Tumour necrosis factor-alpha levels in aqueous humour and serum from patients with uveitis: the involvement of HLA-B27

OBJECTIVE: To study the local and systemic levels of the tumour necrosis factor-alpha in patients with active uveitis and to determine the implication of TNF-alpha in rheumatological uveitis and to observe if this relationship is more significant in the B27 positive patients. PATIENTS AND METHODS: Patients were selected on the basis of a diagnosis of uveitis of any aetiology. Data from 23 patients were stratified into two categories according to the presence or absence of systemic rheumatic disease. The first group comprised nine patients with rheumatic disease; the second group contained 14 patients without rheumatic disease. The patients were also sub-classified into those who were HLA-B27 positive (14 patients) and those who were not. TNF-alpha levels in serum and aqueous humour from a group of 16 patients with uncomplicated cataracts were analysed as a control group. RESULTS: In the control group (n = 16) the serum TNF-alpha concentration was 13.1 +/- 2.9 pg/ml and the aqueous humour concentration of TNF-alpha was 0.56 +/- 1.53 pg/ml. In uveitis patients (n = 23) the serum TNF-alpha concentration was 35.35 +/- 26.77 pg/ml and the aqueous humour concentration of TNF-alpha was 15.1 +/- 1.70 pg/ml (p < 0.01). In HLA-B27 positive patients (n = 9) the serum TNF-alpha concentration was 45.56 +/- 34.17 pg/ml and the aqueous humour concentration of TNF-alpha was 15.89 +/- 0.93 pg/ml. In HLA-B27 negative patients (n = 14) the serum TNF-alpha concentration was 28.79 +/- 19.38 pg/ml and aqueous humour concentration of TNF-alpha was 14.57 +/- 1.91 pg/ml (p < 0.01). CONCLUSIONS: The concentration of TNF-alpha in aqueous humour in patients who are HLA-B27 positive is significantly greater than in those who are B27 negative. No significant differences in the concentrations of TNF-alpha in serum or aqueous humour in patients with or without rheumatic diseases were detected. TNF-alpha is a cytokine that may participate actively in the pathogenesis of clinical uveitis.     

SSP-PCR方法中两种不同引物检测人类白细胞抗原B27的比较研究

目的 用序列特异引物聚合酶链反应(SSP-PCR)方法检测人类白细胞抗原B27(HLA-B27)表达情况,探讨使用不同引物检测HLA-B27用于强直性脊柱炎鉴别诊断的敏感性及特异性.方法 使用两种不同特异性引物测定强直性脊柱炎(AS)患者HLA-B27,比较其检测的阳性率,结合临床诊断判断准确性.结果 HLA-B27新引物应用检测的敏感性、特异性均高于原引物.结论 新设计序列特异性引物的应用更适宜于临床检验和研究工作.     

Improved penetration of aminoglycosides and fluoroquinolones into the aqueous humour of patients by means of Acuvue contact lenses

Objectives: In order to improve the penetration of topically applied drugs in ophthalmology, the suitability of hydrophilic contact lenses (Acuvue, Vistacon, power −1.0 D) as a drug delivery system for antibiotics was tested. A prospective study was undertaken to determine the transcorneal penetration of five topically applied aminoglycosides and fluoroquinolones into the aqueous humour of patients. Methods: Two hundred and sixty-five patients undergoing cataract extraction received 0.3% gentamicin, kanamycin, tobramycin, ciprofloxacin or ofloxacin solution by two different modes of administration: either as eye drops (nine drops every 15 min, starting 2 h prior to surgery) or by means of a drug delivery system (Acuvue contact lenses soaked for 1 h in eye drop solution without preservatives, 1–5 h prior to surgery). At the beginning of cataract extraction, 50–100 μl aqueous fluid was aspirated from the anterior chamber and immediately stored at −80 °C. Antibiotic concentrations were measured using fluorescence polarisation immuno-assays (aminoglycosides) or high-performance liquid chromatography (fluoroquinolones). Results: After soaking for 1 h in 0.3% eye drop solutions, Acuvue contact lenses released about 190–250 μg aminoglycoside and ofloxacin and 1000 μg ciprofloxacin. These amounts are considerably lower or in the same order of magnitude than obtained with application of eye drops (1350 μg). From the aminoglycosides tested, only gentamicin and tobramycin, but not kanamycin, were able to penetrate into the aqueous humour of patients. After the wearing of antibiotic-soaked lenses, mean aqueous humour concentrations were higher than after the use of eye drops. This difference reached significance in tobramycin (1.09 (1.30) μg · ml⁻¹ vs 0.49 (0.79) μg · ml⁻¹), ciprofloxacin (1.23 (0.60) μg · ml⁻¹ vs 0.38 (0.33) μg · ml⁻¹) and ofloxacin (5.55 (2.53) μg · ml⁻¹ vs 0.56 (0.37) μg · ml⁻¹). The percentage of patients with aqueous humour concentration above the MIC₉₀ of Staphylococcus epidermidis, the most common cause of postoperative endophthalmitis, was 92% and 100% after wearing ciprofloxacin- or ofloxacin-soaked lenses, respectively. Conclusion: Gentamicin and tobramycin penetrated into the aqueous humour of patients, whereas kanamycin was not able to overcome the corneal barrier. Acuvue contact lenses soaked in 0.3% eye drop solutions can release sufficient amounts of gentamicin, ciprofloxacin and ofloxacin to produce bacteriostatic concentrations in the humor aquosus. Acuvue contact lenses can be recommended as a drug delivery system for fluoroquinolones. Received: 15 October 1998 / Accepted in revised form: 16 December 1998     

