HBsAg合成肽段的免疫源性和抗肽血清与天然HBsAg的结合测定

我们将三种乙型肝炎表面抗原的合成肽段在联结载体或不接载体的情况下,分别制备成4种免疫原对兔子进行了免疫。大部分兔子都产生了抗肽抗体。其中抗P_(122-48)(auw)与抗P_(122-148)(adw)都能与天然HBsAg反应,而抗P_(122-148)(adw)的抗HBs活力比文献报道的都高,通过结构分析表明:含有天然蛋白上免疫显性区域的免疫原是合成疫苗的理想选者。     

狂犬病疫苗蔗糖密度梯度离心纯化工艺开发

目的:开发冻干人用狂犬病疫苗(鸡胚成纤维细胞)的连续流蔗糖密度梯度离心纯化工艺。方法:分别比较两种不同初始蔗糖浓度和不同上样速度对纯化效果的影响,初步确定纯化工艺;通过多批次实验确定样品收集范围;比较不同浓缩倍数条件下杂质去除和抗原回收情况,确定合适的收获液浓缩比例;比较不同批次样品纯化后的杂质去除率和重复性,判定本纯化工艺的稳定性。结果:选取60%作为初始蔗糖浓度,在上样速度为150～200ml/min时,可以有效地对10倍浓缩的病毒收获液进行纯化;卵清蛋白、牛血清白蛋白和庆大霉素去除率分别达到99%,95%和95%,且工艺具有极好的稳定性。结论:开发的连续流蔗糖密度梯度离心技术可以作为冻干人用狂犬病疫苗(鸡胚成纤维细胞)的产业化纯化工艺。     

一个具有寄生虫病感染和恢复的捕食模型(英文)

本文假设感染的食饵有恢复率和对捕食者有收获,研究了一个对部分食饵和全部捕食者具有寄生虫病感染的捕食模型.用定性理论证明了边界和正平衡点的稳定性.结论表明恢复率和收获率对正平衡点的稳定性有影响.     

苦瓜MAP30基因的毕赤酵母表达载体构建及重组菌株的筛选

目的:从苦瓜中克隆MAP30全长基因,并将该基因连接至表达载体pPIC9中,建立酵母菌落PCR筛选方法。方法采用改良SDS法从苦瓜表皮中提取基因组DNA,设计特异性的引物,通过PCR技术扩增出全长861bp的MAP30基因。该基因经XhoⅠ和EcoRⅠ双酶切,连接至毕赤酵母表达载体pPIC9中。重组载体转化GS115菌株,运用菌落PCR鉴定重组菌株。结果:基因测序表明,该基因已成功插入酵母表达载体pPIC9α-factor分泌信号下游,同源性分析表明该基因与GeneBank(AF284811)的核苷酸同源性达99.9%,氨基酸同源性达100%。菌落PCR显示外源基因已整合入酵母GS115菌株中。结论:成功地克隆了MAP30全长基因,并构建了含MAP30基因的重组毕赤酵母表达载体,并获得了整合菌株,为下一步研究奠定了基础。     

Andrographolide induces cell cycle arrest and apoptosis in human rheumatoid arthritis fibroblast-like synoviocytes

The pseudo-tumoral expansion of fibroblast-like synoviocytes is a hallmark of rheumatoid arthritis (RA), and targeting rheumatoid arthritis fibroblast-like synoviocytes (RAFLSs) may have therapeutic potentials in this disease. Andrographolide, a diterpenoid compound isolated from the herb Andrographis paniculata, has been reported to have potent anti-inflammatory activity. In the present study, we aimed to investigate the effects of andrographolide on human RAFLSs and the underlying molecular mechanism(s). RAFLSs were isolated from patients with RA and treated with or without various concentrations (i.e., 10, 20, and 30 μM) of andrographolide for 48 h. 3-[4,5-Dimethyl-2-yl]-2,5-diphenyl tetrazolium bromide assay revealed that andrographolide treatment decreased the proliferation of RAFLSs in a dose-dependent manner. Cell cycle analysis using propidium iodide (PI) staining showed a G0/G1 cell cycle arrest in andrographolide-treated RAFLSs. Immunoblotting analysis of key cell cycle regulators demonstrated that andrographolide treatment caused a dose-dependent increase in the expression of cell-cycle inhibitors p21 and p27 and a concomitant reduction of cyclin-dependent kinase 4. Exposure to andrographolide-induced apoptosis of RAFLSs measured by annexin V/PI double staining, which was coupled with promotion of cytochrome C release from mitochondria and activation of caspase-3. Moreover, andrographolide-treated RAFLSs displayed a significant decrease in the Bcl-2/Bax ratio compared to untreated cells. In conclusion, our data demonstrate that andrographolide exerts anti-growth and pro-apoptotic effects on RAFLSs, thus may have therapeutic potential for the treatment of RA.     

