Aim To establish a new,multi-parameter,real time and qualitative cell migration evaluation method.Methods Lung cancer cell line A549 was cultured on the glass bottom dish.After treated with different dosages of berberine or Rg3 for 24 hours,several scratching lines at the same dimension were made and observed by Living Cell Working Station.8 observation areas were selected randomly and imaged continuously for 24 hours.Transferred Area(TA),Transferred Distance(TD),Single Cell Transferred Speed(SCTS)and the Cell Division Number among defined area(CDN)were analyzed after getting sequence images.Results The focus stage and the incubation system were sufficient to keep cell proliferation and made it possible for long term observation.Berberine at 25 μmol·L~(-1) and 12.5 μmol·L~(-1) could inhibit the migration of A549(P<0.01).The analysis results of TA,TD,SCTS and CDN were basically coincident.Rg3 at 0.1 mmol·L~(-1) could inhibit SCTS and promote CDN in 6,12 and 24 h(P<0.01),while decrease both TA and TD in 24 hs.Conclusion The method is sensitive,rapid and simple to be applied in the research of tumor metastasis,wound healing and inflammatory response with real time,in-situ and multi-parameters.

Objective To evaluate the effect of comprehensive experiments on multiple trauma emergency rescue in intensive training before nursing practice. Methods 772 students were divided into groups, 16 to 18 students in each group,each team distributed teaching notes, designed experiment scheme toget her, selected and played roles, undertook tasks, practiced independently before the class,then performed comprehensive training in class. Results 94.8% of the nursing students and 100.0% of the training teachers supported this method of comprehensive experiments in intensive training. Conclusions Nursing comprehensive experiments can shorten the gap between the clinical practice and class teaching, improve the students' professional ability and general quality. It is a popular education mode on intensive training before clinical practice.

Objective Oral and Maxillofacial trauma Diagnosis and Treatment Expert system (OMDTES)was used in maxillofacial trauma PBL teaching to improve the quality of PBL teaching,and perfeet the assessment criteria, Methods OMDTES was applied in two procedures of maxillofacial injury PBL teaching activity:the preparation of lesson plan and assessment criteria.Then a questionnaire was designed to assess the effect of this new mode of PBL teaching.Results The new teaching model is welcomed by students and can improve students'interest in learning,the satisfaction of teaching and ameliorate method of assessment of PBL teaching.Conclusion Applying OMDTES in oral and maxinofacial injury PBL teaching has special advantages in raising the leavnevs'activity of learning,training their ability of chnical thinking and analysis.And it is worthy of further research and amelioration.

Objective:To investigate the expression and biological significance of MicroRNA-204 in nasopharyngeal carcinoma (NPC). Methods: qRT-PCR was applied to detect the relative expression of miR-204 in 43 paired nasopharyngeal carcinoma in comparison to the normal nasal mucosa tissues. Pearson chi-square test was used to analyze the relationship between the miR-204 expression and clinical features. The expressions of Bcl-2 and SIRT1 were measured by immunohistochemistry ( IHC ) , Spearman correlation analysis was used to analyze the relationship between miR-204 and Bcl-2,as well as SIRT1. We then transfected the miR-204 mimics into CNE-2 cells,then the Western blot was used to detect the expression of Bcl-2 and SIRT1,which were considered as the potential targets of miR-204. Results:The relative expressions of miR-204 was significantly downregulated in NPC tissues compared to those of the matched normal tumor-adjacent tissues(P<0. 05). Low expression of miR-204 was significantly associated with lymphatic metastasis(P<0. 05) and advanced TNM stage(Ⅲ+Ⅳ,P<0. 05). The expressions of Bcl-2 and Sirt1 in lower miR-204 level group were both higher than in higher miR-204 level group ( P<0. 05 ) . Both the mRNA and protin expression in CNE-2 cells were down-regulated after transfection. Conclusion: Low expression of miR-204 is related to the malignant clinicopathological features in NPC tissues,and miR-204 may through down-regulate Bcl-2 and SIRT1 to suppress NPC genesis and development.

The hapten of Flumequine(FLU) with four carbon atoms spacer arm(FLUABA) was synthesized and coupled to bovine serum albumin(BSA) as immunogen using activated ester method. Balb/c mice were immunized by the artificial immunogen and the splenocytes of immunized mice were fused with Sp2/0 cells to obtain the monoclonal antibody(McAbs). A hybridoma cell line(DB6-E7) secreting anti-flumequine McAbs was obtained by limited dilution method and screened by indirect enzyme-linked immunosorbent assay(ELISA) using heterogenous coating antigen. The results showed that the subtype of the McAb was IgG_1, and the affinity was 8.19×10~8 L/mol. The haptens of FLU, FLUABA and FLUACA, with different space arm, were separately linked to ovalbumin(OVA) for heterologous or homologous coating antigen. The results of indirect ELISA and indirect competitive ELISA(icELISA) indicated that the heterologous coating antigen could improve the sensitivity of ELISA significantly. By heterologous coating antigen(FLU-OVA), the icELISA showed an IC_(50) value of 26.33 μg/L, LOD of 4.0 μg/L, and the workable range of 8-114 μg/L (IC_(20)-IC_(80)). Cross-reactivity studies showed that the McAbs were quiet specific for FLU, no cross-reactivity(<0.1%) was detected between the obtained McAbs and the quinolones compounds or other structural similarity compounds. The developed icELISA for FLU can satisfy the detection criteria of flumequine in animal food-products.

