Targeting allosteric site of AKT by 5,7-dimethoxy-1,4-phenanthrenequinone suppresses neutrophilic inflammation.

BACKGROUND: Acute lung injury (ALI) is a severe life-threatening inflammatory disease. Neutrophil activation is a major pathogenic factor in ALI. Protein kinase B (PKB)/AKT regulates diverse cellular responses, but the significance in neutrophilic inflammation and ALI remains unknown. METHODS: Human neutrophils and neutrophil-like differentiated HL-60 (dHL-60) cells were used to examine the anti-inflammatory effects of 5,7-dimethoxy-1,4-phenanthrenequinone (CLLV-1). The therapeutic potential of CLLV-1 was determined in a mouse model of lipopolysaccharide (LPS)-induced ALI. FINDINGS: CLLV-1 inhibited respiratory burst, degranulation, adhesion, and chemotaxis in human neutrophils and dHL-60 cells. CLLV-1 inhibited the phosphorylation of AKT (Thr308 and Ser473), but not of ERK, JNK, or p38. Furthermore, CLLV-1 blocked AKT activity and covalently reacted with AKT Cys310 in vitro. The AKT309-313 peptide-CLLV-1 adducts were determined by NMR or mass spectrometry assay. The alkylation agent-conjugated AKT (reduced form) level was also inhibited by CLLV-1. Significantly, CLLV-1 ameliorated LPS-induced ALI, neutrophil infiltration, and AKT activation in mice. INTERPRETATION: Our results identify CLLV-1 as a covalent allosteric AKT inhibitor by targeting AKT Cys310. CLLV-1 shows potent anti-inflammatory activity in human neutrophils and LPS-induced mouse ALI. Our findings provide a mechanistic framework for redox modification of AKT that may serve as a novel pharmacological target to alleviate neutrophilic inflammation.

Characterization of papillary thyroid microcarcinomas using sonographic features in malignant papillary thyroid cancer: a retrospective analysis.

The diagnosis of malignant thyroid nodules is still a clinical challenge. This study aimed to determine the ultrasonographic characteristics of papillary thyroid carcinoma. The ultrasonographic and pathological data of 2453 thyroid nodules in a cohort of 1895 Chinese patients who underwent thyroidectomy from January 2010 to December 2012 were retrospectively reviewed. Anteroposterior and transversal (AP/TR) diameters ≥1, solid structure, infiltrative margins, hypoechoic appearance, and microcalcifications were more common in malignant nodules than in benign nodules (P < 0.01). These ultrasonographic features were independent risk factors of malignancy (P < 0.01) as determined by logistic regression analysis. Based on multivariate analysis, these characteristics were also present in large nodules (diameter >10 mm). However, in small nodules (diameter ≤10 mm), only AP/TR ≥1 and infiltrative margins were independent risk factors of malignancy (P < 0.01). Ultrasonography is of high diagnostic value for malignant thyroid nodules and may help to improve the differential diagnosis. Small and large nodules have distinct ultrasonographic features.

Telmisartan increases lipoprotein lipase expression via peroxisome proliferator-activated receptor-alpha in HepG2 cells.

In addition to their hypotensive properties, angiotensin receptor blockers (ARBs) have been shown to exert clinical antidyslipidemic effects. The mechanism underlying these ARB lipid metabolic effects remains unclear. Some ARBs, for example, telmisartan, activate peroxisome proliferator-activated activated receptor-gamma (PPAR-gamma). We hypothesized that PPAR-gamma-activating ARBs might exert antidyslipidemic effects via PPAR-alpha. In this study, we assessed the effect of telmisartan on the expression of PPAR-alpha and lipoprotein lipase (LPL). PPAR-alpha expression was detected by reverse-transcription polymerase chain reaction and Western blot in HepG2 hepatocytes as well as differentiated C2C12 myocytes treated with increasing concentrations of telmisartan (0.1-10 µmol/L) for 48 h. Results showed that 1 µmol/L and 10 µmol/L telmisartan significantly increased the expression of PPAR-alpha mRNA and protein in HepG2 cells (p < 0.01). No effect was shown in differentiated C2C12 cells. Similarly, 1 µmol/L and 10 µmol/L telmisartan significantly increased the expression of LPL mRNA and protein in HepG2 cells (p < 0.01), and this increase was significantly (p < 0.01) inhibited by the PPAR-alpha-specific antagonist MK886. These results indicate that certain of the antidyslipidemic effects of telmisartan might be mediated via increased PPAR-alpha-dependent induction of LPL expression.

