腺苷脱氨酶在人睾丸及附睾的分布

应用本实验室前期制备的特异性的抗人精子膜结合腺苷脱氨酶（ＡＤＡ）同功酶的单克隆抗体（ＭｃＡｂ）和免疫组化技术，我们分析了该同功酶在正常人睾丸和附睾中的分布。实验结果显示：（１）ＡＤＡ在正常人睾丸中无分布，而在附睾头部，附睾管腔上皮细胞中开始分布；（２）ＡＤＡ在正常人附睾管腔上皮细胞中的分布，由附睾头部至尾部，其密度递增。这一研究结果提示：（１）ＡＤＡ分布具有附睾特异性；（２）人精子膜结合ＡＤＡ可能来自附睾管腔上皮细胞。     

腺苷脱氨酶在大鼠睾丸,附睾中的分布

利用本实验室前期制备和鉴定了的抗人精子膜结合脱氨酶单克隆抗体和ＡＢＣ组分技术，我们分析了正常性成熟和性未成熟大鼠睾丸及附睾中的该酶分布情况。实验结果显示：（１）ＡＤＡ在正常性成熟和性未成熟大鼠睾丸和附睾中无分布；在性成熟大鼠中，ＡＤＡ的分布从际睾头部开始出现；（２）ＡＤＡ在正常性成熟大鼠附睾中的分布由附睾头部至尾部其密度未发现变化。     

慢性乙型肝炎患者血清腺苷脱氨酶与肝脏病理的关系

腺苷脱氨酶（ADA）是嘌呤核苷循环中的一种重要的酶,在核酸代谢中有重要的意义。血清中90％以上的ADA来源于肝脏,属于肝细胞的胞浆酶。当肝细胞受损、坏死或细胞膜通透性增加时,血清ADA活性增高。本研究旨在探讨血清ADA与肝脏病理的关系。     

脑脊髓液中腺苷脱氨酶活性对结核性脑膜炎诊断价值的临床研究

目的 探讨脑脊髓液中腺苷脱氨酶（CSF-ADA）活性对结核性脑膜炎的诊断价值及在病程中的动态变化.方法 选择160例患者纳入本研究,76例结核性脑膜炎为病例组;84例非结核性脑膜炎为对照组,其中细菌性脑膜炎36例,病毒性脑膜炎30例,隐球菌性脑膜炎18例.每例患者均于治疗前抽取CSF,采用酶耦联Trinder法测定CSF-ADA活性,结果以（-x）±s表示,组间差异使用Mann-Whitney U检验.47例结核性脑膜炎患者于抗结核治疗后2周和6周时再次检测CSF-ADA,治疗前后差异使用配对t检验.结果 结核性脑膜炎组CSF-ADA活性为（12.9±6.4）U/L,非结核性脑膜炎组为（6.0±4.1）U/L,两组比较差异有统计学意义（U=7.860,P＜0.05）.取CSF-ADA≥9 U/L作为临界值时鉴别结核性脑膜炎与非结核性脑膜炎价值最高,灵敏度为84.21%,特异度为83.33%.随着患者病情好转,CSF-ADA活性逐渐降低.结论 CSF-ADA活性≥9 U/L可作为诊断结核性脑膜炎的一项辅助诊断指标,抗结核治疗后CSF-ADA活性可作为疗效判断的参考指标.     

腺苷脱氨酶在人精子膜上的定位及其单克隆抗体对人精子穿卵能力的影响

利用本实验室制备的抗人精子膜结合腺苷脱氨酶（ＡＤＡ）单克隆抗体（ＭｃＡｂ）和间接荧光免疫技术，分析了该酶在正常人精子膜上的位置。同时，利用该酶单抗分析了该酶活性对人精子穿透去透明带卵的影响。上述两个实验的结果显示：（１）ＡＤＡ结合于人精子中段、尾部膜的外侧；（２）利用抗ＡＤＡＭｃＡｂ抑制ＡＤＡ活性可明显抑制正常人精子穿透去透明带卵的能力。这一研究结果提示，ＡＤＡ作为精子的膜蛋白组分，可能参与对精子生理功能的调节过程。     

Alteration of adenosine deaminase levels in peripheral blood lymphocytes of patients with gastric cancer

The present study was undertaken to determine the adenosine deaminase (ADA) activity of peripheral lymphocytes in patients with gastric cancer, with respect to the cancer progression, the effect of surgery and/or immunotherapy. The gastric cancer patients showed lower lymphocyte ADA activity than did the normal control. The lymphocyte ADA activity did not decrease with the cancer progression. There was a significant correlation between lymphocyte ADA activity and blastogenesis of lymphocyte by phytohemaglutinin or concanavalin A. Six months following gastrectomy, the lymphocyte ADA activity was increased, as compared with the preoperative value. The ADA activity of patients on post-operative OK-432 showed a greater increase, as compared to that of patients not given this treatment. In conclusion, decreased lymphocyte ADA activity in gastric cancer patients might be due to either the cancer bearing status or to the immunological suppression.     

