基于血热证探讨银屑病病证结合动物模型的构建

目的:以中医病因学说为切入点探讨如何构建银屑病血热证大鼠模型。方法:30只雄性SPF级SD大鼠随机分为空白组、银屑病组、银屑病血热证组,每组10只。空白组大鼠不做任何处理,银屑病组大鼠背部剃毛暴露2 cm×2 cm区域,将5%心得安乳剂按照0.3 g/cm2剂量均匀涂抹以建立银屑病模型,3次/d,共21 d。银屑病血热证组大鼠与银屑病组同法造模,同时用紫外线灯模拟日光照射背部皮肤1 h,1次/d,共21 d,以及按照1 mL/100 g剂量灌服干姜、甘草煎液,1次/d,共21 d。从背部皮肤皮损形态、组织病理Baker评分、体质量、肛温、进食量、饮水量、排便量、排尿量、T3、T4及炎症因子TNF-α、IL-17的表达评价动物模型。结果:银屑病血热证组大鼠背部皮肤出现典型的红斑、丘疹、鳞屑,Baker评分高于空白组(P<0.01);银屑病血热证大鼠症状表现有体质量减轻、肛温升高、进食量减少、饮水量增多、排便量、排尿量减少,与银屑病组比较,差异有统计学差异(P<0.05);与银屑病组比较,银屑病血热证组大鼠T3、T4及炎症因子TNF-α、IL-17均升高(P<0.05)。结论:银屑病血热证大鼠模型既具有银屑病皮损病理特征,又具有中医血热证症状。     

依帕司他原料药有关物质的HPLC测定方法改进研究

目的 采用改进的高效液相色谱法测定依帕司他原料药有关物质。方法 采用依利特Hypersil BDS C₁₈色谱柱（250 mm×4.6 mm,5 μm）,以乙腈-pH6.5磷酸盐缓冲液（0.025 mol·L^-1 KH₂PO₄+0.025 mol·L^-1 Na₂HPO₄,用H₃PO₄调节pH至6.5）为流动相,梯度洗脱,流速为1.0 mL·min^-1,检测波长为396 nm和280 nm,柱温为30℃,进样量为20 μL。结果 依帕司他与相邻杂质能较好地分离。立体异构体杂质在浓度0.010~30 μg·mL^-1内线性关系良好,检测限和定量限分别为0.80,2.4 ng·mL^-1。精密度、稳定性、准确度、耐用性均符合要求。结论 该方法可用于依帕司他原料药有关物质的检查。     

Chinese Herbal Formula Ermiao Powder (二妙散) Regulates Cholinergic Anti-inflammatory Pathway in Rats with Rheumatoid Arthritis

Objective: To investigate the effect of Chinese herbal formula Ermiao Powder (二妙散, EMP) on the expression of cholinergic anti-inflammatory pathway in rats with rheumatoid arthritis (RA). Methods: Seventy two rats were randomly ivided into 6 groups according to body weight, including normal control group, collagen induced arthritis (CIA) group, three doses EMP groups, and methotrexate (MTX) group (n=12 per group). All of the rats except for those in the normal control group were given multipoint subcutaneous injection of bovine type Ⅱ collagen to establish a CIA model. Three EMP groups received a high- (4.5 g/kg), medium- (3.0 g/kg), and low- (1.5 g/kg) doses of EMP by intragavage, respectively. MTX group was injected intraperitoneally MTX at 0.9 mg/kg once a week as the positive control. The administration was 3 consecutive weeks. Joint swelling, arthritis index, and body weight changes in different experimental groups of rats were tested. The joint damage was evaluated by masson staining. Quantitative real-time polymerase chain reaction, Western blot, and immunohistochemistry (IHC) were performed to evaluate the expression of CHRNA7, encoding α 7 nicotinic acetylcholine receptor in the cholinergic anti-inflammatory pathway, in different tissues and their localization in the spleen and joints. Results: CHRNA7 expression levels were significantly higher in the joints and spleens of CIA group than those in normal control group (both P<0.05). Moreover, the CHRNA7 mRNA and protein levels in the spleen and joints of MTX and three doses of EMP groups were significantly lower than CIA group (all P<0.05). Compared with the MTX group, treatment with low-dose EMP resulted in significant reduction of CHRNA7 mRNA and protein expression levels (P<0.05 or P<0.01). IHC showed positive signals of CHRNA7 in the white pulp and red pulp of the spleens of rats; CHRNA7 was expressed on fibroblast-like synoviocytes, macrophages, and endothelial cells in the joints of rats, and the expression in the joints of low-dose EMP group was significantly lower than that in the CIA group (P<0.01). Conclusion: Cholinergic anti-inflammatory pathway was involved in the generation of the inflammatory reaction in CIA rats, and EMP exerted therapeutic effect on RA through cholinergic anti-inflammatory pathway.     

