Objective To explore the relationship between microRNA (miRNA)-200c expression,epithelial-mesenchymal transition (EMT) and the sensitivity to gefitinib in colon cancer cells.Methods The inhibitory effects of gefitinib on four types of colon cancer cell lines (HT29,SW620,HCT1116,SW480) were examined by cell counting kit-8 (CCK-8) assay.The expressions of miRNA-200c,epithelial marker (E-cadherin) and mesenchymal markers (vimentin and zinc finger Ebox binding homeobox 1 ZEB 1) at mRNA level in four types of colon cancer cell lines were detected by fluorescence quantitative polymerase chain reaction.The expressions of E-cadherin,vimentin and ZEB 1 at protein level were determined by Western blot.After up-or down-regulated the expression of miRNA-200c,the changes of the expression of EMT related genes and the sensitivity to gefitinib were observed.Results HT29 cells were most sensitive to gefitinib (IC50 =(7.70 ± 0.31) μmol/L),in which the expressions of both miRNA-200c and E-cadherin were the highest,and the expressions of vimentin and ZEB1 were extremely low.HCT116 and SW480 were moderately sensitive to gefitinib (IC50=(11.88±0.97) and (16.63±0.45) μmol/L),and the expressions of miRNA-200c and Ecadherin were moderate.SW620 cell line was most insensitive to gefitinib (IC50 =(26.43 ± 3.68)μmol/L),the expressions of miRNA-200c and E-cadherin were the lowest.The expressions of vimentin and ZEB 1 in SW620 were higher than that of the other three types of cell lines.After upregulated the expression of miRNA 200c,the expression of E-cadherin in SW620 cells increased,the expression of ZEB 1 and vimentin decreased,and the sensitivity to gefitinib increased.After downregulated the expression of miRNA-200c,the expression of E-cadherin in HT29 cells decreased,the expression of ZEB 1 and vimentin increased,and the sensitivity to gefitinib decreased.Conclusion miRNA-200c may up-regulate the expression of E-cadherin through EMT regulation,and then influence the sensitivity to gefitinib in colon cancer cells.

Objective To investigate the effects of interferon-α (IFN-α) and gefitinib on the proliferation and apoptosis of human colon cancer cell line HCT116. Methods Colon cancer cell line HCT116 was selected as research objective. The biological effects of IFN-α and gefitinib alone or combined on the cells were observed at different time point (after worked for 24, 48 and 72 hours). The proliferation inhibition of the medicine on the HCT116 cells was measured by methyl thiazolyl tetrazolium (MTT) assay. Morphologic changes were observed under optical microscope. Apoptosis was measured by flow cytometry (FCM). The results were analyzed with SPSS 13.0 software, two groups compare was tested by t test, and single factor variance analysis was for multiple group data compare. Results IFN-α and gefitinib alone or combined could significantly inhibit the proliferation of HCT116 cells (P<0.05), and there was a time and dose-dependent manner between the degree of inhibition and the working time and concentration of the medicine. With the work of the medicine, apoptosis morphologic changes were observed in the cells. And FCM result indicated that the apoptosis rate significantly increased. After treated with IFN-α and gefitinib alone or combined for 72h, the cell apoptosis rate were 15.6%±0.6%, 13.6%±0.4% and 31.2%±0.3% respectively, which was obviously higher than control group (6.8%±0.3%, P<0.05). Conclusion Both IFN-α and gefitinib were able to inhibit the proliferation and induce apoptosis of HCT116 cells moreover, and a synergistic effect was observed while combine used there two medicines.

OBJECTIVE:To establish a method for the determination of ursolic acid in pedo-stomach-invigorating&digestion-promoting granule.METHODS:The determination was performed by TLC-scanning method in which chloro-form-acetone-glacial acetic acid(18∶2∶0.2)was used as developing solvent,and10%sulphuric acid ethanol solution was taken as developer,which were baked under110℃until limpid prunosus spots were occurred,the scanning method was dual wavelength reflection zigzag scanning with detection wavelength at520nm,reference wavelength at700nm,linearity parameter Sx=3and slit size at0.4mm?0.4mm.RESULTS:Ursolic acid has a good linear relation with peak area score when its sample size was within the range of2?g～10?g(r=0.9996),the average recovery is97.8%(RSD=1.4%).CONCLUSION:This me_ thod can be served as a quality control for pedo-stomach-invigorating&digestion-promoting granule.

