RP-HPLC法测定家兔血浆中甲苯喹哌浓度和药代动力学参数

本文建立了以紫外230nm波长检测的反相高效液相色谱法(RP-HPLC)测定家兔血浆中甲苯喹派浓度。填料使用LiChrosorb RP-C18,流动相为甲醇—水—三乙胺—磷酸(63:37:1:0.8 v/v),血样(或尿样)经碱化后用乙醚提取,再以0.2 mol/L硫酸回提,进样。方法回收率为99.84±3.10(SD)%;天内、天间精密度平均CV为4.12%及3.95%(n=5);最低检测限3ng.经提取的标准线性浓度在25～2000 ng/ml范围内,Y=0.002865X-0.01346,r=0.9999,内源性物质及可能的合并用药不干扰色谱测定。文内用质谱法鉴定血样中甲苯喹哌色谱峰纯度,并由尿样分析对其主要代谢物予以初步验证。本法可应用于药代动力学参数测定。家兔按8mg/kg静注后,药—时曲线符合二室模型T_1/2=4.8008±1.1522(SD)h。     

脱细胞基质骨与成骨细胞和血管内皮细胞的生物相容性

目的:采用自行研制的脱细胞基质猪肋骨作为支架材料,复合兔成骨细胞和血管内皮细胞,对其生物相容性、抗原性和细胞毒性进行观察。方法:实验于2003-04/10在四川大学华西医院人类疾病生物治疗教育部重点实验室完成。用物理、化学方法制备脱细胞基质猪肋骨。体外培养兔成骨细胞、血管内皮细胞。而后将试验分成3组进行。①成骨细胞组:将成骨细胞与脱细胞基质骨材料复合。②血管内皮细胞组:血管内皮细胞与脱细胞基质骨材料复合。③成骨细胞 血管内皮细胞组:成骨细胞 血管内皮细胞与脱细胞基质骨材料复合。④对照组:单纯成骨细胞。于培养1,3,5,7d分别用倒置相差显微镜、组织学切片和扫描电镜观察脱细胞基质异种骨的生物相容性、抗原性;用流式细胞仪检测材料对细胞周期和DNA含量的影响,了解材料对细胞有无毒性的影响。结果:①脱细胞基质骨扫描电镜观察:脱细胞基质骨的骨小梁结构完整,骨陷窝空虚,未见细胞成分残留。②各组细胞的生长、分化和增殖情况倒置相差显微镜观察:3组细胞与脱细胞基质骨材料复合培养12h后,细胞在各组材料表面和孔隙内均可黏附和生长。③组织学观察:MASSON染色显示3组细胞在材料表面和孔隙内大量增殖,并分泌细胞外基质。以成骨细胞 血管内皮细胞组材料表面黏附的细胞数量最多。组织学切片染色显示,各组细胞沿材料边缘紧密附着,细胞与材料的组织相容性良好。④各组细胞黏附、生长、增殖和基质分泌情况扫描电镜观察:3组细胞与材料复合培养5d后,细胞紧密黏附在材料表面。成骨细胞呈梭形或多角形,相邻细胞间有突起相互连接。血管内皮细胞呈椭圆形,有角状突起,与材料附着紧密。其中,成骨细胞 血管内皮细胞组材料表面黏附大量细胞,并有较多胶原形成。血管内皮细胞组材料表面胶原形成很少。⑤细胞周期和DNA含量:成骨细胞 血管内皮细胞组1d,3dDNA合成期高于单独细胞组,无异倍体细胞。结论:脱细胞基质骨具有良好的生物相容性、极低的抗原性、无细胞毒性和致瘤性。成骨细胞与血管内皮细胞联合培养细胞在脱细胞基质异种骨中有很强的增殖潜力。     

