Objective To investigate expression of VEGF and the in vitro angiogenesis ability induced by ovarian cancer cells after RNA interference on expression of matrix metalloproteinase-2 (MMP-2)gene.Methods One specific target sequence of MMP-2 and one non-specific sequence (NC group) were chosen,the DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells,mRNA and protein expression of MMP-2 and VEGF genes were examined by RT-PCR and Western blot analysis,and the angiogenesis ability was detected by in vitro angiogenic assay.Results When compared with the NC group,the mRNA expression of MMP-2 and VEGF were decreased by 78.8 ％ and 75.5 ％ (P ＜ 0.05) in 48 h after transfected,respectively,and protein expression was decreased by 81.2 ％ and 78.3 ％ (P ＜ 0.05) at the same time point.In vitro angiogenic assay suggested that the ability of angiogenesis was inhibited when down-regulated of MMP-2 gene (P ＜ 0.05).Conclusion Down-regulation of MMP-2 gene in ovarian cancer cells by RNA interference could inhibit its VEGF expression and in vitro angiogenesis induced by ovarian cancer cells,which suggestes that the inhibition of MMP-2 gene has an anti-angiogenesis effect,and MMP-2 gene could be a potential target for ovarian cancer gene-therapy.

Objective To observe the clinical efficacy of relatively proximal humeral locking compression plate( LPHP) with the traditional plate for treatment of proximal humeral fractures .Methods The proximal humeral fractures in patients with 58 cases,the patients were divided into two groups ,the treatment group of 32 cases of locking compression plate in the treatment of T plate ,clover plate in the treatment of 26 patients of the control group received traditional.According to the Neer score compared with two groups of patients with postoperative recovery and compli -cations.Results All the 58 patients for 11 ~15 months of follow-up,the treatment group found no fracture ,fracture nonunion,screw and plate humeral head necrosis ,4 cases of shoulder pain was limited;15 cases of excellent score Neer function,good in 12 cases,5 cases,the excellent and good rate was 84.4%.The control group had 3 cases,2 ca-ses of screw loosening of plate fracture ,3 cases of nonunion ,2 cases of humeral head necrosis ,7 cases of shoulder pain limited.6 cases of excellent score Neer function ,good in 9 cases,3 cases,poor in 8 cases,the excellent and good rate was 57.7%.There was significant differences between the two groups (χ2 =5.113,P <0.05) excellent rate. Conclusion LPHP exact treatment of proximal humeral fractures has fewer complications , which can help patients with early rehabilitation exercises .

Objective To comparatively study the features of two reprogramming systems of induced pluripotent stem cells (iPSCs)from human dental origin.Methods Two kinds of reprogramming system,i.e.STEMCCA lentivirus /feed layer and Sen-dai virus /matrigel were used to induce human stem cells from apical papilla(SCAP)into iPSCs,respectively.The induction efficien-cies,workload of generating iPSCs,aneuploidy karyotype ratio,complexities of eliminating exogenous transcription factors and spe-cific markers expression were compared between these two systems.Results The STEMCCA reprogramming system required to prepare the feeder cell MEF.The reprogramming efficiency was 0.1%.Transcription gene-free iPSCs cells were obtained by the Cre-loxp enzyme digestion technique at the later stage.Sendai virus reprogramming system was feeder-free and the preparation of matrigel was quite simple with unified standard.The reprogramming efficiency was 0.7%,which was much higher than that of STEMCCA system(P <0.05).The exogenous virus and transgenes could be gradually eliminated after several passages of natural subclone.Conclusion The Sendai virus/matrigle reprogramming system is much more applicable for the induction of iPSCs from dental origin than the STEMCCA system.

The process of intervertebral disc degeneration,which could result in intervertebral disc structural and functional change,is a chronic one with multiple factors.The pathophysiologic process is still not completely find out.More and more research reports manifest that certain gene polymorphism also lead to increased risk of intervertebral disc degeneration except environmental factors.Discussions about related genetic factors and their pathophysiological role in the process of degeneration could have a further understanding to disease development.Elucidating genetic components which are associated with degeneration could not only provide insights into the mechanism of the process,but also have clinical significance for early diagnosis and prevention.In order to have a thorough understanding of functional role played by different genes,this paper summarize polymorphism and disease correlation by selecting 15 genes after reviewed the related literature published in recent years.Genetic polymorphisms in 15 genes have been analyzed in association with intervertebral disc degeneration,including aggrecan,collagen Types Ⅰ,Ⅸ and Ⅺ,fibronectin,HAPLN 1,CILP,MMP-1,2 and 3,PARK2,IL-1,6 and VDR.Each genetic polymorphism codes for a protein which has a functional role in the pathogenesis of disease.Among the 15 genes analyzed,polymorphisms in aggrecan,Type Ⅸ collagen,MMP3,IL1,IL6 and VDR show the most promise as functional variants.Genetic studies are necessary for understanding the mechanism of the degeneration.Relevant genetic information could be used as a predictive model for determining individuals' risk for intervertebral disc degeneration eventually.

