1.Association between single nucleotide polymorphism of interferon-gamma gene +874 site and susceptibility to ovarian cancer
Guangheng WU ; Jiaying ZHANG ; Wenjing ZUO
Journal of Jilin University(Medicine Edition) 2006;0(03):-
0.05);the frequencies of IFN-? gene +874 site T and A allele showed significant difference between case group and control group(P0.05).Conclusion T allele of IFN? gene+874 site might have a relationship with generation of ovarian cancer,TT genotype might be susceptibility genotype for ovarian cancer.
2.Association between polymorphism protein C inhibitor gene G10877T and male infertility
Peixin LU ; Jiaying ZHANG ; Guangheng WU
Journal of Jilin University(Medicine Edition) 2006;0(03):-
Objective To investigate the association between polymorphism of protein C inhibitor(PCI)gene G10877T and male infertility,and provide theoretical basis for treatment of male infertility.Methods PCR and sequencing technique were applied to detect PCI gene G10877 T polymorphism in 53 normal control and 102 male infertility.Results There were three genotypes of wild type(G/C),hybridization mutation(G/T)and pure mutation(T/T).The analysis of sequencing indicated that in sperums of a proportion of the male infertile patients,TGG in PCI gene G10877T mutated into TGT.The contrast of BLASTB indicated that this mutation made Trp in 271 position change into Cys.Compared with control group,TT genotypic frequency and T allelic frequency in male infertility group had significant differences(P
3.Effect of recombinant human bone morphogenetic protein-4 on cultured bone marrow stem cells
Guangheng LI ; Xiaokui HOU ; Xiangfu WU
Chinese Journal of Orthopaedics 2000;0(11):-
Objective To produce bioactive human bone morphogenetic protein-4 by Escherichia coli genetic engineering and investigate the effect of the product, recombinant BMP-4, on the bone marrow stem cells. Methods cDNA of human morphogenetic protein-4 mature peptide was obtained by RT PCR from tissue of human placenta. The gene was constructed in the pET-22b(+) expression vector and expressed in Escherichia coli BL-21 after transformation and induction by IPTG. The harvested protein was proved to be bioactive by inducing ectopic bone information in mouse thigh. The protein was applied to induce the cultured bone marrow stem cell. Shape change of the cell, ability of ALP (alkaline phosphatase) and concentration of OC (osteocalcin) were investigated. Results SDS-PAGE revealed a new protein band that located in position of 14?103 after 4 hours induction, the new protein made 15% of total bacteria protein, the rhBMP-4 could induce the cultured bone marrow stem cell of New Zealand rabbit to differentiate into osteoblasts and form calcified node. The ability of ALP and concentration of OC of tested group increased significantly than that of the control group. Conclusion The bioactive rhBMP-4 can be produced by Escherichia coli genic engineering, this protein can induce bone marrow stem cells to differentiate into osteoblast cells.
4.Influence of PDTC on expression of LFA-1 mRNA induced with LPS in mouse non-specific keratitis reaction
Guangheng WU ; Jiaying ZHANG ; Lihong YANG ; Le ZHOU
Journal of Jilin University(Medicine Edition) 2006;0(04):-
Objective To analyze the relationship between leukocyte function-associated antigen-1 (LFA-1) mRNA expression and corneal inflammation and discuss the inhibition reaction of nuclear factor-?B(NF-?B)inhibitor on LFA-1 mRNA expression.Methods The corneal suture or combined with subconjunctival injection models were constructed in BALB/C mice.The corneas of each group were excised 1,3,7 and 14 d after operation and LFA-1 mRNA expressions were investigated by reverse transcription polymerase chain reaction(RT-PCR).Results At 1,3 and 7 d after operation, the expressions of corneal LFA-1 mRNA in corneal suture combined with lipopolysaccharide (LPS) (corneal suture+LPs)subconjunctival injection group were higher than those in corneal suture group (P0.05).At 1 and 7 d after operation, compared with corneal suture+LPS group ,the expression of corneal LFA-1 mRNA increased,but it decreased at 3 and 14 d after operation in the group of corneal suture combined with LPS and pyrrolidine dithiocarbamate (PDTC) (P
5.Effects of different immunosuppressive agents on mesangial cell proliferation
Guobiao LIANG ; Guangheng LUO ; Jun SONG ; Li YANG ; Liyuan ZHANG ; Shunwen LUO ; Xianding WANG ; Zhiyuan XIE ; Ke WU ; Youping LI ; Yiping LU
Chinese Journal of Organ Transplantation 2010;31(9):545-548
