Age-specific effects on adverse pregnancy outcomes vary by maternal characteristics: a population-based retrospective study in Xiamen, China.

BACKGROUND: Advanced maternal age (AMA; ≥35 years) is considered to be a major risk factor for adverse pregnancy outcomes. Along with the global trend of delayed childbearing, and in particular, the implementation of China's second and third-child policy leading to a dramatic increase of AMA in recent years, the association between maternal age and pregnancy outcomes requires more investigation. METHODS: A population-based retrospective study was performed. Data were derived from the Medical Birth Registry of Xiamen from 2011 to 2018. Univariate and multivariate logistic regression was used to evaluate the effects of maternal age on pregnancy outcomes. RESULTS: A total of 63,137 women categorized into different age groups (< 25 years, 25-29 years, 30-34 years, and ≥ 35 years) were included in this study. Compared with the mothers aged 25-29 years, the univariate regression analysis showed that mothers aged < 25 years had lower risks of gestational diabetes mellitus (GDM) and cesarean. AMA was associated with higher risks of GDM, hypertension, cesarean, preterm birth, low-birth weight (LBW), large-for-gestational-age (LGA), macrosomia, and stillbirth (all P < 0.01). After adjustment for potential confounding factors, increased risks of GDM, hypertension, cesarean, preterm birth, and LBW remained significantly associated with AMA (all P < 0.05), whereas AMA mothers showed a lower risk of macrosomia than their younger counterparts. Additionally, no significant differences were detected in terms of Apgar score < 7. CONCLUSION: AMA was associated with adverse pregnancy outcomes including increased risks of GDM, hypertension, cesarean, preterm birth, and LBW. This study confirmed the relationship between AMA and certain adverse maternal and fetal outcomes and emphasizes the necessity for women to be cautious about the age at which they become pregnant.

Clinical analysis of the etiological spectrum of bilateral adrenal lesions: A large retrospective, single-center study.

PURPOSE: To investigate the clinical characteristics, endocrinological function, and etiology of bilateral adrenal lesions in hospitalized patients. METHODS: A retrospective study of 777 patients with bilateral adrenal lesions was conducted at the Chinese People's Liberation Army General Hospital between January 2013 and January 2018. Patients' demographic features, hormonal profiles, imaging findings, and histopathological findings were reviewed from database records. RESULTS: Of the 777 patients with bilateral adrenal lesions, 495 were men. The mean age at diagnosis was 52.0 ± 13.0 years. Overall, 511 (65.8%) cases were benign, followed by adrenal metastases (n = 224, 28.8%), pheochromocytoma (n = 26, 3.3%), adrenal lymphoma (n = 9, 1.2%), and adrenal corticocarcinoma (ACC; n = 7, 0.9%). Hormonal evaluation revealed that 34.3% of bilateral adrenal lesions were functional. The primary etiologies of functional lesions were primary aldosteronism (16.6%, 129/777), and primary bilateral macronodular adrenocortical hyperplasia (PBMAH; 8.8%, 68/777). Patients with lymphoma and metastases were significantly older than those with benign nonfunctional lesions (60.4 ± 11.0 years vs. 54.5 ± 10.4 years and 57.9 ± 10.8 years vs. 54.5 ± 10.4 years, respectively; both P < 0.001). Lesions in patients with adrenal lymphoma, ACC, pheochromocytoma, metastases, congenital adrenal hyperplasia, tuberculosis, and Cushing's syndrome were significantly larger than benign nonfunctional lesions (all P < 0.001). CONCLUSION: Benign adrenal lesions and metastases from the lungs are the most common causes of bilateral adrenal lesions. Primary aldosteronism and PBMAH are the most prevalent functional lesions. Moreover, patients with lymphoma or metastases are older and their masses are larger.

Relationship between Obstetric Mode of Delivery and Risk of Overweight/Obesity in 1- to 4-Year-Old Children.

