Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

Objective To study the current incidence of Keshan disease in Yunnan Province,and provide scientific basis for Keshan disease(KD) prevention and control. Methods Based on the Scheme of KD Surveillance, 16 villages in 11 counties were chosen as surveillance sites by the historical data. An survey was made to the residents in the 16 surveillance sites by filling in the questionnaire, inquiry medical history, clinical examination, electrocardiogram and 2 meters post-anterior chest X-ray for suspected cases. KD cases were diagnosed according to the Diagnostic Criteria for Keshan Disease(GB 17021-1997). The prevalence data of KD in the whole province were collected from the KD case report in 2007 and the trace surveys. Results There were 6877 residents in 16 surveillance sites of 11 surveillance counties and totally 39 KD cases were diagnosed with a detection ratio of 0.57% (39/6877). The detection ratio of latent and chronic KD were 0.41%(28/6877) and 0.16%(11/6877), respectively and no acute or subacute cases were found. The cases aged 5 to 14 years old accounting for 66.67% (26/39). Electrocardiogram examination of 6877 residents were made and 5.25% (361/6877) abnormal electrocardiograms were detected in the 16 surveillance sites. Fifty-five people were checked by chest X-ray and there were 31 cases with heart-chest ratio ≤0.50, 16 cases with heart-chest ratio from 0.51 to 0.55 and 8 cases with heart-chest ratio from 0.56 to 0.60. The prevalence rate and incidence rate of chronic KD were 4.24 per 100 000 and 0.50 per 100 000 in Yunnan. No acute or subacute cases were found and the latent cases were listed. The prevalence rate and incidence rate were 7.76 per 100 000 and 1.18 per 100 000 in the 16 surveillance sites. Conclusions The incidence of KD is low incidence in Yunnan Province. Higher ineidence of chronic KD was detected in the some areas and the corresponding control measures need to be adopted.

Objective In order to master the current situation of Keshan disease in Yunnan province and to provide scientific basis for Keshan disease control and prevention. Methods Eighteen villages were selected as the investigation sites in 6 counties across all the Keshan disease wards in Yunnan province,where the residents were investigated. Then,the villages census data was collected,clinical examination aiming mainly on cardiovascular system was carried out,including electrocardiography and X-ray to the suspected patients. Correct diagnose of Keshan disease was made by the Diagnostic Standard of Keshan Disease(GB 17021-1997). At the same time,10 food samples and 10 hair samples for detecting selenium content in every investigation site. Results There were 9818 residents investigated in the 18 investigation sites in 6 counties,and 34 eases of Keshan disease were found,the total incidence rate was 0.35%(34/9818). Among the 34 Keshan disease eases,32 cases were latent Keshan disease,the incidence rate was 0.33%(32/9818); 2 cases were chronic Keshan disease,the incidence rate was 0.02%(2/9818). There was no any acute and sub acute cases be found. Most Keshan disease cases aged from 5 to 14,67.65% (23/34). Abnormal ECG rate was 6.90% (677/9818). Among 56 X-ray films,47 cases had a cardiothoracic ratio less than or equal to 0.50,83.93%(47/56),5 cases from 0.51 to 0.55,8.93%(5/56),4 cases from 0.56 to 0.60,7.14%(4/56). Selenium content was detected in 180 food samples and 180 hair samples. The average food selenium content (mg/kg) was 0.013±0.010,the lowest content in Yongsheng county (0.006± 0.001),the highest content in Tonghai county(0.027±0.009). The average hair selenium eontentwas(0.252± 0.078)mg/kg,with the lowest(0.145±0.043)mg/kg in Yoagsbeng county,the highest (0.297±0.062)mg/kg in Tonghai county. Conclusions The detected ratio of Keshan disease is low in Yunnan province. Most of Keshan disease patients age from 5 to 14. It was presented that the Keshan disease infectious agents were still strong and active. The foodstuffs and hair Selenium content is low in food and hair sample,and varies in different investigation site. It is necessary to supply selenium for prevent Keshan disease in the severe areas.

Objective To investigate the effects of siRNA targeting CD 147 gene on the expression of CD147 in melanoma cell line A375, and on proliferation of these cells. Methods Previously prepared recombinant plasmid pSUPER/CD147 siRNA was used. In this study, the recombinant was transfected into the A375 cells. The mRNA expression of CD147 was measured by semi-quantitive RT-PCR, the proliferation of the cells by MTT assay. Results After the transfection with pSUPER/CD147 siRNA1 and pSU-PER/CD147 siRNA2, the mRNA expression of CD147 in the A375 cells was significantly down-regulated by 90.81% (P

Objective:To investigate the expression profile variation of long non-coding RNA( lncRNA) in the peripheral blood mononuclear cells of rheumatoid arthritis ( RA ) and healthy controls, and explore the role of lncRNA in the pathogenesis of RA.Methods:A total of 12 RA patients and 11 age-matched healthy controls from University Hospital of Hubei University for Nationalities were recruited.Using lncRNA microarray technology to detect differently expressed lncRNAs in 3 cases of RA PBMCs and 3 cases of healthy controls.GO and Pathway analysis was performed.The coding-non-coding gene co-expression networks of lncRNA and mRNA was constructed based on the correlation analysis,and then searched lncRNA in pathogenesis of RA through the cis-analysis and trans-analysis.Results:A total of 1 615 deregulated lncRNAs and 878 deregulated mRNAs were detected in RA patients.GO analysis of different expressed mRNA may involve in metal ion binding,protein kinase binding,nucleotide binding,regulation of transcription,et al.Pathway analysis of different expressed mRNA may involve in TNF signaling pathway,B cell receptor signaling pathway,pancreatic cancer,system lupus erythematosus endometrial cancer,et al.REL,SMAD3 and ETS1 may play an important role in the pathogenesis and development of RA through cis-analysis and trans-analysis.As the NONHSAG027875,FR378506 and NONHSAT031501 also had the similar function,and they may be related to the pathogenesis and development of RA.Conclusion:Differentially expressed lncRNAs may exert a partial role in RA,and may provide potential targets for future treatment of RA.