Objective To investigate the effects of different immunosuppressive agents on mesangial cell proliferation through a mesangial cell injury model in vitro. Methods Mesangial cell line (HBZY-1) in period of proliferation was cultured in vitro with cytochalasin B for 2 h, then HBZY-1 cells were divided into 5 groups: blank (control) group, cyclosporine A (CsA) group, Tacrolimus (Tac) group, mycophelonate mofetil (MMF) group and rapamycin (RAPA) group. Subsequently,the number of HBZY-1 cells at different time points was measured by using the professional image analysis software after treatment for 6, 12 and 24 h, respectively. Results Damaged HBZY-1 cells recovered in all groups. At 6 h, the number of HBZY-1 cells in Tac group was significantly more than that in control group (P<0.05), but the difference had no significance between the other treatment groups and control group (P>0. 05). At 12 h, there was no significant difference in of the number of HBZY-1 cells among the all groups (P>0. 05). At 24 h, there was no significant difference in the cell number between MMF and control groups (P>0. 05). CsA, Tac and RAPA resulted in HBZY-1 cell proliferation, and the cell number in CsA and Tac groups was significantly more than that in the other groups (P<0. 05). As compared with the control group, the cell number in RAPA group was significantly increased (P<0. 05). Conclusion CsA, Tac, MMF and RAPA contribute to recovery of damaged HBZY-1 cells, but CsA and Tac result in over-proliferation of HBZY-1 cells. RAPA and MMF can prevent HBZY-1 cells against over-proliferation, and MMF scarcely results in HBZY-1 cell proliferation.
6.Cell-of-origin for heterotopic ossification induced by bone morphogenetic protein 4 in skeletal muscle
Yangyi YU ; Qiang LIAN ; Jianqun WU ; Xuan ZHANG ; Jinke REN ; Guangheng LI
Chinese Journal of Tissue Engineering Research 2024;28(25):4034-4040
BACKGROUND:Heterotopic ossification of skeletal muscle is a clinically serious complication.For heterotopic ossification of skeletal muscles,the cells involved in the process of heterotopic ossification remain unclear. OBJECTIVE:To investigate the involvement of myocytes,fascia cells,and endothelial cells in the process of heterotopic ossification in skeletal muscle and to observe the cell origin of heterotopic ossification in skeletal muscle induced by bone morphogenetic protein 4. METHODS:Both C2C12 cells and the myotubes formed by the C2C12 cells in the induction medium were cultured,and 500 ng/mL bone morphogenetic protein 4 was added to the medium respectively,and whether the C2C12 cells and myotubes continued to proliferate within 10 days under the treatment were observed under a microscope.Myogenic cells(L6,derived from rats)and fibroblast-derived cells(derived from human)were co-cultured.After treatment with 500 ng/mL bone morphogenetic protein 4 and 10 ng/mL transforming growth factor-β,osteogenic and chondrogenic differentiation potential within 21 days were observed using Safranine O staining and Alcian blue staining.Using transgenic animal FVB/N-TgN(TIE2-LacZ)182Sato mice,15 μL of adeno-associated virus-bone morphogenetic protein 4(5×1010 PFU/mL)were implanted in the thigh muscle space of genetic mice for 10 and 14 days.X-gal staining was used to observe the formation of new blood vessel endothelium in the differentiated bone. RESULTS AND CONCLUSION:(1)Bone morphogenetic protein 4 caused myotube breakdown and increased C2C12 cell proliferation.Compared with other groups,the pure fibroblast-derived cell group had a higher area of positive alcian blue and safarin O staining(P<0.05)and a lower area of alkaline phosphatase staining(P<0.05),while the pure L6 group had a bigger area of alkaline phosphatase staining(P<0.05)but a smaller area of positive alcian blue and safarin O staining(P<0.05).(2)Transplantation of adeno-associated virus-bone morphogenetic protein 4-adsorbed gelatin sponge into FVB/N-TgN(TIE2-LacZ)182Sato mice resulted in heterotopic ossification.(3)X-gal staining results demonstrated that there was no obvious staining in chondrocytes and differentiated bones and Tie2+ endothelial cells did not participate in the formation of the alienated bone.(4)These findings verify that fibroblasts are the primary source of osteoblasts during the adeno-associated virus-bone morphogenetic protein 4-induced ectopic endochondral ossification in skeletal muscle,but myogenic cells are the main source of osteoblasts.Tie2+ endothelial cells might not be the cell source for cartilage and bone.