灯盏花素的研究进展

灯盏细辛[Erigeron breviscapine(vant)Hand MaSS]是产于云南等地的一种具有良好抗心脑血管疾病作用的植物。从70年代开始,灯盏花中一系列化学成分被分离鉴定出来。其中,灯盏花素在治疗脑血栓、脑缺血、脑血管后遗症及对抗血小板凝聚等方面有很好的临床疗效,本文对灯盏花素的提取分离、药理、临床应用及灯盏花乙素的全合成等方面的研究进展进行综述。     

灯盏花素治疗脑梗死50例临床观察

目的：观察灯盏花素治疗脑梗死的临床疗效。方法交１００例急性脑死患者随机分为两组,治疗组５０例,应用灯盏花素治疗;对照组５０例,应用脉络宁治疗。结果治疗组总有效率为９２．００％,对照组照组总有效率为７２．００％,两组比较有显著性差异（Ｐ〈０．０５）。结论灯盏花素是治疗急性脑梗死较为理想的药物,值得临床推广。     

灯盏花素辅助治疗对急性脑梗死患者的疗效分析

目的:观察灯盏花素辅助治疗对急性脑梗死患者血液流变学、血管内皮功能及凝血功能的影响。方法:选择本院确诊为急性脑梗死的患者共144例,包括观察组和对照组各72例,对照组采用常规对症治疗,观察组在对照组基础上加用灯盏花素静脉滴注辅助治疗,比较两组患者治疗前后血液流变学、血管内皮功能及凝血功能的差异。结果:经治疗后,观察组NIHSS评分(3.01±1.40)显著低于对照组(3.96±2.17),观察组总有效率显著高于对照组(94.4%vs.86.1%,P0.05)。两组间治疗前各指标水平差异未发现统计学意义(P0.05);两组治疗后全血黏度(whole blood viscosity,WBV)、全血还原黏度(whole blood reduced viscosity,WBRV)、血浆粘度(plasma viscosity,PV)和红细胞聚集指数(erythrocyte aggregation index,EAI)均显著降低(P0.05);其中观察组各个指标均显著低于对照组(P0.05);两组治疗后血管内皮素-1(endothelin-1,ET-1)水平与循环内皮细胞(circulating endothelial cell,CEC)计数显著降低,而一氧化氮(nitrie oxide,NO)及可溶性血栓调节蛋白(soluble thrombomodulin,s TM)水平显著升高(P0.05),其中观察组ET-1水平显著低于对照组,而NO及sTM水平显著高于对照组(P0.05);两组治疗后凝血酶原时间(prothrombin time,PT)及部分活化凝血酶原时间(activated partial thromboplastion time,APTT)显著升高,纤维蛋白原(fibrinogen,Fib)和D-二聚体(D-dimer,DD)显著降低(P0.05);其中观察组PT和APTT显著高于对照组,DD显著低于对照组(P0.05)。结论:灯盏花素辅助治疗能有效改善急性脑梗死患者血液粘稠状态,保护血管内皮功能及改善凝血功能。     

云南灯盏花遗传变异的RAPD分析

运用RAPD (randomamplifiedpolymorphicDNA)技术对云南灯盏花 (Erigeronbrevis capus) 6个居群进行分析 ,研究其遗传变异及遗传结构 ,用外类群紫菀对比分析其亲缘关系。随机挑选 16个引物在 6个居群中共产生 2 33条DNA片段 ,其中 192条带具有遗传多态性 ,约占 82 4 0 %。多态位点百分比PPB、等位基因数A、有效等位基因数Ae、Nei’s基因多样性指数H和Shannon多样性指数I在物种水平分别为 82 4 0 %、 1 82 4 0、 1 30 0 5、0 1896、 0 30 2 1;居群的平均值分别为 5 4 2 3%、 1 5 40 1、 1 2 6 91、 0 16 0 7、 0 2 4 6 0。灯盏花的遗传多样性较丰富 ,基因分化系数Gst值为 0 346 0 ,有 6 5 4 0 %的变异来自居群内 ,6个居群的遗传一致度较高 ,在 0 90 37～ 0 972 3之间 ,平均为 0 94 10。利用UPGMA对 6居群聚类分析 ,结果表明 :丘北居群 (YB)和文山居群 (YX)亲缘关系最近 ,小哨居群 (Y )和新平居群 (YA)次之 ,丘北居群 (YB)和腾冲居群 (ZA)亲缘关系最远。遗传关系与各居群地理分布区间的距离大致成正相关     

