OBJECTIVE: To establish an cell model of hyperparathyroidism by isolation, in vitro culture, and identification of parathyroid cells from patients with secondary hyperparathyroidism (SHPT). METHODS: The parathyroid gland tissues obtained from 10 patients with SHPT were dissociated by collagenase digestion for primary culture of the parathyroid cells. Morphological changes and growth characteristics of the cells were assessed by microscopic imaging and cell counting. The mRNA and protein expression levels of parathyroid hormone (PTH), calcium-sensing receptor (CaSR), and glial cells missing 2 (GCM2) in the primary and passaged cells were determined by immunofluorescence, qRT-PCR, and Western blotting. RESULTS: Primary cultures of parathyroid cells were successfully obtained. The cells exhibited a high expression of PTH shown by immunofluorescence assay and had a population doubling time of approximately 71.61 h. PTH secretion in the second-passage (P2) cells was significantly lower than that in the primary (P0) and first-passage (P1) cells (P < 0.001). Despite a significant downregulation of CaSR mRNA (P=0.017) and protein (P=0.006) in P1 cells as compared with P0 cells, no significant differences were found in mRNA and protein expressions of PTH or GCM2 between the two cell generations. CONCLUSION Primary cultures of parathyroid cells isolated from SHPT patients by collagenase digestion show similar biological properties to the cells in vivo.
Humans ; Hyperparathyroidism, Secondary/metabolism* ; Parathyroid Glands/metabolism* ; Parathyroid Hormone ; RNA, Messenger/metabolism* ; Receptors, Calcium-Sensing/metabolism*

Calcium and phosphorus balance studies were carried out in subtotally nephrectomized rats (NX), with or without parathyroidectomy (NX-PTX; NX sham-PTX) in order to determine the ability of the remnant kidney to regulate excretion of these elements in the absence of parathyroid hormone. The rats were fed three different phosphorus diets, and calcium intake was also varied. We found that the NX-PTX rats adapted to the three different phosphorus diets in a manner indistinguishable from the NX sham-PTX rats. The per cent of ingested phosphorus excreted in the urine increased as dietary phosphorus increased. When supplementary calcium was added to the diet urinary phosphorus excretion fell and fecal phosphorus increased, in an identical fashion in the NX-PTX and NX sham-PTX animals. Urinary calcium excretion decreased as dietary phosphorus increased, and UCaV increased when supplementary calcium was provided in the diet. Total body calcium and phosphorus balance (intake-(urine+feces)) varied with intake, but was not significantly different between the NX-PTX and NX sham-PTX rats. These experiments demonstrate that subtotally nephrectomized rat have a parathyroid-independent mechanism(s) for regulating both urinary and fecal calcium and phosphorus excretion. The mechanism is not revealed by the present study, but may relate to changes in serum calcium and/or phosphorus which occur following parathyroidectomy.
Animal ; Calcium/metabolism* ; Homeostasis* ; Kidney/physiology ; Male ; Nephrectomy* ; Parathyroid Glands/surgery* ; Parathyroid Hormones/physiology ; Phosphorus/metabolism* ; Rats ; Rats, Inbred Strains

Primary hyperparathyroidism is a generalezed disorder of calcium, phosphorus and bone metabolism due to an increased secretion of parathyroid hormone. Single parathyroid adenoma is the most common cause of primary hyperparathyroidism. Because parathyroid hormone has been proposed as an important inhibitor of renal bicarbonate reabsorption of proximal tubule, proximal renal tubular acidosis is not rare in primary hyperparaphyroidism. After parathyroid resection, significant hypocalcemia and hypophosphatemia requiring prolonged medical management may develop, termed hungery bone syndrome. We experienced a case of primary hyperparathyroidism associated with proximal renal tubular acidosis, and severe hungry bone syndrome after resection of the adenoma of parathyroid gland.
Acidosis ; Acidosis, Renal Tubular ; Adenoma ; Calcium ; Hyperparathyroidism, Primary ; Hypocalcemia ; Hypophosphatemia ; Kidney Tubules, Proximal ; Metabolism ; Parathyroid Glands ; Parathyroid Hormone ; Parathyroid Neoplasms ; Phosphorus

The enlarged parathyroid glands associated with chronic renal failure were recognized during the 1930's. The number of patients on long-term hemodialysis due to chronic renal failure is steadily increasing and the hyperparathyroid state certainly became a clinical problem in the dialysis population. The physiologic mechanisms leading to secondary hyperparathyroidism are multifactorial with renal phosphate retention, skeletal resistance to parathyroid hormone (PTH) action and impairment vitamin D metabolism being some of the known factors. Despite intensive medical management however inadequate control of parathyroid hyperplasia may necessitate surgical intervention. The goal of surgical therapy is to resect sufficient tissue to reverse the hyperparathyroidism without rendering the patient permanently hypoparathyroidism. We experienced a case of secondary hyperparathyroidism and reported its result of total parathyroidectomy, autogenous transplantation and cryopreservation of parathyroid gland.
Cryopreservation ; Dialysis ; Humans ; Hypercalcemia ; Hyperparathyroidism ; Hyperparathyroidism, Secondary* ; Hyperplasia ; Hypoparathyroidism ; Kidney Failure, Chronic ; Metabolism ; Parathyroid Glands ; Parathyroid Hormone ; Parathyroidectomy ; Renal Dialysis ; Renal Osteodystrophy ; Vitamin D

No abstract available.
Aged ; Biological Markers/blood ; Biopsy ; Calcium/blood ; Female ; Humans ; Hyperparathyroidism/blood/diagnosis/*etiology/surgery ; Hyperplasia ; Parathyroid Glands/metabolism/*pathology/radionuclide imaging/surgery/ultrasonography ; Parathyroid Hormone/blood ; Parathyroid Neoplasms/blood/*complications/diagnosis/surgery ; Parathyroidectomy ; Treatment Outcome ; Up-Regulation