5-羟甲基糠醛在中药加工过程中的变化及药理作用研究概况

5-羟甲基糠醛（5-HMF）是中药中广泛存在的化学成分,在中药炮制、贮藏、配伍过程中含量变化明显。现代药理学研究表明5-HMF具有抗氧化、改善学习记忆、抗过敏、保护神经细胞、抗炎等药理作用。结合国内外有关文献,对5-HMF在中药道地性、炮制加工、贮藏方面的含量变化及药理作用进行综述,以期为提升中药材质量标准和临床合理用药提供参考。     

RAW 264.7细胞炎症模型优化及异喹啉类生物碱抗炎活性研究

目的 对RAW 264.7细胞炎症模型进行优化,并考察异喹啉类生物碱的抗炎活性。方法 采用内毒素脂多糖（LPS）诱导RAW264.7细胞炎症反应,以细胞上清液中炎症因子分泌水平为指标,优化LPS诱导浓度、时间及阳性药地塞米松（DEX）孵育时间,并考察异喹啉类生物碱的抗炎活性。结果 RAW 264.7细胞上清液中肿瘤坏死因子-α（TNF-α）分泌水平随LPS诱导浓度、时间增加而升高,但当LPS质量浓度超过100 ng/mL、诱导时间超过12 h后,TNF-α分泌水平趋于平缓;LPS为20 ng/mL、诱导12 h的TNF-α分泌水平明显升高。DEX对TNF-α分泌抑制作用随孵育时间延长而增强,但孵育4 h后,TNF-α分泌抑制作用趋于平缓。异喹啉类生物碱呈现不同程度TNF-α分泌抑制作用,其抑制活性与其季胺型母核结构及R基团烷氧基取代等密切相关。结论 LPS为20 ng/mL、诱导12 h能较好诱导RAW 264.7细胞炎症反应;异喹啉类生物碱的TNF-α分泌抑制作用与其季胺型母核结构、R基团取代等密切相关。     

血清化学与网络药理学关联研究酸枣仁的体内效应成分

目的研究酸枣仁的血清移行成分,并构建入血成分-失眠靶点网络的效应物质研究模式。方法应用RSLC-Q Exactive Orbitrap-HRMS技术建立大鼠口服酸枣仁水提物后含药血清样品的分析方法。通过对比空白血清与含药血清色谱图异同,扣除内源性成分干扰,鉴定血中主要移行成分及其代谢产物。采用Drug Bank数据库检索失眠相关靶点,并通过String数据库和Cytoscape软件绘制失眠蛋白-蛋白相互作用网络;其次在TCMSP和CoolGeN数据库中分别检索酸枣仁入血成分所对应的靶点信息,将以上靶点集合与失眠蛋白网络进行映射,构建入血成分-失眠靶点网络。结果鉴定与表征了大鼠血清中15个移行成分,包括6个黄酮类及1个代谢物、5个三萜皂苷类和2个生物碱类及1个代谢物。将入血成分靶点集合与失眠蛋白-蛋白相互作用网络进行映射,筛选到乌药碱、酸李碱、芹菜素、斯皮诺素和酸枣仁皂苷A 5种入血成分与失眠蛋白靶点具有相关性。3类不同成分所作用的失眠靶点有交集也各有特异性。皂苷类特异性靶点为γ-氨基丁酸受体;生物碱类特异性靶点为褪黑素受体。初步认为上述5种成分为酸枣仁治疗失眠的体内效应成分。结论从体内化学成分群和潜在效应物质群2个层次共同锁定酸枣仁的效应物质,为酸枣仁体内效应成分的研究进行了有益的研究模式探索。     

Developmental validation of STRmix™ NGS,a probabilistic genotyping tool for the interpretation of autosomal STRs from forensic profiles generated using NGS