2种检测灯盏花素血药浓度方法的比较

目的：建立测定Beagle犬血浆中灯盏花素浓度的反相高效液相色谱法和液相-质谱联用法,并将其进行多参数的两两比较。方法：采用液-液萃取法处理血清,反相高效液相色谱法测定,流动相为甲醇-水（pH3．0）=42：58,EclipseXDB-C18为固定相;将同样的样品用液相-质谱法开展平行检测;将所测得的数据做两两对比,采用SPSS统计软件进行分析。结果：2种方法测定的数据具有良好的相关性（r=0．982,P〉0．05）;最低检测限前者为0．05μg／mL,后者为0．01μg／mL,反相高效液相色谱法的最低检测限和灵敏度不及液相-质谱联用法。结论：反相高效液相色谱法可用于灯盏花素毒代动力学研究,与液相-质谱联用法检测结果无显著差异,且费用较低,不失为-种低成本且行之有效的检测方法。     

降纤酶加灯盏花素治疗大面积脑梗死疗效观察

目的　观察降纤酶加灯盏花素治疗大面积脑梗死的疗效。　方法　随机将 85例病人分为对照组40例及治疗组 45例 ,治疗组在对照组用药的基础上加用降纤酶及灯盏花素治疗。 60天后进行疗效评价。　结果　基本治愈率是治疗组为 46.7% ,对照组为 2 5 % ,两组比较 ,治疗组疗效明显优于对照组 (P <0 .0 5 )。　结论　降纤酶加灯盏花素治疗大面积脑梗死疗效好 ,疗程短。降纤酶溶栓治疗以早期 (6h内 )应用效果好 ,但超过 6h仍然有效。     

灯盏花素对脑出血患者氧化应激的影响

目的：探讨中药灯盏花素注射液对脑出血患者氧化应激的影响。方法：实验分成两组,以30例健康人为对照组,以25例早期脑出血患者为实验组,采用灯盏花素进行治疗,观察治疗前后两组血液中SOD、LDH的活性及MDA的含量。结果：与对照组相比,治疗前实验组SOD活性降低,而LDH的活性和MDA的含量升高。采用灯盏花素治疗后,实验组SOD显著升高,LDH的活性和MDA的含量降低;与对照组相比,无明显差异。结论：灯盏花素可通过抑制中性粒细胞产生呼吸爆发,增强机体清除氧自由基的能力,并降低脂质过氧化损伤,可应用于治疗早期脑出血。     

药用植物灯盏花的组织培养

以灯盏花花葶、花盘及叶柄为外植体,MS为基本培养基,通过不同的激素种类和浓度配比,建立灯盏花组培快繁体系。结果如下:在所有实验方案中,花葶的出愈率最高,是理想的快速繁殖材料。较适宜的诱导愈伤组织的培养基为MS+BA1.0mg/L+IBA0.05mg/L+蔗糖3.0%,诱导不定芽的培养基为MS+BA2.0mg/L+IAA1.0mg/L+蔗糖3.0%或MS+Kt3.0+IAA0.5mg/L+蔗糖3.0%,而根的诱导则是在1/2MS+NAA1.0Mg/L+蔗糖3.0%的培养基上进行。同时对组织培养过程中灯盏花植株再生的方式进行了讨论。     

