单克隆抗体ELISA测定血浆富组氨酸糖蛋白的方法

建立用单克隆抗体酶联免疫吸附测定法（ＥＬＩＳＡ）测定血浆富组氨酸糖蛋白（ＨＲＧ）的方法。结果显示：ＭｃＡｂ３Ｃ６Ｆ３包被浓度为１０μｇ·ｍｌ＾－１,单抗酶标结合物作１：２００稀释时标准曲线最理想;最低检出限为１ｎｇ·ｍｌ＾－１,批内ＣＶ８．２％,批间ＣＶ１１．５％,回收率为８９．１％,测定范围为２．５－８０ｎｇ·ｍｌ＾－１;用该法测定４０例正常人血浆ＨＲＧ为（１１０．３±９．７）ｍｇ·Ｌ＾－１,２     

高效液相色谱法测定全血环孢霉素 A

采用 Ｃ１８ 反相色谱柱法测定定值全血（ Ｂ１０ Ｒ Ａ Ｄ） ，３ 个浓度的水平。结果：均在预期范围内，其最低检出浓度为２５ ｎｇ·ｍｌ－ １ ，至少在５０ ～１ ０００ ｎｇ·ｍｌ － １ 范围内呈线性；绝对回收率为９９６ ％ ～１０８８ ％ ；批内和批间变异系数分别为７２ ％ 和９３ ％ 。     

盐酸异丙嗪体外杀精子作用的实验研究

３０例健康育龄男性精液的体外杀精试验显示：盐酸异丙嗪具有快速强力杀精作用,当药物浓度在１８６ｍｍｏ·Ｌ－１（０６ｍｇ／ｍｌ）以上时,在１０ｓ内死亡;在００７５～１２４ｍｍｏｌ·Ｌ－１（０．２５～０．４ｍｇ／ｍｌ）浓度范围内,其杀精作用随药物浓度增加和作用时间延长而增强。当浓度为０３１ｍｍｏｌ·Ｌ－１（０．１ｍｇ／ｍｌ）作用时间为３０ｍｉｎ时,杀精作用可接近或达到１００％;而浓度为１２ｍｍｏｌ．Ｌ－１（０．４ｍｇ／ｍｌ）时,５ｍｉｎ内即可杀灭全部精子     

白细胞介素—1α诱导牛脑微血管平滑肌细胞增殖及药物的拮抗作用

作研究了白细胞介素－１α对培养的牛脑微血管平滑肌细胞增殖的影响。在５～５００ｐｇ～ｍｌ＾－１范围内，ＩＬ－１α可诱导培养的牛脑微血管平滑肌细胞增殖，当用５０ｐｇ·ｍｌ＾－１ＩＬ－１α与该平滑肌细胞共孵育２４ｈ时，刺激作用最强，增殖率达２９．１％。欧芹素乙、异欧芹素乙，６－（α，α－苯基乙酰哌嗪基苯基）－４，５－二氢－５－甲基－３（２Ｈ）哒嗪酮在浓度为１０＾－６～１０＾－４ｍｏｌ·Ｌ＾－１时，可剂     

尿毒症血浆中的中分子物质对猪肾皮质Na^＋,K^＋—ATPa酶的抑制 …

为研究中分子物质（ＭＭ）对Ｎａ＾＋,Ｋ＾＋－ＡＴＰａｓｅ的抑制作用,我们采用凝胶层析技术,分离出尿毒症病人及健康人的ＭＭ,同时从猪肾皮质提纯了Ｎａ＾＋,Ｋ＾＋－ＡＴＰａｓｅ,其比活性２２０μｍｏｌ无机磷／ｍｇ蛋白．小时。尿毒症ＭＭ峰２－３浓度在１．０ｍｇ／ｍｌ以上,峰２－４浓度在５．０ｍｇ／ｍｌ以上时对Ｎａ＾＋－Ｋ＾＋ＡＴＰａｓｅ上时对Ｎａ＾＋,Ｋ＾＋－ＡＴＰａｓｅ有抑制作用,正常人ＭＭ无抑制作用     