In order to establish the optimum condition for purifica tion of the nucleoprotein(NP) of rabies virus by immunoaffinity chromatography, the efficient and non-denaturative eluents(Mg-el) was obtained by using ELISA elution model; furthermore, it didn't damage the activity of NP. Two kind of NPs , expressed by recombinant vaccinia virus (rVac-N) and recombinant baculovirus (BRN), were purified by a Sepharose CL 4B column and a 2C12- Sepharose 4B colum n. By Western-blot and SDS-PAGE, high purity and good antigenical intact NPs w ere identified. The purified ribonucleoprotein (RNP) of rabies virus 5aG strain was also obtained. After immunized with NP and RNP, mice developed a strong anti -nucleoprotein response and were protected against a lethal challenge of rabies virus CVS strain. There were not difference been observed among the mice immuni zed with different purified protein. These data indicate that the NPs are antige nical and immunogenical comparable to the authentic rabies RNP and therefore pre sent a potential source of an effective ,safe and economical subunit vaccine.

0.05);and there were significant diffe-rences between the final diagnosis and pre-hospitalized diagnosis in all patients with VE(?2=47.08 P

To observe BAG-1, EGFR, and PARP-1 expressions in invasive breast cancer and its correlation with clini-cal pathological indicators, as well as to evaluate their clinical significance. Methods:The BAG-l, EGFR, and PARP-1 expressions in a tissue microarray of invasive breast cancer and peritumoral tissues were detected through immunohistochemical staining. The clinical and pathological significance of BAG-1, EGFR, and PARP-1 were evaluated. Results:The BAG-1, EGFR, and PARP-1 expression lev-els are higher in invasive breast cancer tissues than in peritumoral tissues (P<0.05). BAG-1 expression in invasive cancer tissues is not related to age, tumor site, lymph node metastases, and clinical TNM staging of patients, but is related to size, grade, ER, PR, and HER-2 expressions and molecular subtype (P<0.05). EGFR expression is related to size, clinical TNM staging, and molecular subtype (P<0.05). PARP-1 expression is related to grade, lymph node metastases, ER, and molecular subtype (P<0.05). BAG-1 expression is not significantly correlated with EGFR and PARP-1 in all cases, but BAG-1 and PARP-1 expressions are positively correlated in tri-ple-negative breast cancer tissues (P<0.05). Results of the univariate analysis revealed that the BAG-1 and PARP-1 expressions and the molecular subtypes are associated with the prognoses of breast cancer patients. Multivariate analysis revealed that BAG-1 and PARP-1 expressions are factors that are independent of the prognosis. Conclusion: BAG-1, EGFR, and PARP-1 overexpressions in human breast tissues suggest that BAG-1, EGFR, and PARP-1 are related to breast cancer development. BAG-1, EGFR, and PARP-1 are poten-tial biomarkers of breast cancer diagnosis and prognosis.

Aim To construct a subtracted cDNA library o f differentially expressed genes in human gastric carcinoma induced by diallyl dis ulfide(DADS). Methods Differentially expressed cDNA species induc ed by DADS in MGC 803 human gastric carcinoma cell line was determined by using suppression subtractive hybridization (SSH). Then these cDNA species were direct ly inserted into T/A cloning vector to set up the subtractive library. Amplification of th e library was carried out with transformation of E.coli by high voltage electrop erforation. One hundred positive bacteria clones were randomly picked and identi fied using PCR method. Results The amplified library contained more than 1,000 positive bacteria clones. Random analysis of 100 clones with PCR m ethod showed that all clones contained 100～600 bp inserts.Conclusions A subtracted cDNA library of differentially expressed genes in MGC 803 hum an gastric carcinoma cell line induced by DADS is constructed successfully with SSH and T/A cloning techniques. The library is efficient and lays solid foundati on for screening and cloning new and specific tumor correlative genes of human g astric carcinoma, and provides a new idea for further exploring the mechanism of DADS effects on carcinoma cells.

Abstract：Objective To establish a sensitive，rapid and convenient method for the detection of dengue antigen and assist clinical diagnosis of dengue. Methods In this paper，we developed a rapid detection method for dengue antigen based on microfluidic immune magnetic beads. Solidwork software was used to design microfluidic chip，which was prepared by mechanical processing and chemical sealing. Immunomagnetic beads of dengue antibody were prepared by chemical coupling reaction. Using HRP ⁃TMB ⁃H2O2 as color system，dengue NS1 antigen was detected on microfluidic chip carrier by double antibody sandwich method. Finally，57 clinical samples were tested by the novel method and traditional ELISA kit，and the accuracy of the method was analyzed，and the advantages and disadvantages of the two methods were compared. Results 20 minutes was needed to detect dengue NS1 antigen by using the novel ELISA method，and the reaction system only needed 10 μg beads and 10 μL samples. In the verification experiment，the method could distinguish the negative from the positive obviously. The positive sample had color rendering，while the negative and blank samples had no color rendering. In terms of detection performance，the coincidence rate between the new ELISA method and the traditional ELISA method reached 100%. Conclusion The novel ELISA detection platform had the advantages of simple，rapid，reagent and sample saving，high sensitivity，good stability and high accuracy，and could be used for the detection of dengue antigen.