INTRODUCTION: Childhood obesity is an important public health problem, which may lead to increased risk of obesity in adulthood. The relationship between the incidence of obesity and the mode of delivery is not clear. Cesarean section (CS) may be one of the risk factors of obesity in children. We investigated the relationship between the mode of delivery of pregnant women and the risk of overweight/obesity in children of all ages from 1 to 4 years. METHODS: Registered in the maternal and child registration system of Xiamen city, newborns born between January 2011 and December 2012 were followed up to 4 years old. RESULTS: 9,964 cases were included in the study, of which 3,462 cases (34.7%) were cesarean deliveries. From 1 to 4 years of age, BMI Z-scores and the risk for overweight/obesity of children delivered by CS were higher than by the vagina. Longitudinal analysis of anthropometric outcomes assessed during study visits in 1- to 4-year-old offspring exposed to CS showed that after adjustment for kinds of effect factors, the changes in BMI Z-scores were 0.04 (95% CI: 0.01-0.09, p = 0.003), significantly higher than vaginal delivery, and the risk incidence of overweight/obesity by increased 8% in CS offspring; OR = 1.08 (1.01-1.21, p< 0.05). CONCLUSION: The mode of cesarean delivery is related to the risk of overweight and obesity in children aged 1-4 years. When pregnant women choose cesarean delivery without medical indications, they should be concerned that their offspring may have a higher risk of obesity.

Independent associations of thyroid-related hormones with hepatic steatosis and insulin resistance in euthyroid overweight/obese Chinese adults.

PURPOSE: The aim of the study is to explore the independent association of free triiodothyronine (FT3), free thyroxine (FT4) and thyroid stimulating hormone (TSH) with hepatic steatosis and insulin resistance. METHODS: A cross-sectional study of 88 overweight/obese adults who underwent anthropometric measurements [BMI, waist circumference (WC) and waist-to-height ratio (WHtR)], hepatic steatosis assessment (FibroScan) and thyroid-related hormones tests was conducted from 2018 to 2020 in Xiamen, China. RESULTS: Subjects with increasing tertiles of FT3 showed significantly higher levels of controlled attenuation parameter (CAP) ((295.4 ± 44.1, 290.1 ± 68.2 and 331.7 ± 43.6 (dB/m) for tertile 1-3, respectively, p = 0.007) and fatty liver index (FLI) score (47.7 (33.9-60.8), 61.5 (45.1-88.9) and 90.5 (84.5-94.8), respectively, p < 0.001). FT3 significantly and positively correlated with obesity index (BMI, WC, and WHtR), homeostatic model assessment of insulin resistance (HOMA-IR) and hepatic steatosis (CAP and FLI). Multivariable linear regression analyses with adjustment for potential confounding factors showed FT3 was independently associated with BMI (regression coefficient (ß (95%CI): 0.024 (0.004-0.043), p = 0.020), HOMA-IR (ß (95%CI): 0.091 (0.007-0.174), p = 0.034), CAP (ß (95%CI): 25.45 (2.59-48.31), p = 0.030) and FLI (ß (95%CI): 0.121 (0.049-0.194), p = 0.001). Neither FT4 nor TSH was significantly associated with any indicators of obesity, insulin resistance or hepatic steatosis. CONCLUSIONS: Increased FT3, but not FT4 or TSH, was independently associated with higher risks of hepatic steatosis and insulin resistance in euthyroid overweight/obese Chinese adults. Trial registration Registration is not applicable for our study.

Benefits and harms of hemithyroidectomy, total or near-total thyroidectomy in 1-4 cm differentiated thyroid cancer.

OBJECTIVE: For 1-4 cm differentiated thyroid cancer (DTC), current ATA guideline recommended hemithyroidectomy (HT) as an acceptable alternative initial procedure to total or near-total thyroidectomy (TT). The aim of this study was to evaluate benefits and harms of HT, TT in 1-4 cm DTC. DESIGN: Retrospective cohort study. PATIENTS: DTC patients aged 18 years or older who underwent initial thyroidectomy in a tertiary medical centre were included from January 2008 to July 2018. MEASUREMENTS: The structural persistent/recurrent disease, reoperation rates and surgical complications were compared using Cox proportional regression and logistic regression. Propensity score matching was performed to adjust for related clinicopathological variables. RESULTS: Among 1824 DTC patients, 795 patients sized 1-4 cm were included. A total of 286 patients underwent HT and 509 patients underwent TT. In the matched analysis, no significant difference in disease-free survival (DFS) between HT and TT was observed during the median follow-up period of 56.5 months (hazard ratio [HR] 0.86; 95% CI, 0.37-2.00; p = .733). The difference in DFS between two groups was consistent regardless of age, sex, tumour size, follow-up duration. Meanwhile, HT was associated with a decreased risk of surgical complications (odds ratio [OR] 0.47, 95% CI 0.31-0.71, p < .001), as well as lower proportion of levothyroxine replacement (p = .007). Two cases in HT group received reoperation. Further multivariate analysis showed surgical procedure was not associated with structural persistence/recurrence (HR 0.68; 95%CI, 0.29-1.58, p = .367). CONCLUSIONS: For patients with 1-4 cm DTC without clinical evidence of lymph node metastasis or extrathyroidal extension, HT was associated with lower risk of surgical complications than TT while provided similar benefits as TT.

