Background Leguminous Sophora moorcroftiana (SM) is a genuine medicinal material in Tibet. Many research results have reveal the Sophora moorcroftiana alkaloids (SMA), as the main active substance, have a wide range of effects, such as antibacterial, antitumor and antiparasitic effects. However, there are few reports on the inhibition of lung cancer (LC) and its inhibitory mechanism, and the pharmacological mechanism of SMA is still unclear, Therefore, exploring its mechanism of action is of great significance. Methods The SMA active components were obtained from the literature database. Whereas the corresponding targets were screened from the PubChem and PharmMapper database, UniProt database were conducted the correction and transformation of UniProt ID on the obtained targets. The GeneCards and OMIM databases identified targets associated with LC. Venny tools obtained the intersection targets of SMA and LC. R language and Cytoscape software constructed the visual of SMA - intersection targets – LC disease network. The intersection targets protein-protein interaction (PPI) network were built by the STRING database. The functions and pathways of the common targets of SMA and LC were enriched by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Subsequently, molecular docking And A549 cells vitro experiment were performed to further validate our finding. Results We obtained six kinds of alkaloids in SM, 635 potential targets for these compounds, and 1,303 genes related to LC. SMA and LC intersection targets was 33, including ALB, CCND1, ESR1, NOTCH1 and AR. GO enrichment indicated that biological process of SMA was mainly involved in the positive regulation of transcription and nitric oxide biosynthetic process, and DNA-templated, etc. Biological functions were mainly involved in transcription factor binding and enzyme binding, etc. Cell components were mainly involved in protein complexes, extracellular exosome, cytoplasm and nuclear chromatin, etc., Which may be associated with its anti-LC effects. KEGG enrichment analysis showed that main pathways involved in the anti-LC effects of SMA, including pathway in cancer, non small-cell lung cancer, p53, PI3K-Akt and FOXO signaling pathways. Molecular docking analyses revealed that the six active compounds had a good binding activity with the main therapeutic targets 2W96, 2CCH and 1O96. Experiments in vitro proved that SMA inhibited the proliferation of LC A549 cells. Conclusions Results of the present study, we have successfully revealed the SMA compounds had a multi-target and multi-channel regulatory mechanism in treatment LC, These findings provided a solid theoretical reference of SMA in the clinical treatment of LC.

Abstract In this paper，Nostoc commune crude polysaccharide was extracted by hot water and Ultrasonic-assisted extraction, then purified by DEAE-cellulose52 cellulose column chromatography and Sephacryl G-100 gel column chromatography. Homogeneous polysaccharide HNCP3 extracted with hot water and UNCP4 extracted by ultrasound were obtained. The structures of HNCP3 and UNCP4 were characterized by molecular weight determination, infrared spectroscopy, DSC detection, sodium periodate oxidation, smith degradation reaction and methylation analysis. The conformation of the solution was studied by SEM and AFM. The results showed that the Ultrasonic-assisted extraction had an effect on the molecular weight, monosaccharide composition molar ratio and configuration of Nostoc commune. The main chains of HNCP3 and UNCP4 were mainly →6) -D-Glcp (1→ and →2,6) -D -Glcp, but there are 1,2,6-galactose branches and 2,3-Me2-D-Ara in UNCP4 and there is no HNCP3. The results of the composition of monosaccharides showed that both HNCP3 and UNCP4 have the presence of mannose. SEM and AFM showed that the structure of UNCP4 was helical, and the solution conformation of HNCP3 and UNCP4 was different in different solution environments. In experiments on the antioxidant activity and hypoglycemic effect of HNCP3 and UNCP4 in vitro, it was shown that UNCP4 has higher antioxidant activity scavenging ability and UNCP4 is stronger than HNCP3 inhibitory activity. Therefore, UAE increases the NCP extraction rate while also increasing the biological activity in NCP.

