安氏隐孢子虫卵囊分离纯化方法研究

目的 建立获取纯净的安氏隐孢子虫卵囊外培养模型。方法研究改进分离纯化安氏隐孢子虫卵囊的方法,采用蒸馏水稀释Sheather’s蔗糖溶液配制不同密度的蔗糖溶液,比较不同稀释比的Sheather’s蔗糖溶液漂浮卵囊的效果,并首次比较了蒸馏水和PBS稀释的Sheather’s蔗糖溶液分离纯化卵囊的活性,将两种方法分离纯化的卵囊37℃水浴1h进行脱囊率比较。结果2：1蔗糖溶液（2体积Sheather’s蔗糖溶液与1体积的蒸馏水）分离奶牛粪样中卵囊效果最好。漂浮三次即可获得大量卵囊,回收率为83．70％,三次漂浮的卵囊总数较Sheather’s蔗糖溶液高57．53％。蔗糖溶液多梯度密度纯化法纯化后的卵囊杂质较少,回收率为47．52％。蒸馏水稀释的蔗糖溶液分离纯化卵囊脱囊率为87．87％,PBS稀释的蔗糖溶液分离纯化卵囊的脱囊率为84．71％,差异无统计学意义P〉0.05）。结论2：1蔗糖溶液适用于分离安氏隐孢子虫卵囊,且用蒸馏水代替PBS稀释Sheather’s蔗糖溶分离纯化隐孢子虫卵囊其活性不受影响。可用于体外培养研究。     

济宁市部分医院手术室空气洁净度调查

[目的]了解济宁市医疗机构洁净手术室的空气净化状况,促进医疗机构加强洁净手术室的管理。[方法]对济宁市10家医院的36间洁净手术室进行空气化学、微生物指标和相关技术指标现场静态检测。按《医药工业洁净室（区）悬浮粒子的测试方法》、《医药工业洁净室（区）沉降菌的测试方法》和《公共场所卫生检验标准方法》进行检测,按GB 50333-2002《医院洁净手术部建筑技术规范》中的等级标准进行分级评价。[结果]36间洁净手术室中设计等级为Ⅰ、Ⅱ、Ⅲ级的洁净手术室合格率分别为80.0%、83.3%、87.5%;空气中尘粒数与沉降菌菌落数具有相关性,沉降菌菌落数与0.5μm尘粒数的相关系数为0.481（P〈0.01）,沉降菌菌落数与5μm尘粒数的相关系数为0.150（P〈0.01）,参照相关标准进行评价,静压差、照度、噪声、温度、相对湿度、换气次数、风速等技术指标合格率分别为83.3%、94.4%、91.7%、94.4%、88.9%、88.5%、94.4%,其中静压差低于洁净等级合格率。[结论]应在设计和施工阶段重视洁净室相关技术指标的审查和监理,提高静压差等合格率。     

Use of coupled plasma filtration adsorption for septic shock treatment

Coupled plasma filtration adsorption (CPFA) is a new technology proposed for septic shock, removing both pro- and anti-inflammatory mediators. We report the first use of CPFA in France. The patient was 80-years-old and was admitted to the intensive care unit because of septic shock from urinary source. Besides conventional septic shock treatment, four successive CPFA sessions were daily performed and lasted from 6 to 11 hours. At the end of the sessions, a noteworthy hemodynamic and inflammatory improvement was observed. This case confirms the feasibility and the relative simplicity of CPFA. It also illustrates the recent and few scientific data of the medical literature concerning this technology.     

藻兰蛋白分离纯化方法的改进

目的 在藻兰蛋白原有分离纯化方法基础上，探讨对其进行改进的可能性。方法 用５０％（Ｗ／Ｖ）硫酸铵沉淀法粗分离的藻兰蛋白，再用ＤＥＡＥ离子交换柱和羟－基磷灰石柱纯化。结果 藻兰蛋白纯度达到９２％，并在６１９．４ｎｍ处有最大吸收峰。在聚丙烯酰胺凝胶电泳（ＰＡＧＥ）上只汪条带。分子量约为２６ＫＤ。结论 该方法既可达到较高纯度，又适合大量生产。     

Characterization of acetylcholinesterase from Korean electric ray and comparison withTorpedo californica

This study has been undertaken to examine the acetylcholinesterase (AChE) of electric organ from korean electric ray (Narke japonica). Korean electric ray was caughted at Chungmu sea and transported to the laboratory, where electric organs were removed and stored at −70°C until used. Acetylcholinesterase(AChE) of electric organ was purified by affinity column that was prepared with dicaproyl-methylpyridinium linked to Sepharose 4B. Upon purification, the specific activities in Ellman unit were increased by 52 and 39 times for high salt soluble AChE (HSSE, 870.86 ΔOD/min/gram of tissue) and detergent soluble AChE(DSE, 105.42 ΔOD/min/gram of tissue), respectively. Each subunit of AChE separated by SDS polyacrylamide gel electrophoresis(SDS-PAGE) was transferred to immobilon P by western blotting and detected by mAbs raised against each subunit of AChE from electric organ ofTorpedo californica. Collagenic tail of AChE fromNarke japonica were identified by monoclonal antibody specific to collagenic tail of AChE fromTorpedo californica, likewise 103Kd protein of AChE fromNarke japonica was detected by monoclonal antibody specific to 103Kd of AChE fromTorpedo californica. However, molar ratio of three subunits of AChE fromNarke japonica is different from that ofTorpedo californica. Furthermore, catalytic subniit of AChE fromNarke japonica was not identified by monoclnal antibody specific to catalytic subunit of AChE fromTorpedo californica. These results showed differences in molecular structure of AChE fromNarke japonica and AChE fromTorpedo californica eventhough they showed same enzymatic activities.     

