荧光示踪法研究逆行岛状皮瓣静脉回流

目的探讨采用荧光示踪法研究逆行岛状皮瓣静脉回流的可行性，并初步观察静脉回流规律。方法20只新西兰大白兔，每只取耳静脉血0．ImL，分离RBC并用FITC标记。流式细胞仪检测已标记的RBC阳性率及荧光强度，倒置荧光显微镜观察其形态。取20只新西兰大白兔，在动物双侧后肢内侧分别建立4cm×3cm隐动、静脉逆行岛状皮瓣模型（n=10）和顺行岛状皮瓣模型（n=10），血管蒂长3cm。将一侧后肢随机设定为实验组，皮瓣制备后注射已标记的RBC悬液5pL；对侧为对照组，不注射示踪剂。实验组按顺行和逆行皮瓣分成两组，即顺行皮瓣组和逆行皮瓣组，每组10个；再根据注入示踪剂途径不同，分为动脉和静脉2个亚组，每亚组5个皮瓣。注射示踪剂5S后取下皮瓣立即冷冻，取连续的3张冰冻切片（5～7pm），其中2张行HE染色和GENMED染色，另]张不染色直接压片，荧光显微镜观察荧光分布。结果流式细胞仪分析FITC标记的RBC阳性率在99％以上，荧光强度均≥10。；倒置荧光显微镜下标记的RBC呈均匀分布的绿色荧光，荧光强度均匀、稳定。冰冻切片显示实验组皮瓣蒂部均出现荧光，对照组未见荧光。顺行岛状皮瓣组荧光主要分布在静脉腔、静脉壁、动脉内膜和外膜；逆行岛状皮瓣组荧光分布在动脉内膜、外膜和静脉壁。结论荧光示踪剂可用于静脉回流研究，顺行岛状皮瓣静脉主要通过静脉腔、静脉壁、动脉内膜和外膜回流；逆行岛状皮瓣静脉主要通过动脉内膜、外膜和静脉壁的“迷宫式途径”回流。     

In vivo determination of body fat by measuring total body carbon

Total body carbon (TBC) is measured in vivo by neutron inelastic scattering. The fast neutrons needed for the irradiation are produced by a miniature deuterium-tritium (D-T) neutron generator. Body fat and protein are the main contributors to TBC. Bone ash and carbohydrates contribute less than 3%. Fat is calculated from TBC after the subtraction of the carbon contributions from protein, bone, and glycogen. The technique was applied to 14 normal volunteers (8 females, 6 males) aged 24-94 y who underwent neutron inelastic scattering and neutron activation measurements for body carbon, nitrogen, and calcium. The initial results agree with other techniques. Unlike models that evaluate body fat by subtracting lean body mass from body weight, the TBC technique is not sensitive to assumptions on the composition of lean body; therefore, it is appropriate for studies of adults of any age and health condition.     

Inhibitory effects of benzoate on chiral inversion and clearance of N(G)-nitro-arginine in conscious rats.

N(G)-nitro-arginine (NNA) is known to exhibit stereoselective pharmacokinetics in which N(G)-nitro-d-arginine (d-NNA) has a faster clearance rate than N(G)-nitro-l-arginine (l-NNA) in anesthetized rats, and d-NNA undergoes unidirectional chiral inversion. It was postulated that chiral inversion of d-NNA was performed in a two-step pathway by d-amino acid oxidase (DAAO) followed by an unidentified transaminase. Such chiral inversion contributes (at least partially) to the pharmacokinetic stereoselectivity of NNA. This study used the selective inhibitor of DAAO, sodium benzoate, to test the above hypothesis. An i.v. bolus injection of d-NNA (32 mg/kg) and l-NNA (16 mg/kg) in conscious rats exhibited biphasic disposition with different pharmacokinetic parameters in a stereospecific manner (approximately 5-10-fold differences). Unidirectional chiral inversion of d-NNA but not l-NNA was found from these animals. In addition to its similar inhibitory effects on the d-NNA conversion and DAAO activity in kidney homogenates, sodium benzoate completely blocked chiral inversion of d-NNA and led to a smaller stereospecific difference, reflected by a nearly 50% reduction of d-NNA clearance and a 2-fold increase in t(1/2) and area under the curve of d-NNA in benzoate-pretreated rats. The results suggest that DAAO plays an essential role in chiral inversion of d-NNA and chiral inversion contributes mostly to the pharmacokinetic stereospecificity of NNA.     

Design of a bifunctional fusion protein for ovarian cancer drug delivery: single-chain anti-CA125 core-streptavidin fusion protein.