运用PCA模型和代谢组学等方法,研究单次给药后复方藏兽药翼草景香颗粒主要成分在健康靶动物体内的变化规律及时量关系;运用主成分分析模式识别技术,寻找表征特定状态的生物标志物和潜在生物标志物的候选变量,从而揭示翼草景香颗粒在动物体内的生物学本质及代谢作用机制.选取健康西藏牦牛、绵羊,分成高、中、低剂量组,口服投喂复方藏药一次,另设不给药对照组,于给药前和给药后不同时段采集血样与粪样.利用GC-MS/MS对样本进行检测,检测数据分别通过GC-MS Postrun Analysis软件进行处理、NISI MS Search 2.0质谱库检索分析和Origin2018统计软件进行统计分析.由PCA得分可知,复方藏药中剂量组牦牛给药5.5 h、6.5 h和7.5 h时血样中藏药活性成分排名靠前,活性较高;复方藏药中剂量组绵羊给药0.5 h、7.5 h时和给药24.0 h时血样中藏药活性成分可以明显区分、得分最高.给药后牦牛、绵羊血液中检测出安息香醛、3-(2,6,6-三甲基-1-环己烯基)-2-丙烯醛、β-谷甾醇、2-己基-1-癸醇、三花生酸甘油酯、(±)-1,2-二棕榈精、2-乙基己酸乙烯酯、十二烯基丁二酸酐、二氢青蒿素等多种共有活性成分.该复方藏药发挥药效的主要成分包括安息香醛、3-(2,6,6-三甲基-1-环己烯基)-2-丙烯醛、β-谷甾醇等,这些共有成分可能与消炎、解热及抗肿瘤功效有关.

目的:基于成分-靶点-通路的网络药理学探讨野生植物砂生槐的生物活性机制,为野生砂生槐资源充分的开发和合理利用奠定基础.方法:PubChem、PharmMapper数据库筛选砂生槐化学成分药效团潜在靶点,Uniprot数据库对获得靶点进行UniProt ID校正和转化,STRING数据库构建蛋白互作网络,Cytoscape软件构建砂生槐有效活性成分-靶点、靶点互作的可视化网络,DAVID数据库对靶标进行基因本体、京都基因与基因组百科全书功能富集.结果:从砂生槐中共筛选出羽扇醇、亚油酸、β-谷甾醇、猫尾草异黄酮和金雀花碱等15种有效化学成分,药物靶点蛋白互作网络调控中涉及的关键的靶点有MDM2、ERBB2、AR、MTOR等;GO功能富集结果表明,生物过程主要为低氧反应、细胞运动调节;分子功能主要为蛋白结合;细胞组分主要为胞浆、胞外体;KEGG通路结果表明,涉及的通路主要为癌症、代谢、cAMP和FoxO信号通路.结论:砂生槐中的药物成分主要作用于MDM2、ERBB2、AR、MTOR等关键靶点,从而达到对癌症类、代谢类、神经系统类疾病的治疗效果,这不仅有助于临床靶向治疗药物的精准开发,也为野生植物砂生槐生物活性的进一步研究提供多靶点和多途径的参考依据,具有较高的资源开发价值.

优化砂生槐子生物碱的提取工艺,为兽药开发利用提供依据.建立砂生槐子中4种生物碱(氧化苦参碱、氧化槐果碱、苦参碱、槐果碱)含量的高效液相色谱(HPLC)测定方法,以4种生物碱的提取率及干粉得率的总评"归一值"(O D)为评价指标,分别考察乙醇浓度、料液比、超声温度、超声时间、超声功率因素对OD值的影响,并结合Box-Behnken中心组合设计的三因素三水平响应面试验方法来优化砂生槐子生物碱的提取工艺.结果显示,最佳的提取工艺为:乙醇750 m L/L,提取温度55℃,料液比为1 g/m L:30 g/mL,超声时间为50 min,超声功率为150 W,在此条件下,OD值为0.5586,干粉得率为20.37％,4种生物碱总含量为388.48 m g/g.此工艺优化显著提升了砂生槐子生物碱作为兽用药物的开发价值.

苦参碱类生物碱具有多种生物活性和兽医临床应用前景,广泛存在于豆科槐属药用植物中,主要包括氧化苦参碱、苦参碱、槐果碱、氧化槐果碱和槐定碱等成分.常见的提取方法包括水提、醇提、酸水提、浸渍、渗漉等,常见的工艺优化方法包括正交设计、响应面设计等,常见的纯化方法包括大孔吸附树脂纯化、模板分子印迹固相萃取-高效液相色谱-串联质谱方法等.为寻找更加便捷、快速、高效的方法来获取高含量、高纯度的苦参碱类生物碱成分,论文对苦参碱类生物碱的提取方法、提取工艺优化及纯化方法的国内外研究进行综述.

