AIM:To detect the expression of miRNA-363 and SOX4 in osteosarcoma tissues and to investigate the effect of miRNA-363 on the viability and apoptosis of human osteosarcoma cell line MG-63.METHODS: Real-time PCR was used to detect the expression level and the relationship of miRNA-363 and SOX4 mRNA in the osteosarcoma tis-sues and the corresponding paratumorous tissues collected from 63 patients.The expression levels of miRNA-363 and SOX4 in osteosarcoma cell line MG-63 after transfected with miRNA-363 mimics were measured .The cell viability was measured by CCK-8 assay.Flow cytometry was used to monitor the changes of cell cycle and apoptosis .The changes of SOX4 and miRNA-363 expression levels in the MG-63 cells after transfection with SOX4 siRNA or pcDNA/SOX4 was detect by real-time PCR.RESULTS:The expressed level of miRNA-363 was lower , and the expression level of SOX 4 was higher in the osteosarcoma tissues than those in the adjacent normal tissues .A significantly negative correlation between the expression levels of miRNA-363 and SOX4 was observed .The expression of miRNA-363 in the MG-63 cells after transfection with miRNA-363 mimics was significantly up-regulated, while the expression of SOX4 in the MG-63 cells was significantly down-regulated , with significant difference as compared with the cells transfected with miRNA-NC and control cells .The viability of MG-63 cells was inhibited, the cell cycle was arrested in G0/G1 phase, and the cell apoptosis was increased by transfec-tion with miRNA-363 mimics.The relative protein expression levels of SOX 4 in SOX4 siRNA group and pcNDA/SOX4 group were significantly different from those in negative control group , but the relative expression levels of miRNA-363 had no significant difference .Over-expression of SOX4 restored the viability of the MG-63 cells reduced by miR-363.CON-CLUSION:The expression level of miRNA-363 is low in human osteosarcoma tissue .miRNA-363 may inhibits the viabili-ty of osteosarcoma cell line MG-63 and promotes cell apoptosis in vitro via inhibiting the SOX4 expression.

BACKGROUND:The treatment of limbs’ open fracture with severe soft tissue injury is very complicated. There are many wound exudates, high frequency of dressing change, and high infection rate. How to deal with the contradiction between wound treatment and fracture fixation is the considerable problem in clinical therapy. ＜br> OBJECTIVE:To explore the effect of external fixator combined with vacuum sealing drainage on limbs’ open fracture with severe soft tissue injuries. ＜br> METHODS:A total of 34 patients with limbs’ open fracture with severe soft tissue injuries were enrol ed in the First Affiliated Hospital, Xinxiang Medical Col ege from December 2009 to December 2010. There were 20 males and 14 females, with an average age of 36.6 years old. There were 10 cases of upper limb fracture and 24 cases of lower limb fracture. After debridement, cutaneous deficiency accounted for 1%to 6%of surface area. On the basis of external fixation, the wound received vacuum sealing drainage (treatment group;n=17) and conventional dressing (control group;n=17). According to limb swel ing and granulation growth, delayed suture, skin graft or flap to cover the wound were used. After treatment, infection rate of the wound and growth of granulation were observed. In accordance with wound healing and growth of granulation, curative effects were evaluated. ＜br> RESULTS AND CONCLUSION:In the treatment group, there were 11 cases of healing, 5 cases of effective effects, and 1 case of ineffective effect, with a total effective rate of 94%and the infection rate of about 15%. In the control group, there were 7 cases of healing, 6 cases of effective effects, and 4 cases of ineffective effects, with a total effective rate of 76%and the infection rate of about 40%. Significant differences in healing rate, total effective rate and infection rate were detected between the two groups (P＜0.05). The healing time of wound was 8.66-16.23 days in the treatment group and 15.68-22.36 days in the control group. The healing time was significantly shorter in the treatment group than in the control group (P＜0.05). It is thus clear that vacuum sealing drainage has obvious advantages in repair of limbs’ open fracture with severe soft tissue injury compared with the conventional dressing.

Objective: To study the expression and purification of the chimeric virus-like particles displaying epitopes of EV71 as a candidate of enterovirus 71 gene recombined vaccinet. Methods: The fusion protein hepatitis B core (HBc)-SP70 was constructed by inserting SP70 into the main immunogenic region of truncated hepatitis B core antigen (HBcAg) sequence, expressed in E. Coli, and purified through sonication, ion exchange chromatography, CsCl cushion centrifugation and density gradient centrifugation. Then its antigenicity was detected by ELISA and Western blot assay. Results: Recombinant plasmid pHBc-SP70 was successfully constructed. And the soluble fusion protein was efficiently expressed induced by IPTG. The purity of the chimeric virus-like particles (VLPs) was up to 90% after the purification process described in method . The purified fusion protein HBc-SP70 could be spontaneously folded and assembled into empty virus-like particles and react with the monoclonal antibodies against HBc and SP70. Conclusions The chimeric VLPs displaying epitopes of EV71 were efficiently expressed and purified in E. Coli. with excellent antigenicity, which laid a foundation for evaluation of the immune effect evoked by this EV71 gene recombined vaccine.

OBJECTIVETo downregulate the expression of pituitary tumor transforming gene 1 (PTTG1) in osteosarcoma (OS) cells by siRNA technology and to investigate related biological impact on cell proliferation, cell cycle and cell invasion of OS.

METHODSThree OS cell lines and osteoblast hFOB1.19 cell line were used in this study. Control siRNA and PTTG1 siRNA were employed to transfect OS U2OS cells, and PTTG1 protein level was detected by Western blot after the transfection. Effects of PTTG1 siRNA on cell proliferation, cell cycle and cell invasion were investigated by CCK-8, flow cytometry and Boyden chamber, respectively. Finally, activity of Akt and its downstream target gene expression were analyzed by Western blot in U2OS cells upon various treatments.

RESULTSExpression of PTTG1 protein in 3 OS cells (MG-63, SaOS-2 and U2OS) was significantly higher than that in osteoblast hFOB1.19, among which U2OS cells displayed the highest level. PTTG1 siRNA markedly downregulated the expression of PTTG1 protein in U2OS cells, leading to obvious inhibition of cell proliferation, altered cell cycle distribution and reduced ability of invasion of U2OS cells. Moreover, downregulation of PTTG1 reduced the expression of p-Akt (S473 and T308), MMP-2 and MMP-9 proteins, along with enhanced expression of p21 and E-cadherin proteins.

CONCLUSIONSPTTG1 may be tightly linked to the development of OS and therefore may serve as a novel target for precision therapy of OS.

Bone Neoplasms ; metabolism ; pathology ; Cadherins ; metabolism ; Cell Cycle ; drug effects ; physiology ; Cell Movement ; Cell Proliferation ; drug effects ; physiology ; Down-Regulation ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Osteosarcoma ; metabolism ; pathology ; RNA, Small Interfering ; pharmacology ; Securin ; genetics ; metabolism ; Transfection