酚妥拉明和多巴胺治疗难治性哮喘390例临床观察

目的：探讨酚妥拉明和多巴胺治疗难治性哮喘的临床疗效。方法：每次以酚妥拉明（０．５～１）ｍｇ／ｋｇ,多巴胺（０．２５～０．５）ｍｇ／ｋｇ加入１０％葡萄糖溶液（１００～１５０）ｍｌ中用微泵控制滴速,以５ｕｇ．ｋｇ＾－１．ｍｉｎ＾－１静滴,连用３ｄ,观察疗效。结果：用药后１２～２４ｈ,气急渐平,心率减慢,紫绀消失,孝鸣音减少,血气基本恢复正常,总有效率达９３．３％。未发现明显副作用。结论：在抗炎、吸氧、     

四种抗生素对细菌内毒素试验干扰作用的观察

本文按照美国药典１９９５年２３版规定的细菌内毒素限值,考查了硫酸庆大霉素注射液,注射用青霉素钠,注射用氨苄青霉素钠,注射用头孢唑啉钠四种抗生素对细菌内毒素试验的干扰性,结果表明,在浓度分别为６００ｕ．ｍｌ＾－１,１００００ｕ．ｍｌ＾０－１,６．８ｍｇ．ｍｌ＾１和６．８ｍｇ．ｍｌ＾－１时对细菌内毒素试验均无干扰作用,可用细菌内毒素检查法控制热原。     

高效液相色谱法快速直接测定血清苯丙氨酸和酷氨酸

目的：建立快速、直接、同时测定血清苯丙氨酸和酷氨酸的方法。方法：血清标本加高氯酸去蛋白后取上清,采用固定流量洗脱高效液相色谱紫外检测法测定。结果：苯丙氨酸的线性范围为６～１５１２μｍｏｌ．Ｌ＾－１,最低检测浓度为０．７５μｍｏｌ．Ｌ＾－１,平均回收率为９８．６％,批内ＣＶ为３．６８％,批间ＣＶ为４．０１％。酷氨酸的线性范围为５．４～１３８０μｍｏｌ．Ｌ＾－１,最低检测浓度为０．７μｍｏｌ．Ｌ＾－１     

高效液相色谱测定体液乙酰白霉素

为建立一种灵敏度高、特异性和实用性强的体液乙酰白霉素测定方法，采用Ｌｃ－６Ａ高效液相色谱仪、Ｓｈｉｍ－ＰａｃｋＣＬＣ－ＯＤＳ分析法，并根据原药理化性质选用甲醇：０．２ｍｏｌ／Ｌ醋酸铵＝２．８：１为流动相，流速１．５ｍｌ／ｍｉｎ，波长２３１ｎｍ条件下对测定方法进行研究，结果表明：血尿药物浓度在０．０５０～３．０００μｇ／ｍｌ范围内，药物浓度与Ａ５峰高线性关系良好：回收率血、尿药物平均值分别为９６．４３％与９６．２５；变异系数血、尿药物浓度为０．５μｇ／ｍｌ与１．５μｇ／ｍｌ时，两者批内变异系数在４．７５％～５．７５％间，批间系数在６．４９％～６．７０％范围间。     

血浆中芦丁和槲皮素的HPLC检测方法及其在大鼠体内的药物动力学研究

目的：建立血浆中芦丁和槲皮素浓度ＨＰＬＣ分析方法，并用此方法对大鼠静脉 注射及灌胃给药进行药物动力学研究；方法：血浆标本经甲醇－冰醋酸提取，以乙腈－水为流动相，紫外３６０ｎｍ处检测；结果：芦丁和槲皮素线性范围均在１μｇ．ｍｌ＾－１～１５０μｇ．ｍｌ＾－１；最低检测浓度芦丁为０．３μｇ．ｍｌ＾－１、槲皮素为０．５μｇ．ｍｌＡ＾－１；此方法简便、灵敏、可靠，能满足临床药物动力学研究的需要。颈静脉 分别     