Resistance to thyroid hormone caused by heterozygous mutation of thyroid hormone receptor B gene c.G1378A: Report of one Chinese pedigree and literature review.

The same thyroid hormone receptor B gene (THRB) mutation led to thyroid hormone resistance with different clinical manifestations in the kindreds.

Higher Apnea-Hypopnea Index (AHI) and Oxygen Desaturation Index (ODI) Were Independently Associated with Increased Risks of Hypertension in Patients with T2DM: A Cross-Sectional Study.

OBJECTIVE: The current study aimed to explore the relationship between OSAS and hypertension and whether polysomnography (PSG) indices were independently associated with hypertension in patients with type 2 diabetes (T2DM). METHODS: This study recruited 316 T2DM patients. Multivariable logistic regression analyses were performed to determine the independent association of apnea-hypopnea index (AHI) and oxygen desaturation index (ODI) with hypertension with adjustment for potential confounders. RESULTS: Among 316 patients, 130 (41.1%) and 204 (64.6%) had hypertension and OSAS, respectively. T2DM patients with hypertension showed significantly increased levels of AHI ((median (interquartile range)): 17.2 (5.7-34.9) vs. 5.7 (2.1-17.3) events/hour, p < 0.001), nonrapid eye movement AHI (NREM-AHI) (17.6 (5.5-36.5) vs. 5.2 (2.2-16.6) events/hour, p < 0.001), ODI (48.4 (21.9-78.0) vs. 22.6 (10.8-48.1) events/hour, p < 0.001), and severities of OSAS and decreased levels of lowest SaO2 ((mean ± standard deviation): 74.0 ± 10.4 vs. 77.3 ± 9.8, p = 0.004). Multivariable logistic regression analysis showed that higher levels of AHI, NREM-AHI, and ODI were significantly associated with increased risks of hypertension, and the adjusted odds ratios (ORs) with 95% CI were 1.026 (1.008-1.044, p = 0.004), 1.026 (1.009-1.044, p = 0.003), and 1.005 (1.001-1.010, p = 0.040), respectively. Compared with non-OSAS, severe OSAS was significantly associated with the risk of hypertension with the adjusted OR (95% CI) of 3.626 (1.609-8.172, p = 0.002), but associations of rapid eye movement AHI (REM-AHI) and lowest SaO2 with hypertension were not statistically significant. CONCLUSION: Increased AHI, NREM-AHI, ODI, and severities of OSAS were significantly associated with higher risks of hypertension in T2DM patients. Detection and treatment of OSAS are needed to prevent hypertension in T2DM patients.

Primary Adrenal Lymphoma: Two Case Series From China.

