Natural killer cell activity in the peripheral blood of patients with Cushing's syndrome

BACKGROUND: Natural killer (NK) cells are CD3(-)CD16(+)CD56(+) bone-marrow-derived lymphocytes mediating first-line defence by direct cytotoxicity against various types of target cells without prior immunization. NK cell activity is positively regulated by immune interferon (IFN-gamma); among hormones, glucocorticoids are potent in vitro and in vivo inhibitors, whereas ACTH and beta-endorphin in many experimental circumstances enhance NK cytotoxicity. DESIGN: We measured NK cytotoxicity of peripheral blood mononuclear cells (PBMC) obtained at 0800h and 2000h from 26 patients with Cushing's syndrome (12 pituitary-dependent, 12 adrenal-dependent and two dependent on ectopic ACTH secretion). In vitro responsiveness to IFN-gamma or cortisol was also tested. METHODS: NK activity was measured in a 4-h direct cytotoxicity assay using K562 cells as targets. Plasma ACTH, serum and urinary free cortisol were concomitantly measured with commercially available kits. RESULTS: Spontaneous activity and responsiveness to IFN-gamma or cortisol were significantly greater in 15 age- and sex-matched controls than in Cushing's patients at 0800h. In pituitary-dependent Cushing's patients, plasma ACTH correlated positively with mean levels of spontaneous NK activity (r=0.64, P<0.05) and negatively with cortisol-dependent percentage inhibition (r=-0.69, P<0.02). In adrenal-dependent Cushing's patients, a negative correlation was observed between levels of spontaneous NK activity and urinary free cortisol (r=-0.67, P<0.02). CONCLUSIONS: Our data indicate that excess endogenous glucocorticoids affect spontaneous NK cell activity and responsiveness to exogenous IFN-gamma or cortisol. The differential patterns observed between pituitary-dependent and adrenal-dependent groups are compatible with a positive immunomodulatory role of pituitary pro-opiomelanocortin-derived peptides that effectively counterbalance, at least partially, glucocorticoid immunosuppression.     

肝癌患者周围血白蛋白mRNA检测

目的肝细胞性肝癌（ＨＣＣ）预后差，唯有在ＨＣＣ转移前行手术切除才为其最佳的治疗选择，而用形态学方法找血中极其微量的ＨＣＣ细胞是非常困难的。为了更好确定周围血中ＨＣＣ细胞。方法采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）方法检测了２１例ＨＣＣ患者和２２例对照者的周围血白蛋白ｍＲＮＡ。结果发现晚期ＨＣＣ患者阳性率高，且与临床分期有关，有肺或门静脉累及者的阳性率较无肝外转移者高（Ｐ＝０．００３２）；血清甲胎蛋白（ＡＦＰ）＜２００μｇ／Ｌ和＞２００μｇ／Ｌ的ＨＣＣ患者阳性率之间差异无显著性（Ｐ＞０．０５）。结论该方法对ＨＣＣ早期微小转移有预示的可能，对ＨＣＣ患者预后的评估方面起一定的作用，尤其对血清ＡＦＰ阴性或低值的ＨＣＣ患者能起到补充诊断。     

Altered circadian blood pressure rhythm in patients with Cushing's syndrome

The circadian blood pressure rhythm was compared in patients with Cushing's syndrome, essential hypertension, and primary aldosteronism. In patients with essential hypertension or primary aldosteronism, a clear nocturnal fall in systolic and diastolic blood pressure and heart rate was observed. This fall was seen in untreated subjects as well as in patients receiving combined treatment with a calcium antagonist, diuretic, converting enzyme inhibitor, alpha-blocker and beta-blocker, or sympatholytic drug. In these groups, there was a positive correlation between heart rate and systolic or diastolic blood pressure. On the other hand, in patients with Cushing's syndrome, there was no nocturnal fall in blood pressure but in some patients a rise was observed. In all patients there was a nocturnal fall in heart rate. Thus, there was no significant correlation between heart rate and blood pressure in these patients. Exogenous glucocorticoid eliminated the normal nocturnal fall of blood pressure in patients with chronic glomerulonephritis or systemic lupus erythematosus. These results suggest that the changed circadian blood pressure pattern in patients with Cushing's syndrome is not due to antihypertensive treatment or to the mineralocorticoid excess accompanying this disease, but it is attributable to excess glucocorticoid or the associated disturbance in the adrenocorticotropic hormone-glucocorticoid system (or both). This conclusion also implies that the normal circadian rhythm of blood pressure may be regulated at least in part by the adrenocorticotropic hormone-glucocorticoid system.     

