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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.
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Humanos , Hiperparatireoidismo Secundário/metabolismo , Glândulas Paratireoides/metabolismo , Hormônio Paratireóideo , RNA Mensageiro/metabolismo , Receptores de Detecção de Cálcio/metabolismoRESUMO
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.
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Humanos , Hiperparatireoidismo Secundário/genética , MicroRNAs/metabolismo , Hormônio Paratireóideo , RNA Mensageiro , Receptores de Calcitriol/genéticaRESUMO
This study was designed to assess the immune protective effects of the vaccine strain of a precocious line of Eimeria necatrix with different doses and at different immunization times. The immunizations had a negative effect on weight gains of chickens to a certain degree but could be compensated during the "compensatory growth period" after immunity was established in the chickens. The number of oocysts excreted was positively correlated with the immunization dose. All the immunized chickens, whether they were immunized once or twice or immunized with different doses of sporulated oocysts, were able to resist attack from 1x105 virulent sporulated oocysts of E. necatrix. The lesion scoring showed that no significant difference existed in the chicken groups immunized with different doses (300 and 600) of sporulated oocysts. However, a difference existed in the immune homogeneity established in the different immunized groups, and two artificial immunizations were superior to one artificial immunization, indicating that two could extend the duration of oocyst excretion and allow more chances for the immunized chickens to become repeatedly infected.
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AIM:To find an efficient and simple surgical procedure of the external dacryocystorhinostomy ( EXT- DCR) in the treatment of chronic dacryocystitis. METHODS:A total of 270 cases were performed in this retrospective study. A comparison of clinical effects and hemorrhage complications between the experimental group, in which 189 eyes of 167 patients that underwent traditional EXT-DCR without packing, and the control group, in which 121 eyes of 103 patients that underwent traditional EXT-DCR with Vaseline gauze were performed. The postoperative follow-up ranged from 6 to 36mo ( the mean follow-up was 15±3. 6mo). RESULTS: There were no significant differences in the curative rates between the two groups (97. 9% vs 97. 5%, P>0. 05). But the incidence rate of moderate or severe postoperative nasal bleeding was lower in the experimental group (15. 9%, 30 eyes) than the control group (31. 4%, 38 eyes) and PCONLUSION: The traditional EXT - DCR without packingis an effective procedure and is more effective in reducing moderate or severe postoperative bleeding when the intraoperative anastomose and the postoperative drainage are well done and the intraoperative bleeding is well controlled.
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0.05).The expression of NF-?Bp65 protein in epithelial ovarian cancer was significantly correlated with FIGO stage(P0.05).There was obviously negative correlation between KISS- 1 and MMP-9 expression in ovarian cancer(rs=-0.547,P