微透析和高效液相色谱检测万古霉素在清醒兔眼玻璃体内的浓度

背景：万古霉素玻璃体内注射后的药代动力学资料较少。 目的：观察万古霉素玻璃体腔内注射后在清醒兔眼玻璃体内的浓度。 方法：将微透析探针植入清醒正常兔眼和细菌性眼内炎兔眼玻璃体内24 h后,向玻璃体内注射10 g/L万古霉素0.1 mL,利用微透析采样技术联合高效液相色谱法连续检测万古霉素注射后0.5,1,2,4,6,12,24,48,72,84 h,兔眼玻璃体内的药物浓度。 结果与结论：万古霉素在正常兔眼玻璃体内的代谢呈开放式二室模型,玻璃体内的峰值浓度为695.92 mg/L,半衰期51.66 h;在细菌性眼内炎兔眼玻璃体内的代谢呈一室模型,万古霉素的峰值浓度为713.35 mg/L,半衰期为11.91 h。所有动物给药84 h,玻璃体内万古霉素的浓度均高于最小抑菌浓度。实验在动物清醒状态下实时、连续、动态采样,检测结果准确,能满足万古霉素药动学分析的需要。     

家兔静脉注射环丙沙星后眼组织分布及其药代动力学研究

选健康合格一级新西兰家兔24只,雌雄兼用,体重2.5-3.0kg,随机均分为8组,每组3兔6眼,每组动物均由左耳缘静脉注射给药。给家兔静脉注射环丙沙星(Ciprofloxacin CPFX)后,用高效液相色谱仪测定眼内各组织中药物分布的浓度和药代动力学参数。环丙沙星在泪液、角膜房水、虹膜-睫状体、晶体和玻璃体组织内峰浓度值分别为8.92±2.88、142.84±25.     

壳聚糖抑制兔外伤性增生性玻璃体视网膜病变的作用*

背景：在眼科领域,已发现壳聚糖可以抑制青光眼手术瘢痕形成,但还未有明确的报道可以阻止和减缓增生性玻璃体视网膜病变的发生。 目的：观察壳聚糖对兔外伤性增生性玻璃体视网膜病变的防治作用及作用机制。 方法：用自体富含血小板血浆玻璃体内注射制备兔外伤性增生性玻璃体视网膜病变模型,并同时将生理盐水、5%壳聚糖、10%壳聚糖和15%壳聚糖溶液0.1 mL注入兔眼玻璃体内,在不同时间点观察壳聚糖对增生性玻璃体视网膜病变形成的防治作用。 结果与结论：对照组第10天就出现视网膜脱离,增生性玻璃体视网膜病变随着时间延长发展至更严重等级。壳聚糖组也发生增生性玻璃体视网膜病变,但其严重程度均低于对照组。10%和15%壳聚糖组给药5 d后,增生性玻璃体视网膜病变的临床分级显著低于对照组(P < 0.05)。组织学检查壳聚糖组未发现明显视网膜形态改变。结果说明壳聚糖玻璃体腔内注射是安全的,并能有效抑制兔外伤性增生性玻璃体视网膜病变的发生发展。 关键词：壳聚糖;增生性玻璃体视网膜病变;外伤性;自体富血小板血浆;兔 doi:10.3969/j.issn.1673-8225.2012.08.003     

三种方法建立外伤性增生性玻璃体视网膜病变的兔模型研究

目的：比较三种方法建立外伤性增生性玻璃体视网膜病变(TPVR)兔模型的效果。方法：青紫蓝兔24只,共48只眼,在均造成眼外伤的基础上随机分成4组,每组各12只眼：实验组3组,分别为富血小板血浆(PRP)组[玻璃体腔内注射0.3 mL PRP]、转化生长因子β₂(TGF-β₂)组[玻璃体腔内注射0.3 mL TGF-β₂(50μg/L)]和视网膜色素上皮(RPE)组[玻璃体腔内注射0.3 mL RPE细胞(1.5×109/L)],以及对照组(玻璃体腔内注射0.3 mL生理盐水)。术后每周通过眼底照相和B超观察玻璃体及视网膜的增生情况;术后第4周采用Western blot法检测玻璃体内视网膜增生过程中α-平滑肌肌动蛋白(α-SMA)的表达,采用冰冻切片法在显微镜下观察各组视网膜组织结构。结果：术后第4周,对照组及3组实验组TPVR的平均分级分别是0、3.67、2.33和2.50,3组实验组病变级别显著高于对照组(P<0.05),PRP组病变级别高于TGF-β₂组和RPE组(P<0.05),...     

