Objective To explore the impact of basic fibroblast growth factor (bFGF) and parathyroid hormone-related protein (PTHrP) on early and late chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs) induced by transforming growth factor β1 (TGF-β1). Methods BMSCs were isolated from 3 healthy Japanese rabbits (2-month-old, weighing 1.6-2.1 kg, male or female), and were clutured to passage 3. The cells were put into pellet culture system and were divided into 5 groups according to different induce conditions: TGF-β1 group (group A), TGF-β1/bFGF group (group B), TGF-β1/21 days bFGF group (group C), TGF-β1/PTHrP group (group D), and TGF-β1/21 days PTHrP group (group E). At the beginning, TGF-β1 (10 ng/mL) was added to all groups, then bFGF and PTHrP (10 ng/mL) were added to groups B and D respectively; bFGF and PTHrP (10 ng/mL) were added to groups C and E at 21 days respectively. The gene expressions of collagen type I (Col I), Col II, Col X, matrix metalloproteinases (MMP)-13, and alkaline phosphatase (ALP) activity were detected once every week for 6 weeks. The 1, 9-dimethylmethylene blue (DMMB) staining was used to observe the extracellular matrix secretion at 6 weeks. Results The expression of Col I in groups C and E showed a significant downward trend after 3 weeks; the expression in group A was significantly higher than that in groups C and E at 4 and 5 weeks (P lt; 0.05), and than that in groups B and D at 3-6 weeks (P lt; 0.05); and significant differences were found between groups B and C at 3 and 4 weeks, and between groups D and E at 3 weeks (P lt; 0.05). After 3 weeks, the expressions of Col II and Col X in groups C and E gradually decreased, and were significantly lower than those in group A at 4-6 weeks (P lt; 0.05). Groups B and D showed no significant difference in the expressions of Col II and Col X at all time points, but there was significant difference when compared with group A (P lt; 0.05). MMP-13 had no obvious expression at all time points in group A; significant differences were found between group B and groups A, C at 3 weeks (P lt; 0.05); and the expression was significantly higher in group D than in groups A and E (P lt; 0.05). ALP activity gradually increased with time in group A; after 4 weeks, ALP activity in groups C and E obviously decreased, and was significantly lower than that in group A (P lt; 0.05); there were significant differences between groups B and C, and between groups D and E at 2 and 3 weeks (P lt; 0.05). DMMB staining showed more cartilage lacuna in group A than in the other groups at 6 weeks. Conclusion bFGF and PTHrP can inhibit early and late chondrogenic differentiation of BMSCs by changing synthesis and decomposition of the cartilage extracellular matrix. The inhibition is not only by suppressing Col X expression, but also possibly by suppressing other chondrogenic protein.
Objective To study the effect of signal-selective parathyroid hormone (PTH) analogue peptide on Wnt signal ing factors in osteoblasts isolated from neonatal mouse, and provide theoretical basis for the mechanism of PTH’s function in bone metabolism. Methods Osteoblasts were isolated from calvaria of 2-3-day-old C57BL neonatal mouse and identified by alkal ine phosphatase (ALP) staining, and Alizarin red staining. The cells at passage 1 were divided into 4 groups: control group, PTH (1-34) group, G1R19 (1-34) group, and G1R19 (1-28) group. Then the medium was changed to α-MEM supplemented with 1%FBS. After 12 hours, trifluoroacetic acid or three peptides [(10 nmol/L PTH (1-34), 10 nmol/L G1R19 (1-34), and 100 nmol/L G1R19 (1-28)] were added into the culture medium. After 4 hours, the cells were washed gently ithcold PBS 3 times before total RNA was isolated. The expressions of Wnt related genes were measured by quantitative eal-time PCR. Results Most of the cells were polygonal and triangular; the cells were positive for ALP staining with blue cytoplasm at 14 days and the Al izarin red staining showed the formation of red mineral ized nodules in the special mineral ization induction medium at 28 days. The expressions of osteocalcin mRNA and Wnt5b mRNA in PTH (1-34) group, G1R19 (1-34) group, and G1R19 (1-28) group were significantly higher than those in control group (P lt; 0.05); the expression of Wnt2 mRNA was significantly lower than that in control group (P lt; 0.05); the expression of β-catenin mRNA in PTH (1-34) group was significantly higher than that in control group (P lt; 0.05); the expression of Wnt7b mRNA in PTH (1-34) group and G1R19 (1- 34) group was higher than that in control group, and the G1R19 (1-34) group was higher than PTH (1-34) group and G1R19 (1-28) group (P lt; 0.05). Conclusion In the Wnt-related factors, PTH (1-34) and G1R19 (1-34) affect mainly canonical Wnt signal factors, but the G1R19 (1-28) chiefly acts on non-canonical Wnt signal factors.