Objective: Primary adrenal lymphoma (PAL) is a rare form of adrenal mass. We summarize our experience in its clinical presentation, biochemical indexes, radiological features, pathological information, therapy regimens, and outcomes. Methods: This was an institutional review board-approved retrospective review of medical records and surgical pathology specimens of patients with a diagnosis of PAL at the Chinese People's Liberation Army General Hospital and the First Affiliate Hospital of Xiamen University between July 2007 and July 2017. Results: Twenty-six patients were identified. The mean age at presentation was 60.84 ± 13.14 years with a male-to-female ratio of 2.25:1 (18:8). The most common presenting symptoms were loss of appetite (65%, 17/26), weight loss (62%, 16/26), abdominal pain (58%, 15/26), and fatigue (58%, 15/26). The levels of lactate dehydrogenase (75%, 15/20), ß2-microglobulin (100%, 10/10), C-reactive protein (82%, 14/17), and ferritin (88%, 7/8) and the erythrocyte sedimentation rate (83%, 10/12) were elevated. Bilateral involvement was seen in 21 of 26 patients (81%); 12 of 19 evaluated patients with bilateral lesions (63%) were confirmed to have adrenal insufficiency. On computed tomography (CT), the mean tumor diameter was 7.31 ± 3.35 cm and the median Hounsfield density was 37.0 HU (range: 31.0-45.0 HU); 67% (10/15) and 27% (4/15) of lesions presented with mild and moderate enhancement after injection of contrast medium. 18F-fluorodeoxyglucose positron emission tomography (FDG PET)-CT revealed not only an adrenal tumor but also extra-adrenal lesions. Diffuse large B-cell lymphoma (DLBCL) was the most common phenotype (92%, 24/26). Ninety-two percent (24/26) of patients received chemotherapy while 8% (2/26) received unilateral adrenalectomy plus chemotherapy. The prognosis of PAL was poor, with a general survival time of 7.20 ± 5.18 months. Conclusion: PAL is a rare disease. The clinical characteristics of PAL include loss of appetite and weight loss. Endocrine evaluation should be performed to determine whether patients have adrenal insufficiency, especially patients with bilateral lesions. FDG-PET appears to be more accurate than other imaging modalities in revealing extra-adrenal sites. Better therapy is required to improve the poor prognosis of PAL.

Efficacy of Vildagliptin Added to Continuous Subcutaneous Insulin Infusion (CSII) in Hospitalized Patients with Type 2 Diabetes.

INTRODUCTION: The aim of this study was to compare the efficacy of vildagliptin as add-on therapy to short-term continuous subcutaneous insulin infusion (CSII) with CSII monotherapy in hospitalized patients with type 2 diabetes mellitus (T2DM). METHODS: A total of 200 hospitalized patients with inadequately controlled T2DM were randomized into groups, with one group receiving CSII monotherapy (CSII group, n =100) and the other group receiving CSII plus vildagliptin as add-on (CSII + Vig group, n = 100). Of these, 191 completed the 7-day trial (CSII group, n = 99; CSII + Vig group, n = 92) and included in the analysis. The glycemic control and variability of the patients were measured using all-day capillary blood glucose (BG) monitoring. Weight and fasting C-peptide levels were evaluated before and after the interventions. RESULTS: Mean BG concentrations during the whole treatment period were lower and the time to reach target BG was reduced in the CSII + Vig group compared with the CSII group (9.89 ± 3.37 vs. 9.46 ± 3.23 mmol/L, P < 0.01; 129 ± 4 vs. 94 ± 5 h, P < 0.01, respectively). Similarly, the indicators of glycemic variability, namely the standard deviation of BG and the largest amplitude of glycemic excursion, were significantly decreased in the CSII + Vig group compared with the CSII group (2.68 ± 1.05 vs. 2.39 ± 1.00 mmol/L, P < 0.01; 7.19 ± 2.86 vs. 6.23 ± 2.73 mmol/L, P < 0.01, respectively). CONCLUSIONS: Short-term CSII with vildagliptin as add-on therapy may be an optimized treatment for hospitalized patients with T2DM compared with short-term CSII monotherapy.

MATERNAL GESTATIONAL DIABETES MELLITUS AND OFFSPRING BODY MASS INDEX FROM 1 TO 4 YEARS.

