1,25二羟基维生素D_3对兔角膜碱烧伤朗格罕氏细胞影响

目的：研究1,25二羟基维生素D3（骨化三醇）对兔角膜碱烧伤后角膜朗格罕氏细胞分布的影响,并初步探讨其作用机制。方法：在兔角膜制作碱烧伤模型,然后实验组局部和全身给予1,25二羟基维生素D3,分别在第3,7,21天时对正常组,实验组和对照组家兔行角膜共聚焦显微镜,HE染色观察角膜病理改变。结果：正常组角膜中央在三个时间点均未检测出朗格罕氏细胞。实验组和对照组碱烧伤后3、7天角膜中央出现朗格罕氏细胞,对照组密度高于实验组（p〈0.05）;碱烧伤后21天两组朗格罕氏细胞密度相近（p〉0.05）。实验组炎性反应程度在第7,21天时轻于对照组。结论：1,25二羟基维生素D3能够在兔角膜碱烧伤早期抑制朗格罕氏细胞的向心性迁移,并且能在一定程度上抑制炎性反应。     

威廉·哈维和发生论

作为一名杰出的生理学家,威廉·哈维是血液循环的发现者,并著有《心血运行论》一书。他的这一发现也被誉为科学史上的一次伟大革命,并奠定了实验生理学的研究。     

左旋多巴甲酯缓释微球的研究

目的:由于长期服用左旋多巴治疗帕金森病,其药物浓度波动刺激易引起异动症,本实验旨在制备突释小,药物释放浓度稳定的左旋多巴甲酯微球制剂。方法:将左旋多巴甲酯用复乳法包裹于PLGA微球内,采用C18反相色谱研究药物包封率和体外释放行为。结果:通过调节药物浓度和不同高分子组合筛选出突释小,包封率高且缓慢释放的处方。结论:左旋多巴甲酯包裹于PLGA能实现理想的缓释效果,降低药物浓度波动,为后期药效学实验提供基础。     

英国科学家研制一种新塑料其表面能高效培育干细胞

英国格拉斯哥大学组织工程师马修·道尔贝和南安普敦大学肌肉与骨骼科学研究系主任理查德·奥瑞福使用制造蓝光光盘的方法成功研制一种新的塑料,该表面上布满细小的凹坑,这种表面有助于干细胞更有效地生长和扩散成对治疗有用的细胞。在此基础上,有望研制出大规模的干细胞培育工厂,这种干细胞培育工厂将使科学家更有效地治疗糖尿病、关节炎、阿尔茨海默症、帕金森氏病等病症。     

应用缝合锚结合联合腱外侧半肌腱重建喙锁韧带治疗肩锁关节脱位

目的:观察治疗肩锁关节脱位的不同手术方式的治疗效果,以确定最佳的手术方法。方法:将60例肩锁关节脱位病人随机分为治疗组和对照组,分别采用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动的治疗方法及克氏针张力带内固定方法,术后3个月及6个月对患者进行JOA总体情况评定、JOA评分比较、肩锁关节疼痛VAS评定情况比较及安全性评价。结果:应用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位3个月及6个月后JOA总体评定情况结果及.JOA评分结果优于克氏针张力带治疗方法;应用缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位3个月及6个月后肩锁关节疼痛程度均轻于同时期克氏针张力带治疗方法。结论:缝合锚结合联合腱外侧半肌腱反转移位重建喙锁韧带辅助颈腕吊带制动治疗肩锁关节脱位是一种安全有效的临床治疗方法。     

1,25 二羟基维生素D3 对兔角膜碱烧伤朗格罕氏细胞影响

目的:研究1,25二羟基维生素D3(骨化三醇)对兔角膜碱烧伤后角膜朗格罕氏细胞分布的影响,并初步探讨其作用机制。方法:在兔角膜制作碱烧伤模型,然后实验组局部和全身给予1,25二羟基维生素D3,分别在第3,7,21天时对正常组,实验组和对照组家兔行角膜共聚焦显微镜,HE染色观察角膜病理改变。结果:正常组角膜中央在三个时间点均未检测出朗格罕氏细胞。实验组和对照组碱烧伤后3、7天角膜中央出现朗格罕氏细胞,对照组密度高于实验组(p<0.05);碱烧伤后21天两组朗格罕氏细胞密度相近(p>0.05)。实验组炎性反应程度在第7,21天时轻于对照组。结论:1,25二羟基维生素D3能够在兔角膜碱烧伤早期抑制朗格罕氏细胞的向心性迁移,并且能在一定程度上抑制炎性反应。     

LT（K63／R72）, a New Mutant of Escherichia coli Heat-labile Enterotoxin, Exhibits Characteristics More Similar to LT（K63） than LT（R72）