出血热误诊为上呼吸道感染1例

1病例报告患者,男,22岁,因间歇性全身乏力、肌肉酸痛2 wk,发冷1 wk,发热4 d入院.曾在我院查WBC 4.5×109/L,N 0.60,L 0.4,体温波动在38～40℃.初步诊断"上呼吸道感染",用阿莫西林、VC银翘片、清热解毒冲剂等治疗无效.查体:T 38.9℃,BP面性12/8 kPa.全身皮肤无出血点,双眼球结膜轻度充血,咽部充血,软腭未见充血点,心肺腹部未见阳性体征.实验室检查:WBC 7.85 × 109/L,N 0.79,L 0.21,HGB 150g/L,PCL30×109/L,尿蛋白3.2g/L,流行性出血热抗体( ).诊断:流行性出血热.入院后立即按照流行性出血热的治疗原则给予抗病毒、抗渗出、抗出血治疗.具体包括卧床休息,给予高热量,多维生素,易消化饮食;维持水、电解质、酸碱及血浆渗透压平衡;给予大剂量(5 g)Vit.C和Vit.E.同时给予氢化可地松100 mg/d,稀释后缓慢静脉滴注.入院后3 d患者的尿量由450 mL/d增至750 mL/d,肌酐204.6μmol/L,BUN 13.3 mmol/L.5 d尿量增加至4000 mL/d.经综合治疗10 d,肌酐和BUN检查等正常,痊愈出院,随访1 mo未见异常.     

Clinicopathological features of giant cell carcinoma of the pancreas

BACKGROUND: Giant cell carcinoma of the pancreas (GCCP) as a tumor of high malignancy, large size, and inflammatory reaction occupies 2.1%-12.8% of all cases of pancreatic malignancies. This study was to analyze cases of GCCP collected in 8 years at our hospital in an attempt to describe some features of GCCP in Chinese people. METHODS: The clinicopathological features of 19 patients who had been pathologically diagnosed as having GCCP from 1021 patients with pancreatic malignancies collected by Pancreatic Disease Research Group (PDRG) of Changhai Hospital were retrospectively analyzed compared with those of 96 patients with common pancreatic carcinoma (PC) who were randomly selected from 1002 patients with pancreatic carcinoma. The differences of location, clinical symptoms,     

Increased NK Activity Is Responsible for Higher Cytotoxicity to HEF Cells by Lymphocytes of Women With Threatened Preterm Delivery

ABSTRACT: Recently we have shown that lymphocytes of pregnant women with threatened preterm delivery (risk group) exerted significantly higher cytotoxic activity to human embryonic fibroblast (HEF) cells than those of healthy pregnant women. The purpose of this study was to get information on the mechanism of this cytotoxicity. The possibility of prior sensitization to embryonic antigen was excluded, since no difference could be demonstrated between cytotoxic activity of lymphocytes obtained from women with two or more previous pregnancies and that of lymphocytes from never-pregnant women. For determining the effector cell type responsible for cytotoxicity, lymphocytes of 50 healthy pregnant women and those of 50 risk patients were tested in different cytotoxicity tests using HEF and K-.562 target cells. The proportion of NK cells among lymphocytes was determined by counting large granular lymphocytes (LGL), IgG Fc receptor bearing cells, and cells positively stained by NK specific monoclonal antibody. Though no difference in the proportion of NK cells between the two groups was found, risk patients' lymphocytes were significantly more cytotoxic to K-562 target cells than those of healthy pregnant women. Investigations at the single-cell level made it obvious that this higher cytotoxic activity originated from increased target cell lysing ability of their lymphocytes, while their conjugating capacity did not differ significantly from that of lymphocytes obtained from healthy pregnant women. Staining conjugates with monoclonal antibodies showed that NK cells were concentrated on the surface of both HEF and K-.562 cells, so it seems likely that NK activity is responsible for the cytotoxic activity of pregnancy lymphocytes to HEV cells.     

