1.Bone morphogenetic protein 9-induced osteogenic differentiation of dental follicle cells in vitro
Chinese Journal of Tissue Engineering Research 2015;(14):2255-2260
BACKGROUND:Bone morphogenetic protein 9 is proved to promote the osteogenic differentiation of various kinds of stem cel s, but whether it can induce the osteogenic differentiation of dental fol icle cel s in vitro is yet unclear. OBJECTIVE:To investigate whether bone morphogenetic protein 9 can induce the osteogenic differentiation of rat dental fol icle cel s in vitro. METHODS:Purified rat dental fol icle cel s at passage 3 were transfected with bone morphogenetic protein 9 adenovirus. Then, alkaline phosphatase activity, calcium deposition and expression of osteogenesis-related factors at mRNA and protein levels were detected in the dental fol icle cel s. RESULTS AND CONCLUSION:After transfection with bone morphogenetic protein 9, the dental fol icle cel s showed continuously enhanced alkaline phosphatase activities and obviously enhanced calcium deposition. Real-time PCR results demonstrated that the mRNA expressions of alkaline phosphatase, osteocalcin, bone sialoprotein, osteopontin and core binding factor were increased significantly. The western blot assay showed that the expression of osteopontin enhanced in the dental fol icle cel s after transfection with bone morphogenetic protein 9. In summary, bone morphogenetic protein 9 can induce the osteogenic differentiation of dental fol icle cel s.
2.A preliminary study on abnormal expression of long noncoding RNA CTD-2012K14.6 in placentas and its role in inducing macrosomia
Linping YAN ; Lan WU ; Tianying ZHONG ; Lan LIU
Chinese Journal of Endocrinology and Metabolism 2019;35(2):138-142
Objective To explore the role of long noncoding RNA ( lncRNA) CTD-2012K14. 6 in the development of gestational diabetes mellitus (GDM) related macrosomia. Methods The quantitative real-time PCR ( qRT-PCR) was performed to measure the expression of CTD-2012K14.6 in placentas of women with or without GDM, and the quantity of CTD-2012K14. 6 expression and its association with fetal weights were analyzed; Bioinformatic analysis was performed to predict the downstream molecules. CTD-2012K14. 6 over-expressing lentiviral and siRNA was constructed in human trophoblastic cell line HTR-8/SVneo cells, qRT-PCR and Western blot (WB) were used to invest its effect in modulating the expression of downstream molecules. Results The expression of CTD-2012K14.6 in GDM placentas was significantly higher than that in normal controls (1.70 ± 0.63 vs 1.00 ± 0.56,t=3.68,P<0.01), and positively correlated with fetal weight (r=0.8501, P<0.01); on-line analysis showed that CTD-2012K14.6 was located at chr16:67,549,214-67,563,958, which was located in the intron of CCCTC-binding factor( CTCF); Up-regulating CTD-2012K14.6 could significantly reduce the expression of CTCF mRNA and protein, and increase the expression of insulin-like growth factor-Ⅱ( IGF-Ⅱ) mRNA and protein, while down-regulating CTD-2012K14.6 could significantly increase the expression of CTCF mRNA and protein, and reduce the expression of IGF-ⅡmRNA and protein. Conclusion The CTD-2012K14. 6 may play an important role in the pathogenesis of GDM related macrosomia by upregulating the expression of CTCF and IGF-Ⅱ.
3.Search of serum protein biomarkers for systematic lupus eryt.hematosus using protein chip tech- nology
Yan LAN ; Xiusheng TANG ; Jie WU ; Jun QIN ; Jimin CHEN
Chinese Journal of Dermatology 2009;42(8):560-562
Objective To study the changes of serum protein spectrum in patients with systematic lupus erythematosus (SLE) in order to screen specific protein markers. Methods Serum samples from 72 patients with SLE and 85 age- and sex-matched controls were assessed using surface-enhanced laser desorp-tion/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) with weak cation exchange (CM10) pro-rein chip. Forty samples from the patients and 50 control samples were randomly selected to serve as a pre-liminary training set; significantly different protein peaks were automatically chosen for the system training and development of a decision classification tree model. The validity of the model was then challenged with a blind test set (including another 32 samples from patients and 35 from human controls). Results A total of 73 effective protein peaks were detected within the mass/charge ratio (m/z) interval 2000 - 50000, among which, 15 protein peaks significantly differed between patients with SLE and controls (P < 0.01). Three pro-tein peaks with an m/z value of 4001, 6305 and 7356 were automatically chosen as a biomarker pattern in the training set that discriminated patients with SLE from controls with a sensitivity of 90.0% (36/40), speci-ficity of 92.0% (46/50) and accuracy rate of 91.1% (82/90). When the SELDI marker pattern was tested with the blinded test set, it yielded a sensitivity of 87.5% (28/32), specificity of 91.4% (32/35) and accuracy rate of 89.6% (60/67). Conclusions SELDI-TOF-MS protein chip could be used to screen serum protein for SLE, and the decision classification tree model with these biomarkers may favor the diagnosis of SLE.