LT(K63), a non-toxic mutant and LT(R72), a low toxic mutant of E. coli heat-labile enterotoxin are frequently used mucosal adjuvants. In many cases, the adjuvanticity of LT(K63) is lower than that of LT(R72), but LT(K63), which induces a mixed Th1/Th2 response, exhibits a higher level of protection than LT(R72) which induces a polarized Th2-type response. To utilize the advantages of both adjuvants, a doublemutation LT(K63/R72) was generated and purified. The characterization results showed that there was no significant difference in production rate and immunogenicity between wild type LT and LT mutants. The results also showed that the toxicity and the trypsin sensitivity of LT(K63/R72) are between that of LT(K63)and LT(R72). Using HPLC, when samples in an OHpak SB-800 column were eluted by denatural buffer(TEAN containing 10 mg/ml SDS), we found the stability of LT(K63/R72) was higher than that of LT(R72)and lower than that of LT(K63). Through further analyzes, we found that LT(K63/R72) exhibits characteristics more closely related to LT(K63) than LT(R72).     

Prolonged Alzheimer-like tau hyperphosphorylation induced by simultaneous inhibition of phosphoinositol-3 kinase and protein kinase C in N2a cells

Co-injection of wortmannin (inhibitor of phosphatidylinositol-3 kinase, PI3K) and GF109203X(inhibitor of protein kinase C, PKC) into the rat brain was found to induce spatial memory deficiency and enhance tau hyperphosphorylation in the hippocampus of rat brain. To establish a cell model with durative Alzheimer-like tau hyperphosphorylation in this study, we treated N2a neuroblastoma cells with wortmannin and GF109203X separately and simultaneously, and measured the glycogen synthase kinase 3 (GSK-3)activity by y-32p-labeling and the level of tau phosphorylation by Western blotting. It was found that the application of wortmannin alone only transitorily increased the activity of GSK-3 (about 1 h) and the level of tau hyperphosphorylation at Ser^396/Ser^404 and Ser^199/Ser^202 sites (no longer than 3 h); however, a prolonged and intense activation of GSK-3 (over 12 h) and enhanced tau hyperphosphorylation (about 24 h) were observed when these two selective kinase inhibitors were applied together. We conclude that the simultaneous inhibition of PI3K and PKC can induce GSK-3 overactivation, and further strengthen and prolong the Alzheimerlike tau hyperphosphorylation in N2a cells, suggesting the establishment of a cell model with early pathological events of Alzheimer‘s disease.     

创伤弧菌vvhA基因的克隆、原核表达系统的构建及鉴定

目的 克隆创伤弧菌（Vibrio vulnificus,Vv）溶细胞素基因（υυhA）,构建原核表达系统并鉴定其表达产物的免疫性.方法 采用PCR技术从Vv GTC333和WZ01株DNA中扩增全长υυhA基因,T-A克隆后测定其核苷酸序列.采用pET32a质粒构建vvhA基因原核表达载体,在E coli BL21（DE3）宿主菌中用不同浓度的IPTG诱导目的重组蛋白rVvhA表达,采用Ni-NTA亲和层析法提纯rVvhA,SDS-PAGE检测表达和提纯效果.采用兔抗Vv全菌抗体的Western Blot和兔抗rVvhA血清的免疫扩散试验鉴定其免疫反应性和免疫原性.结果 所克隆的vvhA基因核苷酸序列与GeneBank公布的同源性分别为96.09%和98.26%.在0.5 mmol/L IPTG诱导下,rVvhA产量可占细菌总蛋白的18%.提纯的rVvhA经SDS-PAGE后仅显示单一的蛋白条带.重组蛋白rVvhA能与兔抗Vv全菌抗体发生特异性结合,免疫家兔可获得高效价抗体.结论 该研究成功地构建了创伤弧菌υυhA基因高效原核表达系统,所表达的rVvhA具有良好的免疫原性和免疫反应性,可作为Vv免疫检测试剂盒及疫苗的抗原.     

黑曲霉中内切菊粉酶基因及其全合成基因在解脂耶氏酵母中表达的比较

利用酵母密码子偏爱性将黑曲霉(Aspergillus niger)中的内切菊粉酶(Endoinu linase)基因通过基因全合成的方式合成为酵母密码子偏爱性的内切菊粉酶基因。然后将原始和全合成的内切菊粉酶基因克隆到解脂耶氏酵母表达载体PINA1296上,得重组解脂耶氏酵母表达载体pHBM2020、pHBM2021,将两种质粒分别转化解脂耶氏酵母(Yarrowia lipolytica)CLIB725,筛选得到重组解脂耶氏酵母CLIB725(pHBM2020)、CLIB725(pHBM2021),将两种重组酵母摇瓶培养,经SDS-PAGE、测酶活检测表明两种基因在解脂耶氏酵母中都有表达,全合成菊粉酶比原始菊粉酶酶活要高。