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*

Among many causes of calcium containing urinary stones, the hypercalciuria associated with normocalcemia is the most common one. Absorptive hypercalciuria and renal hypercalciuria constitute two major forms of idiopathic hypercalciuria. According to Coe et al., a renal leak of calcium and secondary hyperparathyroidism are present in renal hypercalciuria but not in absorptive hypercalciuria. It has been suggested that fasting urinary cAMP may provide a measure of parathyroid function. So determination of fasting urinary calcium and cAMP may be useful in the differentiation of the two major forms of idiopathic hypercalciuria. We performed the study to evaluate the significance of urinary calcium and cAMP in the determinating the types of hypercalciuria. Calcium metabolism studies and urinary cAMP determination were done in 28 in-patients with urinary stones under usual diet and fasting state. The results were as followings: 1. Among 28 patients, 15 patients had renal hypercalciuria, 6 had absorptive hypercalciuria and 7 had normocalciuria. 2. On usual diet, 24-hour urine cAMP level was 3.815+/-0.560 nmol/mg creatinine in renal hypercalciuria, 3.508+/-1.253 nmol/mg creatinine in absorptive hypercalciuria and 3.202+/-0.980 nmol/mg creatinine in normocalciuric patients. 3. On fasting state, 2-hour urine cAMP level was 4.611+/-2.591 nmol/mg creatinine in renal hypercalciuria, 4.304+/-3.173 nmol/mg creatinine in absorptive hypercalciuria and 2.400+/-1.080 nmol/mg creatinine in normocalciuric patients. 4. urinary cAMP on usual diet and fasting condition did not differ significantly among 15 renal and 6 absorptive hypercalciuric patients. (t-value; 0.82 and 0.22, respectively, p>0.1)
Calcium ; Creatinine ; Diet ; Fasting ; Humans ; Hypercalciuria ; Hyperparathyroidism, Secondary ; Metabolism ; Urinary Calculi*

OBJECTIVE: To identify the miRNAs targeting vitamin D receptor (VDR) gene and their effect on parathyroid hormone (PTH) secretion in secondary hyperparathyroidism. METHODS: Primary parathyroid cells with secondary hyperparathyroidism were isolated by collagenase digestion and cultured. The miRNAs targeting VDR were screened by bioinformatics methods and full transcriptome sequencing, and dual-luciferase reporter assay was used to verify the targeting relationship between VDR and the screened miRNA. The effects of overexpression or inhibition of the candidate miRNA on VDR mRNA and protein expressions and PTH secretion were evaluated using qRT-PCR and Western blotting. The expression levels of the candidate miRNAs and VDR mRNA in clinical specimens of parathyroid tissues were verified by qRT-PCR, and the expression of VDR protein was detected by immunohistochemistry. RESULTS: We successfully isolated primary parathyroid cells. Dual-luciferase reporter assay verified the targeting relationship of hsa-miR-149-5p, hsa-miR-221-5p, hsa-miR-222-3p, hsa-miR-29a-5p, hsa-miR-301a-5p, hsa-miR-873-5p, hsa-miR-93-3p with VDR, and among them, the overexpression of hsa-miR-149-5p and hsa-miR-301a-5p significantly increased PTH secretion in the parathyroid cells. In patients with secondary hyperparathyroidism, hsa-miR-149-5p was highly expressed in the parathyroid tissues (P=0.046), where the expressions of VDR mRNA (P=0.0267) and protein were both decreased. CONCLUSION The two miRNAs, hsa-miR-149-5p and hsa-miR-301a-5p, may promote the secretion of PTH in patients with secondary hyperparathyroidism by down-regulating the expression of VDR gene.
Humans ; Hyperparathyroidism, Secondary/genetics* ; MicroRNAs/metabolism* ; Parathyroid Hormone ; RNA, Messenger ; Receptors, Calcitriol/genetics*

