Objective To investigate the role of bone marrow mesenchymal stem cells (BM-MSCs) in the invasion and metastasis of gastric cancer cells and to explore its mechanism.Methods SGC7901 and KATO-Ⅲ gastric cancer cells were co-cultured with BM-MSCs respectively,and the invasion ability of SGC7901 and KATO-Ⅲ gastric cancer cells were detected by Transwell assay.Secondly,CD133 + and CD133-cells were sorted from KATO-Ⅲ gastric cancers and co-cultured with BM-MSCs respectively to compare their changes in invasiveness.Meanwhile,the expressions of p-AKT and epithelial-mesenchymal transition (EMT) relative factors in gastric cancer cells were detected by Western-blot.The role of CD133 in BM-MSCs affecting the ability of invasion of gastric cancer cells was further vertified by the overexpression of CD133 in SGC7901 cells.SPSS17.0 software was used for statistical processing,and the stand deviation of the measurement data were expressed as the standard deviation,independent sample t test was conducted.Results The invasiveness of co-cultured SGC7901 and KATO-Ⅲ cells was significantly enhanced.The invasive ability of KATO-Ⅲ CD133+ cells co-cultured with BM-MSCs tended to increase more significantly than that of co-cultured CD133 cells[(259.0 ± 24.0)vs (58.0 ±5.6),P ＜ 0.001].The expressions of p-AKT,Snail and N-cadherin were significantly increased in co-cultured CD133+ cells (P =0.003,P =0.003,P =0.002),while the expression of E-cadherin was reduced (P =0.021).After co-cultured with BM-MSCs,the expression of E-cadherin was also reduced in CD133-cells (P =0.005),but the expressions of p-AKT,Snail and N-cadherin were no significantly changes (P =0.744,P =0.277,P =0.295).SGC7901 co-cultured with BM-MSC after overexpression of CD133 showed higher i nvasiveness than blank control group[(239.3 ± 24.0) vs (103.3 ± 15.5),P ＜ 0.001].The expressions of p-AKT,Snail and N-cadherin were significantly increased when co-cultured with BM-MSCs in the group of CD133 overexpression (P =0.001,P =0.001,P =0.001),while the expression of E-cadherin was significantly decreased(P =0.003).The expressions of Snail and N-cadherin were also significantly increased after co-cultured with BM-MSCs in the blank control group (P =0.001,P =0.004),and the expression of E-cadherin was significantly decreased (P =0.018),while the expression of p-AKT was not significantly changed (P =0.193).Conclusions BM-MSCs can enhance the invasion and metastasis of gastric cancer cells by promoting the EMT of gastric cancer cells.CD133 may be involved in the regulation of EMT in gastric cancer cells through the PI3K/AKT signaling pathway.

Objective To investigate the value of endoscopic retrograde appendicitis therapy ( ERAT) in the diagnosis and treatment of atypical acute appendicitis. Methods All the 48 patients suspected of atypical acute appendicitis in Jiangsu Province Hospital from January 2015 to December 2016 were randomly divided into ERAT group and conservative treatment group according to the treatment method. The final appendectomy rate of the two groups was analyzed. Results Only 17 of the 24 patients in the ERAT group received endoscopic treatment because of complex conditions or personal wishes, and 16 cases were diagnosed as acute appendicitis. Surgical resection was performed in 5 cases because of recurrence of the disease after ERAT, and the appendectomy rate was 31. 2％ ( 5/16 ) . In the conservative treatment group, all 24 patients were treated with antibiotics. Twenty of them underwent surgical resection with appendectomy rate of 83. 3％ ( 20/24) , and 1 of them had appendiceal perforation. The appendectomy rate of the ERAT group was significantly lower than that of the conservative treatment group (χ2=11. 111, P<0. 05) . Conclusion ERAT has a high diagnostic and therapeutic value for atypical acute appendicitis.