颈内动脉持续输注丙泊酚构建肌松兔模型的药代动力学变化

背景：经动脉选择性脑内给药方法是现阶段提高脑内药物浓度,降低药物对其他系统功能影响的有效方法之一。 目的：建立颈内动脉持续输注丙泊酚构建肌松兔模型,分析丙泊酚药物浓度变化规律。 方法：颈内动脉置管进行丙泊酚恒速持续输注建立兔颈内动脉持续输注丙泊酚肌松兔模型,在不同时点取得两侧颈内动静脉血及两侧脑组织样本,应用高效液相荧光法检测药物浓度,然后将所得数据进行数学转换,拟合处理,统计学回归分析药代动力学特点。 结果与结论：高效液相色谱荧光法测定丙泊酚浓度方法可行,系统稳定可靠。颈内动脉持续输注丙泊酚药物浓度增长率分析,数据分布呈对数正态分布图形,属于非指数动力学模型,即改良的对数正态分布模型,             ,其中σ代表脑内药物浓度变化波动性的稳定性,与脑组织药物摄取和脑循环等多种因素有关的综合变量。说明颈内动脉持续输注丙泊酚药代动力学模型属于对数正态分布函数,属于非指数函数动力学模型。两侧脑内浓度随时间的变化规律遵循对数正态分布函数规律。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

微透析技术在建立灵长类帕金森病模型中的应用

为了检测单侧注射ＭＰＴＰ制备的帕金森病恒河猴模型纹状体内多巴胺及其代谢产物的含量变化,本研究采用脑内微透析技术和高效液相色谱－ 电化学方法检测了双侧尾状核头部多巴胺及其代谢产物３,４－二羟基苯乙酸和高香草酸的含量。结果证明,ＭＰＴＰ注射侧与注射对侧相比,多巴胺、３,４－二羟基笨乙酸和高香草酸的含量分别降低８５．７％ , ９５．１％ 和６７．８％ 。这与动物呈现单侧帕金森病症状,核磁共振检查显示单侧黑质区域密度减低、面积缩小以及经抗酪氨酸羟化酶免疫组化方法显示脑切片单侧多巴胺能神经元明显减少等现象一致。以上结果表明,脑内微透析技术是活体检测脑内神经递质含量变化的有效方法,可以反映模型的病程变化,可用于评价帕金森病的治疗效果。     

实体肿瘤药物输运与给药方式的集中参数模型及实验研究

目的研究不同给药方式对实体肿瘤抗癌药物疗效的影响,并通过动物实验验证。方法分别建立体内药代动力学和肿瘤内药物输运的集中参数模型,模拟单次快速注射和等时间间隔三次快速注射时肿瘤间质组织药物浓度Ct随时间的变化。实验小鼠分为两组,分别进行抗癌药羟基喜树碱(HCPT)单次快速注射和等时间间隔3次快速注射,观察其肿瘤内药物平均浓度。对数值模拟结果和动物实验结果进行比较。结果数值模拟结果显示,对于相同总量药物,等量等间隔的三次给药,肿瘤间质组织药物浓度Ct高于单次给药。动物实验的实测结果相同。结论3次给药的效果显著优于单次给药。集中参数模型基本能够定量反映不同给药方式的效果。     

卵磷脂络合碘治疗视网膜脱离术后玻璃体混浊

目的：观察口服卵磷脂络合碘--沃丽汀治疗视网膜脱离术后玻璃体混浊的疗效.方法：选择因视网膜脱离行巩膜扣带术,术后不同程度玻璃体混浊的患者216例（216眼）,随机分为两组.试验组给予神经营养剂及口服沃丽汀治疗,分别在给药后的1周,1月及3月随访.对照组仅予神经营养剂治疗.结果：试验组1月内玻璃体混浊减轻、眼底明显清晰、自觉症状明显好转者36例（33.3%）,3月内有明显改善者64例（59.3%）,3月以上改善不明显者11例（10.2%）;所有病例均未发现有药物引起的副作用.对照组1月内玻璃体混浊减轻、眼底明显清晰、自觉症状明显好转者14例,3月内有明显改善者28例（25.9%）,3月以上改善不明显者41例（38%）.术后3月有效率组间比较差异有统计学意义（P＜0.01）.结论：沃丽汀能够有效减轻视网膜脱离术后玻璃体混浊的程度.     