Pituitary stalk interruption syndrome in 58 Chinese patients: clinical features and genetic analysis.

OBJECTIVES: Pituitary stalk interruption syndrome (PSIS) is rare and its clinical features and pathogenesis are poorly understood. This study characterized the clinical and genetic features of PSIS in Chinese patients. DESIGN AND PATIENTS: Clinical data of 58 patients with PSIS and 46 patients with GH deficiency but a normal pituitary stalk (NPS) were retrospectively analysed. HESX1, LHX4, OTX2 and SOX3 polymorphisms were screened in 33 PSIS patients, and GH1 and GHRHR in 4 NPS patients. RESULTS: Deficiency of GH was 100% in both PSIS and NPS groups. Other deficiency rates for PSIS and NPS groups were as follows: ACTH, 77·6% and 23·9%; TSH, 43·1% and 10·9%; LH/FSH, 94·2% and 47·4%; and combined pituitary hormone, 93·1% and 41·3% respectively. In PSIS and NPS patients, the percentages of anterior pituitary hypoplasia were 98·3% and 54·3%, pituitary stalk abnormality were 100% and 0%, and ectopic neurohypophysis were 91·4% and 0%. A novel heterozygous sequence variant (c.142A>T, p.T48S) was found in HESX1 in one PSIS patient, 3 polymorphisms (c.63T>C, p.G21G; c.450C>T, p.N150N; and c.983A>G, p.N328S) in LHX4 in 7, 1 and 31 PSIS patients, respectively, and a hemizygous polymorphism (c.157G>C, p.V53L) in SOX3 in one PSIS patient. No OTX2 abnormality was detected in PSIS patients, and no GH1 or GHRHR polymorphisms in NPS patients. CONCLUSIONS: Compared with NPS, PSIS patients had more severe anterior pituitary hormone deficiency, lower anterior pituitary hormone secretion and higher probability of abnormal pituitary morphology. HESX1, LHX4 and SOX3 polymorphisms may be associated with PSIS.

[The clinical characteristics of multi-detector computed tomography of congenital adrenal hyperplasia].

OBJECTIVE: To explore the imaging features of congenital adrenal cortex hyperplasia (CAH). METHODS: A total of 45 patients clinically confirmed as CAH were retrospectively analyzed to investigate the imaging features and strengthening way of the multi-detector-row Computed tomography. RESULTS: The imaging features of all the cases presented as following: 25 with bilateral adrenal hyperplasia, 6 with unilateral adrenal hyperplasia, 6 with adrenal nodular hyperplasia, 2 with adrenal hyperplasia and unilateral solid cystic lesion, 2 with adrenal hyperplasia and double side real cystic lesion, 1 with adrenal hyperplasia and unilateral cystic changes and 3 with normal adrenal. The unilateral or bilateral hyperplasia adrenal could be homogeneously enhanced, while the enhanced performance of other cases was inequitable. CONCLUSIONS: The adrenal imaging features of CAH by multi-detector-row CT are variable, with the bilateral adrenal hyperplasia as the main form, which could be restored to normal morphology after hormone replacement therapy.No regression of the tumor size is observed in cases with adrenal mass. CT scanning combined with clinical manifestation and biochemical examination could facilitate the diagnosis of CAH.

Medullar thyroid carcinoma in mediastinum initially presenting as Ectopic ACTH syndrome. A case report.

A rare case with ectopic adrenocorticotrophic hormone syndrome (EAS) caused by medullar thyroid carcinoma (MTC) in mediastinum was reported. This 49 year-old male patient initially presented with serious and intractable hypokalemia. Endocrine evaluations showed increased levels of adrenocorticotrophic hormone (ACTH) and urinary free cortisol, which could not be suppressed more than 50% by high-dose dexamethasone suppression test. Computed tomography (CT) scan detected a 5×5×5 cm mass at the bottom of thyroid in anterior mediastinum. The patient underwent total thyroidectomy with central compartment and ipsilateral modified radical neck dissection. Pathological examination showed an infiltrating thyroid medullary carcinoma with abundant amyloid deposition, meanwhile immunohistochemical positive for ACTH. After surgery, serum levels of kalium, as well as cortisol and ACTH returned to normal range. During follow-up, the patient's clinical manifestation of Cushing syndrome relieved.

Spontaneous remission of acromegaly or gigantism due to subclinical apoplexy of pituitary growth hormone adenoma.