胆红素IX_α的提取与纯化研究

用直接水解法从新鲜猪胆汁中提取胆红素IX_α,所得胆红素粗品用改良的氯仿抽提法进行纯化。通过元素分析、红外光谱和质谱等对胆红素的纯度进行确证。胆红素的质谱图上出现清晰的特征谱峰,这些谱峰可望在胆红素异构体和纯度的快速检验方面得到应用。     

Extraction and Purification of Hepatocyte Growth Factor of New-Born Calf

新生小牛肝经组织匀浆、加热和离心制得粗制的肝细胞生长因子（ＨＧＦ），然后经过超滤、ＤＥＡＥＤＥ５２纤维层析，Ｓｕｐｅｒｏｓｅ１２凝胶过滤，ＭｏｎｏＱ离子交换层析和高效液相色谱进行分离提纯，用３Ｈ标记的胸腺嘧啶核苷掺入法在人肝瘤ＳＭＭＣ７２２１细胞株上检测提纯的新生小牛肝细胞生长因子的生物学活性，得到一分子量为（１２～１４）×１０３的有促进肝细胞有丝分裂的有效成分。显而易见此ＨＧＦ的分子量不同于循环在血浆中的ＨＧＦ的分子量。本文对２种ＨＧＦ的不同作用进行了讨论。     

云南产螺旋藻多糖的分离纯化与分析

云南程海湖工业化养殖的螺旋藻（干品）经热水提醇沉，ｓｅｖａｇ法脱蛋白，透析，再经ＤＥＡＥ－ｓｅｐｈａｄｅｘ－Ａ２５与ｓｅｐｈａｄｅｘＧ－２００柱层析纯化，得螺旋藻多糖（ＰＳＰ）。用纸层析，柱层析及电泳检查纯度，表明ＰＳＰ为单一组分。ＰＳＰ经ＵＶ－３００扫描，无核酸（２６０ｎｍ）及蛋白质（２８０ｎｍ）特征吸收峰。ＩＲ－４００扫描，出现典型多糖吸收峰。经纸层析及气相色谱分析表明ＰＳＰ组成单糖为Ｌ－岩藻糖、Ｄ－甘露糖、Ｄ－半乳糖、Ｄ－葡萄糖和葡萄糖醛酸。ＰＳＰ平均分子量约为１６６００。     

大鼠A型精原干细胞的分离和纯化

目的摸索完善A型精原干细胞分离、纯化的方法与技术。方法选用9d的SD大鼠,不连续Percoll梯度液分离纯化精原干细胞,应用选择性贴壁法进一步纯化精原干细胞;台盼蓝排斥实验确定精原干细胞的活力;HE染色法观察细胞形态;c-kit细胞免疫组化鉴定细胞类型并进行细胞纯度分析。结果台盼蓝实验显示细胞活力维持于88.73%±0.85%;c-kit细胞免疫组化结果显示分离得到细胞为精原干细胞;细胞纯度为90.48%±1.78%。结论应用梯度离心及选择性贴壁法获得了纯度较高的A型精原干细胞。     

Epitope selection and their placement for increased virus neutralization in a novel vaccination strategy for porcine epidemic diarrhea virus utilizing the Hepatitis B virus core antigen

Porcine epidemic diarrhea virus (PEDV) is a member of the Alphacoronaviridae genus within the Coronaviridae family. It is the causative agent of porcine epidemic diarrhea, a disease that can have mortality rates as high as 100% in suckling piglets. PEDV causes severe economic loss, and has been in existence for decades. A panzootic starting in 2010 renewed interest in the development of a universal vaccine toward PEDV. This report details several design changes made to a Hepatitis B virus core antigen (HBcAg)-based recombinant vaccine strategy, and their effect in vivo. Initially, several multi-antigen vaccine candidates were able to elicit antibodies specific to three out of four B-cell epitopes inserted into the chimeric proteins. However, a lack of virus neutralization led to a redesign of the vaccines. The focus of the newly redesigned vaccines was to elicit a strong immune response to the YSNIGVCK amino acid motif from PEDV. Genetically modified new vaccine candidates were able to elicit a strong antibody (Ab) response to the YSNIGVCK epitope, which correlated with an increased ability to neutralize the CO strain of PEDV. Additionally, the location of the inserted PEDV epitopes within the vector protein was shown to affect the immune recognition toward the native HBcAg during vaccination.