腰背痛患者520例人类白细胞抗原-B27检测及亚型分型

目的探讨人类白细胞抗原B27(HLA-B27)的检测及其亚型分型。方法采用流式细胞术(FCM)及聚合酶链反应-序列特异性引物(PCR-SSP)法对520例腰背痛患者外周血HLA-B27及其亚型进行检测。结果在520例腰背痛患者中,FCM阳性率为55.4%,PCR-SSP法阳性率为56.7%。两者的阳性率差异无统计学意义(χ2≈3.79,P>0.05)。PCR-SSP法检出HLA-B2704、HLA-B2705、HLA-B2705/07、HLA-B2702、HLA-B2704/05、HLA-B2705/02、HLA-B2704/02、HLA-B2704/05/07等亚型,分别占42.0%、29.2%、23.1%、1.4%、0.7%、0.7%、0.3%。结论 FCM与PCR-SSP法是目前常用的检测HLA-B27的方法,两者阳性率差异无统计学意义,但PCR法可进行亚型分型,这些亚型与强直性脊柱炎等疾病的诊断、治疗及预后判断相关。     

ELISA法检测HLA-B27抗原的表达及意义

目的对不同HLA-B27检测方法之间的差异及各自的优缺点进行评价并探讨其临床意义。方法分别采用微量淋巴毒法（MLCT）及ELISA检测372例患者外周血标本的HLA-B27抗原,将二者结果进行分析比对;并对患者人群中的B27含量进行观察。结果AS患者组和非AS患者组HLA-B27阳性例数及阳性率相比,差异有统计学意义（PELISA〈0.01,P MLCT〈0.01）;两种检测方法有很强的一致性（K=0.757,P=0.034〈0.05）结果符合率为88.2%;以MLCT法为标准,ELISA法检测敏感度和特异度分别为87.6%,88.6%,阳性预测值84.3%,阴性预测值91.1%,误诊率11.4%,漏诊率12.4%,Youden指数0.762;HLA-B27含量与起病年龄之间存在较弱的正相关（r=0.345,P=0.026）。结论两种检测方法均结果基本一致,ELISA法HLA-B27检测对AS的早期诊断与判断预后有价值;B27浓度与发病年龄之间可能有一定联系。     

Development and validation of a reversed-phase liquid chromatographic method for the assay of lidocaine in aqueous humour samples

A simple, fast and reliable reversed-phase liquid chromatographic method was developed for the assay of lidocaine in human aqueous humour samples. The samples were analysed without any preliminary treatment on a C8 column with UV detection at 225 nm. The mobile phase consisted of methanol/sodium dihydrogen phosphate (30 mM) containing sodium pentansulphonate (10 mM) adjusted to pH 2.5 with phosphoric acid (50:50 v/v). Validation of the method showed it to be precise, accurate and linear over the concentration range of analysis with a limit of detection of 0.2 μg ml⁻¹. The limit of quantitation was 2.5 μg ml⁻¹ with a relative standard deviation of 2.5%. Linear regression analysis in the range 2.5–60 μg ml⁻¹ gave correlation coefficients higher than 0.999. No interference from three commonly co-administered drugs was observed. The method developed was applied to the analysis of lidocaine in aqueous humour samples in order to evaluate and compare the efficacy of two different forms of administration of lidocaine for topical anaesthesia in cataract surgery.     