BACKGROUND: Subclinical apoplexy of pituitary functional adenoma can cause spontaneous remission of hormone hypersecretion. The typical presence of pituitary growth hormone (GH) adenoma is gigantism and/or acromegaly. We investigated the clinical characteristics of patients with spontaneous partial remission of acromegaly or gigantism due to subclinical apoplexy of GH adenoma. METHODS: Six patients with spontaneous remission of acromegaly or gigantism were enrolled. The clinical characteristics, endocrinological evaluation and imageological characteristics were retrospectively analyzed. RESULTS: In these cases, the initial clinical presences were diabetes mellitus or hypogonadism. No abrupt headache, vomiting, visual function impairment, or conscious disturbance had ever been complained of. The base levels of GH and insulin growth factor-1 (IGF-1) were normal or higher, but nadir GH levels were all still > 1 µg/L in 75 g oral glucose tolerance test. Magnetic resonance imaging detected enlarged sella, partial empty sella and compressed pituitary. The transsphenoidal surgery was performed in 2 cases, and the other patients were conservatively managed. All the patients were in clinical remission. CONCLUSIONS: When the clinical presences, endocrine evaluation, biochemical examination and imageology indicate spontaneous remission of GH hypersecretion in patients with gigantism or acromegaly, the diagnosis of subclinical apoplexy of pituitary GH adenoma should be presumed. To these patients, conservative therapy may be appropriate.

[The clinical features of ectopic ACTH syndrome: a report of 16 cases].

OBJECTIVES: To improve the diagnostic and therapeutic ability of ectopic ACTH syndrome by analysing its clinical features. METHODS: Sixteen cases of ectopic ACTH syndrome diagnosed in Chinese PLA General Hospital from 2000 to 2009 were analyzed retrospectively. RESULTS: (1) The main causes of ectopic ACTH syndrome were lung tumor and thymic carcinoid; (2) Abnormal glucose metabolism, hypertension, hypokalemia and edema of both lower limbs were the most common clinical symptoms; (3) Laboratory examination showed a significant increase of serum cortisol, adrenocorticotropic hormone (ACTH) and 24 h urinary free cortisol (24hUFC) together with severe hypokalemia and alkalosis; (4) High dose dexamethasone suppressing test, corticotrophin-releasing hormone (CRH) stimulating test and petrosal sinus sampling were the most meaningful diagnostic methods; (5) Most of the primary lesions might be detected with chest film and CT; (6) Resection of the primary lesion was the treatment of first choice. CONCLUSION: The diagnosis of ectopic ACTH syndrome is very hard. Resection of the primary lesion is the best treatment.

[Multiple endocrine neoplasia type 1 presenting as hypoglycemic coma: a report of four cases and review of literatures].

OBJECTIVE: To investigate the clinical characteristics of multiple endocrine neoplasia type 1 (MEN1) patients presenting with hypoglycemic coma as chief manifestation and the related clinical experience in diagnosis and therapy. METHODS: We analyzed the clinical data of 4 patients who were hospitalized because of recurrent hypoglycemic coma and diagnosed as having MEN1 by endocrinological, radiological and pathological examinations. RESULTS: In the 4 cases of Whipple trilogy, radiological examination showed occupying lesion in the pancreas and pathological examination confirmed the diagnosis of insulinoma. In 2 cases the insulinomas were multiple. In this series, one case was complicated with pituitary adenoma, parathyroidoma (recurrent after operation) and adrenocortical adenoma, one case with pituitary adenoma, parathyroidoma (2 tumors) and adrenal nodular hyperplasia, one case with pituitary adenoma and parathyroidoma, and the remaining one with pituitary adenoma and suspectible parathyroidoma. CONCLUSIONS: For patients with insulinoma, MEN1 should be considered. In patients with MEN1, the presence of multiple or ectopic parathyroid adenomas (or hyperplasia) and insulinomas should be inspected during operation. After operation, examinations should be regularly performed to identify whether the diseases relapse or new endocrine neoplasias occur.

A case of relapsed autoimmune hypothalamitis successfully treated with methylprednisolone and azathioprine.

Autoimmune hypothalamitis is a rare autoimmune neuroendocirne disease. A case of a 70-year-old female with autoimmune hypothalamitis was reported. The chief clinical characteristics were diabetes insipidus and adenopituitary function deficiency. Cranial magnetic resonance imaging (MRI) scan indicated a mass in the hypothalamus. The diagnosis of autoimmune hypothalamitis was presumed. After treatment with prednisone, there was a marked reduction in the mass and the hypothalamus-adenopituitary function partially improved. However, after glucocorticoid therapy was withdrawn, the hypothalamic lesion relapsed progressively. High dose methylprednisolone pulse therapy (HDMPT) in combination with azathioprine was initiated thereafter. During follow-up, MRI scan indicated the lesion shrank strikingly, and the patient's clinical condition improved as well. In view of the good response of the hypothalamic lesion to glucocorticoid and immunodepressant, the putative diagnosis of autoimmune hypothalamitis was confirmed. This case report suggested that HDMPT in combination with azathioprine therapy might be an effective trial for autoimmune hypothalamitis treatment.

