BACKGROUND:Induced pluripotent stem cels and mesenchymal stem cels-derived microvesicles have been confirmed in various tissue repairs, which are expected to become more effective and safe therapy for articular cartilage repair. OBJECTIVE:To overal understand the research progress in the use of induced pluripotent stem cels and mesenchymal stem cels-derived microvesicles in articular cartilage repair. METHODS: A computer-based search of PubMed and CNKI was performed by the first author for articles related to stem cel treatment of osteoarthritis published from 2003 to 2015. The keywords were “articular cartilage injury, bone marrow mesenchymal stem cels” in English and Chinese, respectively. In the same field, articles published recently or in authorized journals were preferred. RESULTS AND CONCLUSION:Articular cartilage injury is stil a difficulty in the orthopedics. Many repair methods have been reported, but they al have limitations. Induced pluripotent stem cels and mesenchymal stem cels-derived microvesicles bring a new hope for patients with articular cartilage injury. However, there are stil many problems to be solved, such as extracting and purifying a large amount of cels, proliferation and differentiation potentials, and mechanism underlying cartilage repair.

The carotene content of 63 kinds of plant materials including vegetables, grasses, wild plants and tree leaves were determined by the column chromatography method. Alfalfa, shepherd purse and carrot had the highest carotene content among the tested plants.Four methods were tried for the preparation of carotene concentrates. It was found that percolation with ethyl ether after grinding in ethanol showed the highest yield at low cost.The carotene content of the above mentioned concentrates was almost completely destroyed after 4 months when stored at room temperature and exposed to light. There was appreciable loss of the vitamin after 5 months' storage in the dark at low temperature (below 6℃), however, the loss was much smaller than that stored at room temperature. The loss of carotene was greatly reduced when the carotene concentrates were stored under nitrogen either at low temperature or at room temperature compared with those stored in the air. The addition of heated soy-bean meal to the carotene concentrate also considerably reduced the carotene loss during storage at room temperature.

OBJECTIVETo explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC.

METHODSSix clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot.

RESULTS1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa).

CONCLUSIONThere is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Cytoskeletal Proteins ; genetics ; metabolism ; Female ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; LIM Domain Proteins ; genetics ; metabolism ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; virology ; Osteoporosis ; genetics ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley

Objective To study the correlation between fractional anisotropy(FA)and tumor microarchitecture(MVD,VEGF and celluarity).Methods Fouteen gliomas(5 grade Ⅰ and Ⅱ,4 grade Ⅲ, 5 grade Ⅳ)confirmed histo-pathologically were performed on diffusion tensor imaging(DTI)using a GE Signa Excite Ⅱ 3.0 T MR scanner(8-channel head coil,SE echo planner imaging(EPI),thickness:5 mm, spacing:0,directions:25,B values:0 and 1000 s/mm~2,TR 6000 ms,TE minimum,FOV:240 mm? 240 mm,image matrix 128?128,NEX 2).Postprocessing was done using a DTI specific software to gain FA image.ROIs were drqwn in tumor parenchyma and the value of FA was recorded.The positive expression of VEGF and CD34 was shown using immuno-histochemistry method.The VEGF,MVD,and cellularity of every slices were recorded.Pearson correlation analysis was used.Results FA(which is 0.102?0.080 in grade Ⅰ and Ⅱ,0.171?0.037 in grade Ⅲ,0.200?0.021 in grade Ⅳ)has the trend to raise with the increasing grade of astrocytomas.FA has significant positive correlation to MVD(40/HP in grade Ⅰ and Ⅱ, 86/HP in grade Ⅲ,101/HP in grade Ⅳ),VEGF(8% in grade Ⅰ and Ⅱ,47% in grade Ⅲ,55% in grade Ⅳ),and cellularity(104/HP in grade Ⅰ and Ⅱ,160/HP in grade Ⅲ,265/HP in grade Ⅳ).The correlation coefficients between FA and VEGF,MVD,and cellularity were 0.748,0.668,0.625 respectively.Conclusion As a new imaging method,DTI can reveal the microarchitecture in gliomas and be value of distinguishing gliomas of different grade.DTI provides a new method of precise diagnosis to glioma preoperatively.