灯盏花雄性不育种质鉴定与花药发育的细胞学研究

对云南泸西栽培灯盏花群体进行调查,发现了灯盏花雄性不育种质个体,其出现频率约为1.06×10-4.对所发现的灯盏花不育株形态特征及其花药发育过程进行了观察,并对花粉活力进行鉴定.结果显示:(1)灯盏花不育株根、茎、叶形态与正常可育植株基本相似,管状花小,花丝短,花药瘦小,无花粉粒散出或花粉无活力.(2)灯盏花在其花药发育的小孢子母细胞时期、四分体时期、小孢子时期和单核早期,由于绒毡层细胞液泡化、提前解体,不能为小孢子或花粉发育提供所需物质,导致小孢子母细胞和四分体解体,产生无花粉的花药;或小孢子和单核花粉胞内降解,形成不同形状和外壁纹饰的败育花粉.研究认为,灯盏花花药绒毡层异常是其花粉败育的主要原因.     

Real‐time droplet size analysis using laser micrometer as a process analytical technology tool for continuous dripping process

Process analysis and monitoring during the manufacturing of the dripping pills are essential. However, research on developing sensor‐based technology or process analytical technology (PAT) tools to analyze and monitor the dripping process is minimal. The purpose of this work is to develop a fast and non‐destructive laser detection system for quantitative visualization of droplets, which involves detecting the size of the droplet and calculating the weight of the dripping pills during the dripping process. Several factors influencing the detection performance of the detection system and the detection system capability for quantitation of the pill weight were explored. The laser detection system accurately detects the weight of the dripping pills with the coefficients of determination (R²) higher than 0.99. It was also robust concerning the variation in critical process parameters and critical material attributes. Furthermore, the laser detection system was successfully applied to the production line of Ginkgo biloba leaf dripping pills to monitor the dripping pills weight. The proposed laser detection system can analyze and monitor the dripping process in dripping pill manufacturing with stable performance, high accuracy, and high efficiency.     

Expression of injected HPRT minigene DNA in mouse embryos and its inhibition by antisense DNA

We have used a highly sensitive biochemical microassay to monitor the expression of a cloned minigene for hypoxanthine phosphoribosyl transferase (HPRT, EC.2.4.2.8) in preimplantation mouse embryos. The mouse HPRT promoter and the mouse metallothionein promoter (MT-I) function equally well in embryos at the 2-cell stage whereas the viral SV40 promoter does not allow HPRT expression. Induced HPRT activity from the MT-I HPRT minigene construct occurs in cleavage embryos cultured in the presence of cadmium. In contrast, negation of enzyme expression from the injected minigene DNA is mediated by simultaneous injection of HPRT antisense DNA.     

Localization of injected apolipophorin III in vivo - new insights into the immune activation process directed by this protein

A few years ago, it was shown that intrahemocoelic injection of the insect apolipoprotein apolipophorin III (apoLp-III) stimulates an immune response in larvae of the greater wax moth, Galleria mellonella. Since the mode of action of this activation process is unknown, we followed apoLp-III's pathway in the early phase of the immune-stimulating process, using biotin as a probe. Biotinylated apoLp-III was injected and localized using avidin-coupled horseradish peroxidase. The labeled protein was fully functional; the added amount of biotin per apoLp-III molecule used in this study only slightly decreased its ability to associate with phospholipase C-treated human low-density lipoprotein, as well as the immune-stimulating capability of apoLp-III.Gel electrophoresis with subsequent staining of biotin moieties and lipids revealed that apoLp-III undergoes lipid association in vivo within the first few minutes after injection. After two hours, no biotinylated apoLp-III was detectable in cell-free hemolymph. At this time, a subpopulation of hemocytes showed a distinct peroxidase staining. Control injections of biotinylated bovine serum albumin did not lead to similar results, giving evidence for the specificity of the phenomena observed. The results indicate that lipid association of apoLp-III occurs prior to endocytosis by immune-competent hemocytes, which is followed by the induction of a humoral immune response.     