流式细胞术检测HLA-B27及其临床意义(附112例分析)

目的 通过流式细胞术检测外周血白细胞HLA—B27的表达，并探讨其对强直性脊柱炎（AS）的诊断价值。方法 利用流式细胞仪（FCM）检测33例AS患者、34例背痛及腰腿痛患者及45例健康体检者的外周血HLA—B27表达。结果 33例AS患者中，HLA—B27阳性30例（90．9％）；34例背痛腰腿痛患者中，HLA—B27阳性4例（11．76％）；45例健康人群中，HLA—B27阳性4例（8．9％）。结论 AS患者HLA—B27阳性率明显增高，HLA—B27检测可视为临床支持AS诊断的重要依据。     

HLA-B27/B7抗原检测在诊断强直性脊柱炎中的临床价值

目的探讨HLA-B27/B7抗原检测在诊断强直性脊柱炎（AS）中的临床价值。方法选取144例门诊及住院高度疑似AS的患者,应用流式细胞仪对其标本进行HLA-B27/B7抗原检测。结果HLA-B27抗原阳性率在青少年人群比较高,且男性显著高于女性,受检的144例患者,HLA-B27抗原阳性率为63.9%,低于确诊为AS的阳性率（91.1%）。HLA-B27抗原检测对诊断AS的灵敏度为91.1%（82/90）,特异度为81.5%（44/54）,正确率为87.5%（126/144）。结论 HLA-B27抗原与AS密切相关,对临床诊断AS很有帮助,用流式细胞术进行检测是较好的方法。     

阿托伐他汀对急性冠脉综合征患者白细胞介素6和肿瘤坏死因子的影响

目的 探讨阿托伐他汀对急性冠脉综合征（ACS）患者血清白细胞介素6（IL-6）及肿瘤坏死因子α（TNF-α）水平变化的影响。方法将ACS患者48例随机分为阿托伐他汀组25例和常规治疗组23例,分别于治疗前及治疗4周后采用放射免疫分析方法行血清IL-6及TNF—α检测。结果ACS患者治疗前血清IL-6、TNF-α水平均明显高于对照组（均P〈0．01）。阿托伐他汀组治疗后血清IL-6（0．67±0．05）μg/L、TNF-μ（65．1±13．2）μg／L水平均明显低于治疗前,但仍高于对照组（均P〈0．05）。常规治疗组患者治疗后血清IL-6（0．74±0．08）μg／L、TNF—±（73．6±12．0）μg／L降低不明显（P〉0．05）。结论IL-6、TNF-α水平升高与ACS发病密切相关,阿托伐他汀可降低ACS患者血IL-6、TNF—α水平,具有减轻病变部位炎性反应和保护内皮细胞的作用。     

不确定风湿病患者人类白细胞抗原B27测定

目的调查临床诊断不确定风湿性疾病患者HLA—B27抗原的分布情况，探讨其在风湿性疾病中诊断及鉴别诊断的意义。方法采用标准NIH微量淋巴细胞毒实验方法对747例临床诊断不确定风湿性疾病患者做HLA—B27抗原检测。结果747例中，HLA—B27阳性196例，总阳性率26．24％。HLA—B27阳性者中16岁以下20例(男16例、女4例)，占10．21％；16～40岁162例(男126例，女36例)，占82．65％。大于40岁14例(男9例、女5例)，占7．14％。结论HLA-B27血清学检查对临床诊断不确定风湿性疾病的鉴别诊断具有重要意义．HLA—B27阳性提高了发现AS的可能性。对于诊断不确定的风湿病患者，HLA-B27可作为筛选指标。     

