Objective To study the circadian changes of blood pressure and urinary catecholamine excretion in patients with pheochromocytoma before and after tumor removal. Methods Twenty-seven patients with pheochromocytoma were examined by 24h ambulatory blood pressure monitoring before tumor removal, and 12 of them were examined by ambulatory blood pressure monitoring both before and after operation, meanwhile, their urinary norepinephrine (NE), epinephrine (E) and dopamine (DA) excretions of 10 time-segmental portions of urine at the same day were determined by HPLC method. The circadian changes were analyzed by Cosinor method. Results The circadian changes of catecholamine excretion were observed, and the peak excretion time of E and DA were shifted to 14:00-16:00. The circadian changes of systolic and diastolic blood pressure disappeared in patients with sustained hypertension and existed in patients with paroxysmal hypertension. The circadian changes of blood pressure and urinary catecholamine excretion were returned to normal after surgery. The circadian change of heart rate was observed both before and after operation. Analysis of data from 12 cases with pheochromocytoma showed strongly positive correlationsbetween24hmeansystolicanddiastolic blood pressure and mean urinary NE excretion (r=0.83, 0.91, P

Objective To investigate the potential effects of simvastatin on angiotensin Ⅱ-stimulated secretion and proliferation of adrenocortical carcinoma H295R cells.Methods The H295R cells were divided into control group, Angiotensin Ⅱgroup, simvastatin group and Angiotensin Ⅱ plus simvastatin group.Cortisol in medium was determined by chemiluminescent method , and aldosterone was determined by radioimmunoassay .The mRNA expression of 11 beta-hydroxylase ( CYP11B1 ) and aldosterone synthase ( CYP11B2 ) were examined by RT-qPCR.Cell proliferation was detected by MTS method.Results Compared with control group, angiotensin Ⅱincreased the secretion of cortisol and aldosterone, and the expression of CYP11B1 and CYP11B2.Simvastatin decreased cortisol secretion and CYP11B1 mRNA expression ( P<0.05 ) .Simvastatin also inhibited angiotensinⅡ-induced the secretion of cortisol and aldosterone , and the expression of CYP 11 B1 and CYP11 B2 compared with Angiotensin Ⅱgroup ( P<0.05 ) .Angiotensin Ⅱhad no effect on cell proliferation , while simvastatin significantly inhibited cell proliferation .The inhibitory effect of simvastatin on proliferation was enhanced when simvastatin was prescribed with angiotensin Ⅱ( P<0.05 ) .Conclusions Simvastatin inhibits angiotensin Ⅱ-induced secretion of cortisol and aldosterone in H295R cells.Simvastatin inhibits cell proliferation, which could be enhanced by angio-tensin Ⅱ.

Objective To investigate the effects of Urotensin Ⅱ(UⅡ)on proliferation of rat pheochromocytoma cell line(PC12) and primary cultured human pheochromocytoma cells.Methods We observed the effects of UⅡ on the proliferation of rat PC12 cells and human pheochromocytoma cells in vitro by MTT method.Results 1.UⅡ had no obvious effect on the proliferation of rat PC12 cells.2.UⅡ(at 10-7 and 10-6mol/L) promoted the proliferation of primary cultured human pheochromocytoma cells. Conclusion UⅡ stimulates the proliferation of human pheochromocytoma cells and probably plays a role in the pathogenesis of pheochromocytoma.

Objective To analyze the clinical features of patients with subclinical pheochromocytoma(PHEO).Methods Review of clinical features of 22 patients with subclinical PHEO treated in PUMC hospital from 1997 to 2007.Results All patients were asymptomatic.24hr-urinary catecholamine excretion was detected normal in 10 of 22 cases,while increased in the others.Sixteen patients were prepared with ?-receptor blocker before operation.During the operation,BPmax(maximal blood pressure) before tumor resection,BPmin(minimal blood pressure) after resection and ?BP(BPmax-BPmin) were(163?34)/(86?20)mmHg,(105?12)/(61?10)mmHg and(58?37)/(25?21)mmHg,Respectively,in the prepared group.They were(169?36)/(104?20)mmHg,(97?18)/(56?13)mmHg and(71?48)/(48?29)mmHg in the other 6 cases without ?-blocker preparation.DBPmax and ?DBP in the prepared group were significantly lower than the unprepared group.Conclusion Most patients with subclinical PHEO have increased catecholamine secretion.Blood pressure is fluctuant greatly during operation in some patients.Patients should be treated with ?-receptor blocker preoperatively in order to decreasethe operation risk.

AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.

