Objective To investigate the the effects of chronic intrauterine hypoxia on intimal-media thickening(IMT) and strain rate(SR) in abdominal aorta of male offspring rabbits during adult age.Methods Sixten New-Zealand rabbits were assigned randomly to 2 groups: chronic intrauterine hypoxia group (CIH, 12% O_2, n = 8) and normal oxygen group (NO,21%O_2, n = 8).After delivery,2 male offspring rabbits per litter were selected and breast-fed for 3 months, randomly divided into high-fat diet and normal diet groups.Finally, there were 4 groups in this experiment:chronic intrauterine hypoxia with high fat diet (CIH + HFD, n = 8) ,non-chronic intrauterine hypoxia with high fat diet (NCIH + HFD, n = 8),chronic intrauterine hypoxia with normal diet (CIH + ND, n = 8) and normal control (NC, n = 8).At sixth months of age, Serum levels of total cholesterol(TC) and triglyceride(TG) were assayed.SR in the abdominal aorta of male offspring rabbits was evaluated by ultrasonography.Then, abdominal aorta was taken out and observed by electron microscope and IMT was measured.Results CIH increased the levels of TC and TG (P < 0.01), thickened the IMT (P<0.05) and decreased the SR(P<0.05) of abdominal aorta of male offspring rabbits during adult age.There were relevant pathological changes in different groups.All these above-mentioned profiles were aggravated significantly after feeding high fat diet (P<0.05 or P<0.01).Conclusions CIH increased the IMT and decreased the SR of abdominal aorta of male offspring rabbits during adult age.

Objective To investigate apoptosis of HepG2 ceils after transfecfion with LIGHT gene and interferon-γ. Methods LIGHT gene and interferon-γ were transfected into HepG2 cells by liposome mediated method. The HepG2 cells were divided into group A (transfected with LIGHT gene or interferon-γ), group B (transfeeted with LIGHT gene and interferon-γ) and group C (non-transfection group). The apoptosis rate of the HepG2 cells and the expression of Bcl-2 and Caspase-8 were detected 12, 24, 48 hours after transfeetion. Results (1) The apoptosis rates of HepG2 cells at hour 12, 24 and 48 after transfeetion were 18.8% ± 3.5%, 25.7%± 2.8% and 36.4% ±3.6% in group A, 23.8% ±2.4%, 31.1% ±2.1% and42.5% ±4.5% in group B, and 8.7% ± 2.1%, 9.3% ± 1.6% and 10.9% ± 1.2% in group C. There was a significant difference in apoptosis rate among the 3 groups (F = 15.69, 53.33, 48.28, P < 0.01). (2) The expression of Bcl-2 in HepG2 cells at hour 12, 24 and 48 after transfection was 16.4% ± 5.0%, 13.4% ± 3.5% and 8.6% ± 2.3% in group A, 14.7%±3.8%, 9.1% ±2.0% and 4.6% ±2.0% in group B, and 25.3% ±6. 3%, 19.8% ±4.4% and 10.1% ±3.8% in group C. There was a significant difference in the expression of Bcl-2 among the 3 groups (F = 6.19, 12.29, 5.81, P <0.05). (3) The expression of Caspase-8 at hour 12, 24 and48 after transfection were 19.3% ±2.4%, 27.2% ±1.9% and 33.7% ±3.0% in group A, 22.7% ±2.2%, 30.9% ±3.1% and 38.2% ±3.2% in group B, and 1.2% ±0.8%, 1.8% ±0.6% and 3.2% ±1.5% in group C. There was a significant difference in the expression of Caspase-8 among the 3 groups (F =71.54, 112. 78, I01.61, P < 0.01). Condusions LIGHT gene can signiticanfly promote cell apoptosis through regulating the expression of Bcl-2 and Caspase-8. Interferon-γ enhanced the effect of LIGHT gene on the apoptosis of HepG2 cells.