NGAL对成人心脏术后急性肾损伤的早期诊断价值

目的:探讨血和尿中中性粒细胞明胶酶相关脂质运载蛋白(NGAL)水平对成人心脏术后急性肾损伤(AKI)的早期诊断价值.方法:收集112例心脏手术患者手术前后不同时相的血、尿标本,分别测定血肌酐(Scr)、血NGAL和尿NGAL.根据AKI的诊断标准,将患者分为AKI组和非AKI组,观察两组Scr、血NGAL和尿NGAL的动态变化.用受试者工作特征曲线(ROC)评价血NGAL和尿NGAL对AKI的诊断价值.结果:112例患者中21例术后24 h Scr水平上升至基础值的1.72倍,AKI发生率为18.8%.AKI组术后2 h血NGAL开始显著增高并达到峰值,与基线值比较差异有统计学意义[分别为(82.2±22.3) ng·ml-1和(16.9±3.1) ng·ml-1,P<0.05];尿NGAL较基线值明显上升[分别为(132.4±41.1) ng·ml-1和(30.8±23.1) ng·ml-1,P<0.05],并在术后4 h达到峰值.AKI组术后2 h,血NGAL诊断AKI的ROC曲线下面积为0.866,95%的可信区间为0.756~0.976,以19 ng·ml-1作为AKI的诊断界限时,敏感性和特异性分别为85.7%和88.4%;尿NGAL的ROC曲线下面积为0.914,95%的可信区间为0.86~0.966,以56 ng·ml-1作为AKI的诊断界限时,敏感性和特异性分别为95.2%和88.4%.结论:心脏手术后2 h的血、尿NGAL水平对AKI具有预测价值,其出现变化时间远早于Scr;血、尿NGAL可作为心脏术后并发AKI患者的早期诊断的标记物.     

大剂量萘哌地尔在家犬体内的药物动力学

目的：建立HPLC测定家犬血浆中萘哌地尔浓度的方法，研究大剂量萘哌地尔胶囊在家犬体内的药物动力学，方法：健康家犬5只，单剂量给予萘哌地尔胶囊200mg后，在不同时间点从后肢静脉取血，血浆样品碱化后经乙醚提取，以乙腈：磷酸盐缓冲液（pH6.5)(60:40)为流动相，由ODSC18分析柱分离测定，紫外230nm为检测波长，维拉帕米为内标，血药浓度数据用3p97药物动力学程度处理，结果：线性范围为10－1200ng/ml^-1；方法回收率为98．83％－101。50％，日间RSD≤5．56％，日内RSD≤3．30％，单剂量给予家犬萘哌地尔胶囊200mg后，血药浓度随时间变化规律符合一室开放模型，T1／2Ke为1．91－4．99h,Tmax为1．87－3．21h，Cmax为338．79－414．04ng.ml^-1。结论：本方法简便，回收率高，重现性好，用于大剂量萘哌地尔在动物体内的药物动力学研究，切实可行。     