Objective To investigate the changes of regulatory T cells (Treg) distribution in trinitrobenzene sulfonic acid (TNBS) induced mice colitis and the therapeutic effects of dexamethasone (DEX).Methods A total of 40 mice were evenly divided into healthy control group,ethanol control group,model group and DEX treatment group.The model group was given 2.5 mg TNBS through enema,while healthy control group and ethanol control group were administered with 0.9％ NaCl solution and ethanol solution respectively.DEX treatment group was intraperitoneal injected with DEX (0.6 mg/kg) since the first day after modeling.Mice were executed at 3rd,5th and 7th day after modeling,mice body weight of each group were observed and the rate of both CD25+ forkhead box P3(Foxp3) positive and CD4 positive cells in spleen and mesenteric lymph nodes were measured by flowcytometry.At the 7th day,disease activity index (DAI) and colonic histopathological score were evaluated,the expression of Foxp3 in colon tissue were measured by immunohistochemistry.t-test was applied for each group comparison.Results Compared with healthy control group and ethanol control group,the mice body weight of model group significantly decreased (t=3.604 and 2.422,both P＜0.05); DAI score and colonic histopathologic score increased (t=2.630 and 2.438,both P＜0.05;t=7.910 and 6.600; both P＜0.01).Compared with the model group,DAI score decreased (t=2.076,P＞0.05) and histopathologic severity in DEX treatment group improved significantly (t =2.320,P＜0.05).At the 7th day after modeling,the rate of both CD25+ Foxp3 positive and CD4 positive cells in spleen and mesenteric lymph nodes of model group was 3.320％ ± 0.533％ and 2.888％±0.379％,lower than that of healthy control group (5.379％ ±0.518％ and 4.688％±0.553％; t=2.769 and 2.686; both P＜0.05).After DEX treatment,the rate increased.The expression of Foxp3 in colon tissue of healthy control group,model group and DEX treatment group was 3.000±0.577,7.200±0.800 and 6.000±0.931 per high power field respectively,and the expression of model group and DEX group significantly increased (t=4.257 and 2.739,both P＜ 0.05).Conclusions The distribution of Treg in spleen and colon were abnormal in TNBS-induced mice colitis model.DEX may improve the disease state of the mice model through upregulating Treg.

Objective To study the value of EUS-FNA cytology and fluid carcinoembryonic antigen (CEA) for differential diagnosis of malignant and benign pancreatic cystic lesions.Methods Data of 27 patients who underwent EUS-FNA were reviewed.According to Youden exponent,the optimal cut-off points for cyst fluid CEA were determined by receiver operating characteristic (ROC) curve.Compared with surgical pathology,the accuracy,sensitivity,specificity,positive predictive value (PPV) and negative predictive value (NPV) of the EUS imaging,cytology as well as cyst fluid CEA were determined.Results Of the 27 cases,14 were diagnosed as benign lesions,13 were diagnosed as malignant or premalignant lesions.The accuracy,sensitivity,specificity,PPV and NPV of EUS imaging were 77.8％ (21/27),69.2％(9/13),85.7％ (12/14),81.8％ (9/11) and 75.0％ (12/16).The accuracy,sensitivity,specificity,PPV and NPV of EUS-FNA cytology were 85.2％ (23/27),76.9％ (10/13),92.9％ (13/14),90.9％ (10/11),and 81.3％ (13/16).The corresponding values of fluid carcinoembryonic antigen under the ROCderived ideal cut-off were 74.1％ (20/27),84.6％ (11/13),64.3％ (9/14),68.8％ (11/16) and 81.8％ (9/11) (CEA ＞ 22.24 ng/ml).Conclusion EUS-FNA cytology is highly accurate and specific for differential diagnosis of malignant and benign pancreatic cystic lesions.Cyst fluid CEA shows better sensitivity.EUS-FNA cytology and cyst fluid CEA analysis can basically meet the requirement of differentiating the benign and (pre)malignant pancreatic cystic lesions.