Hypercalcemia in hyperthyroidism role of age and goiter type

Summary Hyperthyroidism is often associated with hypercalcemia which is provoked by osteoclastic activity of the thyroid hormones. These data show that hypercalcemia develops with increasing age and in the presence of a special type of hyperthyroid goiter. Total serum calcium, total protein, and albumin as well as different parameters of thyroid function, namely T₃ RIA¹, T₄ test, ETR and TRH test were determined in a group of 147 patients. The ionized calcium level was estimated from total calcium and albumin.211 measurements were performed. Hyperthyroidism existed in 92 cases. Total calcium was not significantly elevated in hyperthyroidism. Hyperthyroid patients under 61 years of age showed elevated ionized calcium levels in only 2.3% and patients over 60 years of age in 18.8% of cases. Elevated ionized serum calcium levels were observed in 43.8% of hyperthyroid patients with multinodular goiters. The linear correlation between ionized calcium levels and different parameters of thyroid function is much more pronounced in the older group and it was found to be highly significant. 7 of 9 hyperthyroid patients with elevated ionized calcium levels showed multinodular goiters, though no autonomous adenoma. In the hyperthyroid group of patients of over 60 years of age with multinodular goiters the incidence of hypercalcemia was 43.8%. Direct action of thyroid hormone on calcium turnover as well as increasing age and special goiter type seem to be responsible for disturbances in calcium metabolism. A possible calcitonin deficiency in the above mentioned conditions is discussed.

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Abbreviations T₃ RIA triiodothyronine radioimmunoassay - T₄ test thyroxine protein binding assay - ETR effective thyroxine ratio - TRH test stimulation test with thyrotropin-releasing hormone     

微球培养人脂肪间充质干细胞向软骨细胞的诱导分化

目的:观察微球培养的人脂肪间充质干细胞在特定诱导条件下向软骨细胞的分化情况,探讨其作为组织工程化软骨种子细胞的可行性。方法:实验于2006-03/08在湘雅医院中心实验室完成。①实验材料:皮下脂肪组织来源于股骨颈骨折手术患者6例,对本实验均知情同意。②实验方法:取皮下脂肪,分离培养脂肪间充质干细胞,胰酶消化,取传至第6代的细胞,密度调整为1×1010L-1进行接种培养,细胞贴附后加入软骨细胞诱导剂(含10μg/L转化生长因子β1、37.5mg/L维生素C、6.25mg/L胰岛素、6.25mg/L转铁蛋白、10-7mol/L地塞米松、1%新生牛血清的高糖DMEM),倒置显微镜下观察细胞团的变化。③实验评估:诱导14d后,将细胞吹散,细胞爬片,免疫组化染色检测Ⅱ型胶原的表达,甲苯胺兰染色观察细胞内异染情况,RT-PCR检测Ⅱ型胶原蛋白和软骨蛋白聚糖aggrecan mRNA的表达。结果:①脂肪间充质干细胞向软骨细胞诱导后的形态:诱导3d后,细胞团向内呈放射状聚集。②诱导后Ⅱ型胶原的表达及细胞内异染情况:诱导14d后,Ⅱ型胶原在胞浆、胞膜及细胞外基质中均有表达,呈棕黄色;甲苯胺兰染色显示胞浆内呈蓝色异染。③诱导前后Ⅱ型胶原蛋白和aggrecan mRNA的表达:诱导前脂肪间充质干细胞无Ⅱ型胶原蛋白和aggrecan mRNA表达,而诱导14d后二者均有较高表达。结论:在微球状态下可成功诱导脂肪间充质干细胞向软骨细胞分化。     

香豆素衍生物的合成及其活性与相对亲脂水性的关系

4-甲基-7-羟基-6-或-8-烯丙基香豆素是一类新的辐射防护剂,但由于其脂、水溶性都很小,影响药物在体内的吸收及口服效果。为改善其溶解度,从增加有效基本结构的亲水或亲脂性的角度,合成了14个香豆素衍生物,测定抗辐射活性并研究了亲水,亲脂性与活性之间的关系,获得一定的结果。5a和6a能在油酸乙酯中溶解,后者在水中也有一定的溶解性,且二者都保持了活性。