1.Comparative study between mesenchymal stem cells derived form orofacial bone and those from long bone
Lin YUAN ; Han WANG ; Jin SUN ; Zhengyi YANG ; Lu HAO ; Guangsi PAN ; Yina CAO ; Jun QIAN
Journal of Practical Stomatology 2015;(4):526-530
Objective:To compare the proliferation and osteogenic differentiation between human bone marrow mesenchymal stem cells from orofacial bone(OMMSCs)and those from long bone(BMMSCs).Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.BMMSCs were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method.The surface markers of the cells were detected by flowcytometry.Single-colony formation,CCK assay and cell circle analyses were conducted.Osteogenic differentiation ability was evaluated by ALP activity test and Alizarin red staining after osteogenic induction culture.Results:The cell surface markers STRO-1 and CD105 of both stem cells were positive,CD34,CD31 and CD45 were negative. OMMSCs generated significantly higher numbers of colonies than BMMSCs.In addition,OMMSCs had a higher proliferation rate and more cells in proliferative(S +G2 )stage than BMMSCs.After osteogenic induction for 3,5,7 and 10 d,OMMSCs showed higher levels of ALP activity.OMMSCs formed significantly more mineralized nodules than BMMSCs after 21-day ostogenic induction.Conclusion:The proliferation and osteogenic differentiation capacity of OMMSCs are higher than those of BMMSCs.
2.Exosomes serected by mesenchymal stem cells derived form orofacial bone regulate the function of macrophage
Lin YUAN ; Yina CAO ; Zhengyi YANG ; Jin SUN ; Guangsi PAN ; Jun QIAN ; Jingjing SONG ; Han WANG
Journal of Practical Stomatology 2017;33(3):344-348
Objective:To investigate the trait of exosomes serected by mesenchymal stem cells derived form orofacial bone(OMMSCs-Exo) and the communication between the exosomes and macrophages.Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.Their cell surface markers were characterized by flow cytometry.the rate of colony formation and the differentiation potential of OMMSCs were evaluated.Exosomes were prepaired from the culture supernatants of OMMSCs(P4-P6).Transmission electron microscopy(TEM) and western blot were used to identify the exosomes.The expression of miRNAs associated with immunity such as miR-223 and miR-let-7c were determined by Real-time RT-PCR.Human peripheral blood mononuclear cells(PBMCs) were isolated from health donor and co-cultured with OMMSCs-Exo.After co-cultured for 24 h,the communication between exosomes and macrophages was tested using a confocal microscope.Results:Human OMMSCs were proved to have the characteristics of mesenchymal stem cells.The diameter of OMMSCs-Exo ranged from 40 to 160 nm.The OMMSCs-Exo expressed CD63 and CD81 and contained miRNAs associated with immune regulation such as miR223 and miR-let-7c.OMMSCs-Exo could be uptaken by macrophages.After co-culture of OMMSCs-Exo with marcrophages for 72 h,miR223 expression in macrophages increased.Conclusion:OMMSCs-Exo has the potential of immune regulation.
3.Acetyltransferase MORF regulates osteogenic differentiation potential of periodontal ligament stem cells
Lin YUAN ; Jin SUN ; Feng CHENG ; Zhengyi YANG ; Yina CAO ; Guangsi PAN ; Jun QIAN ; Enliang HE ; Han WANG
Journal of Practical Stomatology 2016;32(6):778-782
Objective:To compare acetyltransferase MORF level in periodontal ligament stem cells( PDLSCs) derived from healthy individuals ( H-PDLSCs) with those derived from the individuals with periodontitis ( P-PDLSCs ) . And to determine the effect of MORF on the osteogenic differentiation potential of PDLSCs. Methods: Human H-PDLSCs and P-PDLSCs were cultured and cloned with limited dilution method. H-PDLSCs were stimulated by LPS, TNF-α, IL-β and the mix of the 3 inflammatory factors to imitate inflammatory environment ( IP-PDLSCs ) . Quantitative RT-PCR and Western Blot were applied to examine different expression of MORF in H-PDLSCs and P-PDLSCs. Western Blot was applied to detect expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red staining were applied to determine osteogenic differentiation potential of H-PDLSCs with MORF knock-down. Results:Quantitative RT-PCR and Western Blot showed lower expression of MORF in P-PDLSCs compared with H-PDLSCs( P<0. 05). Western Blot revealed lower expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red stai-ning indicated osteogenic differentiation potential was inhibited in H-PDLSCs with MORF knockdown(P<0. 05). Conclusion: Peri-odontitis can suppress the expression of MORF in PDLSCs and inhibite the osteogenic differentiation potential of PDLSCs.