Objective To assess the efficacy and safety of parathyroid hormone (PTH) on bone mineral density (BMD) and fractures in postmenopausal women with osteoporosis. Methods We searched MEDLINE (1966 to March 2008), EMBASE (1974 to March 2008), The Cochrane Library (Issue 1, 2008), Current Controlled Trials, The National Research Register, CBM (1983 to March 2008) and CNKI (1994 to March 2008). Some related journals were hand searched as well. The quality of included randomized controlled trials (RCTs) was evaluated and meta-analysis was conducted by The Cochrane Collaboration’s software RevMan 4.2.10. Results Twelve studies involving 5550 patients were included. PTH alone or in combination with antiresorptive drugs reduced the risk of vertebral fracture (RR=0.34, 95%CI 0.26 to 0.45, Plt;0.000 01), and increased spine BMD (SMD 0.41, 95%CI 0.17 to 0.65, P=0.0009) and femoral neck BMD (SMD 0.13, 95%CI 0.03 to 0.22, P=0.008). The rate of drop out and loss to follow-up because of adverse events was significantly higher in the PTH group (Peto-OR=1.69, 95%CI 1.39 to 2.05, Plt;0.000 01). Conclusion PTH is effective in the prevention and treatment of postmenopausal osteoporosis, especially in patients with preexisting osteoporotic fractures or with very low bone density. PTH alone or in combination with antiresorptive drugs can reduce the risk of vertebral fractures and increase spine and femoral neck BMD. PTH is more effective than alendronate, but these two should not be used as a combined treatment.
Objective To investigate the threshold value for postoperative parathyroid hormone （PTH） that can predict the development of symptomatic hypocalcemia. Methods The patients received total thyroidectomy and all the operations performed by the same team from June 2010 to December 2011 in the thyroid and breast group of department of general surgery，Xuanwu Hospital of Capital Medical University were enrolled in this prospective study. Serum PTH and calcium were tested at 1-hour postoperatively and thereafter every day during hospitalization. Symptoms of hypocalcemia were recorded. Receiver operator characteristic curve was plotted to test the value of PTH assay to predict symptomatic hypocalcemia. Results There were 45 consecutive patients recruited in this prospective study，9 （20.0%） patients remained normocalcemic and 36 （80.0%） patients developed hypocalcemia postoperatively. There were 19 （42.2%） patients who developed symptoms associated with hypocalcemia and needed calcium supplement. The postoperative PTHs for normocalcemic （n=9），asymptomatic （n=17），and symptomatic （n=19） hypocalcemia patients groups were （40.5±23.7） ng/L，（18.3±9.1） ng/L，and （7.6±4.2） ng/L，respectively （P＜0.001）. The PTH value less than 13.4 ng/L could easily predict who were at high risk to develop symptomatic hypocalcemia with sensitivity of 94.7% and specificity of 76.9%. Conclusions This study suggests that a 1-hour postoperative PTH level at 13.4 ng/L can identify those individuals at risk for developing symptomatic hypocalcemia. Therefore，we recommend early calcium supplementation for these patients to decrease their postoperative morbidity from symptomatic hypocalcemia.
ObjectiveTo evaluate the value of parathyroid hormone (PTH) in predicting hypocalcemia at different time after thyroidectomy. MethodsThe literatures in CBM, WanFang, CNKI, VIP in Chinese, and OVID, PUBMED, EMBASE, and MEDLINE in English were searched. Hand searches and additional searches were also conducted. The studies of predicting hypocalcemia after thyroidectomy by detecting postoperative PTH at different time were selected, and the quality and tested the heterogeneity of included articles were assessed. Then the proper effect model to calculate pooled weighted sensitivity (SEN), specificity (SPE), positive likelihood ratio (LR+), and negative likelihood ratio (LR-) were selected. The summary receiver operating characteristic (SROC) curve was performed and the area under the curve (AUC) was computed. ResultsTwenty-three articles entered this systematic review, 21 articles were English and 2 articles were Chinese. Fifteen of 23 articles were designed to be prospective cohort study (PC) and 8 of 23 articles were retrospective study (Retro). These articles were divided into two groups. Group 1 was the studies of detecting postoperative PTH in 1 hour, which included 2 012 cases (494 of them occurred hypocalcemia). Group 2 was the studies of detecting postoperative PTH between 4-12 hours, which included 693 cases (266 of them occurred hypocalcemia). The publication bias of 2 groups were smaller that founded through the literature funnel. Meta analysis showed that in addition to merge SEN, between the 2 groups with merge SPE, LR+, LR-, and AUC differences were statistically significant (P < 0.01);the forecast effect of group 1 was better than group 2, and the AUC was the largest area when the PTH value in 1 hour after operation was below 16 ng/L. ConclusionDetection of postoperative PTH value is an effective method for predicting postoperative hypocalcemia. The 1 hour after operation for detecting PTH value below 16 ng/L to predict postoperative hypocalcemia have the best effect.