血清CD27和CD27L的检测在胃癌中的临床意义

目的探讨胃癌患者血清CD27和CD27L的表达与胃癌及病理指标的关系。方法应用ELISA方法检测42例胃癌患者血清标本中CD27和CD27L的表达,与40例同期正常对照者的血清标本作对比分析。同时采用单因素方差分析和Spearman相关分析探寻CD27和CD27L的表达浓度与胃癌病理类型、病理分期及TNM分期的关系。结果胃癌患者血清CD27和CD27L的表达显著低于正常对照者(P<0.05)。在胃癌患者中,除血清CD27L的表达在病理分期Ⅱ期和Ⅲ期之间(P=0.017)有显著差异以及血清CD27的表达在N1和N2之间(P=0.022)有显著差异外,其余指标在不同病理类型、分期及TNM分期之间无显著差异。Spearman相关分析显示,CD27和CD27L的表达与病理类型、分期及TNM分期无显著相关性。结论血清CD27和CD27L表达与胃癌的发病密切相关,可作为协同判断胃癌发病的指标。     

慢性酒精中毒血清肿瘤坏死因子α的检测及其临床意义

~~慢性酒精中毒血清肿瘤坏死因子α的检测及其临床意义$遵义医学院第一附属医院神经内科!563003@徐平$遵义医学院第一附属医院神经内科!563003@雷显泽$遵义医学院第一附属医院神经内科!563003@胡泳涛$重庆医科大学第一附属医院神经内科!400016@邹德智~~~~~~~~     

HLA-B27检测在强直性脊柱炎诊断中的临床价值

目的 探讨HLA-B27抗原检测在强直性脊柱炎（AS）早期诊断中的临床价值。方法使用ELISA法对450例健康体检者（非AS对照组）、462例AS疑似患者（AS疑似组）和80名AS患者（AS组）进行HLA-B27抗原检测。结果非AS对照组、AS疑似组和As组的HLA—B27阳性率分别为3．3％、48．9％和92．5％,AS组、AS疑似组的HLA—B27阳性率明显高于非AS对照组（P〈0．01）,AS组的HLA—B27阳性率明显高于AS疑似组（P〈0．01）。结论HLA．B27抗原与AS疾病相关,检测HLA—B27抗原对疑似AS患者的早期诊断有重要意义。     

心力衰竭患者血浆中肿瘤坏死因子及一氧化氮水平的变化

目的探讨肿瘤坏死因子-α(TNF-α)、可溶性肿瘤坏死因子受体(sTNFR)及血浆一氧化氮(NO)在充血性心力衰竭(CHF)发生和发展中的作用与相互关系.方法用断面调查的方法,采用双抗体夹心法测定了68例CHF患者血浆中TNF-α和sTNFR水平,用高效液相法测定了血浆NO的变化,并与30例正常人进行对照.结果检测结果表明,不同程度的CHF患者血清中上述指标均发生明显的改变,与对照组比较存在显著差异(P《0.01),TNF-α,sTNFR和NO三者之间存在明显正相关,并随心力衰竭程度的加重而增加.结论TNF-α及NO水平的变化是影响CHF发生和发展的重要因素.     

Evaluation of tumor necrosis factor-alpha and granulocyte colony-stimulating factor serum levels in lead-exposed smoker workers

Inorganic lead (Pb) is able to modulate the immune response even at low to moderate exposure levels. It inhibits in vitro and in vivo activities of neutrophil leucocytes and influences their blood count in humans. Neutrophil functions are governed by a number of cytokines. Pb has been shown to affect leukocyte production of some of these cytokines in vitro. The objective of this study is to assess serum tumor necrosis factor-alpha (TNF-alpha) and granulocyte colony-stimulating factor (G-CSF) levels of thirty-three male lead-exposed (E) workers at a lead recycling plant as compared with twenty-eight male non-exposed (NE) workers at a food processing plant, whose current smoking habit was known. Serum TNF-alpha and G-CSF levels were measured by a quantitative sandwich enzyme immunoassay. Blood lead levels (Pb-B) were significantly higher in E (geometric mean (GM) 30.7 microg/dl, GSD 1.7; min-max: 9.1-81.6 microg/dl) workers than controls (GM 3.6 microg/dl, GSD 1.7; min-max: 1.0-11.0 microg/dl). E workers had significantly higher serum TNF-alpha (median: 107.1; min-max: 11.1-623.0 pg/ml) and G-CSF levels (median: 53.0, min-max: 31.1-197.0 pg/ml) than NE workers (TNF-alpha: median: 12.0; min-max: 9.4-18.8 pg/ml; G-CSF: median: 34.3, min-max: 25.1-52.2 pg/ml). In particular, the TNF-alpha level was shown to be significantly influenced by lead exposure and smoking habit, as well as by interaction between these two factors. Both serum TNF-alpha and G-CSF levels were correlated with Pb-B and absolute neutrophil count. This study is the first to detect higher serum levels of G-CSF in E over NE workers. Our data confirm that exposure to low to medium doses of lead may interfere in the complex cytokine network involved in inflammation, especially in workers who are current smokers.     