4.UNet fusing patch attention for segmenting knee cartilage on MRI
Xiang WANG ; Cao SHI ; Zhengyi YUAN
Chinese Journal of Medical Imaging Technology 2024;40(5):764-768
Objective To construct a UNet fusing patch attention(PA-UNet)model,and to observe its value for segmenting knee cartilage on MRI.Methods Slice and preprocessing were performed on knee MRI selected from Osteoarthritis Initiative-Zuse Institute Berlin dataset.Taken UNet as the backbone network,a PA-UNet model was constructed based on patch attention mechanism.The effect of PA-UNet model and other models for segmenting both femoral cartilage and tibial cartilage were compared by subjective and objective evaluations.Ablation experiments based on UNet,UNet based on SE with layers 2-4(UNet+SE),+UNet,++UNet,+++UNet,+U-Net+,++U-Net++and PA-UNet models were performed to observe the effect of models for segmenting knee cartilage.Results PA-UNet could accurately segment femoral and tibial cartilage in all simple,medium and difficult samples,which had better segmenting effect on small structures than SegNet,UNet and DeepLabv3+models.The Dice similarity coefficient(DSC)and intersection over union of PA-UNet model for segmenting femoral and tibial cartilage were both higher,while Hausdorff distance of PA-UNet model was lower than those of UNet,DeepLabv3+,SA-UNet,RA UNet and SegNet models.DSC of PA-UNet model for segmenting femoral cartilage and tibial cartilage was 88.97%and 82.72%,respectively,both higher than those of UNet,UNet+SE,+UNet,++UNet,++UNet,+UNet+and++UNet++models.Conclusion PA-UNet could segment knee cartilage completely on MRI,especially for small structures.
5.Role of MDA in the pathogenesis of GERD esophagus mucosal inflammatory injury
Mijiti ABULAJIANG ; Yisireyili MAIMAITI ; Aili AIKEBAIER ; Wulamu WUBULIKASIMU ; Zhengyi CAO ; Yiliang LI ; Alimujiang AZIGULI ; Yuan JIANG ; Abudureyimu KELIMU
Journal of Chinese Physician 2019;21(7):997-1001,1006
Objective To investigate the expression of malondialdehyde ( MDA) in esophageal mu-cosa of different types of gastroesophageal reflux disease ( GERD) patients and its role in the esophageal in-flammation. Methods According to the inclusion and exclusion criteria, 42 patients hospitalized in the the Xinjiang Uygur Autonomous Region People's Hospital from December 2017 to October 2018 were selected as the research group. 8 healthy subjects completed physical examination were set up as healthy control group. GERD completed GERDQ score, 24 h pH monitoring, and taken 3 cm on the dentate line of the esophagus as a specimen. The study group was divided into non-erosive reflux disease (NERD) group (17 cases) and Ero-sive reflux disease [erosive esophagitis (RE)] group (25 cases). Then hematoxylin-eosin (HE) staining, immunohistochemistry, real-time polymerase chain reaction ( qPCR ) , enzyme-linked immunosorbent assay (ELISA) methods were used to detect inflammation, oxidative stress (MDA), antioxidant enzyme [manga-nese superoxide dismutase (Mn SOD), glutathione (GSH), catalase (CAT)], and proinflammatory cyto-kines [monocyte chemotactic protein-1 (MCP-1), interlukin-8 (IL-8), tumor necrosis factorα(TNF-α)]. Results There was no significant difference in body mass index ( BMI ) between the three groups ( P >0. 05). 24 h pH monitoring of esophagus showed that the indexes of weak acid reflux (4
6. Research progress of dynamic regulation of lysine acetylation in heart disease
Yuan ZHAO ; Qianwen NIE ; Jiangyan YIN ; Xuan HUAN ; Meixiang TIAN ; Zhengyi ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2021;26(10):1186-1192
Reversible post-translational modification of proteins is an important process in the physiological regulation of all tissues, including the heart. Lysine acetylation occurs in all organisms, including prokaryotes, and is regulated by a balance between lysine acetyltransferase (adding acetyl to the ε-amino group of lysine) and deacetylase (acetyl group that removes lysine ε-amino group). The heart is an organ rich in acetylated lysine, but the role of acetylated lysine in the heart remains to be elucidated. Therefore, in this paper, we systematically reviewed the gene list of acetyltransferase and deacetylase in mammalian genome and indicated their mRNA expression. The purpose of this study is to discover the research progress of dynamic regulation of lysine acetylation in heart disease and to provide a theoretical basis for the discovery of molecular targets.