清醒大鼠颈静脉采血技术在药代动力学两周期交叉实验中的应用

目的 探讨清醒大鼠颈静脉采血技术应用于药代动力学两周期交叉实验的可行性。方法 24只SD大鼠,雌雄各半,按照简化随机法分成实验组和对照组,每组12只,雌雄各半。实验组大鼠模拟药代动力学两周期交叉实验,生理盐水口服灌胃或尾静脉注射,于给药前后不同时间点采用清醒大鼠颈静脉采血技术采血0.1 mL,每日记录大鼠体质量。对照组大鼠正常饲养不采血,每日记录大鼠体质量。结果 实验组单只大鼠在清醒状态下每周期颈静脉采血达11次,24 h内采血总量为1.1 mL,大鼠一次性颈静脉采血成功率为84.1%,大鼠存活率为100%。实验组大鼠的体质量与对照组大鼠的体质量比较,差异无统计学意义（P>0.05）。结论 采用清醒大鼠颈静脉采血技术不影响大鼠正常生长,可应用于药代动力学两周期交叉实验,能满足实验研究所需的采血次数和采血量要求,并符合动物福利“3R”原则。     

眼前房内抗原注射抑制EAU的发生

目的：探讨眼前房内注射牛视网膜可溶性抗原(S抗原)在诱导眼免疫耐受中的作用。方法：自新鲜牛眼球中提取S抗原，注入Wistar大鼠眼前房内。1周后，皮内注射牛S抗原与弗氏完全佐剂乳化混合物。观察大鼠实验性自身免疫性葡萄膜视网膜炎(EAU)发作时间、临床表现及组织病理学改变；并以ELISA方法检测脾细胞培养上清液中的细胞因子。结果：前房预处理组(groupA)EAU的发生率明显低于阳性对照组(groupB)，所有具有阳性体征者均有组织病理学改变。前房预处理组动物脾细胞培养上清液中的IL-4浓度明显高于阳性对照组；而IFN-γ水平明显低于阳性对照组。结论：前房内注射牛S抗原可以诱导大鼠眼免疫耐受，在很大程度上抑制EAU的发生和发展。     

Subtherapeutic serum vancomycin concentrations during on-line hemodiafiltration

The aim of the study is to study vancomycin serum concentration changes during on-line hemodiafiltration (OL-HDF) and determine whether administration of vancomycin during the last hour of OL-HDF provides therapeutic serum concentrations. Vancomycin was administered intravenously at a dose of 15 mg/kg to 17 chronic hemodialysis patients who were enrolled into two study groups (A and B). In group A patients (n = 11), vancomycin was administered immediately postdialysis. Forty-three hours later, a 4-hour OL-HDF session was performed. Blood samples (S) for vancomycin measurement were drawn before, during, and after the session. In group B patients (n = 6), vancomycin was administered during the last hour of a 4-hour OL-HDF session on Monday. Vancomycin serum concentrations were measured postdialysis, as well as before and after the next two dialysis sessions of the week. In group A patients, mean vancomycin concentrations were reduced by 50% (S(pre): 10.2 ± 1.4 μg/ml, S(post): 5.0 ± 0.9 μg/ml). Vancomycin dialysance was 74.6 ± 29.3 ml/min, while the volume of distribution was 0.42 L/kg. In group B patients, mean vancomycin concentrations before the beginning of Wednesday and Friday dialysis sessions were 3.1 ± 0.7 and 1.4 ± 0.4 μg/ml, respectively. Postdialysis administration of vancomycin followed by OL-HDF resulted in a 50% reduction of vancomycin serum concentrations. Administration during the last treatment hour of OL-HDF results in subtherapeutic vancomycin serum concentrations over the course of the subsequent inter- and intradialytic intervals.     

