以类脂A为靶点应用生物传感器技术筛选抗内毒素中草药

目的以细菌内毒素活性中心类脂A(Lipid A)为靶点,应用生物传感器技术,从42种中草药水煎液中筛选拮抗内毒素作用的中药。方法将Lipid A包被于生物传感器的疏水性样品池,通过生物传感器测定中药与Lipid A的结合能力;对具有高亲和力的中药与内毒素作用,再次测定其与Lipid A的结合能力,并通过定量的内毒素中和实验进一步的对生物传感器的测定结果进行验证。结果在筛选的42种中草药中,观察到赤芍、大黄、青果、栀子等中药能与Lipid A高亲和力结合;通过与内毒素作用后的二次检测,发现赤芍、大黄、青果等中草药含有较高Lipid A结合的有效成分,定量内毒素中和实验的测定结果与生物传感器的定性检测结果一致。结论以Lipid A为靶点,应用生物传感器技术,筛选拮抗内毒素中草药的方法,具有快速、高效、直观、准确的优点。     

白鲜皮抗内毒素活性物质的分离提取与活性研究

目的 筛选与类脂A(lipid A)具有较高结合活性的中药,从中提取具有拮抗内毒素(lipopolysaccharide, LPS)活性的物质.方法 采用生物传感技术,以lipid A为靶点,检测42种中药的水提物与lipid A的结合活性;以白鲜皮为研究对象,通过大孔吸附树脂、溶剂萃取、聚酰胺柱层析和高效液相色谱(HPLC)法,分离提取与lipid A结合活性较高的组分;动态比浊法鲎试验检测HPLC产物DPR-1～3对LPS(0.1 ng/ml)的中和作用;ELISA法检测DPR-2对LPS(100 ng/ml)刺激的RAW264.7细胞释放肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)的影响.结果 8种中药水提物与lipid A具有较高的结合活性(RU＞100 arc/s);从白鲜皮中逐步分离出与lipid A结合活性较高的组分,经HPLC法获得3种物质DPR-1～3;DPR-2对LPS具有一定的中和作用,在8 μg/ml浓度以上可显著抑制LPS诱导的RAW264.7细胞释放TNF-α和IL-6 (P＜0.05),并具有剂量效应关系.结论 从42种中药中筛选出与lipid A具有较高结合活性的中药白鲜皮,并从中分离提取出抗LPS活性物质DPR-2,其抗LPS活性可能与DPR-2对LPS的中和活性有关.     

丹参水提液的纯化工艺研究

目的 优选丹参水提液纯化工艺。方法 以丹参提取液的浊度、鞣质以及有效成分的量为指标,优选两种絮凝澄清剂的用量和絮凝搅拌速度,筛选出较佳参数;对比研究醇水沉淀、絮凝结合乙醇沉淀、絮凝结合膜过滤3种纯化方法。结果 使用壳聚糖进行絮凝澄清处理后的丹参水提液上清液浊度低于用ZTC1+1Ⅱ型絮凝剂处理后的浊度,并能够更好地减轻膜污染程度;壳聚糖结合膜过滤法对丹参水提液的有效成分保留率较高,收膏率相对较高,浊度和鞣质含量无显著差异。结论 壳聚糖作为丹参水提液絮凝澄剂效果优于ZTC1+1Ⅱ型絮凝剂,壳聚糖结合膜过滤法纯化丹参水提液的综合评价指标优于其他两种方法。     

其它

生物化学技术在生药学中的应用根据磷酸二酯酶(PDE)与体内cAMP含量变化有密切关系,而临床上所使用的大多数药物特别是中枢神经系统的治疗药物对PDE活性有显著的抑制作用这一事实,作者提出应用PDE抑制试验作为体外筛选中药有效成分的方法之一.用Brookers方法,用于筛选的浸膏是热水提物的冷冻干燥品,剂量100微克/毫升,筛选了220种中药对PDE的抑制作用.结果表明有30多种药用植物具有生物活性.作者着重阐述三种中药的活性成分对PDE的抑制作用.发现PDE抑制试验方法能反映中     

小桐子果壳抑菌活性部位的分离及其HPLC-MS分析

目的 寻找小桐子〖WTBX〗Jatropha curcas果壳的抑菌活性物质。方法 利用6种不同极性溶剂对果壳进行平行提取,结合抑菌活性测定,筛选出活性最强、提取率大于10%的粗提物;再将筛选出的粗提物,依次利用液液萃取、硅胶柱反复柱色谱,结合PTLC制备分离纯化。同时对各阶段分离产物进行活性追踪,筛选出具有抗菌活性的物质,最后采用HPLC-MS分析,初步鉴定出活性部位主体成分。结果 从果壳中分得抑菌活性强且含量高WA3物质,经HPLC-MS分析,初步鉴定了含量较高的3个化合物,为总量的35.27%,分别为环庚肽、麻疯树素、三萜醇三聚体。结论 小桐子果壳中含有丰富的抑菌活性物质,分别属于肽类、二萜、三萜类化合物。     