BACKGROUND:The adhesion between muscle tendon and surrounding tissue after tendon restoration is one hot topic in clinic.
OBJECTIVE:To construct a deep flexor tendon transection model in the third toe of female Leghorn chicken and to explore the effect of peritenon transplantation on the prevention of tendon adhesions.
METHODS:After the transection models were successful y established, the third toe of left claw was taken as experimental group A. The transected tendon was sutured, the tendon anastomosis end was wrapped with the peritenon of the deep flexor tendon from the ipsilateral fourth toe (experimental group B). The third toe of right claw served as control group A, the transfected tendon was sutured and restored with the peritenon. The fourth toe of right claw was taken as control group B. Gross observation and histological observation of the tendon were performed.
RESULTS AND CONCLUSION:At 28 days postoperatively, gross observation and histological observation of Leghorn chicken were performed and compared using the Kruskal-Wal isH and Nemenyi test, respectively. The results showed that, the therapeutic effect was better in experimental group A than in control group A (P<0.05), but slightly poorer compared with control group B and experimental group B (P<0.05). The postoperative effect was better in experimental group B than in control group A (P<0.05) and showed no significant difference between experimental group B and control group B (P>0.05). The flexor function was evaluated and compared with the least significant difference t-test. The results showed that the postoperative effect was better in experimental group A than in control group A (P<0.05), but slightly poorer compared with control group B and experimental group B (P<0.05). The postoperative effect was better in experimental group B than in control group A (P<0.05) and showed no significant difference compared with control group B (P>0.05). Peritenon transplantation can effectively prevent tendon adhesions and has little impact on normal tendon sliding.

BACKGROUND:Tendon adhesions after tendon repair have been one of the problems difficult to solve in clinical practice. OBJECTIVE:To introduce the current situation and progress of the prevention of postoperative tendon adhesions. METHODS: A computer-based search of CNKI, Wanfang, PubMed databases for articles related to prevention of tendon adhesions published from January 1971 to May 2014 using the keywords of “tendon injury; tendon adhesions; progress” in Chinese and English, respectively. Repetitive content and irrelevant articles were excluded, and finaly 40 articles were included in result analysis. RESULTS AND CONCLUSION: In this paper, we analyze the characteristics of tendons and causes of tendon adhesions, summarize the methods for prevention of tendon adhesions and the therapeutic effects, and thereby analyze the pros and cons of various methods and materials for prevention of tendon adhesions. Prevention of tendon adhesions after tendon repair involves various aspects: thorough debridement, better surgical approach, microsurgical repair of tendon tissues, applications of drugs and barrier materials, physical therapy as wel as appropriate functional exercise after tendon repair. The integrated use of these means can be done to prevent the occurrence of postoperative tendon adhesions.

Objective To investigate whether calcitonin gene-related peptide(CGRP) can influence the proliferation and phenotypic modulation of vascular smooth muscle cells(VSMCs),and what is the relationship between them. Methods Vascular smooth muscle cells(VSMCs) were cultured respectively with rat aorta cultivated for 8 days in vitro and with normal aorta(not culture) through the explant-attached method,and CGRP was added into the culture medium of the experimental groups.The proliferation of cells was labeled by 5-bromodeoxyuridine(5-BrdU) with immunocytochemical method,and the mRNA expression of hypertension-related gene-1(HRG-1) and smooth muscle 22 alpha(SM22?) were determined by RT-PCR. Results The proliferating cells labeled by BrDU from the aorta cultured for 8 days in vitro were increased notablly and the mRNA expression of HRG-1 and SM22? were decreased.While the VSMCs were cultured in the culture medium containing CGRP,the proliferous cells labeled by BrdU were obviously decreased and the mRNA expression of HRG-1 and SM22? were significantly increased.Conclusion It is showed that CGRP could inhibit the proliferation of VSMCs and change the phenotype of VSMCs from synthesize to contractile type.It might be a good cellular model which provides a good experimental platform to research the proliferating vascular disease as well as its prevention and treatment.

Objective To locally inject human umbilical cord blood (HUCB) mesenchymal stem cells (MSCs) to rat traumatic brain injury (TBI) model to investigate expression of neural markers and neurological functional improvement. Methods HUCB-MSCs were labeled by bis-benzimide for over 24 hours and stereotactically transplanted into the brain of the rats. All rats were divided into four groups, ie, sham injury group, TBI group, control (TBI + PBS) group and treatment (TBI + MSCs) group, Im-munohistochemical methods and immanofluorescence staining were used to observe the survival, migration and differentiation of the transplanted cells. The neurological functional improvement was evaluated by u-sing the neurological severity score (NSS). Results There existed a large number of MSCs survived in local region of the brain that received transplants, when some MSCs differentiated into neurons or astro-cytes and expressed the neurocyte markers including NSE and GFAP around the grafted site. Treatment group had significantly improved scores compared with sham injury group, TBI group and control group. Conclusions HUCB-MSCs transplantation can potentially improve neurological functional after TBI and may be a good alternative to bone marrow cells for stem cell transplantation or cell therapy.