Treatment of acute edema attacks in hereditary angioedema with a bradykinin receptor-2 antagonist (Icatibant)

BACKGROUND: In hereditary angioedema, bradykinin is assumed to be the most important mediator of edema formation. OBJECTIVE: To assess whether the selective bradykinin receptor-2 antagonist Icatibant is effective in acute edema attacks of hereditary angioedema. METHODS: In this uncontrolled pilot study, 15 patients with 20 attacks were treated with Icatibant. The attacks were analyzed by using a standardized and validated visual analog scale measurement and compared with historical data of untreated attacks. Plasma bradykinin concentration was measured before and 4 hours after intravenous Icatibant treatment. RESULTS: Symptom intensity decreased within 4 hours after administration of Icatibant; the median time to onset of symptom relief was 1.50, 1.42, and 1.13 hours in the intravenous groups and 0.58 and 0.45 hours in the subcutaneous groups, respectively. The median difference in the 10-cm visual analog scale 4 hours after start of treatment was 4.11 cm (95% CI, 1.72-6.07). Compared with untreated attacks, Icatibant treatment reduced the mean (SD) time to onset of symptom relief by 97% from 42 +/- 14 to 1.16 +/- 0.95 hours (all groups combined). Median bradykinin concentration was 7-fold above the norm during acute attacks at 48.5 pmol/L and decreased to 18.0 pmol/L 4 hours after Icatibant infusion or injection. CONCLUSION: Icatibant was effective in treating acute attacks of hereditary angioedema. CLINICAL IMPLICATIONS: This is the first report demonstrating the clinical usefulness of antagonizing bradykinin binding to bradykinin receptor-2 in hereditary angioedema.     

High-performance liquid chromatography with peroxyoxalate chemiluminescence determination of propentofylline concentrations in rat brain microdialysate

A high-performance liquid chromatographic determination of a neuronal cell protective compound, propentofylline [3-methyl-1-(5-oxohexyl)-7-propyl-7H-purine-2(3H),6(1H)-dione] was performed combining a microdialysis technique with peroxyoxalate chemiluminescence (PO-CL) detection. The microdialysate was subjected to a fluorescent derivatization of propentofylline with 4-(N,N-dimethylaminosulfonyl)-7-hydrazino-2,1,3-benzoxadiazole (DBD-H) without further cleanup, because the membrane used in the microdialysis excluded high-molecular-mass proteins. The proposed method showed a good linearity in the determination range of 0.031 to 1.25 ng/injection; y (microV)=4234 x (ng)- 13.43, r=0.9993 (y=peak height and x=amount of propentofylline). The very low determination limit of 0.031 ng/injection was ca. 200 times more sensitive than that of HPLC-UV determination. The HPLC-PO-CL method was applied for the first time to determine propentofylline concentration in the dialysate obtained from the rat hippocampus after a single oral administration (25 mg/kg). Propentofylline showed its maximum extracellular fluid (ECF) concentration of 125.1+/-15.1 ng/ml (mean+/-SD, n=3) at 0.33 h after administration.     

重组人表皮生长因子联合前列地尔作用于糖尿病溃疡创面

背景：人内源性表皮生长因子的缺乏以及血流动力学的改变会导致创面不愈合的发生。 目的：观察重组人表皮生长因子联合前列地尔作用于糖尿病溃疡动物模型创面的疗效。 方法：Wistar大鼠40只建立糖尿病溃疡动物模型,随机等分为模型组、重组人表皮生长因子组、前列地尔组和重组人表皮生长因子+前列地尔组,分别予以1%碘伏清创、重组人表皮生长因子凝胶外敷、前列地尔静脉滴注、重组人表皮生长因子凝胶外敷和前列地尔静脉滴注联合治疗。 结果与结论：干预后3,7,10,14 d观察发现,相比于模型组,重组人表皮生长因子和/或前列地尔治疗后,糖尿病皮肤溃疡大鼠溃疡面积减小、愈合时间缩短、创面动态愈合率上升(P < 0.01),且两者联合治疗的效果优于重组人表皮生长因子和前列地尔单独治疗(P < 0.01),而重组人表皮生长因子或前列地尔单独治疗的效果接近,提示重组人表皮生长因子与前列地尔联合使用比单纯使用前列地尔或单纯使用重组人表皮生长因子更能显著促进糖尿病溃疡创面的愈合。     