RNA特异性腺苷脱氨酶1与Caveolin 1在脓毒症相关肝损伤中的作用及机制

背景与目的 脓毒症相关肝损伤（SRLI）发病机制尚不清楚,细菌内毒素（LPS）对肝脏血管内皮细胞的炎症损害可能是重要环节。前期研究提示,RNA特异性腺苷脱氨酶1（ADAR1）可能通过调控内皮细胞功能相关蛋白Caveolin 1（Cav-1）在参与血管内皮应激中的局部和全身炎症反应。因此,本研究初步探讨ADAR1与Cav-1在SRLI中的作用,以期为SRLI的早期防治寻找新的方法。方法 取ADARl基因敲除小鼠（ADAR1^ECKO）与野生型小鼠（ADAR1^flox/flox）各20只,腹腔注射LPS（20 mg/kg）诱导脓毒症小鼠模型脓毒症模型,6 h后每组小鼠各取10只,获取肝脏组织,并分离肝窦内皮细胞（LSECs）,通过HE染色观察其肝脏病理学改变,用细胞免疫荧光法观察LSECs中Cav-1及其下游蛋白VE-cadherin的表达,两组其余小鼠用于生存率分析;用ADARl siRNA转染正常野生型小鼠的LSECs后,通过内皮细胞成管实验观察转染后LSECs的增殖情况、Western blot检测Cav-1下游相关蛋白的表达。结果 生存观察结果显示,注射LPS后,ADAR1^ECKO小鼠死亡时间早于ADAR1^flox/flox小鼠,存活率低于ADAR1^flox/flox小鼠（均P<0.05）;组织病理学观察显示,注射LPS 6 h后,ADAR1^ECKO小鼠的肝损伤比ADAR1^flox/flox小鼠更严重;细胞免疫荧光观察显示,注射LPS 6 h后,ADAR1^ECKO小鼠LSECs中Cav-1与VE-cadherin的表达低于ADAR1^flox/flox小鼠。正常野生型小鼠的LSECs转染ADARl siRNA后,成管能力明显减弱,Cav-1下游蛋白VE-cadherin的表达下调,但β-Catenin的表达无明显变化。结论 ADAR1的下调或功能缺失会导致SRLI加重,机制可能涉及其调控Cav-1/VE-cadherin通路的活性。因此,激活ADAR1/Cav-1/VE-cadherin通路可能是防治SRLI的有效策略。     

Urinary excretion of liver fatty acid-binding protein in health-check participants

Background Messenger RNA of liver fatty acid-binding protein (L-FABP) is expressed in proximal tubules of the kidney, and a certain amount is excreted into urine. We analyzed factors relating to the urinary L-FABP excretion in health-check participants.Methods We measured L-FABP in the first morning urine by ELISA in 715 men and 193 women 30–79 years of age who entered a 2-day hospitalized health checkup program. In addition to the routine physical examination and laboratory tests, plasma high-sensitivity C-reactive protein (HSCRP) was assayed.Results In 150 healthy subjects, urinary L-FABP averaged 3.6 ± 0.2µg/g creatinine, whereas the values were significantly increased in patients with hypertension (5.2 ± 0.4, P = 0.010), diabetes mellitus (5.5 ± 0.5, P < 0.001), and chronic hepatitis (5.8 ± 1.0, P = 0.022). Urinary L-FABP excretion was significantly greater in women than in men when the value was related to creatinine. In regression analysis in men, urinary L-FABP was positively correlated with fasting plasma glucose (r = 0.103, P = 0.033) and plasma HSCRP (r = 0.135, P = 0.006).Conclusions It is suggested that renal production and urinary excretion of L-FABP are increased in situations in which arteriosclerosis is promoted, such as hypertension, diabetes mellitus, and cardiovascular inflammation.     

