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。     

Serum adenosine deaminase activity in brain tumours

Summary Adenosine deaminase (ADA) activity in serum was estimated in 86 patients with intracranial tumours and 40 healthy volunteers. Although high ADA concentrations in biological fluids and tumour tissues were observed in several neoplastic conditions, there was no significant difference in the ADA in sera of brain tumour patients when compared to the control values. Therefore, cell-mediated immunity probably does not play a significant role in brain tumours.     

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。     

Sib-pair analysis of adenosine deaminase locus in NIDDM.

Recently, linkage between the ADA gene locus and MODY, a subtype of NIDDM, has been reported. The possibility that the region of chromosome 20q containing the ADA locus also may play a role in susceptibility to NIDDM needs to be investigated. Therefore, we examined the linkage between the ADA locus and NIDDM in affected siblings of 50 European white diabetic pedigrees--21 Italian and 29 British. Departure from independent segregation of the disease and an Alu VpA polymorphism within the 5' flanking region of the ADA locus was tested in the affected sib-pairs with the APM statistical method. After DNA amplification by the PCR and PAGE, five alleles were identified in the ALU VpA tract at the ADA locus in the two populations. Allele frequencies did not differ significantly between the two populations (chi 2 = 2.426, P > 0.05 [NS]). Analysis of the 50 diabetic sib sets, and independently of the Italian and British groups of affected sib pairs, revealed no segregation distortion between the marker locus and NIDDM. We conclude that mutations within or around the ADA locus are unlikely to play a major role in the etiology of NIDDM.     

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。     

Diagnostic value of adenosine deaminase activity in pericardial fluids.

BACKGROUND: The activity of adenosine deaminase (ADA) was determined in serum and pericardial fluid of 70 patients (ages 21 to 71 years) with pericardial effusions of various etiologies and in 15 control subjects. METHODS: The patients were subdivided into five groups on the basis of definite diagnosis: 1) 24 patients with tuberculosis; 2) 22 with malignancies; 3) 12 with uremic pericarditis; 4) 12 with purulent pericarditis; 5) 15 control individuals without pericardial disease. The activity of ADA was determined at the same time in serum and cell-free pericardial fluid according to the method of Karker with minor modification. RESULTS: Mean (+/-SD) ADA activity in pericardial fluid was 66.92+/-4.12 IU/L in group 1; 27.50+/-6.02 in group 2; 28.65+/-4.73 in group 3; 53.05+/-11.14 in group 4; and 5.67+/-1.99 in group 5. Comparing the level achieved in group 1 with all others, the difference is significant at the p<0.001 level. When the cut-off value of 50 IU/L is used the sensitivity of the test for diagnosis of tuberculous effusion is 1, and the specificity is 0.83. Statistical analysis showed that there was no correlation between serum ADA activity and ADA activity in pericardial fluid. CONCLUSIONS: We recommend that determinations of ADA activity in pathologic pericardial fluids seem to be of great value in the early diagnosis of tuberculous pericardial effusions. Levels above 50 IU/L in effusions indicate probable tuberculosis.     

Lymphocyte adenosine deaminase activity in children with idiopathic nephrotic syndrome

Adenosine deaminase (ADA) activity, as a marker of cell-mediated immunity, was evaluated in the serum (S-ADA) and lymphocytes (L-ADA) of 47 children with idiopathic nephrotic syndrome, and 23 healthy controls. The mean S-ADA and L-ADA levels were significantly raised in active nephrotic syndrome (ANS) and in its sub-groups in comparison with controls. The ADA activity was significantly more elevated in relapsers than for the first attack of nephrotic patients, and the frequent relapsers had the highest enzymatic levels both in serum as well as lymphocytes. A significant positive correlation was found between serum and lymphocyte ADA levels (r =0.736, p <0.01). In remission, the S-ADA showed a significant fall in comparison with their corresponding ANS value (p <0.001) and reached the level of controls. The mean L-ADA also showed reduction but the difference was statistically insignificant and the value was significantly raised, when compared with controls. The enzyme activity in serum and lymphocytes normalized in the long-term remission group. Thus, ADA activity was abnormal in ANS cases, and L-ADA demonstrated change both in active as well as remission stage of the disease.     

Malondialdehyde level and adenosine deaminase activity in nasal polyps.