Absorption of chloramphenicol sodium succinate after intramuscular administration in children

Because it is thought that chloramphenicol is poorly absorbed after intramuscular administration, we compared blood levels of chloramphenicol after intramuscular administration with those after intravenous administration in children with a variety of diagnoses. Fifty-seven children were studied on 62 occasions while they were receiving chloramphenicol sodium succinate (25 mg of chloramphenicol per kilogram of body weight) intramuscularly every six hours. The peak level of chloramphenicol was 19.5 +/- 5.99 micrograms per milliliter (mean +/- S.D.) in 11 children after the first dose and 31.4 +/- 12.99 micrograms per milliliter in 51 children after two or more doses. The lowest peak level after intramuscular administration was 13 micrograms per milliliter, which is in the therapeutic range of 10 to 30 micrograms per milliliter. Thirteen children were studied on 17 occasions while they were receiving chloramphenicol sodium succinate (25 mg of chloramphenicol per kilogram) intravenously every six hours. The peak level of chloramphenicol was 19.4 +/- 6.37 micrograms per milliliter in eight children after the first dose and 28.2 +/- 11.09 micrograms per milliliter in nine children after two or more doses. The area under the serum level curve was not significantly different after intramuscular and intravenous administration. We conclude that chloramphenicol sodium succinate is well absorbed after intramuscular administration. This route is cheaper, it demands less staff time, and it does not carry the risks of sepsis and overhydration associated with intravenous therapy.     

衰老大鼠急性肺损伤诱导肾功能受损

目的:观察衰老大鼠的急性肺损伤(ALI)是否可进一步诱导肾功能受损。方法:Wistar雄性大鼠40只, 复制成衰老模型, 然后再随机分成对照组(静脉注射生理盐水), ALI组(静脉注射脂多糖, LPS)及LPS组(左心室内注射LPS), 后两组再分2h及6h组。每组8只。注LPS后2h或6h收集血并取肺、肾。制备肺、肾组织匀浆待测。结果:ALI组在注LPS后仅至6h时血中肌酐(Cr)、尿素氮(BUN)含量才有显著升高(均P＜0.01)。而LPS组Cr、BUN含量均无显著升高。ALI组血中乳酸(LD)、丙二醛(MDA)及一氧化氮(NO)含量在注LPS后2h时均显著升高(P＜0.05, P＜0.01);而肺组织中谷胱甘肽过氧化物酶(GSH-P_X)及Na⁺-K⁺-ATPase活性均显著下降(均P＜0.01);上述变化持续至观察的6h时。ALI组在注LPS后仅至6h时, 肾组织中MDA、NO含量才有显著升高(均P＜0.01)及GSH-P_X、Na⁺-K⁺-ATPase活性显著下降(P＜0.01)。而在LPS组, 除注LPS后2h时的血中和2h、6h的肺组织中NO含量显著升高(均P＜0.05)及2h时的肺组织中Na⁺-K⁺-ATPase活性显著下降(P＜0.05)外, 其它均无显著变化。结论:给衰老大鼠静脉内注射5mg/kg的LPS可致ALI, 并可进一步诱导肾功能受损。     