尼古丁对体外培养人外周血单个核细胞分泌TNF的影响

OBJECTIVE: To observe the effect of nicotine on the secretion of TNF of human peripheral blood mononuclear cells (PBMC) in vitro, and explore the possible mechanism of the high level of TNF caused by smoking. METHODS: Twenty-one non-smokers and 19 smokers were studied. The PBMCs isolated from each volunteer's blood sample were distributed to three groups, and cultured for 72 hours in supplemented RMPI 1640 alone or in supplemented RPMI 1640 containing 50 ng.ml-1 nicotine or 500 ng.ml-1 nicotine respectively. The level of TNF in supernate was quantified with the immuno-radioassay, and PBMC proliferation was observed. RESULTS: The level of TNF spontaneously secreted by PBMCs in smokers was significantly higher than that in non-smokers (P < 0.05). When the concentrations of nicotine were 50 ng.ml-1 and 500 ng.ml-1, the level of TNF increased significantly in non-smokers (P < 0.05, P < 0.01). In smokers, the level of TNF significantly increased when the concentration of nicotine was 50 ng.ml-1; however, the level of TNF significantly decreased (P < 0.05), and the proliferation of PBMCs was inhibited when the concentration of nicotine was 500 ng.ml-1 (P < 0.05, P < 0.05). The level of TNF was positively correlated to the multiple of PBMC proliferation (r = 0.93, P < 0.05). The linear regression equation was Y = 2.913X - 2.955. CONCLUSION: Nicotine can induce PBMCs to secrete more TNF, and the magnitude of the effect is strongly related to the dosage of nicotine, but a great dose of nicotine will inhibit the production of TNF.     

FK506对d-BSA超载的NRK-52E细胞增殖的影响

目的:探讨他克莫司(tacrolimus,FK506)对去脂牛血清白蛋白(denatured bovine serum albumin,d-BSA)超载的大鼠肾小管上皮细胞(normal rat kidney proximal tubular epithelial cell line,NRK-52E)增殖的影响。方法:(1)以不同终质量浓度(1、5、10、20、50 mg·ml-1)d-BSA超载NRK-52E,以正常培养的NRK-52E细胞为对照组,观察不同浓度d-BSA对细胞增殖的影响及d-BSA(20 mg·ml-1)超载不同时间(6、12、24 h)对NRK-52E增殖的影响。(2)以不同浓度(0.1、1、10、20 ng·ml-1)FK506预处理NRK-52E 4 h后加入终质量浓度为20 mg·ml-1d-BSA共孵育24 h,观察不同浓度FK506预处理对d-BSA超载细胞增殖的影响。(3)以10 ng·ml-1的FK506预处理4 h后加入终质量浓度为20 mg·ml-1d-BSA共孵育NRK-52E,观察不同时间(6、12、24 h)对细胞增殖影响。以上细胞增殖测定均采用四甲基偶氮唑盐法(MTT)。结果:(1)终质量浓度为1 mg·ml-1的d-BSA超载NRK-52E 24 h后无明显促进细胞增殖的作用(P>0.05);终质量浓度为5 mg·ml-1的d-BSA能够促进细胞增殖(P<0.05),终质量浓度为10、20、50 mg·ml-1的d-BSA能够显著促进细胞增殖(P<0.01)。(2)以终质量浓度为0.1 ng·ml-1的FK506预处理NRK-52E,细胞增殖与对照组比较差异无统计学意义(P>0.05);终质量浓度为1 ng·ml-1的FK506具有抑制d-BSA诱导的细胞增殖的作用(P<0.05);10、20 ng·ml-1FK506能够显著抑制-d-BSA诱导的细胞增殖(P<0.01)。(3)终质量浓度为10 ng·ml-1的FK506预处理4 h后加入终质量浓度为20 mg·ml-1d-BSA共孵育NRK-52E,6 h后,具有抑制细胞增殖的作用(P<0.05),共孵育12、24 h后,抑制细胞增殖的作用更加显著(P<0.01)。结论:d-BSA超载NRK-52E可以诱导细胞增殖,FK506具有抑制d-BSA促进NRK-52E细胞增殖的作用。     

Target range maximum of cyclosporine blood concentration two hours post dose in stable liver transplant patients

