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.
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.
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 discover the indicators and develop a model for predicting protracted hypoparathyroidism (HPT) after thyroid cancer surgery in order to guide the early therapy for patients with HPT.MethodsThe clinical and postoperative pathological data of patients with thyroid cancer who received surgical treatment in the Xuanwu Hospital and Beijing Pinggu Hospital from January 2019 to December 2020 were retrospectively analyzed. The potential indicators of postoperative HPT and protracted HPT were analyzed by logistic and LASSO regression analysis. A nomogram for predicting protracted HPT was constructed in the training set, and the discrimination and consistency of the nomogram were verified in the training set and the validation set respectively.ResultsAccording to the inclusion and exclusion criteria, a total of 464 patients diagnosed with thyroid cancer were finally included in the study. Among the 100 patients with postoperative HPT (except 1 case of incomplete data), 62 patients showed short-term HPT and 37 patients developed protracted HPT. Multivariate logistic regression analysis showed that the preoperative intact parathyroid hormone (iPTH) level [OR=0.953, 95%CI (0.931, 0.976), P<0.001], lobectomy with contralateral partial lobectomy [OR=3.247, 95%CI (1.112, 9.485), P=0.031], and total thyroidectomy [OR=11.096, 95%CI (5.432, 22.664), P<0.001] were related to postoperative HPT. The multivariant logistic regression analysis revealed that postoperative iPTH level was a predictive factor for protracted HPT [OR=0.719, 95%CI (0.588, 0.879), P=0.001]. The area under receiver operating characteristic curve (AUC) value of postoperative iPTH level in predicting protracted HPT was 0.848 [95%CI (0.755, 0.942)]; The cut-off value was 9.405 ng/L, and its specificity and sensitivity were 0.659 and 0.944, respectively. Moreover, the AUC value of the nomogram model including postoperative iPTH level and other clinicopathologic features (extraglandular invasion, cumulative maximum tumor diameter, and central lymph node dissection) for predicting protracted HPT was 0.900 [95%CI (0.817, 0.982)]; The cut-off score was 118.891, and its specificity and sensitivity were 0.772 and 0.944, respectively; The Hosmer-Lemeshow goodness of fit test indicated good fit of nomogram (χ2=8.605, P=0.377). The AUC value of the nomogram was 0.640 [95%CI (0.455, 0.826)] in the validation set (Pinggu Hospital data). The Hosmer-Lemeshow goodness of fit test also indicated good fit of nomogram (χ2=12.266, P=0.140).ConclusionsThe postoperative iPTH level is an important influencing factor of protracted HPT. The nomogram prediction model based on postoperative iPTH level and other clinicopathologic features has a favorable predictive value for protracted HPT.
ObjectiveTo introduce patients with long-term hypocalcemia and normal parathyroid hormone (PTH) values after total thyroidectomy, and to analyze the possible causes of this phenomenon. MethodsThe medical records of 1 010 consecutive patients with total thyroidectomy treated in the Center for Diagnosis and Treatment of Thyroid Disease, the First Affiliated Hospital of Kunming Medical University from January 2019 to December 2020 were collected. Seven patients with normal PTH and blood calcium before operation and at least 2 times of PTH with hypocalcemia detected more than 6 months after operation were followed-up to understand the symptoms of hypocalcemia, vitamin D level and calcium consumption. ResultsSeven patients with thyroid papillary carcinoma underwent total thyroidectomy without parathyroid autotransplantation, and there were 6 cases with mild deficiency or insufficient of vitamin D before operation. The follow-up time was 12–28 months, and the median follow-up time was 19 months. Seven patients developed hypocalcemia after continuous administration of calcium and calcitriol, and vitamin D levels remained mild deficiency or insufficient, PTH decreased by more than 50% in 6 patients one year after operation compared with that before operation. ConclusionsPatients with long-term normal PTH values and hypocalcemia after total thyroidectomy have obviously lower PTH levels than those before operation. The possible factors are parathyroid damage during operation and vitamin D deficiency. Such these patients should be more properly referred to as “parathyroid insufficiency”.