[Detrimental effects of homocysteine on insulin release and apoptosis of pancreatic beta cells].

OBJECTIVE: To study the effects of homocysteine (Hcy) on the insulin secretion and apoptosis of pancreatic beta cells. METHODS: Clonal mouse pancreatic beta cells of the line NIT-1 were cultured and exposed to Hcy of 50, 100, 250, 500 and 1000 micromol/L for 6, 12, 24, and 48 hours respectively, then glucose of the concentrations of 5.6 mmol/L or 16.7 mmol/L was added for 1 h, and then the insulin concentration in the culture medium was determined by radioimmunoassay, and MTT method was used to detect the survival rate of the cells. NIT-1 cells were exposed to Hcy of the concentrations of 0, 50, 100, 250, 500 and 1000 micromol/L respectively for 24 h, another NIT-1 cells were exposed to Hcy of the final concentration of 250 micromol/L for 0, 6, 12, and 48 h respectively, then flow cytometry (FC) was used to detect the apoptosis of the cells. After exposure to Hcy of the concentrations of 50, 100, 250, 500 and 1000 micromol/L for 72 h DNA agarose gel electrophoresis was performed. RESULTS: Hcy inhibited the basal and glucose-induced insulin secretion in a time- and dose-dependent manner. The insulin secretion amounts of the NIT-1 cells after exposure to 50 micromol/L Hcy for 24 hours and to 100 micromol/L Hcy for 12 hours were significantly lower by 17.1% and 10.8% compared with the control group (all P < 0.01). Incubated with 100 micromol/L Hcy for 12 hours and 24 hours respectively, the survival rates of the NIT-1 cells were 94.56% and 87.93% respectively (P < 0.05, P < 0.01). Incubated with 100 micromol/L Hcy for 24 hours, the apoptosis rate of the NIT-1 cells was 7.21% (P < 0.01); and incubated with 250 micromol/L Hcy for 12 hours, the apoptosis rate of the NIT-1 cells was 12.93% (P < 0.01). Both FC and agarose gel electrophoresis indicated that Hcy induced cell apoptosis time- and concentration-dependently. CONCLUSION: Hcy impairs insulin secretion and induces cell apoptosis of pancreatic beta cells time- and concentration-dependently.

[A survey of factors influencing prognosis and control rate for patients with hypertension in mainland China].

OBJECTIVE: To observe the risk factor stratification and prevalence of target organ damage in hypertensive patients before therapy and blood pressure control rate after 4 or 12 weeks antihypertensive drug therapy. METHODS: In this prospective survey, data on cardiovascular risk factors, target organ damage and concomitant disease were collected in 26 655 hypertensive patients. Among them 26 325 and 3457 patients were recruited for antihypertensive drug therapy for 4 and 12 weeks, respectively and blood pressure control rate was determined. RESULTS: The sedentary lifestyle, smoking, high body mass index, dyslipidemia were found in 52.5%, 34.4%, 31.8%, 24.5%, and microproteinuria, left ventricular hypertrophy, coronary artery disease and diabetes in 21.0%, 23.6%, 20.1%, 26.7% hypertensive patients, respectively. The average systolic and diastolic pressures were 158 +/- 14 mm Hg and 94 +/- 11 mm Hg and 3.2%, 22.2%, 21.1% and 53.3% patients were defined as low, medium, high and very high risk patients in risk stratification to quantify prognosis. There were 77.2%, 20.4% and 2.4% systolic and diastolic, isolated systolic and isolated diastolic hypertensive patients respectively. The goal blood pressure control rate was 50.2% and 56.7% respectively after 4 and 12 weeks antihypertensive drug therapy. The control rate in patients complicated with diabetes and renal disease was significantly lower than patients without them and systolic pressure control rate was remarkably lower than diastolic pressure control rate. Majority patients required 2 or more antihypertensive drugs for effective pressure control (1.5 drug per patients in average in both 4 or 12 weeks groups). CONCLUSION: The prevalence of risk factors, target organ damage and concomitant disease were high in Chinese patients with hypertension and comprehensive interventions were indicated. To reach goal blood pressure control in patients with hypertension, follow up intensifying and drug therapy guidance are required within the context of usual medical care.