Recently, single blood level measurement 2 hours after cyclosporine administration (C2) is taken as a more sensitive indicator of drug exposure in de novo transplant recipients than trough levels (C0). However, few studies focused on the determination of the C2 target range maximum and its associated adverse events in stable liver recipients. This prospective study was designed to assess the relative risk of developing CsA related side effects in patients with high C2-levels. Adverse effects were determined clinically, and by using a specially designed questionnaire. Eventual adverse events as well as C2 levels were determined repeatedly up to 4 times in 3-months intervals (observation period 9 +/- 3 months) in 36 long-term liver recipients (1-13.5 years post-transplant), in addition to conventional C0 levels. Cyclosporine dose was adjusted according to a predefined C0 target level range and clinical status. Totally 103 questionnaires and the corresponding paired CsA blood level records were obtained. C0 levels and C2 levels ranged from 90 to 287 (143 +/- 31) ng/ml and from 212 to 1358 (672 +/- 203) ng/ml respectively. No patient experienced a rejection episode during the observation period, demonstrating the efficiency of the immunosuppressive therapy. However, 33/36 patients (91%) showed symptoms attributable to CsA therapy. C2 levels above 750 ng/ml, determined at least twice in an interval of 3 months, were identified as a relevant risk factor for the presence of multiple adverse effects, which were defined as the combination of hypertension, renal insufficiency and more than two neurological complaints (RR = 3.11, p<0.01). This risk population was not completely identified by determination of C0 level.     

Routine Cyclosporine concentration - C2 level Monitoring. Is it helpful during the early post Transplant Period?

Background

Pre dose or trough blood cyclosporine (CSA) concentration is routinely monitored and the result is used to alter patient''s drug dosing. Patients with identical pre dose blood CSA may have very different systemic exposure to the drug. Recently CSA 2 hour post dose level [C2] has been reported to correlate better with drug exposure. We undertook this study to evaluate the influence of trough and C2, CSA concentration monitoring on short-term renal allograft outcomes.

Methods

25 patients of renal transplant receiving a triple drug regimen of CSA micro emulsion (Panacea Biotec) 8mg/kg, azathioprine 1mg/kg and prednisolone 0.5mg/kg were analyzed prospectively for graft outcomes. CSA levels were monitored in whole blood by radioimmunoassay using monoclonal antibodies, at 72 hours after the transplant.

Results

The mean age of patients was 37.08 + 9.1 years. There were 20 males and 5 females. The mean age of donors was 40.2 + 8.2 years. There were 11 related donors with at least a haplomatch, 4 spousal and 10 unrelated donors with a nil antigen match. The mean pre dose CSA concentration was 289.22 + 171.9ng/ml; range (98.8 + 783.41ng/ml). The CSA concentration at 2 hours after the CSA administration was 838 + 310.87ng/ml (range, 169 + 1268ng/ml). 3 (12%) patients had acute rejection. In these patients the mean pre dose CSA concentration was 328.67ng/ml and the mean C2, CSA concentration was 1006.26ng/ml. CSA induced hemolytic uraemic syndrome was diagnosed in one patient. The trough and C2, CSA concentration levels were 174 and 870.83ng/ml respectively in this patient.

Conclusion

In our study CSA levels, trough and peak showed significant inter patient variability. The trough and C2 concentration levels did not correlate with the episodes of acute rejection. We conclude that in a triple drug regimen with fixed dosing schedules routine trough CSA level monitoring is not helpful in the acute post renal transplant period.Key Words: Cyclosporine levels, Cyclosporine trough levels, C2 levels     

Effect of medical ozone therapy on renal blood flow and renal function of patients with chronic severe hepatitis