Objective: Using the International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria to diagnose gestational diabetes mellitus (GDM), the association between GDM and offspring body mass index (BMI) gains in early childhood in China remains unclear. We aimed to assess the association between GDM diagnosed by the IADPSG criteria and BMI gain and the risk for overweight/obesity in offspring from 1 to 4 years. Methods: This prospective cohort study was based on the healthcare records data from the Medical Birth Registry in Xiamen, China. We included 10,412 mother-child pairs tested for GDM using IADPSG criteria. Results: A total of 1,786 (17.2%) offspring were exposed to GDM. The offspring exposed to GDM had higher mean BMI Z-score (difference, 0.07; 95% confidence interval [CI], 0.02 to 0.12) and risk for overweight/obesity (odds ratio [OR], 1.22; 95% CI, 1.06 to 1.40) compared to those unexposed to GDM from 1 to 4 years of age. However, after adjustment for maternal pre-pregnancy BMI (Model 2), these associations attenuated towards the null (difference in BMI Z-score, 0.02; 95% CI, -0.03 to 0.07; OR for overweight/obesity, 1.09; 95% CI, 0.95 to 1.25). Conclusion: The associations between GDM diagnosed using IADPSG criteria and BMI Z-score and the risk for overweight/obesity in offspring at the age of 1 to 4 years were largely explained by maternal pre-pregnancy BMI. Reducing the prevalence of childhood overweight and obesity in China should focus on maternal weight status before pregnancy, in addition to glycemia during pregnancy. Abbreviations: BMI = body mass index; CI = confidence interval; GDM = gestational diabetes mellitus; IADPSG = International Association of Diabetes and Pregnancy Study Groups; LGA = large for gestational age; MBRX = Medical Birth Registry in Xiamen; OGTT = oral glucose tolerance test; OR = odds ratio.

Independent Association of Serum Fibroblast Growth Factor 21 Levels With Impaired Liver Enzymes in Hyperthyroid Patients.

Fibroblast growth factor 21 (FGF21) is identified as a potential biomarker for liver diseases. However, information is limited regarding serum FGF21 and impaired liver function in hyperthyroidism. We aim to determine the potential association of serum FGF21 levels with impaired liver enzymes in hyperthyroid patients. In this case-control study, 105 normal subjects and 122 overt hyperthyroid patients were included. Among them, 41 hyperthyroid patients who obtained euthyroid status after thionamide treatment received second visit. Serum FGF21 levels were determined using the ELISA method. Compared to the normal subjects, patients with hyperthyroidism had significantly elevated serum liver enzymes, including alanine transaminase (ALT) (p < 0.001), aspartate aminotransferase (AST) (p < 0.001) levels, as well as FGF21 levels (p < 0.001). Further analysis showed serum FGF21 (p < 0.05), as well as thyroid hormone (TH) free T3 (p < 0.05), free T4 (p < 0.05) levels were higher in hyperthyroid patients with impaired liver enzymes than in those with normal liver enzymes. After reversal of hyperthyroid state, elevated serum FGF21 levels in hyperthyroid patients declined significantly (p < 0.001), with a concomitant decrease in serum ALT (p < 0.001), AST (p < 0.001) levels. Correlation analysis showed close correlation between FGF21 and ALT (p < 0.002), AST (p < 0.012), free T3 (p < 0.001), free T4 (p < 0.001). Further logistic regression analysis revealed FGF21 is significantly associated with elevated ALT [Odds Ratio, OR 1.79, (95% confidence interval, CI), (1.30-2.47), P < 0.001], AST [1.59 (1.07-2.34), p < 0.020]. After adjustment of potential confounders, the association between FGF21 and elevated ALT remained significant [1.42 (1.01-1.99), p < 0.043]. In conclusion, serum FGF21 is independently associated with impaired liver enzymes in hyperthyroid patients.

Effect of exenatide after short-time intensive insulin therapy on glycaemic remission maintenance in type 2 diabetes patients: a randomized controlled trial.

Early short-term intensive insulin (STII) therapy can induce drug-free glycemic remission for up to 1 year in half of newly diagnosed type 2 diabetic mellitus (T2DM) patients. Whether exenatide following STII therapy will induce higher long-term glycaemic remission is currently unknown. To assess the effect of STII+ exenatide therapy, compared with STII only, on maintenance of glycaemic remission in newly diagnosed T2DM patients. In this randomized, parallel-group, open-label, controlled trial, 129 patients (66 in STII+ exenatide group and 63 in STII only group) firstly completed 3-week STII therapy, then STII+ exenatide group was treated with exenatide for 12 weeks further. The cumulative probabilities of 1-year and 2-year glycaemic remission in STII+ exenatide group were 68.2 ± 5.7% and 53.0 ± 6.1%, which were significantly higher than STII only group (36.5 ± 6.1% and 31.8 ± 5.9%) (p-values < 0.001). Patients in STII+ exenatide group, compared with STII only group, showed significantly decreased levels of waist (82.2 (81.0, 83.5) cm v.s. 84.2 (82.7, 85.7) cm, p = 0.048) and HbA1c (5.83 (5.60, 6.06)% v.s. 6.49 (6.20, 6.77)%, p < 0.001) after 12-week exenatide treatment, but these differences disappeared after 1-year and 2-year follow-up. As conclusions, Improved effect of sequential exenatide after STII therapy on maintenance of glycaemic remission only occurred during exenatide treatment and lost upon treatment cessation.

