吡酮酸类抗菌药物的研究Ⅷ.1-对氟苯基-6-氟-1,4-二氢-4-氧-7-(1-哌嗪)噌啉-3-羧酸及其类似物的合成和构效关系

为探讨构效关系,合成了1-对氟苯基-6-氟-1,4-二氢-4-氧-7-(1-哌嗪)噌啉-3-羧酸及其喹啉、萘啶、吡啶[2,3-C]哒嗪环系类似物16个。测定了对大肠杆菌的MIC。用Hückel分子轨道理论(HMO)方法计算了四个母体环上电子密度。结果表明:环中氮的位置对药效团——3位羧基和4位羰基氧原子上电子密度的影响较大而影响其抗菌活性。喹啉、萘啶两环系的3位羧基和4位羰基氧原子上的电子密度较高,其体外抗菌活性较高;而噌啉及吡啶[2,3-C]哒嗪两环系的电子密度较低,其体外抗菌活性较低甚至消失。     

Effect of timing of oocyte denudation and micro-injection on survival, fertilization and embryo quality after intracytoplasmic sperm injection

In human in-vitro fertilization (IVF), the oocytes are surrounded by cumulus and corona cells at the time of insemination so that their maturity cannot easily be evaluated. The best IVF results are obtained if the oocytes are inseminated 2-6 h after retrieval. In the intracytoplasmic sperm injection (ICSI) procedure, the oocytes are denuded by enzymatic and mechanical treatment in order to be able to perform the injection. As a consequence, the nuclear maturity of the oocytes can be evaluated and only those that have extruded the first polar body are injected. However, metaphase-II oocytes that have not yet reached cytoplasmic maturity cannot be recognized. The purpose of this study was to investigate the effect of different timing of cumulus- corona cell removal and injection on the outcome of ICSI. For this we allowed the oocytes to complete in-vitro cytoplasmic maturation in two different culture conditions: (i) surrounded by their cumulus and corona cells or (ii) totally denuded. We performed three different studies on sibling oocytes obtained after a standardized buserelin/human menopausal gonadotrophin (HMG) protocol. We investigated the effect of early (1-2 h after retrieval) and late (5-6 h after retrieval) oocyte denudation and injection on the survival and fertilization of the injected oocytes and on embryo cleavage after fertilization. We found no statistically significant differences between early and late injection, indicating that after a standardized buserelin/HMG protocol the metaphase-II oocytes do not need time for further cytoplasmic maturation. Furthermore, a different timing of cumulus-corona cell removal has no effect on the outcome of ICSI, suggesting that the surrounding cells are not necessary for survival, fertilization and cleavage after ICSI.      

Percoll gradient centrifugation can be omitted in sperm preparation for intracytoplasmic sperm injection

Prior to intracytoplasmic sperm injection (ICSI), seminal fluid is currently washed out from the ejaculated semen and further sperm selection is carried out by a discontinuous Percoll gradient. Possible deleterious effects from the sperm-separating substance Percoll on sperm function or embryo cleavage after in-vitro fertilization (IVF) have, to our knowledge, not yet been reported and the use of Percoll has been widely accepted in IVF. In this study, we examined whether the omission of the Percoll step in the sperm preparation has any influence on the outcome of the ICSI procedure. Two methods of sperm preparation for ICSI were compared in a controlled study on sibling oocytes: washing the semen sample once, followed by a Percoll gradient centrifugation versus washing the sperm sample twice without a Percoll gradient centrifugation. The mean fertilization rates were similar for the two sperm preparation methods: 78.2 +/- 21.4 and 75.0 +/- 24.1% respectively of the intact oocytes displaying two pronuclei. Cleavage rates did not differ statistically between the two groups. Whereas in both groups similar percentages of excellent, good and poor quality embryos were obtained, the percentage of fair quality embryos was significantly higher in the group without Percoll (16.3 +/- 20.1 versus 9.1 +/- 15.7%). However, no statistical differences were observed in either the percentage of transferable embryos or in the percentage of embryos actually transferred or frozen in the two groups. In conclusion, spermatozoa from ejaculates that are washed out from the seminal fluid without any further selection can be used for ICSI without any adverse effect on fertilization and embryo cleavage.      