Objective To compare the curative effect of tissue-selecting therapy stapler and procedure for prolapse and hemorrhoids in the treatment of patients with stage Ⅲ to Ⅳ hemorrhoids.Methods The patients with stage Ⅲ to Ⅳ hemorrhoids who underwent prolapse and hemorrhoids or tissue-selecting therapy stapler surgery in the department of General Surgery,Shanghai Ninth People's Hospital and Xinhua Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Chongming Branch,from Jan.2013 to Jun.2014 were accepted and allocated to prolapse and hemorrhoids or tissue-selecting therapy stapler group.The peri-operative parameters about operative time,blood loss,postoperative hospital stay and the time required to return to normal activity were compared by t test,The postoperative complications including pain assessment and the incidence of postoperative bleeding,urine retention,faecal urgency,fecal incontinence,anal stenosis,rectovaginal fistula and recurrence rate were compared by t test and chi-square test.Rank sum test was used to compare the recurrence rate and patient's satisfaction between the two groups.Results The operation time,intraoperative bleeding volum,postoperative hospital stay and the time required to return to normal activity in the procedure for prolapse and hemorrhoids group were signifcantly higher than those in the tissue-selecting therapy stapler group (P =0.021,P =0.003,P =0.001,P ＜0.001).The pain score of procedure for prolapse and hemorrhoids group were all higher than those of the tissue-selecting therapy stapler group in the first post-operative defecation and in post-operative 24 hours and 72 hours (all P ＜ 0.001).The incidence of faecal urgency of the procedure for prolapse and hemorrhoids group in post-operative 1 month (18.6％) was higher than that of the tissue-selecting therapy stapler group (6.6％) (P =0.036).There were no statistically significant differences in the incidence of postoperative bleeding,urinary retention,recurrence rate and patient's satisfaction between two group (P ＞ 0.05).Conclusion Tissue-selecting therapy stapler was superior to the procedure for prolapse and hemorrhoids in operation time,intraoperative blood loss,postoperative pain and the incidence of faecal urgency.Long-term results demonstrate that tissue-selecting therapy stapler and prolapse and hemorrhoids have similar effectiveness.

Objective: To investigate the influence of CD133+expression on patients' survival and resistance of CD133+cells to anti-tumor agents in gastric cancer (GC).
Methods: Influence of CD133 expression on prognosis was analyzed employing sam-ples from patients with GC. GC cell lines were utilized to separate CD133+and CD133?subpopulations by immunomagnetic separation and to analyze the biological features of two subpopulations in vitro and in vivo, especially in resistant to anti-tumor reagents and its apoptotic mechanism.
Results: The lower CD133+group showed a significantly better survival compared with the higher CD133+group. The highest content of CD133+subpopulations for KATO-III cells had stronger proliferative ability than CD133?subpopulations. A single CD133+cell was capable of generating new cell colony and the tumorigenicity rate in nude mice was 100% for CD133+ clonal spheres or for CD133+ cells, but 0% for CD133? cells. Furthermore, the higher expression levels of Oct-4, Sox-2, Musashi-1 and ABCG2 in CD133+ clonal spheres were identified compared with CD133+ cells or CD133? cells. Under the treatment of anti-tumor reagents, CD133+ cells had lower suppression rates compared with CD133? cells while lower level of Bcl-2 and higher level of Bax were found in CD133+cells compared with CD133?cells.
Conclusions: The patients with lower CD133+expression had a better survival. Enriched CD133+ cells in clonal sphere shared the ability to be self-renewable, proliferative, tumorigenic and resistant to anti-tumor agents as probably regulated by Bcl-2 and Bax.

