To measure relative expression level of mRNA in a small number of cultured rat thyroid cells (FRTL-5), we developed a system of a quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay. Human thyroglobulin mRNA in total RNA extracted from a human thyroid tissue was used as an internal control. FRTL-5 cells in a 24 well dish were lysed with denaturing solution containing human RNA. Total RNA was extracted followed by reverse transcription and polymerase chain reaction. After digestion with a restriction enzyme, PCR products were separated by electrophoresis and stained with Sybr Green I, then their fluorescence was measured with fluorescent image analyser. Increase of thyroglobulin mRNA in FRTL-5 cells stimulated by thyroid stimulating hormone (TSH) was observed by this technique. Because this method does not require a large number of cells or radioactive isotopes, it is as useful for the analysis of the relative expression level of mRNAs in the cells as the conservative methods such as Northern Blot.

Animals↗

PubMedDate not supplied

The spectrum of postpartum thyroid dysfunction: diagnosis, management, and long-term prognosis.

OBJECTIVE: To provide an overview of thyroid abnormalities that can occur after delivery. METHODS: We review the diagnosis and management of various types of postpartum thyroid dysfunction. RESULTS: A common problem during the postpartum period, thyroid dysfunction is found in approximately 5% of new mothers in the general population. Such thyroid dysfunction can manifest in various forms (hyperthyroid or hypothyroid, transient or persistent), each of which necessitates different treatment. The most common type of thyroid dysfunction is destructive thyrotoxicosis, which is due to the aggravation of autoimmune thyroiditis. It occurs at 1 to 3 months after delivery as sudden thyrotoxicosis and lasts for 1 to 3 months, after which transient hypothyroidism may ensue in some cases. Simply symptomatic treatment may be sufficient for such transient thyroid dysfunctions. Postpartum Graves' disease is found in approximately 11% of the cases of postpartum thyroid dysfunction. Graves' hyperthyroidism usually occurs at 3 to 6 months after parturition. In the treatment of postpartum Graves' disease, one may choose radioactive iodine, antithyroid drugs, or surgical therapy; however, transient Graves' hyperthyroidism, which often occurs in patients with postpartum Graves' disease, may require neither radioactive iodine therapy nor thyroidectomy. Mothers at high risk for development of postpartum thyroid dysfunction can be screened by testing for antimicrosomal antibodies. Although a strong correlation exists between the presence of antimicrosomal antibodies and postpartum thyroid dysfunction, the cost-effectiveness of screening for postpartum autoimmune thyroid syndrome remains controversial. CONCLUSION: For optimal management of the postpartum conditions of Graves' disease, destructive thyrotoxicosis, and hypothyroidism, the clinician should be aware of diagnostic techniques that will facilitate early intervention or will prompt careful surveillance.

Journal Article↗

PubMedDate not supplied

Cost-effective enzymatic method for simultaneous determination of urinary creatine and creatinine using automatic analyzer.

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Chemistry, Clinical↗