1.The feasibility study of rabbit meniscus tissue-engineering regeneration using the autologous bone marrow-derived mesenchymal stem cells(MSCs) and collagen glycosaminoglycan(GAG) biomaterial template
Qinglei XU ; Haishan WU ; Weijiang ZHOU
Chinese Journal of Orthopaedics 2000;0(02):-
Objective This is a feasibility study of rabbit meniscus regeneration evaluated with the use of autologous bone marrow derived mesenchymal stem cells(MSCs) and collagen glycosaminoglycan(GAG). Methods Autologous MSCs were prepared from the rabbit proximal tibial bone marrow and stimulated in vitro to start fibrocartilage differentiation lineage by bFGF and TGF ? 1. Then, collagen GAG templates enriched with these MSCs were implanted in vivo to the menisci excised rabbit knees as the substitute for the excised meniscus. After 3, 6, 12, 24 weeks postoperatively, the implants were evaluated by the gross, histological and ultrastructural observations. Results The MSCs enriched collagen GAG implants underwent inflammation, degradation, MSCs division and remodeling stages in vivo, and consequently formed a meniscus like fibrocartilage tissue. Special staining and electronic microscope observation proved that the regenerated fibrocartilage were chondrocyte like fibrochondrocytes; in contrast, the results of the control group showed that both collagen GAG implants without MSCs and no substitute had limited regenerating tissue, further evaluations by histological and electronic microscope showed no evidence of fibrochondrocytes, and hence these regenerated tissue were fibrous rather than fibrocartilaginous. Conclusion The inducing of rabbit meniscus regeneration by the autologous bone marrow derived MSCs and porous collagen GAG template is proved to be feasible in this study. However, further studies to improve the biomaterial design, to evaluate the biomechanical properties of the regenerated tissue and to ensure clinical safety etc are needed prior to its clinical application.
2.Preparation and test of type Ⅰ collagen-glycosaminoglycan (GAG) template
Qinglei XU ; Haishan WU ; Weijiang ZHOU ; Dinglin ZHAO
Academic Journal of Second Military Medical University 2001;22(4):337-339
Objective: To synthesize the collagen-GAG template and to evaluate its feasibility to be used as the MSCs vehicle for meniscal tissue engineering. Methods: The collagen-GAG template was synthesized from rat tail type Ⅰ collagen and GAG using Yannas method. Then the post-stimulated MSCs by bFGF and TGF-β1 were added in. The MSCs-enriched collagen sponges were cultured in vitro, two weeks later the histological and ultrastructure detection was performed. Results: The histological and ultrastructure of the collagen-GAG template remained intact after 2 weeks' culture, and the MSCs in it remained viable. Conclusion: The collagen-GAG template synthesized in this experiment is suitable for the meniscal tissue engineering reconstruction as the vehicle for MSCs seed cells.
3.Changes of microRNA-206 in peripheral blood mononuclear cells from children with respiratory syncytial virus bronchiolitis
Hong ZHOU ; Qinglei XU ; Xiaobo MA ; Guoyu WEI ; Jinxiang HUANG
Chinese Journal of Applied Clinical Pediatrics 2015;30(22):1709-1712
Objective To explore the role of microRNA-206 (miR-206) in peripheral blood mononuclear cells (PBMCs) in infantile bronchiolitis caused by respiratory syncytial virus (RSV).Methods Thirty-five cases of infantile bronchiolitis and 25 cases of healthy controls were enrolled into the current study.PBMCs were isolated from the peripheral blood of both healthy subjects and those with infantile bronchiolitis in the acute and the convalescent stages.Total RNAs were extracted from PBMCs which were stimulated by phorbol-12-myristate-13-acetate (PMA) and Ionomycin, and then the RNA was transcribed reversely into cDNA.The expressions of miR-206 and Kruppel-like transcription factor 4 (KLF4) were detected by real-time quantitative polymerase chain reaction (qRT-PCR) method.Plasma interleukin-17 (IL-17) was determined by enzyme linked immunosorbent assay (ELISA).Results There was a significant difference in miR-206 levels of children with RSV bronchiolitis in the acute stage(0.055 ±0.018) and the convalescent stage(0.187 ±0.069) as well as the healthy controls(0.204 ± 0.075).Through pairwise comparison, the miR-206 levels in the children in the acute stage were significantly lower than those in the convalescent stage and healthy