基于清除DPPH·法研究清热解毒中药的体外抗氧化活性

目的:在10种清热解毒药中筛选出抗氧化活性较高的药物,为筛选高效治疗糖尿病的中药和新药提供一定的理论基础。方法:测定清热解毒药醇提液对DPPH·的清除率,并计算半数清除率(IC50),以维生素E为阳性对照,评价其抗氧化活性。结果:不同中药的醇提液在一定浓度范围内,均有较强的体外抗氧化活性,其中连翘的最大效能及强度均最强,且IC50值是0.312 7 g·L-1为最小。结论:连翘抗氧化活性最强。本研究为连翘治疗糖尿病的作用机制以及应用前景提供了新思路,也为临床合理应用提供了参考。     

一种基于荧光图像的体外抗肿瘤药物快速筛选方法

目的:建立一种基于荧光成像的抗肿瘤药物体外快速筛选系统.方法:①采用二乙酸荧光素(FDA)标记测定活细胞数法,评价26个乌药组分对的体外抗肿瘤活性,并对本方法的线性范围和精密度进行了考察;②在活性筛选结果的指导下,应用LC/MS对样品进行分析,初步推断药效活性物质.结果:①该方法在细胞密度为0～ 10 000/孔的范围内荧光强度与细胞个数线性相关(r2 =0.9858),板内的精密度为9.41％;②在26个乌药组分中,筛选得到两个抗肿瘤活性较强的组分C13和C15,抑制率均达到90％以上.结论:本方法快速、稳定、简便、线性范围宽,可用于从复杂中药组分中快速筛选抗肿瘤活性物质.此外,根据LC/MS分析结果并结合文献报道,推断乌药中具有抗肿瘤活性的化合物为异波尔定碱.     

模式生物斑马鱼在中药活性筛选和毒性评价中的应用进展与展望

斑马鱼作为一种非哺乳类模式生物,因其具有体积小、易饲养、发育快速、繁殖力强、便于观察等优势,在医药研究领域得到了广泛应用。近年来,随着技术和方法的不断改进,模式生物斑马鱼在中药领域的应用也逐渐深入,特别是对中药复杂的有效成分、活性物质以及毒性的评价与筛选。斑马鱼作为一种整体动物模型,能够较为客观地对中药活性物质以及毒性进行评价,实现快速有效的高通量筛选。通过对近年来国内外相关文献的查阅与归纳整理,综述了模式生物斑马鱼在中药活性筛选和毒性评价中的应用进展,主要从抗炎、抗血管生成、促血管生成、抗骨质疏松、抗酒精性肝损伤、抗神经性疾病等活性成分筛选以及肝毒性和胚胎毒性等方面进行总结分析,以期为中药安全性与有效性评价提供新的方法和技术。      

中药水提取液对荧光法测定醛糖还原酶活性的干扰

目的：探讨中药水提液采用荧光法检测醛糖还原酶抑制作用的可行性，方法：激发光367nm，发射光410－470nm范围内扫描13种中药水提物荧光强度。结果：所观察13种中药含有较强的荧光物质，干扰体外酶反应荧光测定法检测醛糖还原活性的方法，结论：若水提液不适于采用此种筛选方法，可考察有机溶剂是否有干扰以确定实验条件或选择其它筛选方法。     

细胞膜色谱法在中药应用中的研究进展

细胞膜色谱法(CMC)是将细胞膜结合到活化硅胶表面,制成细胞膜固定相,并利用药物与细胞膜受体之间具有特异性亲和力的特性和色谱学理论来研究流动相中药物与受体相互作用的规律。CMC集合了高效液相色谱技术、细胞分子生物学及受体药理学的优点,并实现了在细胞膜色谱柱内近似动态地模拟药物活性成分在体内的作用过程。这是一种全新的亲和生物色谱技术,其自提出以来迅速在中药研究中成为热点。近年来,随着CMC方法学原理的不断发展和完善,其已大量应用于药物活性成分筛选、药物与受体的亲和作用及中药质量控制等方面,下一步将朝毛细管细胞膜色谱法和微型细胞膜色谱法等更加微型、高效的色谱技术发展。     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Luo Lingjie''''s Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.