优化西藏三颗针中盐酸小檗碱的提取工艺.采用比色法测定盐酸小檗碱的含量,以盐酸小檗碱含量为指标,分别考察pH值、稀酸浓度、溶剂温度、料液比四因素对提取分离盐酸小檗碱的影响,并结合Box-Behnken响应面分析法,优化西藏三颗针中盐酸小檗碱的提取工艺.结果 表明,最佳提取工艺稀酸浓度为0.2％,pH值为10,提取温度50℃,料液比为1:8时,该工艺下盐酸小檗碱的得率最高.说明优化后的提取工艺简便可行,收率高,具有工业化生产推广价值.

确定超临界CO2萃取西藏红花杜鹃叶的最优提取工艺,并初步鉴定分析其提取物的主要化学成分组成.以提取率为评价指标,利用正交优化法对开花期西藏红花杜鹃叶进行超临界CO2萃取,采用气相色谱-质谱联用技术(gas chromatgraphy-massspectrometry,GC-MS)对提取物进行化学成分分析.超临界CO2萃取西藏红花杜鹃叶的最优工艺条件为将红花杜鹃叶粉碎为200目、萃取压力20 MPa、萃取温度为40℃、反应时间2.5h,GC-MS检测分离出了23种含量较高的化合物.经与质谱谱图数据库比对,西藏红花杜鹃叶超临界CO2萃取提取物主要为烷烃(39.54％)、酯(26.94％)、烯(24.48％)、酰胺类(8.78％)等化合物.

The extraction process of Glycyrrhiza soluble polysaccharide (GP) was optimized by RSM, a rat trauma model was established via longitudinal incision on the back skin. The effects of GP combined with microcapsule collagen on the repair of rat injury model were discussed at different levels, Based on the content of hydroxyproline at the whole animal level, the proliferation of granulation tissue stained by HE, the number of microvessels labeled by CD34, the production of collagen fibers stained by Masson, the level of phosphorylation of STAT3 protein and that of VEGF at protein level were investigated. The results showed that after the administration of GP combined with microcapsules, the content of hydroxyproline in granulation tissue increased, the proliferation of capillaries and fibroblasts in granulation tissue became active, and the number of microvessels in wound increased. The formation density of collagen fibers was uniform and orderly. GP combined with microcapsules could activate the expression of p-STAT3 and VEGF proteins and up-regulate the transcription level of VEGF mRNA and miRNA-21 genes. Furthermore, GP combined with microcapsules could accelerate wound healing and promote neovascularization.

为研究藏橐吾在抗菌、抗炎和抗病毒方面的作用,采用乙醇法提取藏橐吾地上部分总黄酮,通过牛津杯法、二甲苯构建小鼠急性炎症模型,腹腔注射法、MTT法和CCK-8法研究藏橐吾在抗金黄色葡萄球菌、消炎和抗羊口疮病毒中的功效.结果表明,藏橐吾地上部分总黄酮浓度达1 g/mL和0.8 g/mL时金黄色葡萄球菌呈中度敏感,0.4 g/mL和0.6 g/mL时呈低度敏感;提取物可显著抑制二甲苯所致小鼠耳部炎症,且与浓度正相关;提取物对羊口疮病毒具有抑制作用,且与浓度正相关.因此,认为藏橐吾具有抗菌、消炎和抗病毒功效.

本研究通过高效液相色谱法测定了川产、藏产三颗针中小檗碱、药根碱和巴马汀的含量,采用微量肉汤稀释法测定三颗针提取物对耐甲氧西林金黄色葡萄球菌(MRSA)USA300的最低抑菌浓度,在亚抑菌浓度下采用结晶紫染色法评估其对MRSA生物被膜形成的抑制和清除作用,同时检测提取物与MRSA共培养上清液的溶血活性.结果表明,三颗针提取物在亚抑菌浓度下对USA300生物被膜的形成具有一定的抑制作用,在较高浓度下三颗针提取物对24 h和48 h形成的生物被膜具有良好的清除作用;不同浓度三颗针提取物对USA300的α-溶血素分泌有一定的抑制作用,其中1/2MIC的抑制作用最强.本研究为临床合理应用抗MRSA药物提供了理论依据.

