Construction of a mutation due to a fourteen base-pair insertion in HNF-1α gene causing maturity-onset diabetes of the young (MODY) in Thai patients.

Objective: The aim of this study is to generate a mutation causing maturity-onset diabetes of the young (MODY) in Thai patients by insertion of a fourteen base-pair (bp) into a HNF-1a gene using a modified site-directed ligase-independent mutagenesis (SLIM) method. Methods: Two pairs of long- and short-tailed primers were designed to amplify a plasmid construct containing a HNF-1a and to insert a 14-bp at a desired position. Long-tailed primers contained the overhanging 14-nucleotide (nt) insert at their termini which were complementary to each other. Polymerase chain reactions (PCR) were performed in two separated tubes using different pairs of primers. After amplifications, PCR products from both tubes were pooled together, denatured and then re-annealed to allow formation of double stranded DNA molecules containing the 14-bp insert within HNF-1a. The pooled and reannealed PCR products without ligation were transformed into competent E.coli cells to generate a ligated recombinant plasmid with a 14-bp insertion. Results: Five of 14 bacterial colonies contained the desired recombinant plasmid with a 14-bp insertion within HNF-1a The efficiency of the method for generation of recombinant plasmid was about 36 percent. Conclusion: This method is simple and rapid to insert a long stretch of nucleotides into a plasmid construct containing a gene of interest at a desired position. A recombinant plasmid containing an insertion mutation in a HNF-1a gene was successfully generated, allowing an opportunity to perform functional study of the mutated gene.

Primary hyperparathyroidism due to cystic parathyroid adenoma: a case report.

Raw Cystic parathyroid adenoma is a rare cause of primary hyperparathyroidism. The authors report one case of cystic parathyroid adenoma, who presented with progressive right hip pain for one year. The patient had severe hypercalcemia at the first presentation and was misdiagnosed as having metastatic cancer at first. An iliac bone biopsy was performed and showed a giant cell tumor. Parathyroid hormone level was evaluated later and was found to be high, 1,555 pg/ml (15-65 pg/ml). An MRI study of the neck was done and revealed a cystic mass 38 x 36 x 40 mm in diameter just below the left lower pole of the thyroid gland. Tc-99m MIBI scan demonstrated increase and retention of radioactivity uptake at the same area. Hyperfunctioning parathyroid gland was considered. Parathyroidectomy was done and histopathology revealed cystic parathyroid adenoma. Serum calcium was normal and hip pain was markedly improved after the surgery.

Thailand diabetes registry project: type of diabetes, glycemic control and prevalence of microvascular complications in children and adolescents with diabetes.

OBJECTIVE: To determine the etiology, glycemic control and prevalence of microvascular complications in Thai diabetic patients who were diagnosed at the age of less than 18 years and who attended diabetes clinics in university or tertiary care hospitals. MATERIAL AND METHOD: A cross-sectional, multi-center, hospital-based diabetes registry was carried out from diabetes clinics of 11 tertiary centers. Demographic data including laboratory results and microvascular complications were recorded. RESULTS: Two-hundred-and-fifty out of the 9419 (2.66%) diabetic patients were diagnosed before the age of 18 years, 78% had Type 1 diabetes (T1DM), 18.4% had Type2 diabetes (T2DM) and 3.6% had other types of diabetes. Mean +/- SD HbAlc of T1DM was 9.3 +/- 2.5, T2DM was 9.7 +/- 2.6 and other types of diabetes were 8.6 +/- 4%. The majority of patients had poor glycemic control according to ADA and WHO guidelines. The percentage of patients who had diabetes for more than 5 years but had not been screened for nephropathy and retinopathy were 57.7% and 16% in T1DM and were 46.4% and 14.2% in T2DM respectively. A significant correlation between microvacular complications (nephropathy and retinopathy) and duration of disease was found in T1DM (p < 0.001). CONCLUSION: The majority of Thai children and adolescents with diabetes had TIDM and unsatisfactory glycemic control. Screening for microvascular complications was under international standard. The national strategic plan for management of this disease especially in this age group should be urgently implemented.