4.Effects of Basic Fibroblast Growth Factor on Proliferation as well as Collagen and Fibronectin Expression on Fibroblasts
Yan WU ; Lan YANG ; Yufei ZHANG ; Xiaohuan YUAN ; Yanhui CHU
Herald of Medicine 2014;(11):1416-1419
Objective To explore the mechanism and effects of basic fibroblast growth factor( bFGF)on skin wound healing. Methods Fibroblasts( FB)were isolated from normal skin and hypertrophic scar and cultivated by direct adherence method. FB were then treated with different concentrations of bFGF(0,0. 1,1,10,100,1 000 ng·mL-1 )and cultivated with serum-free medium for 72 hours. The proliferation and apoptosis of FB in each group were detected by cell counting and trypan blue staining. Content and gene expression of typeⅠand type Ⅲ collagen and fibronectin were determined by ELISA and RT-PCR,respectively. Results bFGF promoted the proliferation of FB at low concentrations promoted apoptosis of FB at higher concentrations. The proliferation of FB from hypertrophic scar was slower than that from the normal skin. bFGF significantly inhibited type Ⅰ collagen production from hypertrophic scar FB but not from the normal skin. Moreover,bFGF up-regulated fibronectin expression in the normal fibroblasts,but not in the hypertrophic scar. No change in type Ⅲ collagen expression and production was observed in FB from either source. Conclusion bFGF has differential effects and mechanisms on FB of the normal skin and hypertrophic scar,suggesting that bFGF may play a role in early phase of skin wound healing and scar formation.
5.Association of interleukin-27 gene polymorphisms with genetic susceptibility to systematic lupus erythematosus in Guangxi Zhuang population
Yan LAN ; Yuanwen JIANG ; Xiusheng TANG ; Jie WU
Chinese Journal of Dermatology 2011;44(8):578-580
Objective To investigate the association between the single nucleotide polymorphisms of interleukin-27 (IL-27) gene and susceptibility to systematic lupus erythematosus (SLE) in Guangxi Zhuang population. Methods In total, 135 patients with SLE and 150 age- and sex-matched human controls of Zhuang nationality were recruited in this study. PCR-restriction fragment length polymorphism (RFLP) analysis and DNA sequencing were performed to analyze the IL-27 gene -964 A/G and -2905 T/G polymorphisms.Results Significant differences were observed in the distribution of IL-27 gene -964 A/G polymorphism (x2 =9.88, P < 0.01 ). The relative risk for SLE in carriers of G allele at position 964 of IL-27 gene was 1.725 times that in carriers of A allele at this position (OR = 1.725,95% CI: 1.227 - 2.425). A significant increase was observed in the frequency of 964G/2905G alleles of IL-27 gene in patients with SLE compared with the controls (10.7% vs. 5.3%, P < 0.01 ), and the 964G/2905G alleles were associated with a significantly increased risk for SLE (OR = 2.351, 95% CI: 1.228 - 4.501 ). Conclusions The IL-27 gene -964 A/G polymorphism is associated with the development of SLE, and the -964 G allele may increase the genetic susceptibility to SLE.
9.Application of high frequency electrical capsulotomy in maturation period cataract surgery
Wen-Hai, WANG ; Xiao-Yan, WU ; Jian-Lan, ZHANG
International Eye Science 2016;16(10):1927-1928
AIM: To explore the clinical effects of high frequency electrical capsulotomy in maturation period cataract surgery.
●METHODS: A total of 68 cases of maturation period cataract were selected and underwent the surgery of continuous circular capsulorhexis using the high frequency electrical capsulotomy.
●RESULTS: The success rate was 91% in 68 cases with the high frequency electrical capsulotomy.
● CONCLUSION: The high frequency electrical capsulotomy in maturation period cataract surgery has significant advantages and brilliant clinical values.
10.Expression of tissue factor in choroidal melanoma and its significance
Meixia, ZHANG ; De, LAN ; Jing, WU ; Junjun, ZHANG ; Mi, YAN
Chinese Ophthalmic Research 2010;28(3):207-209
Background It is well known that tissue factor (TF) is expressed in tumor cells and neovascular endothelial cells of tumor.It plays an important role in the formation of new blood vessels as well as the growth and metastasis of tumor.However,whether TF is expressed or not in choroidal melanoma(CM)is unclear.Objective This study is to investigate the expression of TF in a choroidal melanoma cell line and human choroidal melanoma.Methods The expression of TF was studied in the optimal choroidal melanoma-1 (OCM-1) cell line and ten specimens from CM patients using immunhistochemistry.Ten normal eye specimens from donators were used as controls.Results The TF protein was mainly expressed in the cytoplasm.It is over-expressed in OCM-1 cells with positive rate of 85.33±5.47%.Hyper-expression of TF also was found in human choroidal melanoma with a positive rate of 41.60±14.17%.The integrated optical density (IOD) of positive cells in CM was 33853.67±16445.30,and only 5.65±4.26% of positive cells was found in normal human choroidal tissue.The IOD of TF expression in normal human choroidal tissue was 426.43±316.62.Conclusion The overexpression of TF in CM cells may be a new immunotherapy target for CM treatment.