PURPOSE: It is known that secondary hyperparathyroidism in end stage renal disease (ESRD) patients is associated with vitamin D receptor (VDR) gene polymorphism, but there is no consensus on its genotype. There is lack of data in Ca, P, calcitriol and VDR polymorphism. METHODS: We measured serum Ca, P, alkaline phosphatase, parathyroid hormone (PTH), and 1,25 (OH)2D3 of the 53 hemodialysis patients. The genotypes of VDR were classified BB, Bb, bb according to restriction patterns in PCR of the patients' DNA using Bsm I restriction enzyme. RESULTS: The patients with BB, Bb, bb type were 0 (0%), 15 (28.3%), 38 (71.7%) respectively. Serum PTH levels were 70.0+/-63.3 pg/mL and 146.9+/-184.9 pg/mL in Bb, bb type respectively, and showed significant statistical difference (p<0.05). Serum 1,25 (OH)2D3 levels were 7.68+/-3.41 pg/mL and 6.59+/-2.67 pg/mL in Bb and bb genotype respectively without statistical significance. And there was no significant statistical differences among the serum levels of calcium, phosphorus, alkaline phosphatase. CONCLUSION: Vitamin D receptor gene polymorphism is associated with secondary hyperparathyroidism in hemodialysis patients, and the b allelle is suggestive of poorer bone mineral metabolism.
Alkaline Phosphatase ; Calcitriol ; Calcium ; Consensus ; DNA ; Genotype ; Humans ; Hyperparathyroidism ; Hyperparathyroidism, Secondary* ; Kidney Failure, Chronic ; Metabolism ; Parathyroid Hormone ; Phosphorus ; Polymerase Chain Reaction ; Receptors, Calcitriol* ; Renal Dialysis* ; Vitamin D* ; Vitamins*

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

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

Hypercalcemia rarely develops in patients with acute renal failure associated with rhabdomyolysis. If hypercalcemia occurs, it happens mostly in diuretic phase and rarely in oliguric phase. Secondary hyperparathyroidism, abnormal metabolism of Vitamin D, immobilization of patient, and release of calcium from injured muscle are considered as possible pathogenetic mechanisms of hypercalcemia in acute renal failure associated with rhabdomyolysis. Among them, dissolution of calcium from injured muscle into extracellular fluid is accepted as the major mechanism of pathogenesis of hypercalcemia. A twenty year old male patient, combat policeman, developed syncope in the course of regular physical training. He was diagnosed of acute renal failure associated with rhabdomyolysis at the hospital he initially visited. After being subjected to three sessions of acute hemodialysis, he was transferred to our hospital. During the treatment, laboratory test was performed upon his continuous complaints of lower extremity weakness. It showed severe hypercalcemia with plasma calcium level of 17.6 mg/dL. He was in oliguric phase at the time of this severe hypercalcemia. Patient's lower extremity weakness was gradually improved by hemodialysis using low calcium dialysate. He was discharged after his renal function became normal. He has been followed on regular basis.
Acute Kidney Injury* ; Calcium ; Extracellular Fluid ; Humans ; Hypercalcemia* ; Hyperparathyroidism, Secondary ; Immobilization ; Lower Extremity ; Male ; Metabolism ; Plasma ; Renal Dialysis ; Rhabdomyolysis* ; Syncope ; Vitamin D

PURPOSE: The aims of this retrospective study were to evaluate the sequential changes of parathyroid hormone (iPTH) and calcium metabolism after renal transplantation (RTP) and to identify risk factors for hypertension (HPT). METHODS: Biochemical bone parameters were reviewed in 264 patients at pre-transplant, 6, 12, 36 and 60 months after RTP. RESULTS: iPTH levels fell significantly during the first six months after RTP and remained substantially stable thereafter. The mean total serum calcium level showed significant increase during the first six months and progressive and significant decline after the first year. The mean serum phosphorus level returned to the normal range during the first six months and remained normal thereafter. The serum alkaline phosphatase (ALP) level increased during the first year and gradually decreased after then. The prevalence of persistent HPT was 17.8%. Patients with persistent HPT had significantly elevated serum levels of iPTH at the time of RTP and had spent a longer time on dialysis. Significant positive correlations were observed between the serum iPTH levels on the one hand and the pre-transplant iPTH, serum ALP, and creatinine levels on the other hand. CONCLUSION: The prevalence of persistent HPT after RTP is not uncommon. The patients with long duration of dialysis showing high serum level of iPTH at the time of transplantation are at risk for persistent HPT.
Alkaline Phosphatase ; Calcium ; Creatinine ; Dialysis ; Hand ; Humans ; Hyperparathyroidism, Secondary ; Hypertension ; Kidney Transplantation* ; Metabolism* ; Parathyroid Hormone* ; Phosphorus ; Prevalence ; Reference Values ; Retrospective Studies ; Risk Factors