control group (P < 0.01), but no statistical significance was found between the convalescent stage group and healthy control group(P > 0.05).The levels of KLF4 mRNA of children in the acute stage,convalescent stage as well as the healthy subjects were 0.588 ± 0.161,0.086±0.024,0.075 ±0.019, respectively,which was significantly difference (P < 0.01).The levels of IL-17 were (58.26 ±25.88) ng/L, (9.87 ± 3.01) ng/L, (7.65 ± 2.16) ng/L, respectively (P < 0.01).Compared to the convalescent and the normal control group,both the KLF4 mRNA and IL-17 levels were markedly higher in the acute stage (P < 0.01), but there were no significant differences between children with RSV bronchiolitis in convalescent stage and in the healthy controls (P > 0.05).Furthermore, the result of this study showed a negative correlation between the expression of miR-206 and KLF4(r =-0.624 ,P <0.01)and IL-17 (r =-0.609 ,P <0.01) in children in the acute stage and a positive correlation between KLF4 mRNA and IL-17 in children in the acute stage (r =0.662, P < 0.01).Conclusion The levels of miR-206 may play a role in the onset of RSV associated post-bronchiolitis (PB) and the low expression of miR-206 in children infected with RSV may increase the susceptibility to PB.
4.The change of blood gas, hemodynamics and the expression of tumor necrosis factor in myocardial and pneumonic tissues of aged rats with pneumonia
Lu MA ; Qiao XUE ; Qinglei ZHU ; Shiwen WANG ; Bin XU
Chinese Journal of Geriatrics 2003;0(08):-
Objective To study the mechanism of changes in blood gas and hemodynamics in aged rats with pneumonia. Methods Forty aged male Sprague-Dawley(SD) rats were randomly divided into three experimental groups and the control group. They were contrasted by 40 non-aged male SD rats. Hemodynamic parameters and blood gas were measured, and morphological changes of heart and lung were observed. Plasma levels of tumor necrosis factor-? (TNF-?) was examined by immunohistochemical method. Results PaO 2 and SaO 2 were lower in rats with pneumonia than those in the control rats(P
5.Expressions of granulocyte-macrophage colony-stimulating factor and myeloperoxidase in bronchoalveolar lavage fluid in children with Mycoplasma pneumoniae pneumonia and its clinical significance
Hong ZHOU ; Qinglei XU ; Xiaobo MA ; Peiqi XU ; Gang LIU ; Min ZHANG ; Yue JIN ; Zhengrong CHEN
Chinese Journal of Applied Clinical Pediatrics 2017;32(4):313-315
6.Expression of microRNA-206 in peripheral blood mononuclear cells in asthmatic children
Qinglei XU ; Baolin ZHU ; Xiaobo MA ; Wei ZHANG ; Lingling LIU ; Shengjun WANG ; Jianguo CHEN
Journal of Clinical Pediatrics 2015;(2):105-108
s: Objective To explore the role of microRNA-206 in peripheral blood mononuclear cells (PBMC) in the pathogenesis and development of childhood asthma. Methods Twenty-seven asthmatic children and 25 healthy controls were enrolled in the study. Peripheral blood mononuclear cells were isolated in both healthy subjects and asthmatic children in acute attack and remission stages. Total RNAs were extracted from PBMC stimulated by PMA and ionomycin, and then the RNA was reversely transcribed into cDNA. The expressions of microRNA-206 and Kruppel-like factor 4 (KLF4) and IL-17 mRNA were detected by real-time quantitative PCR (qRT-PCR) method. Results There was signiifcant difference of microRNA-206 levels among asthmatic children in attack stage and in remission stage and normal controls (F=46.58~72.81, P=0.000). Through pair-wise comparison, the microRNA-206 levels of asthmatic children in attack stage were signiifcantly lower than those in remission stage and normal control groups (P<0.01). The KLF4 and IL-17 mRNA levels of asthmatic children in attack stage were signiif-cantly higher than those in remission stage and normal control groups (P<0.01). There was no signiifcant difference of miR-206, KLF4 and IL-17 mRNA between asthmatic children in remission stage and the healthy controls (P>0.05). Furthermore, a negative correlation was found between the expression of miR-206 and KLF4 (r=–0.66, P<0.01) and between the expression of miR-206 and IL-17 mRNA (r=–0.81, P<0.01) in asthmatic children in attack stage. A positive correlation was also found between KLF4 and IL-17 mRNA in asthmatic children in attack stage (r=0.70, P<0.01). Conclusions The expression of miR-206 is decreased in asthmatic children, and miR-206 might be involved in the pathogenesis and development of asthma.