心力衰竭患者血浆中肿瘤坏死因子及一氧化氮水平的变化

目的 探讨肿瘤坏死因子-α（TNF-α）、可溶性肿瘤坏死因子受体（sTNFR）及血浆一氧化氮（NO）在充血性心力衰竭（CHF）发生和发展中的作用与相互关系。方法 用断面调查的方法,采用双抗体夹心法测定了68例CHF患者血浆中TNF-α和sTNFR水平,用高效液相法测定了血浆NO的变化,并与30例正常人进行对照。结果 检测结果表明,不同程度的 CHF患者血清中上述指标均发生明显的改变,与对照组比较存在显著差异（P<0.01）,TNF-α,sTNFR和NO三者之间存在明显正相关,并随心力衰竭程度的加重而增加。结论TNF-α及NO水平的变化是影响CHF发生和发展的重要因素。     

Effects of phycocyanin extract on tumor necrosis factor-alpha and nitrite levels in serum of mice treated with endotoxin

Phycocyanin is a biliprotein which exerts antioxidative and anti-inflammatory effects in various in vivo and in vitro experimental models. In this study phycocyanin effects on tumor necrosis factor-alpha (TNF alpha) and nitrite levels in serum of mice treated with lipopolysaccharide (LPS) was examined. TNF alpha was measured by cytotoxicity on L-929 cells and nitrite by the Griess reaction, after reduction of all nitrates to nitrites by nitrate reductase, 1 h after LPS injection (0.5 mg/kg i.p.) there was a significant increase in TNF alpha levels in mouse serum. Phycocyanin (50-300 mg/kg p.o.), administered 1 h before LPS, reduced dose-dependently the TNF alpha concentration in serum. After 18 h, LPS (30 mg/kg i.p.) also induced a substantial increase in serum nitrite levels, which were reduced dose-dependently by phycocyanin pretreatment (100-300 mg/kg p.o.). The results indicate that phycocyanin exerts inhibitory effects on TNF alpha and NO production which might be ascribed to the antioxidative properties of the biliprotein.     

帕金森病患者外周血肿瘤坏死因子水平变化

目的通过检测帕金森病（PD）患者外周血中肿瘤坏死因子-α（tumor necrosis factor-alpha,TNF-α）,探讨TNF-α在PD患者外周血中的变化。方法采用放射免疫分析方法,对84例（PD组）患者和87例（对照组）健康对照的血TNF-α进行检测;同时对PD患者的年龄、性别、疾病分期和病程进行组内比较。结果PD患者与健康对照者TNF-α含量分别为（14.7±2.3）pmol/L、（16.9±4.4）pmol/L,差异有统计学意义（P〈0.01）;按患者年龄、性别、疾病分期和病程进行组内比较差异无统计学意义（P〉0.05）。结论PD发病过程中存在着免疫系统失调,PD患者外周血TNF-α减少,而且与年龄、性别、疾病分期和病程无关。     

手足口病患儿血清肿瘤坏死因子-α、白介素6和白介素10水平的变化

目的观察手足口病(HFMD)患儿血清肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、白介素-10(IL-10)的变化。方法采用酶联免疫吸附实验(ELISA)抗体夹心法测定15例HFMD患儿(急性期、恢复期)及同期10例健康体检儿童血清TNF-α、IL-6和IL-10的含量,用SPSS软件对所得数值进行统计学分析。结果 HFMD患儿急性期TNF-α、IL-6和IL-10含量均显著高于健康对照组(P<0.05)。恢复期IL-6含量与对照组比较差异无统计学意义(P>0.05),而TNF-α和IL-10水平仍高于健康对照组(P<0.05)。结论 HFMD患儿急性期炎症细胞因子(TNF-α、IL-6、IL-10)水平均显著升高,而抑制性细胞因子IL-10水平和TNF-α水平升高持续到恢复期。