Objective To investigate the possible mechanism of lycopene on protecting against hypoxia/reoxygenation (H/R)-injury.Methods Primary cultured cardiomyocytes,isolated from neonatal mouse,were divided into three groups randomly:control group(C) ; H/R group(4 h H followed by 8 h R) ;lycopene + H/R group(L + H/R),in which the cardiomyocytes were pretreated with lycopene for 4 h before H/R.The survival of cardiomyocytes was counted.Apoptotic cells were detected by TUNEL assays.The release of cytochrome c from mitochondrial matrix into the cytosol,the activity of caspase-3,intracellular ROS levels and the activity of calpain were also determined in these groups respectively at the same time.Results The pretreatment of cardiomyocytes with lycopene significantly improved the survival of cardiomyocytes [C:(89.84 ± 5.15) ％,H/R:(63.59 ± 5.11) ％,L + H/R:(79.25 ± 1.48) ％,P ＜0.05] and reduced the extent of apoptosis [C:(10.37 ± 1.25) ％,H/R:(32.03 ± 4.79) ％,L + H/R:(22.57 ± 3.22) ％,P ＜ 0.05],significantly reduced caspase-3 activation [C:(2.61 ± 0.19),H/R:(5.82±0.92),L + H/R:(3.74 ±0.64) pNA pmol/μg protein,P ＜0.05].To further study the mechanism underlying the benefits of lycopene,interactions between lycopene and calpain activation were examined.Lycopene pretreatment of cardiomyocytes suppressed the activation of calpain (C:272.33 ±300.46,H/R:1156.00 ± 212.02,L + H/R:607.33 ± 166.23,P ＜ 0.05) by reducing the H/R induced increased intracellular ROS levels [C:100％,H/R:(239.79 ± 27.27)％,L + H/R:(188.19 ±17.63) ％,P ＜ 0.05].Conclusion Lycopene may protect against hypoxia/reoxygenation-induced injury by preventing calpain activation.

OBJECTIVETo construct a recombiant lentivirus vector of human LMP-1 and detect the expression of LMP-1 in infected rat bone mesenchymal stem cells.

METHODSLMP-1 gene from the cDNA library were extracted by Polymerase Chain Reaction (PCR). The LMP-1 genes were connected into lentiviral vectors pGC-FU-EGFP which was linearized by Age I enzyme to produce recombiant lentivirus vector called as pGC-FU-LMP-1-EGFP,then packaged by 293T cells. The virus supernant congtaining LV-LMP-1-EGFP was harvested, concentrated and titrated. The rat BMSCs were transfected with recombiant lentivirus LV-LMP-1-EGFP at the most appropriate MOI. The mRNA and protein expression of LMP-1 were detected by RT-PCR and Western blot.

RESULTS1LV-LMP-1I-EGFP was recombined successfully and the titer reached 2x108TU/ml. 2The efficiency of infection was 93.5% ,which was get after LV-LMP-1-EGFP infected rat BMSCs at the most appropriate MOI=100. The expression of LMP-1 gene was confirmed by RT-PCR and Western blot.

CONCLUSIONLentivirus vector containing human LMP-1 gene is constructed successfully,which can transfected efficiently into rat BMSCs,and the infected rat BMSCs can effectively express LMP-1.

Animals ; Female ; Genetic Vectors ; LIM-Homeodomain Proteins ; genetics ; Lentivirus ; genetics ; Male ; Mesenchymal Stromal Cells ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; genetics ; Transfection

Objective To study the optimal waist circumference (WC) cut-off values for identifying metabolic risk factors in middle-aged and elderly subjects in Shandong Province of China.
Methods A total of 2 873 men and 5 559 women were included in this cross-sectional study. Metabolic syndrome (MetS) was diagnosed according to the definition of Chinese Diabetes Society in 2004. The relation between WC and MetS was analyzed by multivariate logistic regression analysis. The optimal WC cut-off values were identified using the area under the ROC curve and the different diagnostic criteria for central obesity were compared.
Results The WC was the risk factor for MetS independent of BMI, blood glucose, blood lipid, and blood pressure. The optimal WC cut-off value was 83.8 cm and 91.1 cm for identifying MetS in women and men, respectively. Compared with 80 cm and 85 cm for women and men, 85 cm and 90 cm had a higher Youden index for identifying all metabolic risk factors and MetS in women and men.
Conclusion The appropriate WC cut-off value is 85 cm and 90 cm for identifying central obesity and MetS in women and men in Shandong Province of China.