Hyperhomocysteinemia in patients with Cushing's syndrome

We evaluated serum homocysteine concentrations and the C677T polymorphism of the gene encoding for methylene tetrahydrofolate reductase, a key enzyme for homocysteine metabolism, in 57 patients with Cushing's syndrome, 41 with active disease, and 16 in remission after successful surgery and 105 blood donors. The patients with active Cushing's syndrome had significantly higher serum homocysteine levels and lower folate concentrations than either the patients in remission or controls. The presence of a statistically significant difference in homocysteine concentrations among the three groups was confirmed after adjustment for confounding variables. In a multiple regression model, homocysteine levels were significantly associated with midnight serum cortisol levels (beta = 0.33, P = 0.01), which is the most sensitive marker of endogenous hypercortisolism, and serum folate levels (beta = -0.32, P = 0.02). The distribution of methylene tetrahydrofolate reductase genotypes was not different between patients and controls. In conclusion, active hypercortisolism is associated with hyperhomocysteinemia and reduced serum folate concentrations, whereas the patients in remission have homocysteine concentrations comparable with healthy subjects. Low serum folate concentrations do not fully account for the increase in homocysteine levels that are positively correlated with cortisol levels. Hyperhomocysteinemia may be key to the prothrombotic state and increased cardiovascular risk of Cushing's syndrome.     

肝衰竭患者外周血单核细胞HLA-DR的表达

目的探讨不同病因所致肝衰竭患者外周血单核细胞HLA-DR基因mRNA表达及其意义。方法随机选取肝衰竭患者40例,其中嗜肝病毒感染10例、药物性肝损伤10例、酒精性肝损伤10例、自身免疫性肝病10例。10名健康对照为本院健康志愿者。应用RT-PCR法检测各组患者外周血单核细胞HLA-DR基因mRNA表达,并动态观察HLA-DR基因mRNA表达的变化,及其与病程发展及临床症状的关系。结果 RT-PCR结果显示嗜肝病毒感染组、正常对照组、药物损伤组、酒精损伤组、自身免疫组HLA-DR基因mRNA表达分别为(137.51±5.22)、(18.14±1.29)、(19.64±2.03)、(11.25±1.22)、(26.61±2.31),且呈逐渐降低趋势,组间差异均有统计学意义(P0.05)。对不同病因组患者单核细胞HLA-DR mRNA表达进行动态观察发现,17例死亡患者HLA-DR mRNA表达持续下降直至患者死亡;另外23例患者,经治疗各组HLA-DR mRNA表达逐渐恢复至接近正常水平,临床症状同步改善。结论外周血单核细胞HLA-DR mRNA表达与肝功能衰竭的严重程度密切相关。     

Circadian blood pressure profile in patients with Cushing's syndrome before and after treatment

The aim of the study was to evaluate the circadian blood pressure (BP) profiles in patients with two forms of Cushing's syndrome, and to compare them to those in patients with essential hypertension. The study included 100 patients with Cushing's syndrome (80 with pituitary adenomas and 20 with adrenal adenomas) and 40 with essential hypertension. Twenty-four-h ambulatory BP monitoring was performed before and after therapy. All 3 groups had similar office-, 24-h-, awake-, and sleep BP mean values. The awake-sleep differences between the patients with two forms of Cushing's syndrome were similar. The night-time BP decline in the patients with Cushing's disease, as well as in those with adrenal adenomas, was significantly lower than that in the patients with essential hypertension. In the patients with both forms of Cushing's syndrome, there was a highly significant decline in the office and ambulatory BP levels after the treatment, and the awake-sleep systolic BP difference became significantly higher. The night-time diastolic BP decline was significantly higher after treatment in patients with adrenal adenomas and not-significantly higher in patients with Cushing's disease. In the patients with Cushing's disease, the duration of hypertension was greater, and lower percentage of normalized BP after treatment was observed in comparison with the patients with adrenal adenomas. The significant negative correlation between duration of the disease and extent of the night-time BP decline suggests that the 'non-dipping' profile is related not only to hypercortisolism itself but also to the severity of hypertension and duration of the disease.     