Objective To investigate the way to culture human parathyroid cells and to investigate its secretory function. Methods After digested by collagenase, parathyroid cells were isolated to get the original generation cells, then the cells were cultured and passaged, and morphological changes of original generation cells and passage cells were observed on every day. The parathyroid hormone(PTH) level secreted by the original generation cells and passage cells were measured on the 1st, 5th, 10th, 15th, and 20th day(original generation cells only) respectively. Results The cellular morphology was complete after digestion. On the 2nd day, most of the parathyroid cells had adhered and spreaded, on the 3rd day, all cells had spread. There was no very obvious changes on these cells after cultured for 4-15 days. From 16 to 20 days, some parathyroid cells went senescence. On the 1st day, all of the passage cells, which were fusiform and little bigger than those of the original generation cells, had adhered and spreaded. From 2 to 15 days, there was no very obvious changes. The concentration of PTH in original generation cells begin to decreased significantly on the 10th day (P < 0.01). The concentration of PTH in passage cells were all lower than those of original generation cells at the same corresponding time, but there were no significant difference on the PTH level on 5th day and 1st day, 10th day and 5th day, 15th day and 10th day in passage cells (P > 0.05). Conclusion Parathyroid cells which were cultured within 10 days possess well morphologic structure and have the strongest secretory function. Although the passage cells still possess secretory function, it is greatly inferior to original generation cells. At last, we consider that original generation cells cultured within 10 days can be regarded as the source of allogeneic cell transplantation.
ObjectiveTo investigate the significance of 99Tcm-MIBI SPECT dual-phase imaging, B-ultrasound and CT in the diagnosis of primary hyperparathyroidism, and its relationship with the level of serum calcium (CA). MethodsThe clinical data of 73 patients with parathyroid dysfunction (serum parathyroid hormone (PTH) > 130 pg/mL) were retrospectively analyzed. The 99Tcm-MIBI SPECT double phase imaging were performed in 73 cases, 63 cases underwent cervical B-ultrasound examination, and 16 cases underwent CT examination. According to the serum calcium (CA) levels, the patients were divided into CA < 2.7 mmol/L group and CA > 2.7 mmol/L group, and the postoperative pathological examination and followed-up results were as the standard, the sensitivity, specificity, positive predictive value, and negative predictive value of 99Tcm-MIBI SPECT dual-phase imaging, B-ultrasound and CT for diagnosis of PHPT in different serum CA levels were compared. ResultsThe sensitivity of 99Tcm-MIBI SPECT dual-phase imaging, B-ultrasound and CT examination for diagnosis of PHPT was 87.6%, 81.8% and 35.7%, respectively; the specificity was 87.5%, 75.5% and 50.0%, respectively; the positive predictive value was 98.2%, 93.7% and 83.3%, respectively; and the negative predictive value was 46.7%, 33.3% and 10%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of 99Tcm-MIBI SPECT dual-phase imaging and B-ultrasound examination for diagnosis of PHPT were significantly higher than those of CT examination(P < 0.05). The sensitivity, specificity, positive predictive value and negative predictive value of 99Tcm-MIBI SPECT dual-phase imaging for diagnosis of PHPT were higher than those of B-ultrasound examination, but the difference was not statistically significant (P > 0.05). In the CA < 2.7 mmol/L group, the sensitivity of 99Tcm-MIBI SPECT dual-phase imaging, B-ultrasound and CT examination for diagnosis of PHPT was 91.1%, 84.7% and 37.9%, respectively; the specificity was 80.2%, 72.9% and 49.7%, respectively; the positive predictive value was 96.8%, 96.0% and 79.4%, respectively; and the negative predictive value was 50.0%, 37.5% and 10.0%. In the CA > 2.7 mmol/L group, The sensitivity of 99Tcm-MIBI SPECT dual-phase imaging, B-ultrasound and CT examination for diagnosis of PHPT was 87.9%, 83.9% and 42.8%, respectively; the specificity was 83.3%, 79.2% and 50.0%, respectively; the positive predictive value was 96.9%, 94.1% and 75.0%, respectively; and the negative predictive value was 50.0%, 40.5% and 20.0%. There were no significant difference in the diagnostic accuracy between the 3 methods and the level of serum CV in different levels. ConclusionsThe diagnostic accuracy of 99Tcm-MIBI SPECT dual-phase imaging and B-ultrasound examination for diagnosis of PHPT patients with PTH > 130 pg/mL (especially parathyroid adenoma) were higher than that of CT examination, and it is not associated with the serum CA concentration.
ObjectiveTo summarize the variation of parathyroid hormone (PTH) after thyroidectomy and the influence factors of postthyroidectomy hypocalcemia (PHC). MethodsClinical data of 95 patients who underwent thyroidectomy in Affiliated Shengjing Hospital of China Medical University from Jan. 2015 to Dec. 2015 were analyzed retrospectively. ResultsOf the 95 patients, there were 27 patient (28.42%) suffered from PHC (PHC group), and levels of serum calcium in the other 68 patients (71.58%) were normal (normal group). There was no significant difference in levels of serum calcium and PTH between the PHC group and normal group before operation (P > 0.05), but levels of serum calcium and PTH in PHC group were both lower than corresponding index of normal group after operation (P < 0.05). The levels of serum calcium and PTH both decreased in PHC group after operation (P < 0.05), and only PTH level decreased in normal group after operation (P < 0.05). PHC was related with type of operation, who underwent two-side operation had higher risk of PHC (P < 0.05), but there was no significant relationship between PHC and gender or age (P > 0.05). ConclusionsPTH is an important factor for PHC. In addition, it is easier to occur PHC when the operative range become bigger.