Pycnodysostosis with novel gene mutation and sporadic medullary thyroid carcinoma: A case report.

RATIONALE: Pycnodysostosis is a rare autosomal recessive skeletal dysplasia caused by a mutation in the cathepsin K encoded by cathepsin K gene (CTSK). Medullary thyroid carcinoma (MTC) is also a relatively rare type of primary thyroid carcinoma. PATIENT CONCERNS: A 31-year-old woman presenting a short stature and a palpable nodule in the front of her neck that had gradually increased in size during the last 2 years was referred to our department. She has experienced multiple fractures at lower limbs in the last 2 decades. DIAGNOSES: The patient's clinical examination revealed short stature, underweight, a prominent forehead, stubby fingers, and a fixed nodule in the right thyroid lobe. Intraoral examination revealed multiple clinically malposed and missing teeth, as well as chronic periodontitis with a narrow and grooved palate. Radiographic examination revealed typical widely separated cranial sutures and an open anterior/posterior fontanel with an obtuse gonial angle, acroosteolysis, and osteosclerosis with narrowed medullary cavities. Ultrasonography of the thyroid gland showed a marked hypoechoic solid nodule in the right lobe in which tumor cell clusters were confirmed by ultrasound-guided fine needle aspiration biopsy and was suspected to be MTC. Laboratory tests revealed dramatically elevated serum calcitonin >2000 pg/L (reference range: 0-5 pg/L) and carcinoembryonic antigen (CEA) 134.37 ng/mL (reference range: 0-5 ng/mL). Genotypic screening revealed compound heterozygous mutations in the CTSK gene (c.158delA, P.Asn53Thr/c.C830T, P.Ala277Val) but no mutation associated with the familial forms of MTC. INTERVENTIONS: The patient underwent a total thyroidectomy with right-sided functional neck dissection. OUTCOMES: CEA and serum calcitonin decreased significantly postthyroidectomy, and no further fracture has been reported by the patient so far. LESSONS: The present study is the first to report a rare case of the coexistence of pycnodysostosis with a compound CTSK gene mutation and sporadic MTC. Radiological techniques and gene analysis play key roles in the definitive diagnosis.

Elevated Serum Fibroblast Growth Factor 21 Levels in Patients With Hyperthyroidism.

CONTEXT: Recent evidence from animal studies indicates that fibroblast growth factor 21 (FGF21), an endocrine hormone that regulates glucose, lipid metabolism, and energy homeostasis, is regulated by T3. However, the role of FGF21 in hyperthyroid patients is unknown. OBJECTIVE: The objective was to study serum FGF21 levels in hyperthyroid patients and the association of serum FGF21 levels with hyperthyroidism. DESIGN AND SETTING: This was a case-control study. PATIENTS AND INTERVENTIONS: A total of 119 hyperthyroid patients and 108 healthy subjects were recruited. Of them, 41 hyperthyroid patients received thionamide treatment for 3 months until euthyroidism was obtained. MAIN OUTCOME MEASURES: Serum FGF21 levels were determined using the ELISA method. RESULTS: Serum FGF21 levels were significantly elevated in hyperthyroid patients as compared with normal subjects [median 290.67 (interquartile range, 156.60-502.33) vs 228.10 (169.85.25-320.10) pg/mL; P < .001]. After thionamide treatment, serum FGF21 levels in hyperthyroid patients declined markedly from 249.10 (139.10-444.00) to 106.90 (38.70-196.15) pg/mL (P < .001). Logistic regression revealed that FGF21, basal metabolic rate, low-density lipoprotein cholesterol, and alanine transaminase were significantly associated with hyperthyroidism. With adjustment for potential confounders, serum FGF21 remained independently associated with hyperthyroidism, with an adjusted odds ratio of 3.123 (95% confidence interval, 1.306-7.468) (P = .010). CONCLUSION: Serum FGF21 levels were elevated in patients with hyperthyroidism and declined after thionamide treatment. And serum FGF21 level was independently associated with hyperthyroidism.