7.5.0T MRI measurement of hippocampal formation subfield volumes in healthy adults
Zhensong WANG ; Jianxian LIU ; Zhangzhu LI ; Xiangsen JIANG ; Zudong YIN ; Lei YU ; Yuan GUO ; Zhengyi LI ; Qian ZHANG ; Jie GAN
Chinese Journal of Medical Imaging Technology 2024;40(5):648-652
Objective To observe the value of 5.0T MRI in measuring the volumes of hippocampal formation(HF)subfields in healthy adults.Methods Cranial high-resolution TIWI were prospectively obtained in 23 healthy adult volunteers using 5.0T and 3.0T MR scanners,respectively.HF was divided into 38 subfields,and then the volume of each subfield was measured using FreeSurfer software.The volumes of HF subfield based on 5.0T and 3.0T MR T1WI were compared,and the correlations of the outcomes were analyzed.Results Significant differences of the volumes of bilateral hippocampal tails,left parasubiculum,left molecular layer of hippocampal head,left granular cell-molecular layer-dentate gyrus head,right granular cell-molecular layer-dentate gyrus body,left cornu Ammonis(CA)1 head,left CA3 head,left CA4 head,right fimbria of hippocampus and left hippocampus amygdala transition area were found between 5.0T and 3.0T T1WI(all P<0.05).The volumes of HF subfields measured based on 5.0T and 3.0T T1WI were moderately to highly positively correlated(all r>0.5,all P<0.01).Conclusion 5.0T MRI could be used to measure the volume of HF subfield in adults.
8.Comparison of osteogenic differentiation abilities of mesenchymal stem cells from different sources of hBMSCs
YUAN Lin ; QIAN Jun ; YANG Zhengyi ; WANG Han ; GUO Wucheng ; CHENG Jieli ; SONG Jingjing ; HE Enliang ; ZHANG Yi
Journal of Prevention and Treatment for Stomatological Diseases 2017;25(9):554-559
Objective:
To compare the osteogenic differentiation abilities of human bone marrow mesenchymal stem cells (hBMSCs) from different sources, and to provide basis for choosing a new source of seed cells in bone tissue engineering.
Methods:
Jaw bone-marrow-derived mesenchymal stem cells (JMMSCs) were isolated from orthognathic surgical sites and cultured by limited dilution for single cell clone. Long bone-marrow-derived mesenchymal stem cells (BMMSCs) were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method. Flow cytometry was used to detect the surface markers of both cells. Osteogenic ability was assessed by PCR and Western Blot after osteogenic differentiation for the following molecules: Runx2, COL-1 and OCN. Alizarin red staining was used for determining the ability of cell mineralization after osteogenic differentiation.
Results :
The expressions of cell surface markers CD90 and CD105 were positive in both type of cells, while CD34, CD14 and CD45 were all negative. After 21 days of osteogenic induction, JMMSCs formed significantly more mineralized nodules than BMMSCs. After 7, 14, 21 days of osteogenic induction, JMMSCs expressed more osteogenic-related molecules than BMMSCs.
Conclusion
The osteogenic differentiation capacity and mineralization ability of JMMSCs are significantly higher than BMMSCs. Jaw bone might be a more suitable source of seed cells in bone tissue engineering compared with long bone.
9.K (lysine) acetyltransferase 2A affects the osteogenic differentiation of periodontal ligament stem cells through the canonical Wnt pathway.
Guo WUCHENG ; Cheng JIELI ; Yang ZHENGYI ; Zhang YI ; He ENLIANG ; Qian JUN ; Song JINGJING ; Sun JIN ; Yuan LIN
West China Journal of Stomatology 2018;36(1):39-45
OBJECTIVE:
This study aims to investigate the mechanism of K (lysine) acetyltransferase 2A (KAT2A) regulation and control on the osteogenic differentiation of periodontal ligament stem cells (PDLSCs).
METHODS:
The expression levels of KAT2A in PDLSCs were compared from each generation of the normal (H-PDLSCs) and periodontitis tissues (P-PDLSCs). The influences of KAT2A gene interference on the osteogenic differentiation of PDLSCs were also detected. In addition, the influences of the KAT2A gene interference to the canonical Wnt pathway and ligands were detected. The upstream and down-stream relationships between KAT2A and canonical Wnt pathway were also determined.
RESULTS:
The decreased expression of KAT2A in PDLSCs from the inflammatory tissue in each generation was compared with that in PDLSCs from the healthy tissue, and the difference was statistically significant (P<0.05). When the KAT2A gene was disrupted, the osteogenesis ability of PDLSC was declined, and the difference was statistically significant (P<0.05). The canonical Wnt pathway was activated, and the antagonist Dickkopf-1 (DKK-1) was reduced. After the DKK-1 addition, the osteogenic differentiation of the disturbed PDLSCs was recovered, and KAT2A was unaffected.
CONCLUSIONS
The KAT2A expression in PDLSCs was decreased because of perio-dontitis. The classical Wnt pathway was activated to inhibit the osteogenic differentiation of the cells.
Acetyltransferases
;
Cell Differentiation
;
Cells, Cultured
;
Histone Acetyltransferases
;
metabolism
;
Humans
;
Lysine
;
Osteogenesis
;
Periodontal Ligament
;
metabolism
;
Periodontitis
;
metabolism
;
Stem Cells
;
Wnt Signaling Pathway