We describe the developmental validation of the probabilistic genotyping software – STRmix™ NGS – developed for the interpretation of forensic DNA profiles containing autosomal STRs generated using next generation sequencing (NGS) also known as massively parallel sequencing (MPS) technologies. Developmental validation was carried out in accordance with the Scientific Working Group on DNA Analysis Methods (SWGDAM) Guidelines for the Validation of Probabilistic Genotyping Systems and the International Society for Forensic Genetics (ISFG) recommendations and included sensitivity and specificity testing, accuracy, precision, and the interpretation of case-types samples. The results of developmental validation demonstrate the appropriateness of the software for the interpretation of profiles developed using NGS technology.     

绵马贯众不同萃取部位的抑菌活性

目的:研究绵马贯众不同萃取部位的抑菌活性。方法:以70%乙醇冷浸72 h制备绵马贯众粗提物,液-液分离法制备石油醚、三氯甲烷、乙酸乙酯、正丁醇和水5个部位萃取物,用K-B纸片扩散法对大肠埃希菌、金黄色葡萄球菌、枯草杆菌、酵母菌、黄曲霉和黑根霉6种供试菌株的抑菌活性进行研究。结果:绵马贯众5个萃取部位均表现出一定的抑菌效果,且随着用药量的增加其抑菌活性总体上呈现出增强的趋势,其中以三氯甲烷萃取部位和乙酸乙酯萃取部位的抑菌活性较大,水萃取部位的抑菌活性最小。结论:不同部位对供试菌株的抑菌活性不同。     

补阳还五汤含药鼠血清对血管平滑肌细胞Rho激酶及TFmRNA、基质金属蛋白酶MMP-2mRNA、MMP-9mRNA表达的影响

目的通过运用益气活血法的代表方剂补阳还五汤对血管平滑肌细胞的干预,观察各组血管平滑肌细胞Rho激酶及TFmRNA、基质金属蛋白酶MMP-2mRNA、MMP-9mRNA的表达等指标的变化。方法将30只SD大鼠随机分为3组,一组为正常对照组,另两组为补阳还五汤低剂量对照组(10 g·kg^-1)和补阳还五汤高剂量对照组(20 g·kg^-1);AS模型大鼠30只随机分为模型组,补阳还五汤低剂量治疗组(10 g·kg^-1)和补阳还五汤高剂量治疗组(20 g·kg^-1)。给SD正常大鼠和AS模型大鼠口服不同浓度的补阳还五汤以制备含药大鼠血清,并在用含药大鼠血清保护体外培养的血管平滑肌细胞之后,检测血管平滑肌细胞Rho激酶mRNA、TFmRNA、MMP-2mRNA、MMP-9mRNA的表达。结果与模型对照组相比,各组补阳还五汤高剂量治疗组血管平滑肌细胞Rho激酶、TFmRNA、MMP-2mRNA、MMP-9mRNA的表达均降低。差异均有统计学意义(P<0.05,P<0.01)。结论补阳还五汤含药鼠血清可以通过抑制Rho激酶的活性,下调ROCK1、TF、MMP-2、MMP-9的mRNA的表达水平抑制血管平滑肌细胞增殖,从而抑制AS病变的形成和发展。     

高斯分峰拟合用于不同产地石膏热解过程分析

目的:探究6个产地中药石膏在恒定速率升温过程中热分解过程。方法:用热重法模拟石膏的热分解过程,采用高斯分峰拟合得到各个热分解反应阶段的DTG曲线。运用Coats-Redfern法计算6个产地石膏的热力学参数。结果:6个产地石膏的DTG曲线可分峰拟合为两个彼此交叉的高斯拟合峰,当升温速率为10℃/min,第一失水过程的温度区间为80~220℃,活化能E₁范围为78.98~96.48kJ/mol;第二失水过程的温度区间为145~190℃,活化能E₂范围为312.05~457.95kJ/mol。6个产地石膏中E₁大小顺序为山西>安徽>湖北>河南>山东>河北,E₂大小顺序为山西>安徽>河南>湖北>河北>山东。结论:使用Coats-Redfern和高斯分峰拟合法可以得到6个产地石膏两个热解阶段的活化能和失重导数曲线,揭示中药石膏复杂的热解过程,为石膏的煅制提供可参考的工艺参数。