1,4-Dihydroindeno[1,2-c]pyrazoles as potent checkpoint kinase 1 inhibitors: extended exploration on phenyl ring substitutions and preliminary ADME/PK studies

A study on substitutions at the four open positions on the phenyl ring of the 1,4-dihydroindeno[1,2-c]pyrazoles as potent CHK-1 inhibitors is described. Bis-substitution at both the 6- and 7-positions led to inhibitors with IC(50) values below 0.3nM. The compound with the best overall activities (36) was able to potentiate the anti-proliferative effect of doxorubicin in HeLa cells by at least 47-fold. Physicochemical, metabolic, and pharmacokinetic properties of selected inhibitors are also disclosed.     

Synthesis and cytotoxicity of novel fatty acid-nucleoside conjugates

N(3)-Hydroxyethyltegafur (3) was synthesized in 91.0% yield under a new condition. A series of novel fatty acid esters of 3 were synthesized. These fatty acid-nucleoside conjugates have shown cytotoxicities against Ec9706 cells and A549 cells, and the structure activity relationship was discussed.     

Dumbbell-shaped circular oligonucleotides as inhibitors of human topoisomerase I

A dumbbell-shaped circular oligonucleotide containing topoisomerase I-binding sites and two mismatched base pairs in its sequence has been designed and synthesized. Our further studies demonstrate that this particularly designed oligonucleotide displays an IC(50) value of 9 nM in its inhibition on the activity of human topoisomerase I, a magnitude smaller than that of camptothecin, an anticancer drug currently in clinical use.     

Size-fractionated chlorophyll a and primary productivity in the Chukchi Sea and its northern Chukchi Plateau

Investigations on chlorophyll a and primary productivity were carried out in the Chukchi Sea and its northern Chukchi Plateau during the 2^nd Chinese National Arctic Research Expedition in the summer of 2003. The results showed that chlorophyll a concentrations were 0.009–30.390 μg/dm³ at the surveyed waters; the surface chlorophyll a concentrations were 0.050–4.644 μg/dm³ and the average value was (0.875±0.981) μg/dm³ in the surveyed area. In the Chukchi Sea Shelf, chlorophyll a concentrations at the depth from 10 m to bottom were higher than that in the surface water, and the concentrations were lower at the depth below 75 m in the Chukchi Plateau. Chlorophyll a concentrations descended in 3 sequential samplings on Transect R, with average values of (2.564±1.496) μg/dm³, (1.329±0.882) μg/dm³ and (0.965±0.623) μg/dm³, respectively. The potential primary productivity ((2.305± 1.493) mgC/(m³·h)) in the Chukchi Sea was higher than that ((0.527±0.374) mgC/(m³·h)) in the Chukchi Plateau. The results of the size-fractionated chlorophyll a and primary productivity showed that microplankton accounted for the majority of the total chlorophyll a (63.13%) and primary productivity (65.16%) at the survey stations. The contributions of the nanoplankton and picoplankton to the total chlorophyll a and primary productivity were roughly the same.     

学习经历在平腹小蜂寄主选择过程中的作用

研究了已经交配与未经交配的平腹小蜂在羽化后不同时间对中间寄主柞蚕卵信息记忆的持效性,比较了经学习训练与未经学习训练的平腹小蜂在不同时间对其自然寄主荔枝蝽卵和中间寄主柞蚕卵的选择性。结果表明：交配过的平腹小蜂在大量羽化后96h检验柞蚕卵的时间和数量都显著增多,而未经交配的平腹小蜂在此时搜索寄主的时间也显著延长,说明平腹小蜂在此时对柞蚕卵信息的记忆开始明显减弱。经学习训练的和未经学习训练的平腹小蜂在大量羽化后96h对荔枝蝽卵的第一选择数和刺卵数都明显提高,并且前者较后者在各个时段对荔枝蝽卵的第一选择数、检验卵数以及刺卵数都多,说明经学习训练的平腹小蜂对荔枝蝽卵的趋性较高,并且随着时间的延长受柞蚕卵信息的干扰逐渐减小,对荔枝蝽卵的学习效果不断增强。     

大黄酚和大黄素甲醚的制备性分离

经系统分离得到大黄酚和大黄素甲醚的混合物后,取该混合物约1g,用少量的氯仿溶解,加入少量的层析用硅胶（100～200目）拌匀,减压使氯仿蒸干。将吸附有大黄酚和大黄素甲醚的硅胶装在已装好的硅胶层析柱的上端,然后进行洗脱。通过实验,得到了最佳分离大黄酚和大黄素甲醚的条件：硅胶与混合物的质量比=150：1;层析柱长与直径的比=23：1;洗脱剂比例：石油醚（60～90℃）：乙酸乙酯（V/V）=17：1;减压淋洗。     