Objective To investigate the inhibitory effects of RNA interference (RNAi) on the expression of matrix metalloproteinase-2 (MMP-2) gene and growth, adhesion,invasiveness and migration of ovarian cancer cells. Methods One specific target sequence of MMP-2 gone and one non-specific sequence (NC group) were chosen,the medium DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells, the RT-PCR and Western blot were used to detect mRNA and protein expression of MMP-2 gene, the growth ability was detected by MTT assay, the abilities of adhesion was detected by cell adhesion assay, the invasion and migration were detected by Matrigel invasion assay and wound healing assay. Results By contrast to the NC group,the mRNA expression was decreased by 73.8 ％,78.8 ％ and 78.4 ％(P＜ 0.05) in 24 h,48 h and 72 h after transfection and protein expression was decreased by 72.6 ％,81.2 ％ and 76.4 ％(P＜ 0.05) respectively at the same time. The 48 h group had the most efficient inhibitory effect. Cell growth curve revealed that cell growth was not significantly inhibited (P＞ 0.05). Adhesion was significantly reduced,the inhibitory rate was 55.0 ％ at 60 min and 44.8 ％ at 90 min (P＜ 0.05),respectively. Invasion and migration were significantly reduced as well,the inhibitory rate on invasion and migration were 29.7 ％ and 35.8 ％(P＜0.05), respectively. Conclusion siRNA mediated MMP-2 down-regulation in ovarian OVCAR-3 cells can inhibits its adhesion,invasion and migration,but do not significantly affect its growth,suggesting a important target to ovarian cancer gene-therapies.

Objective To investigate the association between serum sex hormone levels as well as body mass index and breast cancer in postmenopausal women .Methods We selected the cases of postmenopausal women who accepted surgical treatment for the first time which from March 2013 to December 2013 in the depart-ment of breast surgery the Affiliated Tumor Hospital of Harbin Medical University ,of which 118 cases of patients with breast cancer and 60 cases of benign breast lesions as control group .Meanwhile,information of body height and weight.were collected.Serum estradiol(E2),estrone(E1),testosterone(TSTO)and androstenedione(AED) concentrations were measured in 118 postmenopausal women with breast cancer patients and 60 matched control subjects by enzyme-linked immunosorbent assay(ELISA),then the results were estimated.Results The levels of serum E2,E1,AED of breast cancer group were significantly higher than those in control group (P<0.05). The serum level of TSTO in cases group was higher than the control group ,but the results were not statistically significant(P>0.05).The levels of body mass index(BMI)were not statistically significant(P>0.05)between breast cancer group and the control group .The level of E1 in breast cancer group and levels of E 1,TSTO,AED in overall of overweight group were significantly higher in overweight (P<0.05),yet we have not found association between levels of hormone and BMI .Conclusion The level of serum sex hormones is higher with breast cancer in postmenopausal women ,while high serum levels of E2,E1,AED maybe associated with breast cancer in postm-enopausal women .Sex hormones are higher in postmenopausal women with high BMI .

OBJECTIVETo compare the expression of pinopodes, the marker of endometrial receptivity, during the implantation window in Kunming mice stimulated with two different doses of raloxifene (RAL).

METHODSForty-eight 8-week-old female Kunming mice were randomly divided into 4 groups (n=12), namely saline group, clomiphene citrate (CC, 18 mg/kg) group, RAL (33 mg/kg) group and RAL (44 mg/kg group). In each group, the mice received intragastric administration of 1 mL of normal saline containing CC or RAL at the specified doses or saline only as indicated for ovulation induction, once daily for 2 days. The mice received then injection with 5 IU human chorionic gonadotropin (HCG) and mated and on day 4.5 of gestation, the pregnant mice were sacrificed for examination of the uterus with scanning electron microscopy.

RESULTSAbundant and well developed pinopodes were observed in the endometrium of the mice in the 2 RAL groups and in the saline control group. The mice in CC group showed obviously reduced endometrial pinopodes with poor development.

CONCLUSIONSRAL at two different doses does not obviously affect the expression of pinopodes in the uterine epithelium of mice, suggesting the safety of RAL at these two doses for ovulation induction without causing adverse effects on endometrial receptivity.

Objective:Astragalus polysaccharide (APS) was used in combination with ionizing radiation (IR) to investigate the mechanism of APS on the radiosensitivity of human nasopharyngeal narcinoma CNE-1 cells and the epithelial-mesenchymal transition (EMT). Method:Cell counting kit-8 (CCK-8) was used to detect the cytotoxicity of different concentrations of APS (0，6.25，12.5，25，50，100，200 g·L^-1) on CNE-1 cells. Colony formation assay was used to calculate the survival fraction (survival fraction, SF) of CNE-1 cells treated with 12.5 g·L^-1 APS combined with different radiation doses (0，2，4，6 Gy). The linear quadratic equation mathematical model (LQ) was used to draw the radiosensitivity curve according to SF value. Cell scratch and transwell chamber test were used to detect the migration and invasion ability of cells in each group. The apoptosis of cells in each group was detected by flow cytometry, Western blot was used to detect the expressions of EMT markers, apoptosis markers and protein kinase B/extracellular regulated protein kinases (Akt/ERK) pathway proteins in each group. Result:The results of colony formation assay and radiosensitivity curve showed that the combination of non-toxic dose of 12.5 g·L^-1 APS and radiation dose of 4 Gy could significantly increase the radiosensitivity of CNE-1 cells. Compared with blank group and IR group, APS combined with IR could significantly inhibit the migration and invasion of CNE-1 cells (P<0.05), and increase the rate of apoptosis (P<0.05). In addition, compared with the blank group and the IR group, APS combined with IR could significantly down-regulate the expressions of N-cadherin, p-Akt and p-ERK, and significantly up-regulate the expressions of E-cadherin, Bax and Caspase-3 (P<0.05). Conclusion:APS combined with IR can inhibit the migration and invasion of CNE-1 cells, and increase the apoptosis induced by radiotherapy, which may be related to the inhibition of EMT and Akt/ERK pathway.