ObjectiveTo investigate the expression of CXCR4 and CD133mRNA in primary lesion of primary gastric adenocarcinoma and the relation with clinicopathological features,and to explore the correlation of CXCR4 and CD133.MethodsThe primary lesion of primary gastric adenocarcinoma and normal tissues adjacent to gastric cancer were obtained from 50 patients.The diction of CXCR4 and CD133 protein expression was detected by the immunohistochemical staining,and the relative gray scale values of CXCR4 and CD133mRNA by semi-quantitative RT-PCR (Fisher' s exact probability method).Their relationship with clinicopathological features was also investigated ( Spearman relation analysis).ResultsThe positive rates of CXCR4 and CD133 protein in gastric cancers were 76.0％ and 66.0％ respectively,which were significantly higher than that in normal tissues adjacent to gastric cancer ( 16.0％ and 10％ ; P =0.000,P =0.000).The increment of relative gray scale values of CXCR4mRNA was associated with the larger tumor diameter,the later TNM stage and the occurrence of lymphatic metastasis( P ＜ 0.05 ).And the larger diameter of tumor,the later TNM stage were associated with the higher relative gray scale values of CD133mRNA (P ＜0.05).The levels of the relative gray scale values of CXCR4 mRNA and CD133mRNA were positively related(r =0.453,P ＜ 0.01 ).ConclusionsThe higher expression of CXCR4 and CD133mRNA correlateswith tumour diameter,TNM stage and lymphatic vessel invasion. The relative gray scale values of CD133mRNA increase with the increment of the relative gray scale values of CXCR4.

Objective To compare the inhibition effects of three synthesized fragments used in small interfering RNA(siRNA) against CD133 gene in KATO-Ⅲ gastric cancer cells,and to study effects of suppressed CD133 on the proliferating ability of intervened cells.Methods Three fragments of siRNA were designed and synthesized targeted at the mRNA of CD133.Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133FITC-siRNA.The knock-down effect of the CD133 gene was detected by RT-PCR and Western blotting.CCK-8 (cell counting kit-8 assay) was performed to measure the variation of the cell proliferative viability after the above-mentioned treatment.Results The transfection efficiency of siRNA was (85 ± 8) ％ in KATO Ⅲ Gastric cacer cell.All these three fragments of CD133 siRNA effectively inhibited the expression of CD133 gene,the inhibition rate being (11 ± 2) ％,(19 ± 2) ％,(24 ± 3) ％respectively.Compared with the control group,the cell proliferation viability was restrained (42 ± 4)％ in CD133siRNA-3 group (P ＜0.05).Conclusions CD133siRNAs were successfully transfected into KATO Ⅲ Gastric cacer cells and repressed the expression of CD133.Meanwhile,the CD133siRNA fragment 3 was screened from three CD133 siRNA,which has the best inhibition effect.The results provide preliminary evidence for the intereference of CD133+ gastric cancer cells subsequently.

Objective To investigate the biological effect of epithelial-to-mesenchymal transition (EMT) under the treatment of transforming growth factor (TGF)-β1 on the human gastric cancer cell line SGC7901 in vitro,and to observe whether the TGF-β1 can generate the tumor initiating cells ability in SGC7901 or not.Methods SGC7901 cells were cultured with TGF-β1.The morphological change was observed.The effect on proliferation of SGC7901 cells was detected by CCK-8.The invasion assay was used to investigate the motility and the invasion ability of SGC7901 cells.Immuofluorescence was used to detect the expression of E-cadherin and N-cadherin.The mRNA and protein's expression levels of EMT-related factors and CD44 were analyzed by RT-PCR and Western blotting respectively.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of SGC7901 cells was inhibited,and the ability of motility and invasion of SGC7901 cells were greatly enhanced after being treated with TGF-β1.RT-PCR and Western blotting showed that the expression of Snail (P ＜ 0.05),N-cadherin (P ＜ 0.05) and CD44 (P ＜ 0.05) were significantly increased while the expression of E-cadherin was decreased (P ＜ 0.05).Conclusions TGF-β1 can generate the EMT.The CD44 expression was up-regulated.TGF-β1 can inhibit the proliferation and promote the motility and invasion ability of SGC7901 cells.