岩黄连茎基腐病的分离鉴定及防治

对引起药用植物岩黄莲茎基部腐烂的病原进行分离 ,获得纯的活体病原 ,然后进行病原菌的致病性测定 ,确定致病病原。根据病原形态 ,初步鉴定为 :无性态属于半知菌类葡萄孢属 (Botrytissp.) ,有性态属于子囊菌门葡萄核盘菌属 (Botrytinia sp.)。并提出岩黄连人工栽培中该病的防治方法。     

Concerning the stimulation by injected cyclic AMP of the sodium efflux in barnacle muscle fibres and its transient nature

The efflux of 22Na in ouabain-poisoned barnacle muscle fibres and its transitory response to injected cAMP has been studied by using a new xanthine derivative, 1-propyl-3-methyl-7-(5-hydroxy-hexyl)-xanthine (PMX). Injection of PMX prior to cAMP fails to significantly alter the behaviour of the ouabain-insensitive Na efflux towards the nucleotide. By contrast, injection of PMX following peak stimulation by injected cAMP stops the rate constant for 22Na efflux from falling. This effect of PMX is not mimicked by injected HEPES. (a). Injection of Mg2+ following PMX brings about almost complete reversal of the sustained stimulatory response. (b). Injection of trace metals, e.g. Fe and Zn, following PMX brings about complete reversal of the sustained stimulatory response. (c). Injection of RI or RII subunits following PMX brings about partial reversal of the sustained stimulatory response. Partial reversal is also seen with externally applied imipramine (50 microM). These results support the view that the transitory nature of the response of the ouabain-insensitive Na efflux to injected cAMP is due to high phosphodiesterase activity in these fibres and that the major portion of the response itself is due to activation by cAMP of cAMP-dependent protein kinase.     

Capsicum frutescens L. in Southeast and East Asia, and its dispersal routes into Japan

Isozyme analyses were conducted to study the geographic variation ofCapsicum frutescens L. in Southeast and East Asia, and to investigate its dispersal routes into Japan. Eight enzymes (EST, EM, G6PD, GR, ME(A), PGI, PGM, ShDH) were variable among accessions ofC. frutescens in Southeast and East Asia. Accessions from the Ryukyu Islands were closely related to those in Indonesia, whereas accessions from the Bonin Islands showed exactly the same isozyme patterns as those from Indonesia and Northern Thailand. Accessions in the Ryukyu Islands were different from those in the Bonin Islands, suggesting that there may be two independent dispersal routes into Japan. One route was from Indonesia via the Philippines or Taiwan, or directly to the Ryukyu Islands, and another was from Indonesia via the Mariana Islands, or other islands in the Pacific, to the Bonin Islands.     

湖州市非细菌性急性胃肠炎暴发中诺如病毒的分子生物学特点初步研究

本文初步研究了湖州市非细菌性急性胃肠炎暴发中检出的诺如病毒的分子生物学特征。收集湖州市2008年和2009年2起非细菌性急性胃肠炎暴发疫情中采集的患者粪便标本,采用荧光定量RT-PCR方法对其进行诺如病毒核酸检测,并对核酸阳性标本进行RNA多聚酶部分区域的RT-PCR扩增。选取阳性扩增产物进行纯化及序列测定,结合诺如病毒GI、GII各基因型参考株进行核苷酸序列遗传进化分析。结果2起疫情均同时检出了GI和GII型诺如病毒。随后对其中4份RNA多聚酶区扩增阳性的标本进行测序及序列分析,结果发现2008年检出的2株GI型诺如病毒均为GI/2基因型;而2009年检出的1株GI型诺如病毒为GI/3基因型,另一株GII型诺如病毒则与近些年欧洲和亚洲相继出现的遗传组GII的新基因型GIIb的各代表株同源性最高,为GIIb基因型。这说明湖州地区流行的诺如病毒存在很高的遗传多样性,并且不同时间流行的基因型也存在一定差异。这也是国内首次在急性病毒性胃肠炎暴发中检出诺如病毒GIIb变异株。     

Mitochondrial damage and its role in causing hepatocyte injury during stimulation of lipid peroxidation by iron nitriloacetate.

