BACKGROUND: Cyclooxygenase 2, aggrecanase 1, and insulin-like growth factor 1 are involved in pathological injury of the articular cartilage. OBJECTIVE:To observe the expression of shRNA vectors carrying cyclooxygenase 2, aggrecanase 1 and overexpression vectors carrying insulin-like growth factor 1 in bone marrow mesenchymal stem cels. METHODS:Lentiviral vectors carrying the silencing gene cyclooxygenase 2, aggrecanase 1, the over-expressing gene insulin-like growth factor 1 and binding green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect passage 3 human bone marrow mesenchymal stem cels culturedin vitro (experimental group). The human bone marrow mesenchymal stem cels transfected with no target gene lentivirals were used as negative control group. The human bone marrow mesenchymal stem cels transfected with no treatment served as blank group. RESULTS AND CONCLUSION:Cyclooxygenase 2 and aggrecanase 1 transfected in human bone marrow mesenchymal stem cels were significantly inhibited at gene and protein levels, while the expression of insulin-like growth factor 1 was increased significantly at gene and protein levels. We confirmed that cyclooxygenase 2 and aggrecanase 1 were successfuly silenced while insulin-like growth factor 1 overexpressed by using lentiviral vectors in human bone marrow mesenchymal stem cels, which brings a new hope for the systemic gene treatment of arthritis.

Objective:To study the alveolar bone morphology in the anterior teeth area of the skeletal Class Ⅱ malocclusion subjects with different vertical skeletal types.Methods:64 patients with skeletal Class Ⅱ malocclusion and 15 subjects with normal occlusion were included.The alveolar bone structure of the anterior teeth were observed using CBCT.Results:The labial and lingual alveolar bone height and the alveolar bone thickness of the incisors of the patients were much lower than those of the normal controls.The height of labial and lingual alveolar bone and the alveolar bone thickness of anterior teeth in high-angle subgroup were lower than those in low-angle subgroup.Conclusion:The thickness of the anterior teeth alveolar bone of skeletal Class Ⅱ malocclusion is low,espe-cially in the high-angle group.

Aim To investigate the effects of DGK in-hibitor R59022 on ET-1-induced myocardial hypertro-phy and autophagy, and explore the possible mecha-nisms. Methods Myocardial hypertrophy was in-duced by ET-1 in cultured rat neonatal cardiomyo-cytes. Western blot was used to detect the expression of microtubule-associate protein 1 light chain 3 ( LC3 ) , beclin-1, p62, p-Akt and Akt. mRNA expression of brain natriuretic peptide ( BNP) and beta mysion heav-y chain (β-MHC) and the cell size of cardiomyocytes were detected by RT-PCR and immunofluorescence, respectively. Results Treatment cardiomyocytes with ET-1(10 -7 mol·L-1 ) for 24 h induced the myocardi-al hypertrophy in cultured neonatal rat cardiomyocytes with the activation of autophagy as evidenced by the in-creased expression of autophagy-related proteins LC3-II/I and beclin-1 , as well as the increased p62 degra-dation. While, myocardial hypertrophy induced by ET-1 , including the increased myocardial cell size and the mRNA expression of fetal gene BNP and β-MHC, could be reversed by autophagy inhibitor 3-methyl ade-nine (3-MA) and chloroquine ( CQ) ,but promoted by autophagy agonist rapamycin ( RAPA ) . Pretreatment cardiomyocytes with R59022, an inhibitor of DGK, en-hanced ET-1-induced myocardial hypertrophy by en-hancing autophagy in cardiomyocytes. Furthermore,ET-1 treatment inhibited the activation of Akt by the down-regulation of the Akt phosphorylation, and R59022 en-hanced the effect of ET-1 on the activation of Akt. Conclusions Enhanced autophagy contributes to car-diomyocyte hypertrophy. R59022 deteriorate ET-1-in-duced myocardial hypertrophy by activating autophagy. The possible mechanism may be related to the inhibi-tion of activation of mTOR signaling pathway by inhibi-ting the activation of Akt.

Objective Preparing platelet rich gel through two-times centrifugal technique and co-culturing chondrocytes with PRG, then observing the proliferation and gene expression of chondrocytes, in order to provide a favorable way to prepare tissue engineering cartilage. Methods Centrifugating venous blood of rabbit through two-times centrifugal technique to obtain platelet rich plasma( PRP) ,then detecting the concentration of various growth factor in PRP. Admixing PRP with chondrocytes of rabbit and activating them with activator. After co-culti-vation,the proliferation of chondrocytes through MTT method and expression of ACAN,CollagenⅡand SOX-9 through realtime-PCR were ob-served,and compared with common cultured chondrocytes. Results The concentrations of PDGF-AB,TGF-β1,IGF-1 and VEGF in PRG were significantly higher than those in blood(P<0. 05). After co-cultivation, the proliferation rate of chondrocytes and the expression of ACAN,Collagen Ⅱ and SOX-9 were significantly higher than that of common cultured chondrocytes(P<0. 05). Conclusion Co-culturing chondrocytes with PRG is able to promote the proliferation and gene expression of chondrocytes. We considered that it is a excellent method to construct tissue engineering cartilage.

Objective To observe the curative effect of Modified Gongwaiyun Ⅱ on extrauterine pregnancy.Methods 69 patients of extranterine pregnancy were recruited into a treatment group and a control group randomly. 34 patients in the treatment group were treated with modified Gongwaiyun Ⅱ and 35 patients in the control group were treated with Gongwaiyun Ⅱ, with therapeutic course being one week. Values of β-HCG of serum at the beginning of the treatment,age, days of menelipsis, effective rate, average value of serum β-HCG decreasing and percentage at every course of the treatment, and the number of therapeutic courses were observed. Results There are significant statistic differences between the treatment group and the control group at the number of therapeutic courses, and average value of serum β-HCG decreasing and percentage at every course of the treatment, with P＜0.05. Conclusion The curative effect of modified Gongwaiyun Ⅱ had superiority to traditional Gongwaiyun Ⅱ.

