1.The relationship between thyroid autoantibodies and thyroid function in different populations
Zuoliang DONG ; Laixiang LIN ; Yina SUN ; Kaibin SHI ; Yuqin YAN
Chinese Journal of Endocrinology and Metabolism 2013;(4):324-327
To explore the relationship between thyroid autoantibodies and thyroid function in school children aged 8-10 years,adults,pregnant women,and lactating women in China,in order to provide reference for the prevention and monitoring of thyroid disease.Healthy 8-10 years old school children (693 cases),adults (698 cases),pregnant women(325 cases),and lactating women(332 cases) from six iodine sufficient areas were enrolled.Serum TSH,FT4,and FT3 were determined by chemiluminescent immunoassay,while antithyroid antibody by radioimmunoassay.The positive rate of thyroid autoantibodies in females was significantly higher than that in the male (5.6% vs 2.0% in school children,and 22.8% vs 3.2% in adults) ; while positive rate of autoantibodies in pregnant and lactating women (8.9%,8.7%) were significantly lower than that in the other healthy adult women (22.8%).The incidence of abnormal thyroid function in antibody-positive people was higher than that in negative ones in all groups,and abnormal thyroid function showed mainly as subclinical hypothyroidism.In addition,lactating women with negative autoantibodies presented a higher incidence of abnormal thyroid function,mainly as low FT4.The abnormal thyroid function is related with the positive thyroid autoantibodies,indicating that it is essential to follow-up these people with positive antibodies in order to facilitate prevention,early diagnosis,and treatment of thyroid disease.Reference data for thyroid hormones in lactating women should be establisbed as soon as possible.
2.Mechanism of miR-26a-5p/cAMP response element binding protein 1 molecular axis regulating osteogenic differentiation of adipose-derived mesenchymal stem cells.
Sanfu LIN ; Shoubo CHEN ; Kaibin FANG ; Jinnan SHI ; Wenhua WU ; Wenhuai WANG
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(5):615-621
OBJECTIVE:
To investigate the regulatory effects of miR-26a-5p on the osteogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs) by regulating cAMP response element binding protein 1 (CREB1).
METHODS:
The adipose tissues of four 3-4 weeks old female C57BL/6 mice were collected and the cells were isolated and cultured by digestion separation method. After morphological observation and identification by flow cytometry, the 3rd-generation cells were subjected to osteogenic differentiation induction. At 0, 3, 7, and 14 days after osteogenic differentiation induction, the calcium deposition was observed by alizarin red staining, ALP activity was detected, miR- 26a-5p and CREB1 mRNA expressions were examined by real-time fluorescence quantitative PCR, and CREB1 protein and its phosphorylation (phospho-CREB1, p-CREB1) level were measured by Western blot. After the binding sites between miR-26a-5p and CREB1 was predicted by the starBase database, HEK-293T cells were used to conduct a dual-luciferase reporter gene experiment to verify the targeting relationship (represented as luciferase activity after 48 hours of culture). Finally, miR-26a-p inhibitor (experimental group) and the corresponding negative control (control group) were transfected into ADSCs. Alizarin red staining, ALP activity, real-time fluorescent quantitative PCR (miR-26a-5p) and Western blot [CREB1, p-CREB1, Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN)] were performed at 7 and 14 days after osteogenic induction culture.
RESULTS:
The cultured cells were identified as ADSCs. With the prolongation of osteogenic induction culture, the number of calcified nodules and ALP activity significantly increased ( P<0.05). The relative expression of miR-26a-5p in the cells gradually decreased, while the relative expressions of CREB1 mRNA and protein, as well as the relative expression of p-CREB1 protein were increased. The differences were significant between 7, 14 days and 0 day ( P<0.05). There was no significant difference in p-CREB1/CREB1 between different time points ( P>0.05). The starBase database predicted that miR-26a-5p and CREB1 had targeted binding sequences, and the dual-luciferase reporter gene experiment revealed that overexpression of miR-26a-5p significantly suppressed CREB1 wild-type luciferase activity ( P<0.05). After 7 and 14 days of osteogenic induction, compared with the control group, the number of calcified nodules, ALP activity, and relative expressions of CREB1, p-CREB1, OCN, and RUNX2 proteins in the experimental group significantly increased ( P<0.05). There was no significant difference in p-CREB1/CREB1 between the two groups ( P>0.05).
CONCLUSION
Knocking down miR-26a-5p promoted the osteogenic differentiation of ADSCs by up-regulating CREB1 and its phosphorylation.
Animals
;
Female
;
Mice
;
Cell Differentiation
;
Cells, Cultured
;
Core Binding Factor Alpha 1 Subunit/metabolism*
;
Cyclic AMP Response Element-Binding Protein/metabolism*
;
Mesenchymal Stem Cells
;
Mice, Inbred C57BL
;
MicroRNAs/metabolism*
;
Osteocalcin/metabolism*
;
Osteogenesis/genetics*
;
RNA, Messenger/genetics*
3.Visualizing Seizure Propagation in Freely-moving Mice via Miniature Two-photon Microscopy.
Zhuoran ZHANG ; Shihe JIANG ; Kaibin SHI ; Yan LI ; Wei-Na JIN ; Qiang LIU ; Ting ZHAO ; Heping CHENG ; Fu-Dong SHI
Neuroscience Bulletin 2022;38(12):1593-1597
Animals
;
Mice
;
Microscopy
;
Seizures
;
Brain