7.The significance of peripheral blood CD4 + T cell-derived leptin in asthmatic children
Qinglei XU ; Baolin ZHU ; Xiaobo MA ; Wei ZHANG ; Lingling LIU ; Shengjun WANG ; Jianguo CHEN
Journal of Clinical Pediatrics 2014;(2):122-125
Objectives To explore the role of CD4+T cell-derived leptin in peripheral blood mononuclear cells (PBMC) in asthmatic children. Methods Peripheral blood mononuclear cells were isolated from peripheral blood of both healthy subjects and asthmatic children in attack and remission stages. CD4+T cells were purified from PBMCs by mag-netic beads and were cultured in vitro. Supernatants were used to detect the levels of leptin by ELISA. The expression of the orphan nuclear receptor (ROR)γt was detected by real-time quantitative PCR (qRT-PCR) method. Results There was significant difference in CD4+T cell-derived leptin levels of asthmatic children in attack stage (68.46±13.08 pg/ml), remis-sion stage (36.73±6.13 pg/ml) and normal controls (32.82±5.79 pg/ml) (P<0.01). Through pairwise comparison, the leptin levels in children in attack stage were significantly higher than those in remission stage and normal control groups (P<0.01). But no statistical significance was found between remission stage group and normal controls (P>0.05). The plasma leptin of children in attack stage and remission stage, as well as in normal subjects were 16.64 ± 3.53, 14.91 ± 3.24 and 13.72 ± 5.79 ng/ml respectively with no significant differences (P>0.05). The levels of RORγt mRNA were 0.341 ± 0.175, 0.089±0.028 and 0.068±0.018 in children with asthma during attack stage, remission stage and in normal children respec-tively (P<0.01). Compared to remission and normal control groups, the RORγt mRNA level of children in attack stage was markedly higher (P<0.01), but there was no significant difference between asthmatic children in remission stage and the healthy controls (P>0.05). Furthermore, the result of this study showed CD4+T cell-derived leptin positively correlated to RORγt in asthmatic children in attack stage (r=0.681, P<0.01). Conclusions CD4+T cell-derived leptin is elevated in asthmatic children in attack stage and its level is closely related to the pathological process of asthma.
8.Level of MicroRNA-2 0 6 in Peripheral Blood Mononuclear Cells from Rheumatoid Arthritis and Their Significance
Qinglei XU ; Baolin ZHU ; Xiaobo MA ; Wei ZHAN ; Lingling LIU ; Shengjun WANG ; Jianguo CHENG
Journal of Modern Laboratory Medicine 2014;(5):64-66
Objective To explore the role of microRNA-206 (miR-206)in peripheral blood mononuclear cells (PBMC)from rheumatoid arthritis (RA)patients.Methods 27 patients with RA and 25 healthy controls were enrolled into the current study.Human peripheral blood mononuclear cells (PBMCs)were isolated by standard density-gradient centrifugation over Ficoll-Hypaque solution in rheumatoid arthritis and healthy control volunteers.Total RNAs were extracted from PBMCs which were stimulated by PMA and ionomycin,then the RNA was transcribed reversely into cDNA.The expression of mi-croRNA-206 and Kruppel-like factor 4 (KLF4)and the orphan nuclear receptor RORγt mRNA was detected by real-time quantitative PCR (qRT-PCR)method.Student’s unpaired t-test and spearman correlation were used for statistica1 analysis. Results The expression of miR-206 in the PBMCs of RA patients was significantly decreased compared with that of the healthy controls (0.056±0.019 vs 0.138±0.057,t=3.103,P<0.01),The levels of KLF4 and RORγt mRNAin the PB-MCs of RA patients were increased significantly verves those of the healthy controls(0.604±0.183 vs 0.098±0.027,t=6.651,P<0.01;0.583±0.271 vs 0.069±0.018,t=7.438,P<0.01),Furthermore,a negative correlated between the ex-pression of miR-206 and KLF4 or RORγt mRNA in RA patients (r=-0.639,P<0.01;r=-0.842,P<0.01).Conclusion These results indicated that the augmented expression of KLF4 mRNA may be caused by the attenuated expression of miR-206,and the high level of KLF4 mRNA evokes the proportion of Th17 cells in RA patients.