Objective To construct an efficient eukaryotic expression recombinant vector of human interleukin-1O(hIL-lO),and observe its expression in rabbit synoviocytes(RSCs).Methods Total RNA was extracted from peripheral blood mononuclear cells(PBMCs)of a patient with drug allergy.Specific Drimers for full-length open reading frames(ORFs)of hIL-10 were designed according to GeneBank(NM 000572).Withtotal RNA as the template,full-length ORFs of hIL-10 were amplified by reverse transcription Dolymerase chain reaction(RT-PCR).RT-PCR products were digested by restrictive endonucleotidase.then inserted into plasmid pcDNA4/HisMaxA.Both restrictive endonucleotidase analysis and DNA sequencing Were carried out for inserts verification.RSCs were transfected with recombinant plasmid expression vector PcDNA4 HisMaxA-hiL10 by liposome-mediated gene transfer methods,then cultured in vitro.The supernatants were collected af-ter transfection for 12 hours,24 hours,48 hours,72 hours,7 days,14 days respectively for IL-10 measure-ment by enzyme linked immunosorbent assay(ELISA).Results Full-length ORFs of hIL-1o(0.54 kb)had been successfully cloned from PBMCs through RT-PCR.The inserts and insert location of pcDNA4 HisMaxA were in a fight way verified by enzyme analysis and DNA sequencing.ELISA results showed that exogenous hIL-10 gene had expressed in the transfected RSCs from 12 hours to 7 days after transfection,and hIL-10level of transfection group significantly higher than that of the control group.Conclusion pcDNA4 HisMaxA-hiL10,the hIL-10 efficient eukaryotic expression vectors,has been suecessfully constructed.

BACKGROUND: It has been successful to repair articular cartilage defects by using solid carrier as cytoskeleton. We tried to transplant liquid or gel carrier materials combined cells into the body of animals, and investigated its feasibility.OBJECTIVE: To investigate the feasibility of homo-transplatation with liquid or gel carrier materials of Pluronic F-127-recombinant human bone morphogenetic protein-2 (rhBMP-2) engineered chondrocytes for the repair of full-thickness rabbit articular cartilage defect.DESIGN: A controlled experiment.SETTINGS: Department of Orthopaedics, Weihai Municipal Hospital;Shandong Institute of Orthopaedics and Traumaology.MATERIALS: The experiments were carried out in the laboratory of Shandong Institute of Orthopaedics and Traumaology from November 2001 to September 2003. Thirty-six healthy adult New Zealand rabbits of 2.5-4.5 kg, either male or female, were divided into four groups according to the method of random number table: Pluronic F-127-rhBMP-2 engineered chondrocytes group, Pluronic F-127-rhBMP group, Pluronic F127 engineered chondrocytes group and blank control group, with 9 rabbits in each group.METHODS: After grouping, the 36 rabbits were made into models of articular cartilage defects. Pluronic F-127-rhBMP-2 was used as a vector of chondrocytes which were obtained from New Zealand rabbits after cultured and amplified in vitro. The mixture of Pluronic F-127, Pluronic F-127-rhBMP-2 and cultured chondrocytes was transplanted into the defects of articular cartilage that had been made previously with φb3.5 mm drill.There was not any treatment in the blank control group. At 4, 8 and 12 weeks postoperatively, the repairing conditions of the defects were evaluated with gross observation and histological observation under light microscope and under electron microscope. The repaire quality was assessed blindly according to the Wakitani scoring standard.MAIN OUTCOME MEASURES: ① Healing of cartilage defects; ② Property and morphology of the chondrocytes, characteristics, number and arrangement of collagens in matrix.RESULTS: ① In the Pluronic F-127-rhBMP-2 engineered chondrocytes group, the transplanted chondrocytes could grow better than those in other groups, the defected areas were completely filled at 4 weeks. The regenerated tissues at 8 and 12 weeks had similar appearance with the surrounding normal cartilage tissue, but vague. Delimitation. The histological examination showed that transparent cartilages formed, and the defects were healed. ② Under electron microscope at 8 and 12 weeks, there were mature transparent cartilages in the repaired tissues, and there were irregularly arranged slight, even and non-periodical collagen Ⅱ in surrounding. In the blank control group, only fibrous repair was observed, the regenerated tissue lacked elasticity with rough surface. ③ Repairing quality score: The scores at each time point in the Pluronic F-127-rhBMP-2 engineered chondrocytes group were significantly different from those in the other groups.Those in the Pluronic F-127-rhBMP-2 engineered chondrocytes group and Pluronic F-127-rhBMP-2 group and Pluronic F-127 engineered chondrocytes group were significantly different from those in the blank control group [4 weeks: (3.93±1.91), (4.56±1.07), (4.78±1.09), (8.44±1.13) points:8 weeks: (2.80±1.45), (3.24±1.00), (3.33±1.00), (8.44±1.13) points; 12 weeks (2.22±1.10), (3.01±0.69), (3.00±0.71), (9.00±0.87) points, P ＜ 0.001],but there were no significant differences between the two groups (P ＞ 0.05).CONCLUSION: The mixture of Pluronic F-127-rhBMP-2 and cultured chondrocytes can repair successfully the cartilage defects of femoral condyle of rabbit knees by means of hyaline cartilage than simple application of Pluronic F-127-rhBMP-2 or Pluronic F-127 engineered chondrocytes.