Background Medical ozone therapy system was reported to have certain effects on the treatment of severe hepatitis, but its mechanism is not very clear. One of the causes of death of severe hepatitis is complication of renal damage or hepatorenal syndrome. The present study aimed to observe effects of medical ozone therapy system on plasma renin activity （PRA）, angiotensin II （All）, aldosterone （ALD）, renal blood flow and renal function of patients with chronic severe hepatitis and explore mechanisms of medical ozone therapy in the treatment of severe hepatitis. Methods Eighty-five cases with chronic severe hepatitis were randomly divided into ozone therapy group （43 cases） and control group （42 cases）. The patients in the ozone therapy group were treated with basic treatments plus ozone therapy system. Basic autohemotherapy was used. One hundred milliliter venous blood was drawn from each patient, and was mixed with 100 ml （35 pg/ml） medical ozone and then was returned the blood to the patient intravenously, once every other day for 20 days. Only the basic treatments were given to the control group. PRA, All, ALD, renal blood flow and damage to renal function of the two groups before treatment and 20 days after treatment were compared. Survival rates were also compared. Results Twenty days after the treatment, in ozone therapy group, PRA was （1.31±0.12） ng.m^-1.h^1, All （111.25±17.35） pg/ml, ALD （251.31±22.60） pg/ml, which decreased significantly compared with those before treatment （PRA （2.23±0.13） ng.ml^-1.h^-1, All （155.18±19.13） pg/ml, ALD （405.31±29.88） pg/ml, t=4.67-14.23, P 〈0.01 ）, also lower than those of control group 20 days after the treatment （PRA （2.02±0.11） ng.ml^-1.h^-1, All （162.21±15.32） pg/ml, ALD （401.20±35.02） pg/ml, t=4.97-15.61, P 〈0.01）; renal blood flow was （175.15±28.20） ml/min, which increased compared with that before the treatment （（125.68±21.25） ml/min） and was higher than that of control group 20 days after the treatment （（128.59±23.15） ml/min, t=4.78, 4.61, P 〈0.01）. Renal damage occurred in 2 cases （5%） in ozone therapy group, less than that in control group （9 cases, 21%） （X^2=5.295, P 〈0.05）. Thirty-three cases （77%） in ozone therapy group vs. 16 cases （38%） in control group survived （X^2=12.993, P 〈0.01 ）. Conclusions Basic treatment plus medical ozone therapy for patients with chronic severe hepatitis could decrease PRA, All and ALD levels significantly increase renal blood flow, prevent renal damage to certain extent and improve survival rate of the patients.     

慢性心力衰竭患者血浆肿瘤坏死因子浓度变化

The plasma levels of tumor necrosis factor (TNF) were measured by enzyme-linked immunoabsorbent assay in 109 patients who were grouped as "non-heart failure" (n = 36), "heart failure" (n = 36) and "cachexia" (n = 37). The results showed that there was no obvious difference in caloric intake among the three groups. Mean plasma levels of TNF were increased in patients with heart failure (0.51 +/- 0.2 ng.ml-1, P < 0.05) and even higher in patients with cachexia (6.19 +/- 2.76 ng.ml-1, P < 0.01) than in patients with non-heart failure (0.085 +/- 0.025 ng.ml-1). In the group with enhanced plasma levels of TNF (TNF > 100 pg.ml-1), the incidence of cachexia (61.0%) was obviously greater than that in the group with normal levels of TNF (17.6%). These findings indicate that plasma levels of TNF are elevated in patients with heart failure; high levels of TNF may play an important role in the pathogenesis of cardiac cachexia.     

替米沙坦血药浓度高效液相—荧光检测方法的研究

目的建立测定血浆中替米沙坦浓度的高效液相—荧光色谱分析方法。方法血浆样品以乙醚为提取溶剂处理,分析柱为KromsailKR100-5C18色谱柱（5μm,150 mm×4.6 mm）。流动相为0.01 mol/L KH2PO4水溶液（pH3.5）：乙腈=54：46（V/V）,激发波长Ex=305nm,发射波长Em=365 nm,流速1.0 ml/min,柱温为室温,进样量20μl。结果替米沙坦在5.0～500 ng/ml范围内线性良好（r=0.999 8）,最低检测浓度为2.5 ng/ml（S/N〉3）。该方法的相对回收率为（100.7±4.2）%（n=5）,绝对回收率为（88.4±3.9）%（n=5）,日内RSD为（3.1±1.0）%（n=5）,日间RSD为（4.0±1.5）%（n=5）。结论本方法准确、灵敏、特异性强、重显性好,适用于血浆中替米沙坦浓度测定和体内药代动力学研究。