We have developed a universal ovarian cancer cell targeting vehicle that can deliver biotinylated therapeutic drugs. A single-chain antibody variable domain (scFv) that recognizes the CA125 antigen of ovarian cancer cells was fused with a core-streptavidin domain (core-streptavidin-VL-VH and VL-VH-core-streptavidin orientations) using recombinant DNA technology and then expressed in Escherichia coli using the T7 expression system. The bifunctional fusion protein (bfFp) was expressed in a shaker flask culture, extracted from the periplasmic soluble protein, and affinity purified using an IMAC column. The two distinct activities (biotin binding and anti-CA125) of the bfFp were demonstrated using ELISA, Western blot and confocal laser-scanning microscopy (CLSM). The ELISA method utilized human NIH OVCAR-3 cells along with biotinylated bovine serum albumin (B-BSA) or biotinylated liposomes, whereas, the Western blot involved probing with B-BSA. The CLSM study has shown specificity in binding to the OVCAR-3 cell-line. ELISA and Western blot studies have confirmed the bifunctional activity and specificity. In the presence of bfFp, there was enhanced binding of biotinylated antigen and liposome to OVCAR-3 cells. In contrast, the control EMT6 cells, which do not express the CA125 antigen, showed minimal binding of the bfFp. Consequently, bfFp based targeting of biotinylated therapeutic drugs, proteins, liposomes, or nanoparticles could be an alternative, convenient method to deliver effective therapy to ovarian cancer patients. Peritoneal infusion of the bfFp-therapeutic complex could also be effective in locally targeting the most common site of metastatic spread.     

包皮过长和包茎组织中触觉小体观察

目的观察包茎和包皮过长组织中触觉小体差异情况。方法收集21~25岁年龄组包皮标本44例,按包皮解剖形态分为:①包茎组21例;②包皮过长组23例。用免疫组化方法对两组包皮标本中的触觉小体进行染色,在放大100倍的视野下观察和统计两组包皮标本中触觉小体总数以及视野的总数,并用卡方检验对两组间的差异进行统计学分析。结果两组标本中,分别有8个(8/21包茎组)和10个(10/23包皮过长组)标本中未发现触觉小体,其差异无显著性(P=0.717)。包茎组和包皮过长组中触觉小体的密度分别为33.1%和21.1%,差异具显著性(P=0.022)。结论包茎中触觉小体密度较包皮过长明显升高可能是一种生理性代偿机制,而两种包皮组织中触觉小体的消失可能遵循同一规律。     

经直肠超声引导前列腺穿刺活检术诊断前列腺癌

目的　总结和评价经直肠超声引导下前列腺穿刺活检术对前列腺癌诊断的准确率。方法　222 例直肠指检阳性或 PSA>4μg/L的患者应用经直肠超声引导下前列腺6点系统穿刺活检以明确诊断。结果　222 例受检者中病理证实前列腺结节性增生41例、前列腺炎24例、前列腺肉瘤3例、前列腺癌 154 例,其中低分化癌 74 例、中分化癌 58 例、高分化癌 22 例。术后血尿15例、发热6例,其中高热1例,经抗生素治疗后体温恢复正常、尿检阴性。结论　经直肠超声引导下前列腺穿刺活检无需麻醉,患者痛苦小、安全性高,是诊断前列腺癌的可靠方法。     

Development and optimization of an indirect enzyme-linked immunosorbent assay for the determination of Hexoestrol

An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of hexoestrol (HES) was developed, optimized and validated for the analysis of HES in pork. Many parameters, such as the volume ratio of solution A and solution B, colour developing time, pH value, incubation time, the volume ratio of the standard solution and diluted antiserum, blocking solution, blocking condition, coating solution and coating condition were studied and optimized in the paper. The regression equation of the final inhibition curve is y = - 0.3345x + 1.4955, R²=0.9913. The linear range is 0.1-8.1 ng/ml, and the IC₅₀ is 0.671 ng/ml. The specificity of the assay was evaluated by the cross-reactivity rates of six compounds, of which two structurally related compounds had a relatively higher cross-reactivity rate of 25% and 6%. HES was added into a pork sample at 5 ng/g level and then the sample was extracted. The recovery is between 49.6% and 79.2%, and the variation coefficient is 0.164.     

给药系统

近年来，伴随着药物治疗的快速发展．以药理活性非常强的药物为代表，开发出许多用药时必须给予足够关注的药物，“用药最适化”这一概念逐渐成为人们非常关注的问题。这主要是为了实现用药时的最佳形式。尽可能地根据选择或者需要控制药物的浓度．时间模式。输送到药物的作用表达部位。实现治疗用药的安全性。但是，历来的方法却很难实现这一点，通过利用新的技术和方法调控药物在体内的动态变化，以获得最好治疗效果为中心的药物使用方法和药剂正在开发之中。这就是给药系统，使用可以表达自身概念的语言来命名。虽然都是以各种药物在体内的动态过程作为调控对象，但是还分为①controlled release（调控释放给药）；②开发新的给药途径、克服药物吸收障碍；③靶向给药（targeting）等三个途径。