Objective To analysis causes of reoperation for primary hyperparathyroidism and its clinical characteristics. Method The clinical data of the patients with primary hyperparathyroidism who had undergone reoperation from January 1993 to May 2017 were retrospectively analyzed. Results A total of 11 patients underwent reoperation were collected in the 226 patients with primary hyperparathyroidism. Of the 11 cases, 8 cases underwent twice operations, 2 cases underwent thrice operations, 1 case underwent quintic operation. After the initial operation, 3 cases were persistent diseases and 7 cases were recurrent diseases, 1 patient was not defined as the persistent or recurrent disease. The main clinical manifestations before the reoperation were fatigue, pain in joints, bones, or muscle. The reasons for reoperation included 3 cases of ectopic parathyroid lesions, 3 cases of recurrent parathyroid carcinomas, 1 case of enlarged operation extent for parathyroid carcinoma, 2 cases of regrowth of double parathyroid aedomas, 1 case of missing adenoma, 1 case of parathyroid hyperplasia. Among the location examinations, the 99Tcm-MIBI was most sensitivity (8/9). Eight cases were received reoperation on the original incision, and the remaining 3 ectopic parathyroid lesions on the new incision. After the reoperation, 2 patients were lost of follow-up, 1 patient died, and the remaining 8 patients had no recurrences during follow-up period. Conclusion A comprehensive approach with multiple imageology examinations which attribute to accurate location of lesions, experienced surgeons and well knowledge of parathyroid anatomy and embryology help to descend reoperation ratio and improve success rate of reoperation.
Parathyroid hormone (PTH) exerts multiple effects such as regulating bone remodeling, promoting angiogenesis, etc., and it is an active factor with great application potential for bone repair. In recent years, with the development of scaffold material loading strategies and parathyroid hormone-related peptides (PTHrPs), in situ loading of PTH or PTHrPs on scaffold materials to promote bone defect healing gradually becomes possible. Based on the current status and challenges of intermittent PTH (iPTH) for bone tissue engineering, the review summarizes the in-situ application strategies of PTH and the construction of PTHrPs as well as current problems and further directions in this field, with a view to propel the clinical application of scaffold materials loaded with PTH or PTHrPs in situ.
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.
ObjectiveTo discuss the clinical significance of intact parathyroid hormone (iPTH) detection of postoperative serum and surgical drainage in predicting parathyroid function. MethodsThe clinical data of 50 patients with thyroid cancer performed total thyroidectomy from January 2013 to July 2013 in the PLA General Hospital and the First Affiliated Hospital of PLA General Hospital were analyzed retrospectively.In this cohort, there were 49 pappillary carcinomas and 1 medullary carcinoma.All the patients received total thyroidectomy andⅥlymph nodes dissection, and 18 of these patients received neck lymph nodes dissection additionally.The negative pressure drainage was placed in the operated area, and the drained fluid and blood were tested for iPTH level on the first day and the third day postoperatively. ResultsThe preoperative iPTH levels were within the normal range for all the patients.The postoperative serum iPTH level was (12.85±10.50) ng/L (4.64-13.15 ng/L) and (17.45±11.33) ng/L (7.33-26.50 ng/L) on the first day and the third day, respectively, there was no significant difference of the postoperative serum iPTH levels between the first day and the third day (P=0.293).The postoperative surgical drainage iPTH level was (1 550.29±1 678.18) ng/L (5 000-112 ng/L) and (566.73±753.99) ng/L (2 065-2.81 ng/L) on the first day and the third day, respectively, there was no significant difference of surgical drainage iPTH between the first day and the third day too (P=0.060). ConclusionsBy detecting the postoperative surgical drainage iPTH level combined with serum iPTH level, it could better predict the function of parathyroid after total thyroidectomy, and correctly assess the prognosis of patients.
ObjectiveTo analyze the changes of perioperative thyroid hormone in patients undergoing cardiac surgery under cardiopulmonary bypass, and to provide guidance for postoperative cardiac management.MethodsThe clinical data of 72 patients receiving cardiac surgery under cardiopulmonary bypass in our hospital from January to May 2019 were collected, including 35 males and 37 females, aged 19-72 (52.35±10.40) years. The changes of thyroid hormones before operation, 2 hours and 24 hours after operation were analyzed.ResultsThere was a statistical difference in thyroid stimulating hormone (TSH), triiodothyronine (T3), tetraiodothyronine (T4) and free tetraiodothyronine (FT4) between postoperative 2 hours and preoperation (P<0.05). There was a statistical difference in TSH, T3, free triiodothyronine (FT3), T4 and FT4 between postoperative 24 hours and preoperation (P<0.05). There was a statistical difference in TSH, T3, FT3 and T4 between postoperative 24 hours and 2 hours (P<0.05). Postoperatively T3 and FT3 decreased, TSH increased and then decreased while T4 and FT4 were within the normal range. Repeated measures analysis of variance showed a statistical difference of time effect in TSH, T3, FT3, T4 and FT4.ConclusionPatients with cardiac surgery under cardiopulmonary bypass have different thyroid hormones postoperatively compared with preoperatively. T3 and FT3 decrease, TSH increases and then decreases, while T4 and FT4 are in the normal range. The results require further large-scale, multi-center, high-quality clinical studies to be confirmed.