确定超临界CO2萃取西藏白花杜鹃花的最优工艺条件和初步鉴定其提取物的化学成分.以提取率为评价指标,利用正交试验对西藏白花杜鹃花超临界CO2萃取有效成分条件进行了优化,采用气相色谱-质谱联用技术(gas chromatgraphy-mass spec-trometry,GC-MS)分析了萃取物的化学成分.结果表明,超临界CO2萃取西藏白花杜鹃花的最优提取工艺为开花期白花杜鹃的花粉碎为200目、萃取压力25 MPa、萃取温度为40 ℃、反应时间2.5 h,GC-MS检测分离出24种含量较高的化合物.由此可得,经与质谱谱图数据库比对,初步鉴定主要为烷烃及其含氧衍生物(62.32 ％)、烯和醇(8.09 ％)、酯(4.87 ％)等化合物,其中含有提高机体免疫力和抗肿瘤作用的角鲨烯(含量达4.79 ％)等药用活性成分,具有很好的研究开发前景.

为研究白花杜鹃水提液对多房棘球蚴——原头蚴的体外杀灭作用,以及对多房棘球蚴感染小鼠的治疗作用,通过在体外原头蚴培养体系中分别添加白花杜鹃水提液生药质量浓度分别为10、20和40 mg/mL,以阿苯达唑伊维菌素培养基(阿苯达唑的质量浓度为20 mg/mL、伊维菌素的质量浓度为0.83 mg/mL)、阿苯达唑伊维菌素溶剂培养基和常规培养基分别作为对照组进行原头蚴的培养,分别经第0、12小时和第1、2、3、4、5、6、7天观察记录各药物对原头蚴的杀灭效果,并于第7天通过台盼蓝染色比较各组死亡率;将50只感染多房棘球蚴1个月的雌性BALB/c小鼠随机分为5组,每组10只,依次为白花杜鹃水提液低剂量(10 mg/mL,RL组)、中剂量(20 mg/mL,RM组)、高剂量(40 mg/mL,RH组)治疗组,阿苯达唑伊维菌素治疗组(C组),感染不治疗组(IT组),另包括10只健康小鼠作为空白对照组(B组),各组小鼠连续灌胃给药后第7天,第30天后剖检,对小鼠脏器及棘球蚴组织固定并做石蜡切片进行病理学观察.结果 显示,白花杜鹃水提液低、中、高剂量组,阿苯达唑伊维菌素溶液组,溶剂对照组以及空白对照组原头蚴培养后第7天后,原头蚴的死亡率分别为54.00％、56.53％、89.27％、57.60％、19.27％和15.80％,白花杜鹃高剂量组与对照组相比差异极显著(P＜0.01).多房棘球蚴感染小鼠腹部明显肿胀,剖检可见腹腔内有大量大小不等的包囊.石蜡切片结果表明,RH组小鼠脏器病变程度最低,且RH组和C组小鼠体内少量包囊发生钙化.研究表明,白花杜鹃水提液对体外原头蚴具有杀灭作用,且40 mg/mL白花杜鹃水提液对小鼠感染多房棘球蚴有一定的治疗作用.

藏鸡弧菌性肝炎是由空肠弯曲杆菌感染而引发的一种传染病.主要引起肝脏的坏死.本文将从病原、流行特点、临床表现、病理变化、诊断及防治方面对该病进行分析和总结.

研究翼草景香水煎液对多杀性巴氏杆菌的体外抑菌活性及小鼠感染模型的防治效果.利用传统水煎法制备翼草景香水煎液,用HP LC方法测定翼草景香水煎液中红景天苷浓度,并用红景天苷浓度标定翼草景香水煎液药物浓度,分别采用牛津杯法和试管2倍稀释法测定翼草景香水煎液对多杀性巴氏杆菌抑菌圈直径和最小抑菌浓度(MIC),通过建立死亡率为75％的多杀性巴氏杆菌体内感染小鼠模型,测定翼草景香水煎液浓度为0.12316 mg/mL时,在感染前和感染后给药方式下,分别给与0.1、0.3、0.5 mL药物时对小鼠的体内感染防治作用.结果表明,当翼草景香水煎液浓度为0.49263 mg/mL时,抑菌圈平均直径为28.5 mm;MIC值为0.00385 mg/mL,经病理解剖、细菌镜检、全自动微生物鉴定及药敏系统(VTEK 2 COMPACT)系统细菌鉴定,表明死亡率为75％的多杀性巴氏杆菌体内感染小鼠模型建立成功,当水煎液浓度为0.12316 mg/mL时,0.1、0.3、0.5 mL预防组和治疗组小鼠死亡率分别依次为50％,41.7％,81.8％,66.7％,50％和75％.因此,翼草景香水煎液对多杀性巴氏杆菌具有良好的体内、外抑菌活性,且其预防作用较治疗作用明显.