"Rickets" is the term applied to impaired mineralization at epiphyseal growth plate, resulting in deformity and impaired linear growth of long bones. Rickets may arise as a result of vitamin D deficiency or abnormality in metabolism. Vitamin D-dependent rickets (VDDR) is rare autosomal recessive disorder in which affected individuals have clinical features of vitamin D deficiency. In 1961, Prader first described this disorder including severe clinical features of rickets, such as hypophosphatemia, hypocalcemia, muscle weakness and seizure. Two distinctive hereditary defects, type I VDDR and type II VDDR have been recognized in vitamin D metabolism. Type I VDDR may be due to congenital defects of renal 1 alpha-hydroxylase, the enzyme responsible for conversion of 25 (OH) D3. These patients have low to detectable 1,25(OH)2D3 in presence of normal to raised 25 (OH) D3. In type II VDDR, renal production of 1,25(OH)2D3 is intact but 1,25(OH)2D3 is not used effectively and target organ resistant to 1,25(OH)2D3 is respectively derived from the abnormality in the vitamin D receptor. We report a case of a 25 month-old girl with typical clinical features of VDDR type I rickets, hypocalcemia, increased alkaline phosphatase and secondary hyperparathyroidism.
Alkaline Phosphatase ; Child, Preschool ; Congenital Abnormalities ; Female ; Growth Plate ; Humans ; Hyperparathyroidism, Secondary ; Hypocalcemia ; Hypophosphatemia ; Metabolism ; Muscle Weakness ; Receptors, Calcitriol ; Rickets* ; Seizures ; Vitamin D ; Vitamin D Deficiency ; Vitamins*

BACKGROUNDHyperphosphatemia in renal failure has been identified as a major role in the pathogenesis of hyperparathyroidism that is independent of changes in serum calcium and 1,25(OH)(2)D(3). The aim of this study was to evaluate the expression of parathyroid Pit-1 in hyperphosphatemia-induced secondary hyperparathyroidism (SHPT) of chronic renal failure (CRF) rats.

METHODSWistar rats with CRF induced by 5/6 nephrectomy were ramdomly fed with diet containing 1.2% inorganic phosphate (Pi, high phosphate (HP) group, n = 9) or 0.2% Pi (low phosphate (LP) group, n = 9) for 10 weeks starting from the fourth week after the surgery. Another 7 nephrectomy rats with HP diet were intraperitoneally injected with phosphonoformic acid (PFA, the specific inhibitor of Pit-1, HP + PFA group) 0.15 g/kg every other day for 10 weeks starting from HP diet. Another 6 HP rats injected with the same amount of normal saline as the control of the HP + PFA group (HP + saline group). At the same time, 9 rats with sham surgery received HP diet as the controls. At the 4th week and 14th week, blood was taken for measurement of serum creatinine (SCr), serum calcium (SCa), serum phosphorus (SPi), 1,25(OH)(2)D(3) and intact parathyroid hormone (iPTH). At the 14th week, two parathroid glands (PTGs) of each rat were removed by microsurgery, one gland for immunohistochemistry analysis of proliferating cell nuclear antigen (PCNA), the other one for detection of Pit-1 by Western blotting, and for the measurement of Pit-1 mRNA and PTH mRNA by real-time quantitative polymerase chain reaction.

RESULTSIn nephrectomy rats, high dierary phosphate induced a marked increase in serum phosphate, iPTH, PTH mRNA and PCNA parathyroid cells, accompanying Pit-1 and its mRNA in parathyroid gland increased significantly. However, serum Ca and 1,25(OH)(2)D(3) remained unchanged. PFA decreased Pit-1 and its mRNA levels to reduce intact PTH, PTH mRNA and PCNA-positive parathyroid cells.

CONCLUSIONSExpression of parathyroid Pit-1 in hyperphosphatemia-induced SHPT of CRF rats was upregulated. Pit-1 may mediate the stimulation to parathyroid gland by hyperphosphatemia.

Animals ; Blotting, Western ; Hyperparathyroidism, Secondary ; etiology ; metabolism ; Hyperphosphatemia ; complications ; Immunohistochemistry ; Kidney Failure, Chronic ; complications ; Male ; Polymerase Chain Reaction ; Rats ; Rats, Wistar ; Sodium-Phosphate Cotransporter Proteins, Type III ; genetics ; metabolism