Objective To evaluate the diagnostic value of endoscopic ultrasound guided fine needle aspiration (EUS-FNA) in combination with flow cytometry (FCM) in lymphoma.Methods From January 2011 to December 2011,the cases of suspicious lymphoma with EUS-FNA examination at Shanghai Ruijin Hospital were retrospectively analyzed.The final diagnosis was according to pathological diagnosis of specimen from the surgery and follow up results.The sensitivity,specificity and accuracy of EUS-FNA combined with FCM in lymphoma diagnosis were initially analyzed.Results A total of 14 suspicious lymphoma patients were collected,eight cases were diagnosed as lymphoma by pathological examination of specimen from the surgery or.tissue from aspiration,four cases were non-lymphoma lesions and two cases still had no final diagnosis.The sensitivity and specificity of FCM alone in lymphoma diagnosis were 4/8 and 4/4 respectively.Six cases of lymphoma were detected by EUS-FNA with FCM.The sensitivity,specificity and accuracy of EUS FNA combined with FCM were 6/8,6/6 and 10/12 respectively.Conclusion EUS-FNA combined with FCM has better diagnostic value in lymphoma,especially for gastrointestinal lymphoma and those surrounding deep lesions.

Objective To evaluate the diagnostic value of endoscopic ultrasonography (EUS ) combined with detection of immunoglobulin (Ig ) and T‐cell receptor (TCR ) gene rearrangements in primary gastrointestinal lymphoma (PGIL) .Methods From 12nd January ,2012 to 23rd May ,2014 ,the clinical data of 24 patients with suspicious PGIL under endoscopy (regular biopsy negative and without treatment) and underwent further EUS examination was retrospectively analyzed .All patients received EUS‐guided biopsy or EUS‐guided fine needle aspiration (FNA) and the tissue specimens were detected for Ig and TCR gene rearrangements .Considering biopsy result ,surgical pathological diagnosis and follow‐up result as gold standard ,the clinical significance of sensitivity ,specificity ,positive predictive value (PPV) , negative predictive value (NPV ) and accuracy of EUS combined with Ig /TCR gene rearrangements in PGIL were explored .Results Among 24 patients ,19 patients were finally diagnosed as PGIL ,which were all non‐Hodgkin′s lymphoma (NHL ) ;monoclonal gene rearrangement was found in 13 cases of the 19 cases .The left five cases were not lymphoma lesions (three cases of gastritis ,one case of leather stomach and one case of malignant melanoma) with no monoclonal gene rearrangement .In cases diagnosed as PGIL , 14 were B cell NHL ,which included eight cases of muscosa‐associated lymphoid tissue (MALT) lymphoma and six cases of diffuse large B cell lymphoma .Of which ,immunoglobulin heavy chain (IgH)/immunoglobulin kappa (IgK) gene rearrangement was found in 11 cases .A total of five cases were T cell NHL including one case of anaplastic large cell lymphoma , one case of NK /T cell lymphoma and three cases of enteropathy‐associated T‐cell lymphomas . TCR gene rearrangement was found in two cases .Because theoretically ,there was no gene rearrangement in natural killer (NK )/T cell lymphoma ,so NK /T cell lymphoma was excluded from statistical analysis .The sensitivity ,specificity ,PPV ,NPV and accuracy of EUS combined with Ig /TCR gene rearrangements detection in PGIL were 72 .2% ,100 .0% ,100 .0% , 50 .0% and 78 .3% ,respectively .Conclusion The detection of monoclonal gene rearrangement in tissues from EUS‐guided biopsy or EUS‐guided FNA had better diagnostic value in PGIL ,which improved the objectivity and accuracy of lymphoma diagnosis .

0.05).CONCLUSION:The excellent mechanical properties of composite ligament can meet the mechanical requirements of appropriate ligament tissue engineering scaffolds.