Incubation of isolated rat hepatocytes with 0.1 mM iron nitrilotriacetic acid (FeNTA) caused a rapid rise in lipid peroxidation followed by a substantial increase in trypan blue staining and lactate dehydrogenase release, but did not affect the protein and non-protein thiol content of the cells. Hepatocyte death was preceded by the decline of mitochondrial membrane potential, as assayed by rhodamine 123 uptake, and by the depletion of cellular ATP. Chelation of extracellular Ca2+ by ethylene glycol bis(beta-aminoethyl ether) N,N'-tetraacetic acid or inhibition of Ca2+ cycling within the mitochondria by LaCl3 or cyclosporin A did not prevent the decline of rhodamine 123 uptake. On the other hand, a dramatic increase in the conjugated diene content was observed in mitochondria isolated from FeNTA-treated hepatocytes. Oxidative damage of mitochondria was accompanied by the leakage of matrix enzymes glutamic oxalacetic aminotransferase (GOT) and glutamate dehydrogenase (GLDH). The addition of the antioxidant N,N'-diphenylphenylene diamine (DPPD) completely prevented GOT and GLDH leakage, inhibition of rhodamine 123 uptake, and ATP depletion induced by FeNTA, indicating that Ca(2+)-independent alterations of mitochondrial membrane permeability consequent to lipid peroxidation were responsible for the loss of mitochondrial membrane potential. DPPD addition also protected against hepatocyte death. Similarly hepatocytes prepared from fed rats were found to be more resistant than those obtained from starved rats toward ATP depletion and cell death caused by FeNTA, in spite of undergoing a comparable mitochondrial injury. A similar protection was also observed following fructose supplementation of hepatocytes isolated from starved rats, indicating that the decline of ATP was critical for the development of FeNTA toxicity. From these results it was concluded that FeNTA-induced peroxidation of mitochondrial membranes impaired the electrochemical potential of these organelles and led to ATP depletion which was critical for the development of irreversible cell injury.     

Lysis of a red-tide causing alga,Alexandrium tamarense,caused by bacteria from its phycosphere

There are bacteria coexisting in xenic cultures of Alexandrium tamarense, a red-tide causing alga. However little is known concerning the interactions between the alga and the bacteria in its phycosphere. The objective of the current study was to determine the effect of the bacteria in its phycosphere on the growth of the alga. We added one percent (v/v) Zobell 2216E medium to A. tamarense culture to alter bacterial growth and the results showed that algal cells were all lysed within 14 h. After adding the medium, both the abundance and the extracellular enzyme activity of the bacteria increased by 50–100 times from the 4th to the 10th hour which resulted in lysis of the algae. The 16S rRNA gene fragments of the bacteria were amplified from the DNA extracted from A. tamarense cultures and analyzed by denaturing gradient gel electrophoresis and sequencing. The structure of the bacterial community in phycosphere changed significantly during algal lysis. Two bacterial genera, Alteromonas sp. and Thalassobius aestuarii sp. are key factors that caused the lysis, and the β-glucosidase and chitinase produced by the bacteria in the phycosphere could directly cause the lysis. These experiments provide evidence that bacteria in its phycosphere play a key role in the culture of A. tamarense, and may provide insights into the future biocontrol of red-tides.     

The study of gene GJB2/DFNB1 causing deafness in humans by linkage analysis from district Peshawar

Families with at least 2 or more individuals having hereditary hearing loss were enrolled from different areas of Khyber Pakhtoonkhwa, mainly from district Peshawar. Detailed history was taken from each family to minimize the presence of other abnormalities and environmental causes for deafness. Families were questioned about skin pigmentation, hair pigmentation, and problems relating to balance, vision, night blindness, thyroid, kidneys, heart, and infectious diseases like meningitis, antibiotic usage, injury, and typhoid. The pedigree structures were based upon interviews with multiple family members, and pedigrees of the enrolled families were drawn using Cyrillic program (version 2.1). All families showed recessive mode of inheritance. I studied 8 families of these 10. For linkage analyses, studies for DFNB1 locus, 3 STR markers (D13S175, D13S292, and D13S787) were genotyped using polyacrylamide gel electrophoresis (PAGE) and haplotypes were constructed to determined, linkage with DFNB1 locus. From a total of 8 families, a single family-10 showed linkage to DFNB1 locus.