Functional interleukin-17 receptor A are present in the thyroid gland in intractable Graves disease.

Th17 lymphocytes and its main cytokine, IL-17, play an important role in autoimmune thyroid diseases, such as intractable Graves disease (GD). IL-17 signals are transmitted through its receptor, IL-17RA. The intrathyroid expression of IL-17RA in intractable GD is not understood. In this study, ELISA was used to measure serum IL-17 levels in patients with untreated GD, intractable GD or GD in remission and healthy controls. Real-time PCR, flow cytometry and immunofluorescence staining evaluated IL-17RA mRNA and protein expression in thyrocytes. IL-6, chemokine ligand 10 (CXCL10) and intercellular adhesion molecule (ICAM)-1 expression was measured in IL-17-stimulated thyrocyte cultures to evaluate the functional status of IL-17RA. Our data indicates that serum IL-17 levels are significantly increased in intractable GD and affected thyrocytes show functional IL-17R expression. These changes facilitate the IL-17-mediated upregulation of IL-6, CXCL10, and ICAM-1. The IL-17/IL-17R interaction could be a potential target for therapeutic interventions in intractable GD.

385 C/A polymorphism of the fatty acid amide hydrolase gene is associated with metabolic syndrome in the Chinese Han population.

INTRODUCTION: The endocannabinoid system participates in food intake, energy balance and lipid and glucose metabolism. The biological effects of cannabinoids are limited by the activation of the endocannabinoid degrading enzyme fatty acid amide hydrolase (FAAH). This study aims to analyse whether 385 C/A polymorphism of FAAH is associated with metabolic syndrome (MetS) in the Chinese Han population. MATERIAL AND METHODS: A total of 112 subjects at risk for MetS and 80 healthy controls from Fuzhou, China were genotyped for 385 C/A polymorphism of FAAH using TaqMan assay. Anthropometric measurements and biochemical assessments such as BMI, waist circumference, blood pressure, serum triglycerides (TG), serum total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, fasting plasma glucose, and plasma insulin levels were performed. RESULTS: CA and AA genotypes of FAAH had higher incidence in MetS subjects than in control subjects. CA and AA genotypes of FAAH in subjects with MetS had relatively elevated levels of waist circumference, body mass index, homeostasis model assessment of insulin resistance (HOMA-IR) and serum triglycerides, and lowered level of high-density lipoprotein cholesterol (HDL-c) compared with CC genotype in MetS subjects. CONCLUSIONS: Results suggest that 385 C/A polymorphism of the FAAH gene may confer an increased risk of MetS in the Chinese Han population.

Identification of HLA class I dependent immunogenic peptides from clonotypic TCRbeta expressed in cutaneous T-cell lymphoma.

The clonotypic T-cell receptor (TCR) is a potential target antigen for specific immunotherapy of cutaneous T-cell lymphoma (CTCL). We identified T-cell epitopes from the rearranged TCR beta chain of the malignant T-cell population by the "reverse immunology" approach. Peptide-specific T-cell lines were generated against predicted epitopes and tested for the recognition of tumor cells and cells transfected with the full-length DNA coding for TCRV beta chain. Two peptides derived from the clonotypic TCRVbeta of a HLA-A2 positive patient could induce peptide-specific T cells from peripheral blood mononuclear cells of healthy donors and the patient as assessed by IFN-gamma ELISpot assay. Furthermore, the reactive CTLs efficiently recognized autologous Sézary tumor cells, as well as HLA-A2 positive 293 cells transfected with recombinant plasmid expressing the corresponding TCRVbeta29 protein. Similar results were obtained in a HLA-A3+ patient for TCRVbeta7-Jbeta2.7. In conclusion, our experiments show that the TCR beta chain harbors epitopes suitable as targets for specific vaccination which might be a promising approach for the specific immunotherapy of cutaneous T-cell lymphoma patients.

High cell surface expression of Newcastle disease virus proteins via replicon vectors demonstrates syncytia forming activity of F and fusion promotion activity of HN molecules.