Evaluation of antibiotic-induced nephrotoxicity in preterm neonates by determining urinary α1-microglobulin

α₁-Microglobulin (α₁-m, protein HC), a relatively low molecular weight protein of about 31,000 daltons, was measured in urine of three groups of 34 preterm neonates: group A consisted of 9 healthy preterm neonates; groups B (n = 13) and C (n = 12) consisted of preterm neonates with suspected or confirmed bacterial infections. Immediately after birth, all group B neonates were treated with ampicillin and aztreonam in combination, and all group C neonates were treated with oxacillin and amikacin in combination. To optimize amikacin administration, computerized individually tailored doses were administered. Urine samples were obtained from a short collection in sterile bags on the 1st, 4th, and 7th day after delivery in all infants. Urinary α₁-m concentrations were measured by a turbidimetric method (latex agglutination photometric immunoassay) and results were expressed as a ratio to urinary creatinine. In group A, urinary α₁-m concentrations were stable after birth. In group C, α₁-m excretion increased immediately within the 1st day of treatment, and over the 1st week of life urinary α₁-m levels were significantly higher than in group A (P = 0.033). These data support the conclusion that amikacin administration was the most important factor inducing renal tubular dysfunction in the neonates of group C. Received March 6, 1995; received in revised form and accepted January 19, 1996     

RNA结合基序蛋白6-RNA结合基序蛋白5融合基因在乳腺组织中的表达

目的 观察RNA结合基序蛋白6-RNA结合基序蛋白5(RBM6-RBM5)融合基因的存在及其在乳腺肿瘤细胞中的表达.方法 采用巢式聚合酶链反应(PCR)法对RBM6-RBM5融合基因进行检测,分析RBM6-RBM5融合基因在肿瘤细胞及正常组织中的表达.结果 4例乳腺肿瘤组织RBM6-RBM5融合基因的表达为阳性,相对表达量分别为:0.52±0.02、0.61 ±0.04、0.58 ±0.05、0.65±0.03;2例乳腺肿瘤组织RBM6-RBM5融合基因的表达为阴性,相对表达量为0;6例正常乳腺组织RBM6-RBM5融合基因的表达为阴性,相对表达量为0.结论 RBM6-RBM5融合基因存在于乳腺肿瘤组织中,RBM6-RBM5融合基因的表达与乳腺肿瘤的发生明显相关.同时,该融合基因在不同乳腺癌患者提供的乳腺癌肿瘤组织中的表达结果也不尽相同.     

环腺苷酸效应元件结合蛋白在学习记忆中的作用及机制

背景 环腺苷酸效应元件结合蛋白(cAMP response-element binding protein,CREB)是新发现的学习记忆正调控因子.近年来的研究提示CREB可能通过多种途径调控学习记忆. 目的 阐述CREB在学习记忆中的作用及机制,为认知障碍相关疾病的防治提供思路. 内容 阐述CREB的来源、结构及其在学习记忆中的作用.分析其调控学习记忆的相关机制. 趋向 CREB的确切上游调控机制及自身的表达调控需要进一步明确,对其深入研究可为多种认知障碍疾病的防治提供新的靶点与思路.     

Estramustine binding protein in human benign prostatic hypertrophy

Using a highly sensitive radioimmunoassay (RIA) of estramustine binding protein (EMBP) with the antibody raised against rat-EMBP, the concentration of EMBP in human benign prostatic hypertrophy (BPH) tissue was evaluated. Using mechanical separation procedure, EMBP was found significantly higher in the epithelium (10.7 ng/mg protein) than in the stroma (2.9 ng/mg protein). When human BPH was classified into glandular, fibromuscular and mixed type, the concentration of EMBP was the highest in glandular type (15.7 ng/mg protein) and the lowest in fibromuscular type (10.8 ng/mg protein).     

Open follow-up study of tobramycin nebuliser solution and colistin in patients with cystic fibrosis

A previous study of tobramycin nebuliser solution (TNS) compared with colistin [C] in cystic fibrosis (CF) patients, chronically infected with pseudomonas, showed benefit for the TNS treated patients over one treatment cycle only. The current report is of an extension of that study. An open randomised cross-over study of TNS compared with C was conducted on 21 patients who had previously been on the 1 cycle study. They continued for a further 5 months and then crossed over to the alternate treatment. There was an advantage for TNS over C in FEV(1) % predicted change over time. The C slope was -0.88% per month and the TNS slope 0.35% per month (p=0.0002). This suggests advantages of TNS over C in a study with a small number of patients. Larger studies are required.     