Adult ; Aged ; Female ; Humans ; Hyperparathyroidism, Secondary ; metabolism ; pathology ; Immunohistochemistry ; Male ; Middle Aged ; Parathyroid Glands ; chemistry ; RNA, Messenger ; analysis ; Receptors, Calcium-Sensing ; analysis ; genetics ; Uremia ; metabolism

Unlike most secretory cells, high extra cellular calcium inhibits rather than stimulates hormonal secretion in several cells such as parathyroid cells, Juxtaglomerular cells and osteoclast. To gain further insight into the common but unique stimulus-secretion coupling mechanism in these cells, bovine parathyroid slices were incubated in various conditions of Krebs-Ringer (KR) solution containing essential amino acids. Parathyroid cells showed the inverse dependency of secretion on extra cellular calcium concentration as we expected. Ammonium acetate overcame the inhibitory effect of 2.5 mM of calcium and the maximum effect was as much as the five times of the basal value, while there was a little additive effect under 0 mM CaCl2. PTH secretion was biphasic according to the change of extra cellular osmolarity and the lowest response was observed at 300 mOsm/l. In Na-rich KR solution, high concentration of nigericin (> 10(-4)M) completely overcame the inhibitory effect of 2.5 mM CaCl2 and the maximum stimulatory effect was 8 times greater whereas it was only 2 times greater without CaCl2. In K-rich KR solution that abolished the K-gradient between the extra cellular solution and the cytoplasm, the rate of PTH secretion increased, and furthermore the addition of nigericin increased the rate of secretion significantly. The results above suggested that the osmotic swelling of the secretory vesicle in parathyroid cells might promote exocytosis as in Juxtaglomerular cells. We propose that the swelling of the vesicle is also prerequisite for secretion in several cells inhibited paradoxically by Ca++, whatever the signal transduction pathway for swelling of the secretory granules induced by the lowering of Ca++ in cytoplasm are.
Acetates/pharmacology ; Animal ; Body Fluids/*metabolism ; Cattle ; Cell Membrane Permeability ; Ionophores/pharmacology ; Mannitol/pharmacology ; Nigericin/pharmacology ; Osmosis ; Parathyroid Glands/drug effects/*secretion ; Parathyroid Hormones/*secretion ; Support, Non-U.S. Gov't

Parathyroid hormone related protein (PTHrP) has important biological functions in calcium metabolism. The aim of this study was to silence the expression of PTHrP by RNA interference and recombinant adenovirus, and to provide a material to investigate the relative functions of PTHrP in goat mammary gland epithelial cell. The Block-iT shRNA interference system was used in this experiment. We designed and synthesized two pairs of complementary single-strand DNA oligonucleotides (shRNA-322/357) targeting two different sites of PTHrP mRNA. Then the oligonucleotides were inserted into shuttle vector pENTR/CMV-GFP/U6. After detection of the interference efficiency by Western blotting, we chose pENTR/CMV-GFP/U6-322 and adenovirus backbone vector pAD/PL-DEST to produce recombinant vector pAD/PL-DEST/CMV-GFP/U6-322. The first generation recombinant adenovirus particles (AD-PTHrP-322) were produced and further amplified by transfecting HEK-293 cells. The titer of the recombinant adenovirus reached 2.0 x 1(9) PFU/mL determined by TCID50 assays. The result of real-time quantitative PCR indicated that mRNA expression levels of gene were reduced 29.2%, 68.1% and 82.6% (P < 0.05), respectively, when goat mammary gland epithelial cells were infected with AD-PTHrP-322 after 24, 48 and 72 h, in which PTHrP. Western blotting also showed that the expression of PTHrP was reduced by infecting the cells with AD-PTHrP-322. AD-PTHrP-322 has been proved with significant interference effect on expression of PTHrP.
Adenoviridae ; genetics ; metabolism ; Animals ; Epithelial Cells ; metabolism ; Female ; Genetic Vectors ; genetics ; Goats ; HEK293 Cells ; Humans ; Mammary Glands, Animal ; cytology ; Parathyroid Hormone-Related Protein ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Transfection

Multiple endocrine neoplasia type 1 (MEN1) syndrome includes varying combinations of endocrine and non-endocrine tumors. There are also a considerable number of atypical MEN1 syndrome. In this case, a 68-yr-old woman was referred to the Department of Endocrinology for hypercalcemia. Five years ago, she had diagnosed as primary hyperaldosteronism and now newly diagnosed as parathyroid hyperplasia with laboratory and pathologic findings. Hurthle-cell thyroid cancer was also resected during the parathyroid exploration and small meningioma was found on brain MRI. Her general condition has markedly improved and her adrenal mass and meningioma are being closely observed now. We could find the loss of heterozygosity of the MEN1 locus in parathyroid glands, suggesting a MEN1-related tumor, but not a germline mutation. Considering a variety of phenotypic expression and a limitation of current molecular analysis, periodic follow up will be needed in patients with a MEN1-like phenotype.
Aged ; Base Sequence ; Brain/radionuclide imaging ; Female ; Humans ; Hyperaldosteronism/complications/*diagnosis ; Hyperparathyroidism, Primary/*diagnosis/etiology/pathology ; Loss of Heterozygosity ; Magnetic Resonance Imaging ; Meningeal Neoplasms/complications/*diagnosis/radionuclide imaging ; Meningioma/complications/*diagnosis/radionuclide imaging ; Mutation ; Parathyroid Glands/pathology ; Proto-Oncogene Proteins/genetics/metabolism ; Sequence Analysis, DNA ; Thyroid Neoplasms/complications/*diagnosis/pathology ; Tomography, X-Ray Computed