盐酸戊乙奎宁在水溶液中的降解机理

Penequine hydrochloride (Ⅰ) is a new potent anticholinergic drug, the degradation mechanism of Ⅰ in aqueous solutions is reported in this paper. Ⅰ is an ethereal compound, stable in neutral and alkaline solutions, but it decomposes in strong acidic solutions. Its main degradation products have been separated and identified by means of TLC, MS, GC/MS and GC/FTIR. With reference to the general degradation rule of ethereal compound and the structure of the degradation products, we deduced that the ether linkage of Ⅰ splits in acidic solutions by the catalysis of hydrogen ion, producing 3- quinuclidinol (DP1) and 1-phenyl- 1-cyclopentyl glycol. The latter turns into 1-phenyl-l-cyclopentyl acetaldehyde through dehydration and rearrangement. In addition, small amounts of other degradation compounds, e. g. benzaldehyde, acetophenone, phenylcyclopentyl ketone, etc. have also been found, but the mechanism remains to be further studied.     

PLS—紫外分光光度法同时测定复方阿斯匹林片中三组分含量

本文研究了复方阿斯匹林片中阿斯匹林、非那西丁和咖啡因含量同时测定的紫外分光光度法的最佳实验条件,并简述偏最小二乘法(PLS)在多组分同时测定中的基本原理和应用。三组分模拟试样回收率平均值的置信区间分别为100.1±0.23%,100.0±0.25%和100.1±0.33%(置信度95%)。PLS法是一种理想的多组分测定方法,计算速度较快,结果更准确可靠,尤其适用于成批试样的分析,为微机控制的紫外可见分光光度计提供了一种新方法。     

肝脏可溶性复合物不同剂量、不同途径移植对肿瘤细胞增殖抑制的体内外实验

目的:肝脏可溶性复合物具有保护肝脏、刺激肝组织再生等生物学活性,观察天然物质肝脏可溶性复合物对肿瘤细胞生长增殖的抑制作用.方法:实验于2006-05/2007-02在四川大学华西医院生物治疗国家重点实验室实验肿瘤研究室完成.①分离人胚胎、成年及新生小鼠肝脏组织,生理盐水清洗、剪碎、筛网过滤,用生理盐水制备混悬液,3 000 r/min离心,收集上清,制备肝脏可溶性复合物.②体外实验:用上述不同来源的肝脏可溶性复合物体外处理肿瘤细胞,四甲基偶氮唑盐比色法测定其对乳腺癌细胞EMT6增殖的影响.③体内实验:观察成年鼠肝脏可溶物质对乳腺癌细胞EMT6体内生长的抑制作用及其对荷瘤鼠生存状况的影响,包括不同给药剂量及不同给药途径两个实验,给药途径包括在接种肿瘤细胞部位的对侧腋下、同侧腋下、腹腔注射及灌胃等.结果:①体外实验显示不同来源的肝脏可溶性复合物能明显抑制肿瘤细胞EMT6增殖率,肿瘤增殖抑制率均显著高于血清白蛋白处理组(P＜0.05),并呈剂量依赖性.②成年鼠肝脏可溶物质8mg/L组抑瘤率高于2,4 mg/L组(P＜0.05),未观察到明显毒副效应.③比较不同给药途径,成年鼠肝脏可溶物质同侧注射组的抑瘤率较其他3组的抑瘤率高(P＜0.05),各成年鼠肝脏可溶物质给予组的体质量增长率比相应生理盐水对照组高(P＜0.05).④与相应生理盐水对照组比较,在同侧腋下注射成年鼠肝脏可溶物质的小鼠生存期明显延长(P＜0.05).结论:肝脏可溶性复合物具有抑制肿瘤细胞生长的作用,并且呈一定的剂量依赖性.不同的给药途径中,在接种肿瘤细胞部位的同侧腋下给药抑瘤效果最好.     

丘脑卒中患者认知功能损害及其临床判别

目的:探讨丘脑卒中后认知损害的特点,尝试应用回归函数方程推测和判别这种损害程度。方法:①选择2004-04/12大连市第二人民医院和大连市海港医院收治,并经CT确诊的丘脑卒中患者21例为丘脑卒中组;按1∶3匹配原则,将63例年龄与性别与丘脑卒中组匹配的非脑部疾病者入选为对照组。②丘脑卒中组住院后进行影像学和生理、生化学指标检查,对所有入组人员进行神经心理学测查其认知功能,内容包括4个纬度(智能纬度、执行功能纬度、注意机能纬度和记忆纬度)13个认知因子,以韦氏成人智力量表、韦氏记忆量表和威斯康星卡片分类实验等为测查工具。③将对照组每项认知因子均数根据其在认知中的作用不同,加或减1.64标准差为界值,以判断丘脑卒中组认知损害程度,并建立回归方程和认知损害程度表达式。结果:84例均进入结果分析。①丘脑卒中组在13个认知指标中有9个指标与对照组相比差异显著(P<0.05),4个纬度均有涉及。②建立丘脑组认知损害程度函数表达式,计算出每例认知损害程度函数值(F0)。以丘脑组每例认知损害程度函数值为应变量,以临床影响因子为自变量建立了回归方程Y=43.679-2.304*β。以Y值推测F0值。当Y<0.98,可考虑为轻度认知损害,当Y=0.98～8.15时可考虑为中度认知损害,当Y>8.15时可考虑为重度认知损害。结论:①丘脑卒中可引起广泛的认知损害。②临床可以根据影响因素建立回归方程,计算出Y值,以Y值推测F0值。     