采用植物化学成分系统预试法对藏橐吾地上部分的化学成分进行研究,结果显示藏橐吾地上部分含有萜类化合物、甾体类化合物、挥发油、油脂、强心苷类化合物、酚类化合物、有机酸、生物碱、醌类和黄酮类化合物.

研究菪可轮蒲水煎液对多杀性巴氏杆菌的体内外抑菌活性作用.用传统水煎法制备菪可轮蒲水煎液,用HPLC法测定菪可轮蒲水煎液中龙胆苦苷浓度,并用龙胆苦苷浓度标定复方药物浓度,分别采用牛津杯法和宏量肉汤稀释法测定菪可轮蒲水煎液对多杀性巴氏杆菌的抑菌圈直径和最小抑菌浓度(MIC),通过建立死亡率为75％的多杀性巴氏杆菌体内感染小鼠模型,测定浓度为0.0232 mg/mL的菪可轮蒲水煎液,在感染前和感染后给药方式下,分别给予0.1、0.3、0.5 mL药物时对小鼠多杀性巴氏杆菌感染的预防和治疗作用.结果显示,菪可轮蒲水煎液浓度为0.0464 mg/mL时,对多杀性巴氏杆菌的抑菌圈平均直径为13.5 mm,MIC值为0.0029 mg/mL,经病理剖检、细菌镜检、全自动微生物鉴定及药敏系统(VITEK 2 COMPACT)细菌鉴定,表明死亡率为75％的多杀性巴氏杆菌体内感染小鼠模型建立成功,且预防组各组和治疗组各组小鼠死亡率依次为58.3％、41.7％、83.3％、66.7％、58.3％和91.7％.综合以上试验结果,菪可轮蒲水煎液对多杀性巴氏杆菌具有良好的体内外抑菌活性,且其预防作用较治疗作用明显.

The study was aimed to determine the efficacy and toxicity of Aconitum extract on Melophagus ovinus and it's skin toxicity,irritation and allergy in vitro,in order to provide a theo-retical basis for the development,utilization and clinical application of Tibetan A conitum.1%,2%,4%,8% and 10% Tibetan Aconitum extract were dropped in plates lined with absorbent pa-per and Melophagus ovinus to observe the killing efficacy on Melophagus ovinus at 1,2,4,8 h,re-spectively.The same dose of 1%,2% Tibetan Aconitum extract was taken to apply evenly in rab-bits intact and damaged skin to detect the skin toxicity,irritation and allergy of Tibetan A conitum extract.The results showed that at 1 h after administration,all drug groups and positive control group showed a kill effect on Melophagus ovinus in the different levels,and at 4 h after adminis-tration,the killing rate of positive control group was 100%,while that was 70% of 1% and 2%Tibetan Aconitum extract groups at 8 h with 2% drug dose showed the better effect.The skin toxicity test showed that rabbits skin had no erythema and edema,the body weight was normal, and the skin irritation test showed that the extract had no irritation to rabbits skin,while the results of skin allergy showed the it had no allergy to rabbits.In conclusion,Tibetan Aconitum extract had the good killing effects on Melophagus ovinus in vitro,and the skin medication was safe.

对细粒棘球蚴病的诊断与防治进行了综述,以期为该病的进一步研究提供参考.

沙门菌病是目前危害养禽业最常见的细菌疫病之一[1],在公共卫生学上有着重要的意义.随着进藏旅游的盛行,从而带动高原藏鸡、藏蛋等需求增加,近年来西藏养禽业在养殖规模和数量上得到了快速的发展,随之而来鸡病发生也更加复杂化.由于有良好的疫苗和完善的免疫程序,目前病毒性传染病基本得到了控制,而细菌性疾病尤其是鸡沙门菌病呈现出扩张流行的趋势[2].为了解鸡沙门菌在西藏流行情况,项目组选择在养禽区拉萨市5个县(区)12个养殖场不同品种不同来源的鸡进行采样分析,为临床上科学防治本病提供一定的依据.