OBJECTIVE: Although there are many reports on adenosine deaminase (ADA) activities in different tissues, no information is available about the enzyme activity in nasal mucosa and polyp tissues. Whereas ADA is related to the production of free radicals by neutrophils, malondialdehyde (MDA) is an indicator of lipid peroxidation that is a general mechanism of tissue damage by free radicals. This study is aimed at determining and comparing the ADA activity and MDA level in nasal polyps and normal mucosa. STUDY DESIGN AND SETTING: Twenty-three patients with nasal polyps and a control group consisting of 14 patients with septal deviation and lower turbinate hypertrophy were included in the study. Tissue MDA level was measured by the method of Okawa with modification and tissue ADA activity by the method of Giusti. RESULTS: In patients with nasal polyp, mean tissue MDA level and ADA activity were 2.43 +/- 0.38 nmol/mg protein (Pr) and 0.235 +/- 0.055 U/mg Pr, respectively, which were significantly higher than those of control nasal mucosa (1.03 +/- 0.41 nmol/mg protein and 0.056 +/- 0.011 U/mg Pr, respectively) (P < 0.05). In addition, tissue MDA level was positively correlated to ADA activity in nasal polyps (r = 0.701, P < 0.001). CONCLUSIONS: The present study showed the presence of detectable ADA activity in nasal mucosa, and also significant increases in both tissue MDA level and ADA activity in NP tissue when compared to normal turbinate tissue. EBM rating: B-2b.     

骨标志物检测在预测糖皮质激素诱发自身免疫疾病骨质疏松中的意义

目的自身免疫疾病患者长期应用糖皮质激素可以诱发骨质疏松,但是往往在应用早期尚无显著骨密度变化,为寻找在应用激素早期即可观察到的骨代谢变化,我们对98例自身免疫病患者进行了骨标志物检测,并研究其与激素应用的关系。方法对2011年4月1日至2013年1月31日于我院住院的98例自身免疫病患者进行骨标志物——血清骨钙素、I型原胶原N端前肽(PINP)β胶原降解产物(βCTX)的检测,采用电化学发光法检测。并对该98例患者的临床资料进行收集。应用SPSS13.5软件进行统计分析。结果 98例患者RA52例,SLE14例,SPA10例,其他自身免疫疾病22例。其中入院前已经接受糖皮质激素治疗患者39例,尚未接受激素治疗患者59例,两组患者的性别构成比及平均年龄均无统计学差异。激素治疗组患者血清骨钙素、βCTX和PINP均显著低于未接受激素治疗组,分别为[(7.57±4.46)μg/L vs(20.06±10.40)μg/L,P0.01]、[(0.31±0.22)μg/L vs.(0.52±0.26)μg/L,P0.01]和[(22.71±13.98)μg/Lvs.(49.86±35.46)μg/L,P0.01],有显著统计学差异。而两组间血钙、血磷、血尿素氮、血肌酐和C反应蛋白水平,以及股骨颈和腰椎骨密度值均无统计学差异。其中RA52例患者中,有应用糖皮质激素治疗组患者的骨钙素、βCTX和PINP亦均显著低于未接受激素治疗组,且βCTX和DAS28正相关(r=0.456,P0.01),提示βCTX和疾病活动度相关。结论糖皮质激素治疗可以导致自身免疫疾病患者骨代谢异常,而骨标志物的降低可以较早提示骨代谢的变化,对早期预防骨质疏松治疗有指导意义,而RA患者βCTX的水平升高可提示潜在疾病活动度升高。     

肾病综合征患者腺苷脱氨酶活性的变化及意义

通过检测对激素不同反应、不同病情的肾病综合征（NS）患者的血清及淋巴细胞内腺苷脱氨酶（ADA）的活性变化，观察ADA活性与临床疗效的关系。     

Mucormycosis in systemic autoimmune diseases

Mucormycosis is an emerging infection in systemic autoimmune diseases. All published cases of systemic autoimmune diseases complicated by mucormycosis were reviewed. The clinical features, diagnostic procedures and the main principles of treatment were analyzed. Twenty-four cases of mucormycosis have been reported in systemic auto-immune diseases, of which 83% in systemic lupus erythematosus, all occurring during immunosuppressants. In most cases, the infection was disseminated or rhinocerebral and it had mimicked a flare of the underlying connective tissue disease. A fatal outcome was reported in 58.3% of these patients. In conclusion, mucormycosis often mimics a flare of the underlying systemic disease and is associated with a high mortality rate. Systemic lupus erythematosus is by far the most common associated systemic autoimmune disease. A high degree of awareness is warranted to rapidly rule out infection, of which mucormycosis, in immunocompromised patients with systemic autoimmune disease before a disease flare is conclusively diagnosed.     

Aldolase and adenosine deaminase activity in lymphocytes of patients with glomerulonephritis

Studies on the adenosine deaminase (ADA) and aldolase activities in lymphocytes were performed in 67 patients with glomerulonephritis (gn) and in 20 healthy individuals from the control group to get an insight into the lymphocyte metabolism. Statistically significant decrease of ADA activity was found in the groups of patients with chronic proliferative gn, membranoproliferative gn, membranous gn and lupus nephritis in comparison with the healthy individuals from the control group. As far as decrease of aldolase activity is concerned it has reached statistical significance in patients with mesangial gn, membranoproliferative gn, membranous gn and lupus nephritis. The lymphocyte metabolism did not show any abnormalities in the enzymatic indicators only in patients with acute proliferative gn and submicroscopic gn. The activity comparison between both enzymes in the lymphocytes, contrasted on the basis of high and low clinical dynamics of gn, revealed a tendency to lower ADA and aldolase activities in patients with high clinical dynamics. However, this difference was at the limit of statistical significance (p less than or equal to 0.10).     