体外制成复合软质再生颅骨修复材料填充犬颅骨缺损

目的:目前颅骨修补材料有很多种,但都为异源性无机骨替代物,并且应用该方法又要给患者再次行开颅手术,实验拟开展新型颅骨再生材料的研究。方法:实验于2006-05/11在解放军第一五七医院动物中心及中山大学附属第三医院动物实验室完成。①实验动物:30只犬随机分为实验组20只,对照组10只。②实验方法:应用纳米级羟基磷灰石为支架和成骨细胞培养,加入脱矿的犬类骨基质为载体的重组人类骨形成蛋白2,制成复合软质再生颅骨。实验组犬在右侧颅骨缺损中填补藻酸钙凝胶、成骨细胞、纳米级骨粉的复合材料,左侧颅骨缺损中填补藻酸钙凝胶、成骨细胞、纳米级骨粉和重组人类骨形成蛋白2的复合材料。对照组犬在右侧为单纯颅骨缺损,左侧颅骨缺损中填补藻酸钙凝胶、成骨细胞、纳米级骨粉和重组人类骨形成蛋白2复合材料。实验过程中对动物处置符合动物伦理学标准。③实验评估:手术后1,2,3,6个月X射线片检查颅骨缺损修复情况,对再生的颅骨组织标本进行茜素红S染色,观察成骨能力及再生材料骨膜组织细胞体外培养情况。结果:实验动物均进入结果分析。术后1个月,成骨活跃,骨端新生骨小梁基本覆盖骨断端,缺损区可见较多新生骨小梁形成,骨端新生骨小梁向缺损区长入;术后2个月可见较多散在骨岛形成;术后3个月可见成熟骨,并有髓腔形成,缺损区大量新骨形成。而各对照组骨断端处有散在骨岛,或被增生的纤维结缔组织占据,可见大量纤维组织及毛细血管长入,植入的基质材料基本被吸收,无新骨生成。结论:应用纳米级羟基磷灰石为支架和成骨细胞培养,加入脱矿骨基质为载体的重组人类骨形成蛋白2,制成的复合软质再生颅骨能自身代谢并逐渐骨化,形成新的颅骨。     

应用三维有限元法评估聚甲基丙烯酸甲酯和磷酸钙骨水泥铸件的力学特征

目的:聚甲基丙烯酸甲酯和磷酸钙骨水泥作为经皮椎体后凸成形术填充物,应用三维有限元分析法比较两种材料治疗骨质疏松性椎体压缩骨折的生物学效应。方法:实验于2003-03/09在南通医学院第一附属医院骨科和上海大学上海生物力学工程研究所合作完成。①利用WD-5型万能材料机测量聚甲基丙烯酸甲酯和磷酸钙骨水泥铸件力学性质。②选取骨质疏松患者1例(患者知情同意),排除其他疾患,应用CT扫描技术、图形数字化方法获取胸腰段的三维坐标,利用ANSYS5.0有限元分析软件等工具建立骨质疏松患者胸腰段的三维有限元模型。③模拟L1经皮椎体后凸成形术,对其和相邻椎体及椎间盘的负荷传递、应力、位移等进行分析。结果:①获取了精确的T11￣L3的多排螺旋CT扫描断层影像数据,经ANSYS5.0软件处理后得到4718个节点,1642个薄极单元,4495个八节点等参单元的骨质疏松患者胸腰段三维有限元力学模型。②采用聚甲基丙烯酸甲酯作为经皮椎体后凸成形术的填充物,比骨质疏松椎体应力增加了近15%。后部结构的应力平均增加13.2%,其中椎弓根增加5.9%,峡部增加6.25%,关节点增加27.6%。聚甲基丙烯酸甲酯在稳定椎体、恢复强度和刚度的同时,可能使其后部结构及相邻腰椎出现应力集中现象。③采用磷酸钙骨水泥作为椎体成形术填充物,比骨质疏松椎体应力平均增加了7%,应力集中现象明显小于聚甲基丙烯酸甲酯。结论:三维有限元力学分析表明两种材料均提高了椎体的抗变形能力,有利于椎体功能的重建。与聚甲基丙烯酸甲酯相比,磷酸钙骨水泥能减少成形椎体和相邻椎体之间的应力梯度,从长远看,能减少椎体退变和相邻椎体骨折的机会。