Objective To improve the recognization of bilateral adrenal tumors from different tissues and to discuss the treatment.Methods One case of bilateral adrenal tumors from different tissues:pheochromocytoma in one side and adrenocortical adenoma in the other side was reported and the data of syptoms,physical features,auxillary examination,diagnosis and treatment were retrospectively reviewed.Results The patient was diagnosed as adrenocorticotropic hormone (ACTH) -independent Cushing's syndrome in other hospital and received laparoscopic adrenalectomy for right adrenal tumor.During the operation the blood pressure fluctuated seriously and was even more than 200 mm Hg.The pathological report showed the mass was pheochromocytoma.After the operation,the patient's clinical manifcstation was not changed.Half a year later the lab test showed blood ACTH was still less than 5 pg/ml,24 h urinary-free cortisol (UFC) was 571.32 μg,and 24 h urinary catecholamines (U CA) was:noradrenalin (NE) 22.80 μg,epinephrine (E) 2.55 μg,dopamine (DA) 92.92 μg.CT detected a mass in left adrenal gland.The patient received laparoscopic adrenalectomy for left adrenal tumor after being transferred to Peking Union Medical College Hospital.The tumor was proved as adrenocortical adenoma by postoperational pathology.One week after the operation,the 24 h UFC was 56.2 μg.Conclusions Bilateral adrenal tumors from different tissues are very rare in clinic.Adrenalectomy for tumors from both sides and remaining the normal adrenal glands are recommended.

ObjectiveTo detect mRNA and protein expression of steroidogenic factor-1 ( SF-1 ) and DAX-1 in human adrenocortical tumors and normal adrenal cortex,and to investigate the effect of SF-1 and DAX-1 on the steroidogenesis and development of adrenocortical tumors.Methods Total RNA and protein was extracted from angiotensin Ⅱ unresponsive aldoterone-producing adenomas ( A Ⅱ -U-APA,n =12 ),angiotensin Ⅱ responsive aldoterone-producing adenomas ( AⅡ -R-APA,n =5 ),cortisol-producing adenomas ( CPA,n =10 ),adrenal nonfunctional adenomas ( NFA,n =10 ),aldosterone-producing carcinoma ( APC,n =2 ) and normal adrenal cortex ( NAC,n =8).To analyze gene expression of SF-1,DAX-1,ACTH receptor(ACTHR),and β-actin by real-time quantitative PCR in different tissues.The protein expression of SF-1,DAX-1,and β-actin in the same tissues by Western blot.To study the relationship of ACTHR,SF-1,and DAX-1 with clinical data in adrenocortical tumors.ResultsThe expression of SF-1,DAX-1 mRNA and protein was different in NAC,AⅡ -U-APA,A Ⅱ -R- APA,APC,CPA,and NFA tissues [ relative expression of SF-1 mRNA:24.58±2.45,23.89±3.17,21.59±3.00,(38.75,44.16),14.17±2.80,and 36.38±3.50; DAX-1 mRNA:0.57±0.06,0.37±0.05,0.43±0.05,( 1.52,1.21 ),0.39 ±0.04,and 0.83 ±0.08 ; SF-1 protein:0.76 ±0.11,0.76 ±0.10,0.73 ±0.07,(1.24,1.40),0.55±0.04,and0.98±0.10; DAX-1 protein:0.65±0.14,0.39±0.13,0.43±0.14,(1.18,1.02),0.56±0.04,and 1.03±0.13 ; all P＜0.05 or P＜0.01 ].There was negative correlation by higher SF-1/DAX-1 ratio and tumor size in AⅡ -U-APA tissues.The mRNA and protein expression of SF-1 was lower in CPA and there was the positive correlation with tumor size.Conclusion SF-1 and DAX-1 might play a key role in the development of the adrenocortical tumorigenesis and steroidogenic tissues.