[Lymphocytic hypophysitis: a report of 3 cases].

OBJECTIVE: Lymphocytic hypophysitis is a rare inflammatory lesion of pituitary gland. To enhance the knowledge of lymphocytic hypophysitis, herein we reported 3 cases of lymphocytic hypophysitis and reviewed the literature. METHODS: The clinical data of 3 patients diagnosed as having lymphocytic hypophysitis were analysed. RESULTS: All the three patients were young females, one of them (case 1) was affected in the postpartum period, however, the others were neither pregnant nor in postpartum period. Clinically, the most frequently seen symptoms and signs were attributable to pituitary hypofunction, headache and diabetes insipidus. Pituitary MRI revealed enhanced mass with pituitary stalk enlargement expanding the pituitary fossa, extending into suprasellar area and compressing optic chiasm. Typically the lesion appeared hypointense or isointense on T1-weighted imaging, but hyperintense on T2-weighted imaging. Histopathological examination showed extensive destruction of anterior acini of the pituitary with a dense infiltration of lymphocytes, plasma cells, histocytes and other inflammatory cells. Two patients (case 2 and 3) were successfully treated for mass reduction of pituitary gland and restoration of pituitary function with high dose methylprednisolone pulse therapy (HDMPT). CONCLUSIONS: Lymphocytic hypophysitis is a rare autoimmune endocrinopathy which can affect young woman in the postpartum period, or in the peripartum period, characterized by focal or extensive lymphocytic infiltration of anterior pituitary acini. It may cause pituitary expansion and a varying degree of hypopituitarism mimicking the features observed in pituitary adenoma. HDMPT was proved to be effective for mass reduction of pituitary gland and restoration of pituitary function.

[The expression of G proteins alpha-subunit mRNA in different thyroid diseases].

OBJECTIVE: To investigate the expression of G proteins alpha subunit mRNA in different thyroid diseases. METHODS: Twenty-three thyroid specimens were obtained during surgery, 5 from patients with Graves' disease (GD), 7 from patients with multinodular goiter (MNG), 6 from patients with eufunctioning thyroid adenoma (EFTA) and 5 from patients with thyroid papillary cancer (TPC). The expression of stimulating and inhibiting G proteins alpha subunit mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: (1) The expression of G(s)alpha mRNA in TPCs (1.67 +/- 0.25) was significantly higher than that in normal thyroids (1.10 +/- 0.14) and MNGs (0.96 +/- 0.31), P < 0.05 and P < 0.01. The expression in GDs (1.47 +/- 0.11) and EFTAs (1.36 +/- 0.28) was significantly higher than that in MNGs (P < 0.05), but comparable to that in normal thyroids. (2) The expression of G(i)alpha-2 mRNA in GDs (0.68 +/- 0.26) was lower than that in MNGs (1.15 +/- 0.35), P < 0.05, but there was no difference in the expression of G(i)alpha-1 and G(i)alpha-3 mRNA among different thyroid diseases. CONCLUSION: The results indicated that G(s)alpha could play an important role in the pathogenesis of thyroid papillary cancer and G proteins had different expression in different thyroid diseases.

Saturated free fatty acids, palmitic acid and stearic acid, induce apoptosis by stimulation of ceramide generation in rat testicular Leydig cell.

In men, obesity has generally been associated with reduced plasma testosterone levels and with elevation of the plasma free fatty acids (FFAs). In this study, we investigated the effects of saturated FFAs including palmitic acid (PA) and stearic acid (SA), and polyunsaturated FFA arachidonic acid (AA) on the survival of rat testicular Leydig cell cultured in vitro. PA and SA markedly suppressed Leydig cell survival in a time- and dose-dependent manner. In contrast, AA stimulated the cell proliferation at 5-10 times of physiological concentration. The suppressive effect of PA and SA on cell survival was caused by apoptosis evidenced by DNA ladder formation and Annexin V-EGFP/propidium iodide staining of the cells. The apoptotic effect of PA was possibly mediated by ceramide generation because it could be completely blocked by ceramide synthase inhibitor fumonisin B1 and exogenous ceramide itself could directly induce apoptosis in vitro. Surprisingly, the apoptosis induced by PA could be partly prevented by AA. These results indicate that PA and SA induce apoptosis in testicular Leydig cells by ceramide production and these apoptotic effects may be a possible mechanism for reproductive abnormalities in obese men, and AA can partly prevent the apoptotic effect induced by saturated FFA.