0.05), and the total score were similar in the two groups. Type Ⅱ collagen in the two groups was strongly positive by immunohistochemistry staining. CONCLUSION: Cryopreserved allogenic osteochondral pillars transplantation can repair small full-thickness articular cartilage defects. The chondrocytes are alive in short time, and they can secret cartilage matrix without obvious rejection. It has similar efficacy in histology with autogenic osteochondral pillar transplantation.

Objective:To explore the possible mechanism of exacerbation of anxiety-like behavior in db/db mice after distal middle cerebral artery occlusion (dMCAO).Methods:The db/db mice was used to establish a type 2 diabetes mellitus model. Meanwhile, heterozygous db/+ mice and C57 wild-type (WT) mice were chosen as double control groups. Then a permanent distal middle cerebral artery occlusion model was employed as an acute ischemic stroke model. The blood glucose levels before and post-dMCAO surgery on day1, day3, and day5 were detected. The brain tissue loss at 35 days after stroke was measured by immunofluorescent staining of MAP2. The open-field test was performed to estimate anxiety-like behavior and general motor and exploring ability of the animals. Axons and myelin were immunostained with non-phosphorylated neurofilaments (SMI32) and myelin basic protein (MBP), respectively, to evaluate differences in white matter integrity in WT, db/+ and db/db mice 35 days after stroke. The correlation between SMI32/MBP and open field test parameters (time in center and corner) was analyzed. Flow cytometry was employed to detect the amount of T cells and B cells, including regulatory T cells (Tregs) in the brain tissue.Results:Blood glucose levels in db/db mice were significantly higher than db/+ mice and WT mice in both sham and dMCAO groups ( P<0.01). There was no significant difference in brain tissue loss 35 days post-stroke among db/db mice, db/+ mice, and WT mice. In the open field test, there were significant differences in the total distance of db/db mice, db/+ and WT mice in the sham and dMCAO groups. Db/db mice shorter than db/+ mice ( P<0.05), WT mice ( P<0.01), and db/+ mice shorter than WT mice ( P<0.05). There were significant time differences in the center among db/db, db/+, and WT mice in sham and dMCAO groups. In both the sham and dMCAO groups, db/db mice spent less time in the center area of the open field than WT mice ( P<0.01). In the sham group, db/+ mice spent less time in the center area than WT mice ( P<0.05). In dMCAO group, db/db mice spent less time in the center area than db/+mice ( P<0.05), and db/+ mice spent less time in the center area than WT mice ( P<0.01). For the time in the corner, in both the sham and dMCAO groups, db/db mice and db/+ mice consumed more time than WT mice ( P<0.01 or <0.05). In the dMCAO group, db/db mice spent more time in the corner than db/+ mice ( P<0.05). Referring to white matter injury, an increased SMI32/MBP ratio in EC area and CTX area (data was not shown in this article) after dMCAO in db/db, db/+ and WT mice were detected. In EC area, db/db mice have a higher SMI32 ratio than db/+ mice and WT mice: 4.24 ± 0.37 vs. 1.96 ± 0.37, 1.80 ± 0.36, both have significant differences ( P<0.01). For db/db mice and WT mice, the SMI32/MBP ratio negatively correlates with time in center and positive correlation with time in the corner. Three days after dMCAO, the total cells of CD 3+ T cells, CD 8+ cells, Tregs, in db/db mice group have significantly decreased compared to WT group: 4 079 ± 1 345 vs. 70 055 ± 3 374, 141.30 ± 28.36 vs. 2 714.00 ± 463.20, 148.00 ± 61.15 vs. 3 007.00 ± 639.90 ( P<0.01), while B cell has no change between two groups. Conclusions:By comparing the severity of anxiety-like behavior of db/db mice, the severity of white matter injury, and the number of T cells and B cells in brain tissue after dMCAO, immune-mediated brain white matter injury may aggravate db/db mice′s post-dMCAO anxiety-like behavior. Due to the gene dose effect, db/+ mice are not suitable as a control group for db/db mice in animal experiments involving anxiety-like behavior assessment.