胃癌患者外周血CK20mRNA的表达及临床意义

采集55例胃癌、46例良性胃病及正常献血员50例，应用逆转录多聚酶链反应（RT－PCR）技术研究外周血CK20mRNA的表达。结果：46例良性胃病及正常献血员50例外周血CK20mRNA均呈阴性，55例胃癌中有35例外周血CK20mRNA表达阳性，I、Ⅱ期20例中有7例外周血CK20mRNA表达阳性，表达率低于Ⅲ、Ⅳ期（28／35）；低、未分化腺癌38例中，30例外周血CK20mRNA表达阳性，高于高、中分化腺癌（5／17）。认为PT－PCR检测外周血CK20mRNA敏感性和特异性较高，可作为胃癌患者术前评估的指标之一。     

原发性肝癌患者外周血中甲胎蛋白mRNA的意义

目的由于ＰＣＲ技术的应用,血循环中癌细胞的检测近有很大进展．本文用ＲＴＰＣＲ检测肝癌（ＨＣＣ）和其他慢性肝病患者外周血中ＡＦＰｍＲＮＡ,藉以反映ＨＣＣ细胞的存在,并与其他血清标记物比较．方法ＨＣＣ患者２２例,肝硬变和慢性乙型肝炎患者各１０例,健康成人受试者（对照）５例．取患者和对照的外周血,分离单核细胞,提取总ＲＮＡ并作电泳鉴定,用合成的两对引物进行巢式ＲＴＰＣＲ扩增ＡＦＰｍＲＮＡ,同时分析血清ＡＦＰ和乙肝标记物．结果ＡＦＰｍＲＮＡ在１３例ＨＣＣ（５９１％）,２例肝硬变（２００％）患者外周血中测到,其余标本均为阴性．ＡＦＰｍＲＮＡ阳性的１３例患者肿瘤均大于５ｃｍ,为晚期患者．在该１３例患者中仅有６例（４６１％）在血清中测到ＡＦＰ,但有１２例（９２３％）ＨＢｓＡｇ,抗ＨＢｅ,抗ＨＢｃ全阳性,而ＡＦＰｍＲＮＡ阴性的５例该３标记物全阴性．结论ＲＴＰＣＲ扩增ＡＦＰｍＲＮＡ是检测ＨＣＣ和肝硬变患者循环肝癌细胞的敏感方法．患者外周血中的ＡＦＰｍＲＮＡ有可能作为肿瘤转移和复发的标记对ＨＣＣ诊断、随访观察和疗效评定有较大临床意义．     

CK20 mRNA在胃癌病人外周血中的表达及意义

目的探讨胃癌病人外周血中CK20 mRNA的：表达及临床意义。方法采用巢式RT-PCR技术检测70例胃癌病人手术前、后外周血CK20 mRNA的表达情况，并以20例健康志愿者为阴性对照，以胃腺癌细胞株BGC-823作阳性对照。结果CK20 mRNA术前阳性表达率为32．9％，阳性表达率与组织学类型具有相关性（P〈0．01），而与TNM分期、肿瘤的部位、病人性别无明显相关性（P〉0．05）。术后CK20 mRNA阳性表达率明显高干术前（P〈0．05）；20例健康志愿者无一例阳性表达，胃腺癌细胞株BCK-823均为阳性表达。结论CK20 mRNA作为标志物用于检测胃癌的血循环微转移是可靠的；手术操作可促使肿瘤细胞释放入血。     

Adipokine levels in Cushing's syndrome; elevated resistin levels in female patients with Cushing's syndrome