Diagnosis of Renal Proximal Tubular Injury by Urinary Immunoassay for a Proximal Tubular Antigen, the Adenosine Deaminase Binding Protein

A sandwich ELISA assay has been formatted from two commerciallyavailable murine monoclonal antibodies, URO-4 and URO-4a, directedagainst a 120 000 dalton glycoprotein, the adenosine deaminasebinding protein (ABP), found on the brush border of the renalproximal tubular epithial cell. Untimed urine samples from 37normal individuals and urinary ABP <0.1 AU; 37 patients withpure glomerular disease had ABP <0.4 AU (with 29, or 76%<0.2 AU); 10 patients with pre-renal azotaemia had ABP <0.6(with 8, or 80% <0.3 AU). In contrast, 79 patientswith post-ischaemicacute tubular necrosis had ABP >0.6 AU. Acute renal failuredue to myoglobinuria, contrast dye, and aminogly coside toxicitywere all associated with urinary ABP >1.0 AU. In addition,all six patients with acute bacteraemic pyelonephritis had ABP>0.7 AU, as opposed to ABP <0.2 AU in the urines of 12women with acute cystitis. We conclude that this monoclonalantibody based urinary assay is a sensitive measure of renalproximal tubular injury, reliably distinguishes acute tubularfrom glbmerular disease, and may be helpful in differentiatingforms of urinary tract infection.     

Serum protein binding of propofol in critically ill patients

Background : Disease-induced modifications in the level of serum proteins may change the degree of binding of drugs highly bound to serum proteins. Methods : The serum protein binding of propofol, an intravenous anaesthetic agent, was studied in vitro in samples of serum from healthy volunteers (n = 28) and from critically ill patients (n = 17). The free fraction was obtained by the ultrafiltration technique and was measured by high-performance liquid chromatography. Concentrations of serum albumin, α₁acid-glycoprotein and free fatty acids were also measured in all samples. Results : The percentage of free propofol was significantly increased (P < 0.001) in critically ill patients (1.31 (1.06–2.25)%) vs control subjects (1.07 (0.49–1.47)%). Albumin levels were significantly decreased (P<0.001) in patients (16.3 (8.8–24.6) g–1^-1 vs 45.8 (31.4–55.5) g–1“¹), while levels of α₁-acid-glycoprotein were increased (P<0.001) (1.9 (0.9–2.8) g l^-1 vs 0.9 (0.5–1.4) g 1^-1), as were levels of free fatty acids (0.68 (0.50–1.14) mmol I^-1 vs 0.37 (0.11–1.05) mmol I^-1; P < 0.05). No correlation was found between levels of α^1–acid–glycoprotein or free fatty acids and the bound/free ratio of propofol. However, a linear relationship was found between levels of albumin and the bound/free ratio (r²=O.25; P < 0.001). Conclusion : In conclusion, in these critically ill patients, an increase in the percentage of free propofol occurs. The significance of this observation remains uncertain, but may be validated in future studies. However, the observation supports the common idea that potent drugs should be given with great care in critically ill patients.     

Determination of tobramycin in saliva is not suitable for therapeutic drug monitoring of patients with cystic fibrosis

BACKGROUND: Tobramycin, used in the treatment of infections caused by Gram-negative bacteria, requires therapeutic drug monitoring (TDM) due to its narrow therapeutic index. The collection of blood for these assays may cause pain and trauma to the child and/or be difficult because of limited access to appropriate blood vessels. We undertook an evaluation of the role of saliva concentrations in the TDM of once-daily tobramycin therapy in patients with cystic fibrosis. METHODS: Fourteen patients (mean age 15 years) with cystic fibrosis were enrolled at Women's and Children's Hospital, Adelaide (WCH). All patients received once-daily dose of intravenous tobramycin for 2-3 weeks and had plasma levels measured once a week. At the same time of blood sampling at 1 and 6 h after initiation of tobramycin infusion, the patients also provided saliva samples. For collection of saliva, the Salivette (Sarstedt Laboratories) system was used which was developed specifically for saliva sampling. Concentrations in blood and saliva were measured by the Beckman Synchron CX system, which is utilized for routine assays of plasma tobramycin. RESULTS AND CONCLUSION: Tobramycin could not be detected in saliva within the first 6 h after a once-daily dosing. Therefore, plasma cannot be substituted with saliva for the TDM of tobramycin using the clinical routines at WCH.     