9.Bone mineral density change in patients with hyperthyroidism after 131I therapy: a Meta-analysis
Xue SHEN ; Qinglei XU ; Junyu ZHAO ; Huanjun WANG ; Jinming YAO ; Jianjun DONG ; Lin LIAO
Chinese Journal of Nuclear Medicine and Molecular Imaging 2016;36(4):350-354
Objective To review the influence of 131I therapy on bone mineral density (BMD) in patients with hyperthyroidism.Methods Published articles of prospective randomized controlled study,clinical controlled study or case-control study on BMD change in patients with hyperthyroidism after 131I therapy were selected from PubMed,the Excerpta Media Database (Embase),Cochrane library,Chinese Journal Full-text Database,Wanfang Database,Vip Database and Chinese Biomedical Literature Database.Data from the date of database establishment to October 2015 were all reviewed.The languages were restricted to English and Chinese.Meta-analysis was performed with RevMan 5.3.Results Thirteen trials with a total of 668 hyperthyroidism patients were included.The meta-analysis showed that BMD of the lumbar spine,hip joint,femoral neck and osteocalcin were significantly improved after 131I therapy.The weighted mean difference (WMD) for BMD of the lumbar spine was 0.07 (95% CI:0.04-0.11),P=0.O00 2;that of the hip joint and the femoral neck was 0.13(95% CI:0.09-0.16) and 0.05(95% CI:0.03-0.06),respectively(both P<0.01).The standardized mean difference (SMD) of osteocalcin was-1.20(95% CI:-1.43--0.97) with P<0.01.Furthermore,the improvements were time dependent within the 2 years' follow-up.Conclusions 131I therapy improves the BMD and osteocalcin in patients with hyperthyroidism in a time dependent manner within 2 years' follow-up.
10.Detection and clinical significance of peripheral blood CD 4+T cell-derived leptin in patients with Hashimoto′s thyroiditis
Qinglei XU ; Baolin ZHU ; Xiaobo MA ; Wei ZHANG ; Lingling LIU ; Shengjun WANG ; Jianguo CHEN
International Journal of Laboratory Medicine 2014;(5):531-532
Objective To detect the level of CD4+ T cell-derived leptin(LP) in peripheral blood mononuclear cells(PBMC) from the patients with Hashimoto′s thyroiditis(HT) and to investigate its role in the occurrence and development of HT .Methods The peripheral blood samples in the patients with HT and the healthy controls were collected and separated for obtaining PBMC .The CD4+ T cells magnetic beads was adopted to separate CD4+ T cells for culturing in vitro .The LP level in supernatant was detected by ELISA .The relative expression amount of the orphan nuclear receptor RORγt was detected by real-time quantitative PCR (qRT-PCR) method .Results The LP level of CD4+ T cell culture liquid in the HT patients was markedly higher than that in the healthy control group ,difference was statistically significant (P<0 .05) .Furthermore ,the LP level in CD4+ T cell culture liquid was posi-tively related with the relative expression amount of RORγt in HT patients .Conclusion The CD4+ T cell-induced leptin level is in-creased in HT patients ,which is closely related with the occurrence and development of HT .