一种改良的二甲基亚硝胺肝纤维化模型诱导方法及其病理特点

目的探讨旨在降低死亡率的二甲基亚硝胺(DMN)肝纤维化改良造模方法及其病理特点。方法大鼠分为常规方法造模组(常规组)与改良方法造模组(改良组),每组再分为模型组与正常对照组,均设染毒后4周和8周共2个时间观察点。常规组造模以DMN10μL/kg大鼠体重的剂量,腹腔注射,每周连续3d,连续4周,共12次;改良组同上剂量的DMN腹腔注射,每2d一次,连续4周,共14次。观察大鼠体重、肝体比与脾体比,试剂盒测定血清肝功能,HE染色与天狼猩红染色分别观察肝组织炎症与胶原沉积,盐酸水解法测定肝组织羟脯氨酸含量。结果2种造模方法均在造模4周时形成典型的肝纤维化,模型大鼠的体重、肝体比、脾体比、血清肝功能等均有相应的变化趋势。与常规组比较,改良组模型大鼠造模8周后死亡率显著降低,肝细胞炎症坏死稍轻,而肝窦毛细血管化明显。结论隔日1次腹腔注射较之连续3d腹腔注射DMN是一种较为稳定的肝纤维化改良造模方法,具有肝窦毛细血管化与小叶内纤维化的肝纤维化病理特征。     

组织工程静脉血管种子细胞的培养

目的获取可应用于静脉血管组织工程的种子细胞内皮细胞。方法选取北京地区雄性杂种犬,取其双侧颈外静脉,采用翻转后酶消法,获取原代内皮细胞,对其进行原代培养和传代培养,冻存、复苏和鉴定,并利用光学显微镜进行观察。结果获取犬的颈外静脉后,实验采取了翻转后酶消法,所获得的内皮细胞纯度和数量得到了提高;经过鉴定确实为内皮细胞来源;活性好,增殖快,在较短的时间内能够达到后期实验所需数量。结论经体外培养的犬的颈外静脉内皮细胞可以作为组织工程血管的种子细胞。     

T淋巴细胞在小鼠肾缺血再灌注损伤中的作用

目的研究T淋巴细胞在肾缺血再灌注损伤(IRI)导致的急性肾损害中的作用。方法BALB/c小鼠和BALB/c裸小鼠各24只,分别随机分为A1-4组和B1-4组,每组6只。双肾蒂阻断45 min后恢复血流建立肾IRI模型,假手术对照组I、RI后24、48和72 h时检测Scr、尿蛋白定量及肾病理学,A组检测脾T细胞亚群;对比BALB/c小鼠和BALB/c裸小鼠的肾功能下降、组织学损害程度以及脾T淋巴细胞亚群变化。结果A2-4组和B2-4组均有Scr和尿蛋白定量明显升高(P<0.05),且A组损害程度明显重于B组(P<0.05);A2-4组出现典型的IRI组织损害表现(P<0.05),B2-4组无明显IRI组织损害(P>0.05);A2-3组脾CD3 T细胞百分比较A1组升高(P<0.05),而CD4 /CD8 比值无明显变化(P>0.05)。结论T淋巴细胞是小鼠肾IRI导致急性肾损害的重要病理生理学因素。     

芽胞杆菌苎麻脱胶酶-甘露聚糖酶的纯化及酶学性质

研究了苎麻高效脱胶菌株Bacillus subtilisNo.16A甘露聚糖酶的产生条件,纯化过程以及酶学性质。其优化的培养基为魔芋胶30 g/L,蛋白胨9 g/L,酵母膏2 g/L,KH2PO45 g/L,MgSO4.7H2O 0.25 g/L。优化的培养条件为起始pH 8.0,装样量50 mL,接种量10 mL,发酵72 h。进一步通过硫酸铵沉淀、阴离子交换层析、凝胶过滤3步从Bacillus subtilisNo.16A发酵液中纯化了甘露聚糖酶。结果表明,该酶分子量约为38.5ku,最适作用pH为7.0,最适作用温度为60℃,在55℃以下、中性pH(6.0~8.0)范围内稳定,Ca2 、Mn2 和A l3 、Mg2 对该酶有激活作用,Cu2 、Zn2 有抑制作用。