湖北泽泻的HPLC分析及有效成分含量测定

采用高效液相色谱方法,以乙腈和水(65?35)为流动相,图谱收集波长为210 nm。用指标成分24-乙酰泽泻醇A、23-乙酰泽泻醇B的标准品作回归曲线(利用浓度和峰面积的线性关系),对药店出售的片状干燥泽泻的含量进行测定和图谱分析,并与资料中展示的不同泽泻制品含量相比较,从而对泽泻的制作方法、有效成分含量的影响及泽泻质量进行评价,建立起成熟的质量评价的定量方法体系。     

整合分析氮磷添加对土壤酶活性的影响

本文通过整合分析(Meta-analysis)的方法分析了氮、磷添加对土壤碳、氮和磷素循环水解酶以及土壤氧化酶活性的影响.结果表明: 氮添加显著增加了碳、氮和磷循环水解酶的活性,增幅分别为6.9%、5.6%和10.7%;与氮添加相比,在氮磷同时添加下,3类土壤酶的活性增加更为显著,增幅分别达13.4%、37.4%和13.3%.然而,对于土壤氧化酶,氮以及氮磷的添加都使其活性降低,分别降低了6.1%和0.4%.不同生态系统类型、氮肥类型、施肥速率和施肥试验时间都对土壤酶活性具有影响.在全球大气氮沉降与磷添加逐渐增加的背景下,土壤微生物活性和酶的变化将会对土壤生物地球化学循环过程和土壤生态系统功能产生重要影响.     

玛纳斯河流域土壤斥水性及其影响因素

土壤斥水性(SWR)阻碍入渗及再分布过程.虽然斥水土壤在世界广泛存在,但其产生机理并不明确.本文采用滴水穿透时间(WDPT)法、酒精摩尔浓度(MED)法和接触角(θ)法测定了新疆玛纳斯河流域典型土壤的SWR指标,其中θ的测定采用高度法和质量法.对WDPT是否受烘干及不同容重的影响进行对比,探讨各SWR指标之间的联系,并分析了SWR的关键理化性质影响因子,探讨土壤斥水的机理和原因.结果表明: 高容重土壤的WDPT值更大,烘干土壤的WDPT值比风干土壤大;3种SWR指标(WDPT、MED及θ)之间有一定的相关关系,但MED与θ的相关性并不明显,表明各SWR指标之间虽有一定联系,但表征方面具有差异性;采用高度法和质量法测量水与土壤的θ值时,高度法的θ值大于质量法,高度法之间重复差异小于质量法;使用正辛烷作为参比溶液测量水与土壤的θ值时,其重复的差异较小,低于无水乙醇;在多种理化性质中,黏粒含量比其他土壤属性更显著地影响WDPT和θ;[K⁺]、[Na⁺]均与θ呈正相关;蒙脱石含量与WDPT、MED呈负相关.综上,土壤SWR的测定方法中,WDPT法操作简单但易受影响;MED法缩短了入渗时间但试验过程耗时;接触角法操作复杂,结果相对精确,采用正辛烷作为参比溶液的高度法更可靠.建议采用多种指标综合表征土壤斥水性.     

Preliminary Study on Transformation of Wheat Apical Point

Apical points of young seedlings of wheat (Triticum aestivum) cultivar "Jing 411” and somatic calli of cultivar "FK8” were transformed with plasmids pBI121 and (or) pBIAH-A+ by using microprojectile bombardment. Histochemical assay of GUS activity showed positive reaction on some of the transformation processed apical points and calli. This demonstrated that foreign genes were introduced into the apical meristematic cells as well as the callus cells. The plantlets of cv. "Jing 411” survived after apical point transformation with pBIAH-A+ were transplanted into the field and the progenies were screened with kanamycin. 4 % of the screened seeds germinated into green seedlings with kanamycin resistance. Dot hybridization of total DNA from kanamycin resistant plants showed the existence of foreign DNA in some of the detected plants.     

ERβ对子宫内膜异位症小鼠模型的影响

目的通过建立子宫内膜异位症小鼠模型探讨雌激素β受体对子宫内膜异位症的影响。方法利用雌激素β基因敲除小鼠建立自体子宫内膜异位模型;应用人不同的组织在SCID小鼠建立子宫内膜异位症模型后,注射雌激素β受体激动剂WAY-200070,观察其对异位病灶生长的影响。结果比较30只雌激素β受体基因敲除小鼠及22只同源未敲除小鼠异位病灶生长及组织细胞形态无明显差异（P〉0.05）;雌激素β受体激动剂WAY-200070对不同类型的SCID小鼠内异症病灶生长影响无明显差异（P〉0.1）。结论雌激素β受体对子宫内膜异位病灶的形成影响作用微弱。