BACKGROUND:Many studies have showed that enough blood supply is an essential condition of bone repair and regeneration. OBJECTIVE:To construct the endothelial progenitor cells/bone marrow mesenchymal stem cells (EPCs/BMSCs) composite sheet. METHODS:After isolation and culture, EPCs and BMSCs were co-cultured directly to form EPCs/BMSCs sheet by cellsheet-inducing medium. After 10 days of induction, the sheet was investigated by gross observation, inverted microscope and hematoxylin-eosin staining. The distribution and communication of EPCs and BMSCs during the process of cellsheet induction were observed after the fluorescence labeling separately. Alkaline phosphatase assay and alizarin red staining were applied to examine the ability of osteogenic differentiation of EPCs/BMSCs sheet.
RESULTS AND CONCLUSION:EPCs/BMSCs sheet was harvested after 10-day induction. Cel-cellcontact between EPCs and BMSCs could be observed during the process of the cellsheet preparation. The harvested sheet was composed of multiple layers of cells and cel-produced extracellular matrix. Alkaline phosphatase assay and alizarin red staining both demonstrated that EPCs/BMSCs sheet had good osteogenic differentiation ability. These results suggested that EPCs/BMSCs sheet can be constructed successful y, and the sheet has strong osteogenic differentiation capability in vitro, providing the foundation for the repair of bone defects.

Objective To investigate the expression and resistant gene of integron in multidrug-resistant Acinetobacter baumannii (MDR-Ab).Methods 51 strains of MDR-Ab isolated from a hospital in August-October 2012 were collected, antimicrobial susceptibility testing was performed.Class I(Int I),II (Int II)and III (Int III)of integrase genes and inte-gron variable region gene cassettes were detected by polymerase chain reaction (PCR),and the homology of integron varia-ble region was analyzed by detection results of restriction fragment length polymorphism (RFLP)and DNA sequencing. Results Positive rate of integrase gene in MDR-Ab was 78.43%(40/51).All genes belonged to Int I,while IntⅡand IntⅢ were not found.Variable region cassettes were detected in 97.50% (n=39)of Int I,there were 5 types of integron gene cassettes:aacA4 in 14 strains,aacA4+catB8 in 22 strains,arr-3 +aacA4 in 1 strain,dfrA15 in 1 strain and arr-3 in 1 strain.Conclusion MDR-Ab isolated from this hospital may be related with Int I expression.Int I carried gene cassettes as follows:aacA4,aacA4+catB8,arr-3+aacA4,dfrA15 and arr-3.

BACKGROUND:Extracellular matrix can simulate microenvironment and make the stem cells proliferate maintaining the characteristics of stem cells wel in vitro. OBJECTIVE:To prepare the extracellular matrix from human bone marrow cells and to analyze its microstructure and composition preliminarily. METHODS:Human bone marrow cells of passage 4 were cultured for 14 days, and the induction medium was used during the last 8 days. After decellularization, cells were removed to prepare human bone marrow cells-derived extracellular matrix. The surface morphology of human bone marrow cells-derived extracellular matrix was observed by inverted microscope and scanning electron microscope. Changes of col agen I and biglycan before and after decellularization were observed by immunofluorescence staining. Human periodontal ligament stem cells were seeded onto human bone marrow cells-derived extracellular matrix, fibronectin coated 6-wel plate and normal culture plate to compare the influence of different matrix on cellmorphology and adhesion. RESULTS AND CONCLUSION:We obtained intact human bone marrow cells-derived extracellular matrix by chemical combined with physical decellularization. The structure and amount of col agen I and biglycan had not been compromised dramatical y after decellularization. Human periodontal ligament stem cells growing on the human bone marrow cells-derived extracellular matrix developed in accordance with the orbit of the extracellular matrix, differing from the original cellmorphology. There were more human periodontal ligament stem cells adhering to the extracellular matrix during the same time. These findings indicate that effective decellularization can produce intact the extracellular matrix membrane without destroying its microstructure. Extracellular matrix protein is not compromised due to decellularization. The extracellular matrix affects cellmorphology and promotes celladhesion. We can use the extracellular matrix model to simulate stem cellmicroenvironment and thereafter, acquire a large number of adult stem cells with high quality in vitro.

BACKGROUND:Human bone marrow cells-derived extracellular matrix can promote proliferation of human periodontal ligament stem cells and maintain stem cellproperties.
OBJECTIVE:To preliminarily investigate the effect of human bone marrow cells-derived extracellular matrix on the proliferation of human periodontal ligament stem cells.
METHODS:Human periodontal ligament stem cells and bone marrow cells were separately derived from human periodontal tissue and jaw bone marrow, and human bone marrow cells-derived extracellular matrix was prepared. Human periodontal ligament stem cells were cultured and purified using limited dilution cloning method, and transmission electron microscope was used for ultrastructure observation. Human periodontal ligament stem cells at passage 2 were cultured with human bone marrow cells-derived extracellular matrix and normal culture medium (control group). The cellcounting kit-8 and flow cytometry were used to determine the proliferation potential of human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix.
RESULTS AND CONCLUSION:Compared with the control group, human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix had a superior capacity of proliferation (P<0.05), and the cells met their morphological and biological characteristics, and grew in good conditions. Human bone marrow cells-derived extracellular matrix is a promising matrix for large-scale expanding human periodontal ligament stem cells for future use in stem cel-based therapy.