Pathogenetic significance of aberrant glycosylation of IgA1 in IgA nephropathy

IgA nephropathy (IgAN), the most common form of primary glomerulonephritis worldwide, is defined by predominant IgA1 deposits in the glomerular mesangium. Among abnormalities of the IgA immune system reported so far in IgAN, aberrant O-linked glycosylation in the hinge region of IgA1 is the most consistent finding. IgA1 molecules bearing abnormal glycosylation have been found in serum, in tonsillar lymphocytes, and in eluate from mesangial deposits, and characterized by decreased O-linked N-acetylgalactosamine residues with or without alteration in the terminal sialylation of the O-linked sugars. IgA1 with incomplete galactosylation has a tendency to accumulate in glomerular mesangium by self-aggregation or immune complex formation. Glomerular mesangial cells exposed to immune complexes of these IgA1 can proliferate and secrete cytokines, chemokines, growth factors, and extracellular matrix components promoting inflammatory reactions in the glomeruli. Although genes encoding enzymes involved in the O-glycosylation process, such as C1GALT1, have been reported to be responsible for susceptibility to IgAN, recent evidence suggests that the abnormality is restricted to a small fraction of B cell populations and arises from dysregulated IgA1 production and secretion in mucosal immune system. This review will focus on and discuss the role of incompleteness of IgA1 O-galactosylation in the pathogenesis of IgAN and propose a possible mechanism in which abnormal IgA1 occurs in IgAN. Presented at the 37th Eastern Regional Meeting of the Japanese Society of Nephrology.     

Evaluation of adenosine deaminase activity in patients with head and neck cancer

ADA is a key enzyme in the mammalian purine salvage pathway. The lack of ADA activity has been linked to a lack of cellular immunity in various immune and myeloproliferative disorders. Data on its role in patients with solid tumors are scant and inconclusive. In this report, we have evaluated the activity of this enzyme in the peripheral lymphocytes of patients with head and neck squamous cell cancer (HNC). The mean ADA activity in Stage IV patients (0.57 +/- 0.08 SEM, n = 12) was significantly lower than that of controls (1.55 +/- 0.25 SEM, P less than 0.05, n = 14) and also significantly lower that the mean ADA activity in patients with Stages I, II, and III (1.14 +/- 0.10 SEM, P less than 0.05, n = 17). Fourteen out of 19 controls, Stage I, II, and III patients had positive skin tests compared to 7 out of 10 Stage IV patients. These differences were not statistically significant. There was also no correlation between ADA activity and the absolute lymphocyte counts. Our results indicate that ADA activity in lymphocytes of patients with advanced HNC is lower than that of controls or patients with earlier stages of the disease. ADA may be a more sensitive indicator of suppressed cellular immunity than delayed cutaneous hypersensitivity reactions, or monitoring absolute lymphocyte counts.     

Cerebral oedema after subarachnoid haemorrhage. Pathogenetic significance of vasopressin

Summary The authors report the frequency, characteristic clinical symptoms, laboratory alterations and diagnostic criteria of the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) after subarachnoid haemorrhage.The data on 290 patients with subarachnoid haemorrhage (SAH) during a period of years at the Division of Neurosurgery, University Medical School, Szeged, are analysed. Twenty-seven (9.3%) patients developed SIADH. Thirteen (4.5%) patients had severe and 14 (4.8%) had mild SIADH. The problems of the treatment are discussed in detail and the different therapeutic methods are listed: Nad infusion, water withdrawal and administration of Dilantin, diuretics, mineralocorticosteroids, lithium and demeclocycline. The undesirable side-effects observed accompanying various therapeutic regimen are analysed. The introduction of V₂ antagonists into clinical practice appears to be a most perspective procedure.For study of the pathogenesis of SIADH following SAH, the possibility of treatment with V₂ antagonists on an experimental model of SAH in rat was created. A significant water retention and increases in brain water and sodium content were observed in rats with SAH. Plasma AVP levels were also elevated after SAH. AVP plays an important role in the development of antidiuresis following water loading and disturbance of the brain water and electrolyte balance after SAH. Water retention and the higher brain water and sodium accumulation could be totally prevented by administration of a V₂ antagonist. These results demonstrate that cerebral oedema generated by artificial cerebral bleeding in rats is significantly reduced following the administration of a highly specific V₂ antagonist, suggesting a new approach to the treatment of SIADH.Some parts of this work were presented at the Vth International Symposium on the Neurohypophysis (Hanover, USA, 1992) and published in Neurosurgery 9: 394–397.