肾移植中霉酚酸血浓度的监测

背景：作为抗代谢免疫抑制剂,霉酚酸酯以其低毒副作用的优点已经在实体器官移植中得到广泛应用。然而,临床上移植患者常联合用药,霉酚酸酯与其他免疫抑制剂(如环孢素A和他克莫司)联合使用时的药代动力学特点尚不清楚。 目的：动态监测霉酚酸浓度,比较霉酚酸酯与不同免疫抑制剂联用时药代动力学特点,以探讨其最佳治疗浓度窗,指导临床个体化用药,从而提高疗效,降低其毒副反应。 方法：应用 SyvaEmit-2000药物浓度检测分析仪检测肾移植受者服药(霉酚酸酯)前(C0)、服药后0.5 h(C0.5)和服药后2 h(C2) 霉酚酸血浓度,并根据3个时间点的浓度值来计算0-12 h的霉酚酸血浆浓度-时间曲线下面积值。 结果与结论：入选340例肾移植受者,共检测霉酚酸3点血浆浓度-时间曲线下面积820例次,霉酚酸酯+他克莫司联合用药组574例次;霉酚酸酯+环孢素A联合用药组246例次。霉酚酸酯与环孢素A联合应用时,0-12 h的霉酚酸血浆浓度-时间曲线下面积值显著低于霉酚酸酯与他克莫司联合应用时,差异有显著性意义(P < 0.05)。霉酚酸酯联用他克莫司组服药前、服药后0.5 h霉酚酸浓度值高于霉酚酸酯联用环孢素A组(P < 0.05),霉酚酸酯联用环孢素A组服药后2 h霉酚酸浓度值高于霉酚酸酯联用他克莫司组(P < 0.05)。霉酚酸酯联用他克莫司组服药后0.5 h出峰的病例数高于霉酚酸酯联用环孢素A组(P < 0.05)。结果可见霉酚酸酯与不同免疫抑制剂联用时应根据相应的药代动力学特点给药,如与他克莫司联用时,应该更早地减低他克莫司剂量。三点霉酚酸血浆浓度-时间曲线下面积监测相对简便易行,在临床应用更具有可操作性     

Pharmacokinetic and Pharmacodynamic Profile of New Biosimilar Filgrastim XM02 Equivalent to Marketed Filgrastim Neupogen®

Objective

Filgrastim XM02 is a biosimilar non-glycosylated recombinant methionyl form of human granulocyte colony-stimulating factor (r-MetHuG-CSF) expressed in Escherichia coli for subcutaneous and intravenous administration in the treatment of different forms of neutropenia and stem cell mobilization. This study was conducted to compare the pharmacokinetic and pharmacodynamic characteristics of the new biosimilar filgrastim XM02 with the marketed filgrastim (Neupogen®).

Methods

Two filgrastim doses (5 and 10 μg/kg) of the new biosimilar filgrastim XM02 and the marketed filgrastim were administered either as intravenous infusion or subcutaneous injection in four single-dose, crossover, randomized substudies, conducted in 36 subjects each. Serum concentrations of filgrastim were determined using an enzyme-linked immunosorbent assay test kit on samples taken at intervals up to 48 hours after administration. The CD34+ stem cell count up to 15 days after administration was determined by flow cytometry using a validated CD34+ cell enumeration kit, and the absolute neutrophil count (ANC) up to 96 hours after dosing was determined by the Beckman Coulter AcT differential automated hematology analyzer. The primary pharmacokinetic endpoint was the AUC_48h (area under the serum concentration-time curve) of filgrastim serum concentration determined by the linear trapezoidal rule. Equivalence (biosimilarity) between the two filgrastim products was assessed by 90% confidence limits obtained from analyses of variance of log-transformed pharmacokinetic and pharmacodynamic endpoints, applying 80–125% equivalence intervals.

Results

The mean serum concentration profiles of filgrastim, ANC and CD34+ cells over time were similar for the two filgrastims. The 90% confidence intervals for all test/reference ratios for pharmacokinetic and pharmacodynamic endpoints lay within the accepted bioequivalence range of 80–125%. Both filgrastims showed similar safety profiles and were well tolerated.

Conclusions

Equivalence of the two filgrastims was clearly demonstrated for all four dose/route of administration groups. Equivalence could be demonstrated for the serum concentration profile, for the ANC profile and, even more importantly, for the CD34+ cell count, which is a marker for the ability of the granulocyte colony-stimulating factor to mobilize stem cells.     

Prophylaxis of Gram-positive infections after bone marrow graft. Controlled study of the 6-day administration of vancomycin. Intermediate analysis of 60 patients

We prospectively evaluated after randomization in 60 consecutive recipients of bone marrow transplantation the efficiency of a short administration of vancomycin (10 mg/kg I.V. q 6 H, day -5 to +1). Thirty-three patients were randomized as controls and 27 were designed to receive vancomycin. Both groups were similar in terms of age, diagnosis, status, allogeneic or autologous graft, in vitro marrow treatment or not. Eighteen patients (30%) experienced bacteremias: 15/18 bacteremias were due to Gram positive organisms, including 11 streptococci. These infections occurred with a similar frequency in both groups, the relatively low incidence of bacteremias in this study needs further evaluation in new patients to insure the prophylactic effects of a course of vancomycin in bone marrow transplant recipients.