BACKGROUND: Cushing's syndrome (CS) is associated with central adiposity, insulin resistance and impaired glucose homeostasis. Adipose tissue is thought to regulates glucose homeostasis via circulating adipokines, such as resistin, leptin and adiponectin, although their role in the insulin resistance associated with CS has not been established. DESIGN: We examined the relationship between insulin resistance and adipokine levels in CS patients. We compared plasma levels of resistin, leptin and adiponectin in 10 women and four men patients with CS, with 14 health subjects matched for age, gender and body mass index. A subgroup of three women and four men with pituitary-dependent CS were re-examined at least 9 months after curative surgery. RESULTS: CS patients had significantly more truncal fat and less lean body mass as assessed by DEXA compared to control subjects. Total cholesterol, triglycerides and insulin resistance, as calculated using the homeostasis model assessment of insulin resistance (HOMA-R), was significantly increased in CS patients. Of the adipokines measured, only resistin was significantly different between female CS patients and female control subjects (5.05 +/- 0.56 vs. 2.91 +/- 0.39 micro g/l, P = 0.015). Curative surgery significantly reduced total body fat and truncal fat, leptin, total and low-density lipoprotein (LDL) cholesterol, glucose and HOMA-R. A reduction in both resistin and adiponectin was also observed but the differences between pre- and post-treatment levels did not achieve statistical significance. CONCLUSION: Here we report for the first time that resistin levels are significantly elevated in CS patients and may be important in the insulin resistance associated with glucocorticoid excess.     

长期维持性血液透析患者外周血单个核细胞中MMP—9mRNA表达及其意义

为探讨慢性肾功能衰竭长期维持性血液透析患者外周血单个核细胞（PBMC）基质金属蛋白酶－9（MMP－9）的mRNA表达情况及其临床意义。采用半定量RT－PCR法检测了18例正常人（对照组）、20例慢性肾功能衰竭非透析患者（非透析组）、21例慢性肾功能衰竭长期维持性血液透析患者（透析组）PBMC中MMP－9的mRNA表达。结果透析组MMP-9mRNA表达比对照组和非透析组显著增加。认为长期维持性血液透析患者PBMC中MMP－9的mRNA表达水平显著增加,提示MMP－9增高与透析患者动脉硬化的形成和高血压血管重塑有关。     

大肠癌患者外周血人类斯钙素基因的检及其意义

目的探讨靶基因人类斯钙素(hSTC-1)在大肠癌患者外周血中的表达以及与肿瘤恶性程度的关系.方法采用RT-PCR方法检测15例健康成人、5例妊娠期妇女、14例消化道炎症疾病患者、57例大肠癌患者外周血的hSTC-1 mRNA.结果57例大肠癌患者外周血中hSTC-1 mRNA的阳性率为49.12%(28/57),并与大肠癌的临床分期有显著相关性(P＜0.05).而健康成人、妊娠期妇女、消化道炎症疾病患者外周血则无一例出现阳性.结论应用RT-PCR方法检测大肠癌患者外周血中的hSTC-1 mRNA具有高度敏感性和特异性,它有望成为判断大肠癌生物学行为恶性程度、转移和复发以及疗效观察的客观指标.     

Evaluation of haemostatic and fibrinolytic markers in patients with Cushing's syndrome and in patients with adrenal incidentaloma.

It is known that either chronic glucocorticoid administration or endogenous hypercortisolism frequently induce an hypercoagulable condition. Since little is known about the evaluation of markers of haemostatic and fibrinolytic systems in other adrenal disorders, we studied plasminogen activator inhibitor (PAI-1), tissue-plasminogen activator (t-PA), fibrinogen and von Willebrand factor antigen (vWF-Ag) levels in 11 patients with Cushing's syndrome and in 12 patients with adrenal incidentaloma. In patients with Cushing's syndrome mean PAI-1, t-PA and vWF-Ag levels did not significantly differ from those found in 50 age- and sex-matched controls, while mean fibrinogen levels were significantly higher in patients (337.0+/-39.1 mg/dl) than in normal subjects (278.9+/-8.4 mg/dl). Patients with adrenal incidentaloma showed PAI-1, t-PA and vWF-Ag mean levels superimposable to those in controls, while fibrinogen (319.7+/-27.9 mg/dl) was slightly, although not significantly, higher than in normals. Considering the limits of normal values (as mean+/-2 SD) obtained in the control group, high PAI-1 levels were found in 2 patients with Cushing's syndrome and in 3 patients with incidentaloma. An elevation of fibrinogen levels was found in 3 patients with Cushing's syndrome and in 3 with incidentaloma. Increased vWF-Ag levels were found only in 1 patient with Cushing's syndrome. An increased t-PA level was occasionally observed only in the patient with adrenal carcinoma. On the whole, an alteration of at least one of haemostatic and fibrinolytic parameters was detected in 55% of the patients with Cushing's syndrome and in 42% of those with adrenal incidentaloma. In conclusion, early alterations of coagulation and fibrinolytic systems may be found in some patients with adrenal disorders, thus suggesting the opportunity of an accurate follow-up in order to identify possible risk factors for cardiovascular disease and thromboembolism.     

