰,且分析方法验证与重现性考察均合格。洋艾素和金丝桃苷分别在2.59~55.47μg/mL和4.02~51.68μg/mL范围内线性关系良好,回归方程分别为Y₁=43.24X-12.39(R=0.9996)和Y₂=63.82X-30.04(R=0.9994),准确度试验中,平均加样回收率分别为100.06%和100.14%,RSD值分别为2.36%和2.74%(n=9),其余方法学考察均合格,洋艾素和金丝桃苷的平均含量分别为2.41μg/mL和9.95μg/mL。结论建立的质量标准稳定可行。