Objective To investigate if TGF-β1 induces epithelial-mesenchymal transition (EMT) and promotes the obtaining of stemness characteristics in gastric cancer cell lines.Methods After KATO-Ⅲ cells were cultured with or without 5 ng/mL TGF-β1,the morphological change was observed and compared under phase-contrast microscopy.At the same time,the effect of TGF-β1 on the proliferation of KATO-Ⅲ cells was detected by CCK-8.On the other hand,the mRNA and protein' s expressions of EMT-related factors,ESC markers and TICs markers were analyzed by RT-PCR and Western blotting methods too.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of KATO-Ⅲ cells was inhibited after treated with TGF-β1 (P ＜ 0.05).After treated with TGF-β1,the relative mRNA expression levels of Snail (0.5219 ±0.0147) and N-cadherin(0.6640 ±0.0124) were higher than that in control group(0.2049 ±0.0214,P =0.004,0.2722 ± 0.0098,P =0.001),the relative protein expression levels of Snail (0.4769 ± 0.0234) and N-cadherin (0.5014 ± 0.0216) were higher than that in control group (0.2534 ± 0.0345,P =0.02,0.2026 ± 0.0268,P =0.009),while the relative E-cadherin mRNA and protein levels in TGF-β1 treated group (0.4701 ± 0.0215,0.1349 ± 0.0258) were lower than that in control group (0.6792 ± 0.0157,P =0.01 ; 0.6055 ± 0.0227,P =0.004),while the relative mRNA expressions of ESC markers such as Sox2,OCT4,Nanog in TGF-β1 treated group (0.594 ± 0.039、0.438 ± 0.033、0.489 ± 0.037) were higher than that in control group (0.143 ± 0.013,P =0.001,0.156 ± 0.025,P =0.001,0.325 ± 0.046,P =0.03),the relative mRNA expression levels of CD44 (0.437 ±0.037) and CD133(0.543 ±0.028) were higher than that in control group (0.247 ±0.024,P =0.000,0.139 ± 0.016,P =0.000),the relative protein expression levels of CD44 (0.429 ± 0.034) and CD133 (0.316 ±0.027) in TGF-β1 treated group were higher than that in control group (0.152 ± 0.014,P =0.000,0.110 ±0.010,P =0.000),cloning sphere-forming capacity was greatly enhanced after treated with TGF-β1 (P ＜ 0.01).Conclusion TGF-β1 can induce EMT in KATO-Ⅲ cells and promote the obtaining of stemness characteristics in gastric cancer cell lines.

OBJECTIVETo research the absorbed character of piperine in Erxiekang plaster, and piperine by transdermal absorption was determined.

METHODThe percutaneous absorption of piperine in vitro at different times was conducted by Franz osmosis and diffusion apparatus as well as high-performance liquid chromatography.

RESULTIt showed that the piperine through skins of mice gradually increased within the experiment time. After 52 h, the penetration of piperine was 78.51%, and remained basically unchanged.

CONCLUSIONThe method is reliable, and can be used for Erxiekang plaster of determination of transdermal absorption.

Alkaloids ; metabolism ; Animals ; Benzodioxoles ; metabolism ; Linear Models ; Male ; Piperidines ; metabolism ; Polyunsaturated Alkamides ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Skin Absorption

Objective To determine the relationship between acceptance of disability and posttraumatic stress response in patients with brachial plexus injury. Methods Total 160 patients with brachial plexus injury were recruited and investigated with the Impact of Event Scale-Revised (IES-R) and the Acceptance of Disability Scale (ADS). Results The total score of ADS was (79.07±11.99) which showed medium level of acceptance of disability. The total score of IES-R was 4-66 (33.51±14.41), which showed that most of the patients suffered from posttranmatic stress response. Significantly negative correlation was found between acceptance of disability and posttraumatie stress response(r=-0.480, P<0.001). Conclusions Nurses should pay more attention to the acceptance of disability and posttranmatic stress response in patients with brachial plexus injury, and provide appropriate health education and effective psychological intervention to improve the patients' mental health and quality of llfe.