Glucose metabolism in patients with subclinical Cushing's syndrome

This clinical review will summarize the available data regarding the effect of either physiological or increased glucocorticoid concentrations on glucose metabolism and insulin-sensitivity, in order to clarify the role, if any, of subclinical Cushing's syndrome (SCS), a status of altered hypothalamic-pituitary-adrenal axis secretion in the absence of the classical signs or symptoms of overt cortisol excess, in patients with adrenal incidentalomas (AI) and diabetes mellitus type 2. Focusing on patients with SCS associated to AI, while there is convincing evidence in the literature that even a mild hyper cortisolemia is associated with alterations of glucose metabolism, evidence is insufficient to conclude that the simple correction of chronic, even mild, hypercortisolism can completely revert metabolic, mainly glycemic alterations. At the same time, considering the variability of the prevalence of Cushing's syndrome in patients with diabetes mellitus type 2 reported in the literature, no agreement does exist whether screening for CS can be useful and recommended in those patients.     

PNS外周血单核细胞趋化因子与RANTES mRNA的表达及意义

目的 探讨原发性肾病综合征(PNS)患者外周血单核细胞趋化因子(MCP-1)与受激活调节的正常T细胞表达和分泌因子(RANTES)mRNA的表达及临床价值。方法 PNS患者54例，分别于治疗前及治疗后3个月采用逆转录聚合酶链反应(RT—PCR)方法检测外周血MCP-1及RANTES mRNA的表达水平。结果 PNS患者外周血MCP-1、RANTES mRNA的表达显著高于正常对照组。治疗后显著降低；其中膜性肾病患者表达水平较低，系膜毛细血管性肾炎患者表达水平较高；伴有肾小管间质损害者表达水平显著高于无肾小管间质病变者，治疗后未完全缓解者显著高于完全缓解署。MCP-1、RANTES mRNA表达水平与血肌酐水平呈显著正相关，与24小时尿蛋白定量、血清白蛋白水平无显著相关性。结论 PNS患者外周血MCP-1、RANTES mRNA表达水平与病理类型、肾小管间质损害、血清肌酐水平及疗效有一定关联，对其检测有助于PNS病情及预后的评估。     

Thyroid axis in patients with Cushing's syndrome.

Thyroid hormone values and serum thyrotropin (thyroid-stimulating hormone [TSH]) responses to the intravenous administration of 400 micrograms of protirelin were determined in ten patients with Cushing's syndrome and in ten matched normal subjects. In patients with Cushing's syndrome, the serum thyroxine (T4) level was mildly depressed and free T4 level was normal. The mean (+/- SD) concentrations of serum triiodothyronine (T3) and free T3 were both reduced in patients compared with normal subjects (P less than .001). At 20 and 60 minutes after protirelin administration, serum TSH levels were, respectively, 3.3 +/- 2.7 microU/mL and 2.6 +/- 2.3 microU/mL in patients with Cushing's syndrome and 12.3 +/- 5.4 microU/mL and 10.7 +/- 5.4 microU/mL in normal subjects (P less than .001). The reduced serum T3 and free T3 levels are due to a glucocorticoid suppressive effect on the peripheral conversion of T4 to T3. The protirelin test is of limited value in assessing the thyroid status because the response of TSH is frequently blunted or absent due to glucocorticoid excess.