Serum protein binding of propofol in patients with renal failure or hepatic cirrhosis

Background: Serum protein binding is a limiting factor in the access of drugs to the central nervous system. Disease-induced modifications of the degree of binding may influence the effect of anesthetic drugs. Methods: The protein binding of propofol, an intravenous anaesthetic agent which is highly bound to serum albumin, has been investigated in serum samples from healthy volunteers, from patients with chronic renal failure not undergoing hemodialysis, from patients with chronic renal failure included in a regular hemodialysis program, and from patients with hepatic cirrhosis. Protein binding was determined by the ultrafiltration technique using an Amicon Micropartition System, MPS-1. Results: The percentage of unbound propofol (mean(SD)) in healthy volunteers (n=16) was 0.98 (0.48) % showing a high interindividual variability. Chronic renal failure did not significantly modify serum protein binding of propofol. In the chronic renal failure group not undergoing regular hemodialysis (n=>9), unbound propofol was 0.92 (0.34) %. In addition, patients in periodic dialysis did not show changes in propofol binding either compared before (1.11 (0.33) %; n=13) or after hemodialysis (0.87 (0.38) %; n=12). A slight decrease in albumin concentration was found in all renal patients (P<0.05) in comparison to healthy volunteers. Creatinine and urea concentrations were higher in these patients (P<0.01) but in the postdialysis group, the differences in urea levels were not significant when compared with those of volunteers. No changes in the degree of propofol binding were observed in patients with hepatic cirrhosis (0.97 (0.30) %; n=14) when compared with the group of healthy volunteers. Significant differences were observed in albumin (P<0.01) and bilirubin (P<0.05) concentrations. Considering all subjects, the degree of binding did not correlate with biomedical data. Conclusion: Due to the the absence of significant changes in the protein binding it is unlikely that there will be an exaggerated pharmacological response in patients with renal and hepatic disease following the administration of a standard propofol dose, although due to interpatient variability careful titration can be recommended.     

Partial characterization of a proacrosin binding protein.

All of the acid (pH 4.0) extracted proacrosin from porcine epididymal spermatozoa was found to be tightly associated with a specific protein referred to as the binding protein. A combination of gel filterations and gel electrophoresis revealed that the binding protein is composed of a major 28 kd and a minor 29 kd protein. Both of the proteins were shown to be nonproteolytic by gelatin SDS-PAGE analysis and the amino acid composition analysis of the purified 28 kd protein revealed that it is not related to the proteolytic component of the proacrosinacrosin system.     

Ovarian cancer-associated lymphocyte recognition of folate binding protein peptides

Background: Tumor-associated lymphocytes (TAL) isolated from ovarian cancer patients contain cytotoxic T lymphocytes (CTL) capable of recognizing specific HLA/peptide complexes on tumor cells leading to tumor cell lysis. Currently, HER2/neu, overexpressed in only 30% of breast and ovarian cancers, is the only known source of CTL-recognized peptides in epithelial cancers. Therefore, we have investigated peptides derived from folate binding protein (FBP), which is over-expressed in more than 90% of ovarian cancers and in the majority of other epithelial tumors. Methods: TAL were isolated from the malignant ascites of four consecutive HLA-A2⁺ ovarian cancer patients and incubated in IL-2. Initial chromium-release assays were performed within 1 week. T2 cells, incubated with peptide, were used to reconstitute T cell epitopes. The FBP sequence was interrogated for HLA-A2 binding peptides, and five were synthesized (E37–41). Results: Freshly cultured, unstimulated ovarian TAL recognize peptides derived from FBP. These peptides are presented in the context of HLA-A2, and are specifically recognized in a HLA class I-restricted fashion. TAL recognition of these reconstituted T cell epitopes is concentration dependent. Furthermore, the FBP peptides are shown by cold target inhibition studies to be naturally processed and presented antigens. Conclusions: FBP peptides are recognized by freshly isolated TAL from ovarian cancer patients, suggesting in vivo expression and sensitization. Because FBP is over-expressed 20-fold in most adenocarcinomas, these peptides may be used in a widely applicable peptide-based vaccine for epithelial tumors.Presented at the 51st Annual Cancer Symposium of The Society of Surgical Oncology, San Diego, California, March 26–29, 1998.