Objective To evaluate the value of captopril challenge test (CCT) in the diagnosis of primary aldosteronism (PA).Methods A total of 674 patients [(45.0±13.7) years, men 341, women 333] admitted to Peking Union Medical College Hospital from 2000 to 2015 were analyzed.Among them, 222 subjects were with essential hypertension (EH), 28 were with pheochromocytoma (PHEO), 246 were with idiopathic hyperaldosteronism (IHA) and 178 were with aldosterone producing adenoma (APA).All patients received CCT.24 h urine sodium was measured in partial patients.Plasma renin activity (PRA), aldosterone (ALD) were detected.Results Compared with EH [PRA: before 0.5(0.2,0.9) μg·L-1·h-1, after 0.8(0.4,1.5) μg·L-1·h-1;ALD: before (393±122) pmol/L, after (360±97) pmol/L] and PHEO [PRA: before 0.3(0.1,0.9) μg·L-1·h-1, after 0.4(0.1,1.6) μg·L-1·h-1;ALD: before (396±108) pmol/L, after (374±114) pmol/L], lower levels of PRA and higher levels of ALD before and after CCT were observed in PA patients [PRA: before 0.1 (0.1,0.2) μg·L-1·h-1, after 0.1 (0.1,0.2) μg·L-1·h-1;ALD: before (468±216) pmol/L;after (457±199) pmol/L].After CCT, the suppression rate of ALD [2.8% (-8.8%,15.4%) vs 6.6% (-4.3%, 17.6%)] and increasing rate of PRA [0(0,50%) vs 50%(0, 200%)] in PA patients were lower than those in EH patients.The ALD/PRA ratio (ARR) were higher in PA than that in EH or PHEO patients.In the EH subjects, ALD levels of seated posture were higher than those of recumbent posture both before and after receiving captopril, but with no changes in ARR after CCT.No significant differences in ALD and ARR (before and after receiving captopril) were observed between seated and recumbent position in the PA group.The ARR after CCT tended to decrease in EH subjects with elevated urine-sodium compared with those with normal urine-sodium.No changes could be viewed in ALD and PRA levels between normal urine-sodium and elevated urine-sodium groups among APA, IHA and EH patients either before or after CCT.Among patients with APA, the ALD levels before CCT and the ARR after CCT were lower in the patients with AngiotensionⅡ(AngⅡ) reactive than those without.A ROC curve analysis suggested that the optimal cutoff value was 46.2 (ALD unit:ng/dl;PRA unit:μg·L-1·h-1) for ARR after challenge in diagnosing PA, with the sensitivity of 88.7% and specificity of 84.8%.Conclusions ARR after 25 mg captopril had high sensitivity and specificity in diagnosis of PA with the cutoff of 46.2.Seated CCT could replace recumbent CCT as a more confirmatory test.The PRA increasing rate should be taken into consideration when diagnosis of PA.

Objective:To analyze KCNJ5 mutation of adenomas in patients with aldosterone-producing adenoma (APA) companying with hypokalemia, and to compare the clinical characteristics of patients with and without KCNJ 5 mutations.Methods:Clinical data of 144 APA patients were retrospectively analyzed. DNA were extracted from adenoma tissues, and amplified and sequenced for KCNJ5 gene. The serum potassium level and cardiac complications in patients with and without KCNJ5 gene mutation were compared.Results:Among 144 tumors, 131 tumors (91%) had KCNJ5 mutation, including 68 tumors with G151R, 56 tumors with L168R, 5 tumors with E145Q, and two tumors with novel mutations, V156_K160delITE and G151delinsVR. Compared with patients without KCNJ5 mutation, patients with KCNJ5 mutation had lower preoperative serum potassium levels, more cardiac complications, lower postoperative systolic blood pressure, and better postoperative hypertension relief. There were no statistical differences in age, gender, blood pressure, serum potassium level, plasma renin activity or plasma aldosterone concertration.Conclusion:91% adenomas in patients with APA and hypokalemia had KCNJ5 mutation, suggesting that KCNJ5 mutation is the main cause in these patients.

Objective To summarize the clinical phenotype profiles and mitochondrial DNA mutation in maternally inherited diabetes and deafness ( MIDD ) , and to improve the diagnosis and treatment of this disease in clinical practice. Methods Sixteen patients with MIDD in six families from Peking Union Medical College from 2007 to Dec 2014 were confirmed as carrying the mitochondrial ( mt) DNA 3243 A to G mutation. Sanger sequencing was used to detect the mt DNA 3243 A to G mutation. The peak height G/A ratio was used to determine mutation heteroplasmy levels. Results The patients with early onset of diabetes (35. 0 ± 14. 6 years), deafness, normal or lower body mass index ( BMI) , and maternal hereditary tendency suggested the diagnosis of MIDD. The peak height G/A ratio was significantly different according to the onset age of MIDD [≤25 years (61. 6 ± 20. 17)%;25-45 years (16.59±8.64)%;>45 years(6.37±0.59)%;P<0.01]. The peak height G/A ratio was negatively correlated with the onset age of MIDD(r=-0. 785,P=0. 001). Conclusion Early onset of diabetes with deafness, normal/lower BMI, and maternal hereditary tendency strongly suggests the diagnosis of MIDD. The peak height G/A ratio might provide a simple prediction regarding the onset age and severity of MIDD.