MicroRNA-29a and microRNA-122 expressions and other inflammatory markers among obese children with diabetes.

OBJECTIVES: This study was conducted to study the expression of both microRNA-29a and microRNA-122, and serum levels of sestrin-2, interleukin-6 (IL-6), and other inflammatory markers among obese children with/and without diabetes mellitus. METHODS: One hundred obese children with diabetes in addition to 100 age- and sex-matched obese children without diabetes, and 100 age- and sex-matched apparently healthy children were included in the study. Expressions of both microRNA-29a and microRNA-122, and serum levels of sestrin-2, IL-6, tumor necrosis factor-α (TNF-α), and high sensitive-CRP (hsCRP) were measured for all included study populations. RESULTS: Study results showed that the expressions of both microRNA-29a and microRNA-122, serum levels of IL-6, TNF-α, and hsCRP were significantly higher among obese children with diabetes in comparison to both obese children without diabetes and healthy children. In contrast, serum sestrin level was significantly low among obese children with diabetes in comparison to the other study populations. Expressions of both microRNA-29a and microRNA-122 were correlated with waist circumference, BMI, total cholesterol, triglycerides, LDL-cholesterol, HbA1c, c-peptide, glucose, insulin, homeostatic model assessment-insulin resistance (HOMA-IR), IL-6, hsCRP, and TNF-α among obese children with diabetes. However, serum sestrin-2 level was correlated inversely with these parameters. Higher expressions of both microRNA-29a and microRNA-122 among obese children either with or without diabetes mellitus (DM) can suggest their roles in the development of obesity among children. CONCLUSIONS: The study results can hypothesize that down-regulation of these micro-RNAs may solve this health problem with its sequelae, a hypothesis that needs more studies.

Efficacy of Scintigraphy, Ultrasound and Both Scintigraphy and Ultrasonography in Preoperative Detection and Localization of Primary Hyperparathyroidism.

OBJECTIVE: The aim of our study was to evaluate the efficacy of preoperative dual-phase 99mTc-methoxyisobutylnitrile (MIBI) parathyroid scintigraphy (PS), and ultrasound (US) in primary hyperparathyroidism (pHPT) diagnosis and compare the results with the surgical findings. METHODS: Forty-five patients were enrolled in this study. Preoperative serum parathyroid hormone (PTH) levels, calcium (Ca), phosphate (P), and alkaline phosphatase (AP) levels were measured. All parathyroid patients were evaluated by ultrasonography, dual phase 99mTc-MIBI. Surgical findings were used as a reference standard. RESULTS: Of the 45 patients included in this study, 30 were females (66.7%) with an age range between 30 years and 70 years (mean age 41± 13). The sensitivity and specificity of 99mTc-MIBI scintigraphy was 97.4% and 71.4%, respectively, while the sensitivity of ultrasound was 94.4% and specificity 44.4%. The sensitivity, specificity, and accuracy of combined scintigraphy and ultrasound was higher-97.4%, 83.3%, and 95.6%, respectively. CONCLUSIONS: The combination of MIBI and US appears promising for localizing parathyroid pathology in patients with primary hyperparathyroidism. The concordance rate is high together with a lower chance of missing concomitant thyroid pathology, which might alter the surgical approach.

Pediatric Differentiated Thyroid Cancer: Our Experience.

OBJECTIVES: To report our experience in the management of thyroid cancer in children and adolescents in a tertiary referral hospital and regional cancer institute as compared to previously published data. METHODS:  A retrospective study was conducted for patients diagnosed with differentiated thyroid cancer (DTC) who received treatment during the period from January 2014 to August 2018. Medical reports from our hospital database were extracted and information of those under 18 years old were discussed regarding their demographics, treatment received, and follow-up outcomes. RESULTS:  Out of 300 patients with DTC diagnosed in the period of study, 12 were 18 years old or less (4%). Female to male ratio was 5:1. Their ages ranged from nine to 18 years old (average: 13.1 years). One patient had a positive family history for DTC, and one patient had lung metastasis. Total thyroidectomy and postoperative 131I were performed for all patients. The median follow-up period was 1.75 years (range: six months to four years). Eleven patients have shown complete remission after treatment (91.6%), and one case has had persistent disease. CONCLUSIONS:  Pediatric thyroid cancer is not uncommon. Despite its aggressiveness in this age group, outcomes are more favorable than in adults. We report our experience in the diagnosis and management of pediatric DTC in our community with satisfactory outcomes and comparable results to literature reports. Future studies are needed to evaluate the long-term complications of radioiodine therapy.

Tetracycline-regulated expression enables purification and functional analysis of recombinant connexin channels from mammalian cells.

Intercellular coupling mediated by gap junction channels composed of connexin protein underlies numerous physiological processes, such as cellular differentiation, tissue synchronization and metabolic homoeostasis. The distinct molecular permeability of junctional channels composed of different connexin isoforms allows cellular control of coupling via regulation of isoform expression. However, the permeability properties of most connexin isoforms have not been well characterized due to the difficulty of manipulating and measuring the diffusible concentrations of cytoplasmic messenger molecules and metabolites, and to a lack of control over channel isoform composition, in vivo. Here we present a method to express and purify active connexin hemichannels of a single isoform or a consistent ratio of two isoforms from cultured cells using the Tet-On inducible expression system and one-step anti-haemagglutinin immunoaffinity purification. The procedure yields 10-20 microg of pure connexin protein from 2.5x10(8) HeLa cells. The purified channels are shown to be useful for in vitro permeability analysis using well established techniques. This method has substantial advantages over existing methods for heterologous connexin expression, such as the ease of co-expression of two isoforms at a constant ratio, consistently high expression levels over many passages, and the ability to study channel properties in situ as well as in purified form. Furthermore, the generic cloning site of the new pBI-GT vector and the commercial availability of anti-haemagglutinin (clone HA-7)-agarose make this affinity tagging and purification procedure easily applicable to other proteins.