For functional studies, the hemagglutinin-neuraminidase (HN) and the fusion protein (F) of Newcastle disease virus (NDV) were expressed in BHK cells using two vectors which are based on the Semliki Forest virus (SFV) replicon. The first system of high protein expression works by transfection of RNA which before has been in vitro transcribed from a vector containing the gene for the SFV self-amplifying replicase (REP) and a foreign gene using the SP6 promoter. A high level of protein (HN or F) expression was detected 18-20 h after transfection. To study the host range of this expression system, a panel of different cell lines were compared for transfections with SFV RNA. A wide range of expression efficiency was observed, the highest being BHK cells. The second system is based on a DNA plasmid in which the SFV-REP and a foreign gene are expressed in cells under the transcriptional and translational control of the cytomegalovirus immediate-early enhancer T7 promoter. DNA-electroporated BHK cells expressed also high levels of the recombinant proteins but at a delayed time point (40-48 h) as compared with the corresponding RNA. Co-expression of the two NDV proteins, HN and F, via this DNA vector in the same cells led to syncytium formation in the cell monolayer, showing that both proteins expressed in this way, were functionally active. F alone, expressed via this vector, displayed residual fusion activity suggesting its proteolytic cleavage and its functional independence from HN.

Importance of serine 200 for functional activities of the hemagglutinin-neuraminidase protein of Newcastle Disease Virus.

Newcastle disease virus (NDV) is an avian paramyxovirus with replication competence in human tumor cells and interesting anti-neoplastic and immune stimulatory properties. In order to increase tumor selectivity of replication, we prepared mutants from the avirulent strain Ulster with monocyclic replication cycle and adapted them for multicyclic replication in human melanoma cells. Two mutants (M1 and M2) showed interesting functional differences: while M2 showed T cell co-stimulatory effects in a tumor-specific cytotoxic T lymphocyte (CTL) assay, M1 did not. A distinct difference of these 2 virus mutants appeared also when testing their capacity to induce interferon-alpha and -beta as well as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) molecules in human monocytes. Sequence analysis of the hemagglutinin-neuraminidase (HN) molecules of the 2 virus mutants showed 7 non-silent mutational differences. Upon cloning of the HN mutant genes into an expression vector and transfection of cells, only HN derived from M2 (HN-M2) was detected at the cell surface by immunostaining with specific antibodies and showed hemadsorption and neuraminidase activity. In order to define which amino acid was responsible for the loss of functional activity of HN derived from M1 (HN-M1), distinct HN mutants were generated via site-directed mutagenesis and tested. Substitution of serine 200 by a proline abrogated HN expression and its hemadsorption and neuraminidase activities. Molecular modeling revealed that proline 200 in HN influences flexibility of a loop near the entrance to the neuraminidase active site, a function that may be crucial for the functions of this viral protein.

Two ways to induce innate immune responses in human PBMCs: paracrine stimulation of IFN-alpha responses by viral protein or dsRNA.

In order to study mechanisms of induction of IFN-alpha by Newcastle disease virus (NDV), we used two replicon systems which are based respectively on DNA and RNA of the Semliki forest virus (SFV) and transfected these into baby hamster kidney cells (BHK) which do not produce interferon-alpha. Co-incubation of BHK cells which were transfected with the two vector systems, with human PBMCs, showed that production of IFN-alpha takes place by two different ways. When using the DNA-based SFV vector, only transfectants expressing cell surface HN molecules of NDV (and not the mock-transfected cells) elicited such a response via interaction of these HN molecules with viral receptors on PBMCs. In contrast, BHK cells transfected with RNA which had been in vitro transcribed from the RNA-based SFV vector without foreign gene as insert (mock-transfected) elicited a comparable IFN-alpha response. Transfer by transfection of poly(I:C), an analogue of double stranded RNA (dsRNA), into the BHK cells induced also by itself the production of IFN-alpha. Therefore induction of "danger signals" (as double-strand RNA replicative intermediates) might be responsible for this discrepancy observed in IFN-alpha induction in PBMCs between the two studied SFV vector systems based on transfection of DNA and on RNA. These observations highlight two ways of IFN-alpha induction which additively may explain the high interferonogenic capacity of NDV as virus: i) via cell-surface expressed HN after transfection of the DNA-based SFV replicon and ii) via transfection of self-amplifying RNA.