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AIM:To investigate the effect of microRNA-429 (miR-429) on the expression of occludin (Ocln) in intestinal epithelial cells ( IECs) and intestinal epithelial barrier function in diabetic mice.METHODS:Diabetes mel-litus mouse model was induced by intraperitoneal injection of streptozocin.The expression of miR-429 in IECs of diabetic mice was inhibited by antagomiRNA-429 injected via tail vein.The expression of miR-429 and mRNA expression of Ocln were detected by real-time PCR.The protein expression of Ocln was determined by Western blotting and immunohistochem-istry.The urinary lactulose/mannitol ratio was measured by gas chromatography.The plasma LPS concentrations in the mice were measured by chromogenic end-point TAL kit.RESULTS:The results of real-time PCR confirmed that the ex-pression of miR-429 in IECs of diabetic mice was remarkably inhibited by tail-vein administration of antagomiRNA-429, and resumed to similar level of normal mice on the 6th day after the administration.After suppressing the level of miR-429, the expression of Ocln in IECs of diabetic mice increased significantly (P<0.05), while the urinary lactulose/mannitol ra-tio and the plasma LPS concentration decreased obviously ( P<0.05 ) .CONCLUSION:AntagomiRNA-429 effectively suppresses miR-429 expression in IECs of diabetic mice, and then enhances the expression of Ocln and partially resumes the intestinal epithelial barrier function.
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Objective To investigate the clinical characteristics and prognosis of patients with systemic lupus erythematosus (SLE) complicated with acute pancreatitis (AP) .Methods From January 1999 to December 2013 ,the clinical data of 16 patients with SLE complicated with AP among the total 2 526 cases of SLE was collected .A retrospective analysis was performed and the clinical data of patients was classified and documented ,which included general information ,past history ,clinical symptoms , laboratory findings ,imaging findings ,treatment and outcome .The rank sum test was performed for analysis of non‐normal distributed measurement data ,and the Fisher′s exact test was used for count data analysis .Results The incidence of SLE complicated with AP was 0 .63% (16/2 526) .Among them ,ten patients were mild acute pancreatitis (MAP) and six patients were severe acute pancreatitis (SAP) .All patients were treated with fasting ,gastrointestinal decompression ,nutritional support ,anti‐acid ,anti‐inflammatory ,glucocorticoid and somatostatin and so on . Six patients were cured , seven patients improved and three patients died (two SLE complicated with SAP ,one SLE complicated with MAP) . Compared with the SLE patients complicated with SAP ,the SLE patients complicated with MAP were more easily to have lupus nephritis(6/6 versus 5/10 ,Fisher′s exact test) ,hematological system injuries (6/6 versus 5/10 ,Fisher′s exact test) ,liver injuries (5/6 versus 0/10 ,Fisher′s exact test) ,more organs involved (mean 7 versus 3 ,Z= -3 .225) and higher SLE disease active indexes (DAI) score (mean 13 .5 versus 6 .5 ,Z= -2 .876);the differences were statistically significant (all P<0 .05) .Compared with the cured and improved SLE patients complicated with AP ,lupus encephalopathy (2/3 versus 1/13 ,Fisher′s exact test) ,more organs involved (mean 7 versus 5 ,Z= -2 .276) and higher SLE DAI score (mean 21 versus 12 ,Z= -2 .195) was more common in dead SLE patients complicated with AP;the differences were statistically significant (all P< 0 .05) .Conclusions SLE patients complicated with SAP are more easily to get lupus nephritis ,hematological system injuries ,liver injuries ,activity of SLE and multiple‐organ systems involved . The prognosis of SLE patients complicated with AP was poor in those with activity of SLE ,multiple‐organ involved and lupus encephalopathy .
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[ ABSTRACT] AIM:To study the process of promoting mouse embryonic stem cells ( ESC) to specify to definitive endoderm by up-regulating of Nodal signal pathway in order to find the best cultivated systems of differentiation of mouse ESC to definitive endoderm cells.METHODS:The cells were divided into different groups based on the culture medium:ESC group ( serum-free medium +LIF) , natural differentiation group ( serum-free medium) and activin A group ( serum-free medium +50μg/L activin A).The cells and the sterilized coverslips with cells were collected at 1, 3, 5 and 7 d of the cultivation.The proportion of CXCR4 +cells was detected by flow cytometry.The expression of CXCR4 was determined by immunocytochemical method, and the protein expression of OCT4 and CXCR4 was detected by Western blot.RE-SULTS:The proportion of CXCR4 +cells showed no dramatic change in ESC group along with the extending of cultivation day, while there were gradually increased in natural differentiation group and activin A group and the highest level was ob-served at 5 d.Among the 3 groups, the proportion of CXCR4 +cells at 5 d was the highest in activin A group.The brown or tan staining in the cells observed under microscope was considered as positive CXCR4 by immunocytochemistry.The pro-tein levels of OCT4 and CXCR4 in ESC group along with the extending of cultivation days was observed.The expression levels of OCT4 were gradually decreased in the cells in natural differentiation group and activin A group, while those of CX-CR4 were gradually increased with the highest level at 5 d.It was highest in the cells in activin A group.CONCLUSION:The proportion of definitive endoderm was the highest at 5 d of the induction during in vitro mouse ESC differentiation.Up-regulation of Nodal signal pathway by adding activin A at the early stage of mouse ESC differentiation promotes mouse ESC to specify to definitive endoderm with CXCR4 molecular marker.
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AIM:To observe the inhibitory effect of siRNA targeting to Wip1 gene on the Wip1 gene expression in the colon cancer cells and to investigate the influence of Wip1 gene silencing on the chemotherapy sensitivity of colon cancer cells.METHODS:Wip1-811 siRNA targeting to Wip1 gene was transfected into RKO colon cancer cells with high expression of Wip1 gene.The mRNA expression of Wip1 was measured by real-time PCR.The protein level of Wip1 was detected by Western blotting.The viability of RKO colon cancer cells was measured by MTS assay.The cell apoptosis and cell cycle were analyzed by flow cytometry.RESULTS: Wip1-811 siRNA efficiently inhibited the expression of Wip1 at mRNA and protein levels.The enhanced chemotherapy sensitivity of RKO colon cancer cells was observed after inhibition of Wip1 gene expression.The viability of RKO colon cancer cells was decreased from (89.4 ±6.6)%to (74.7 ±3.9)%af-ter treated with 5-fluorouracil (P<0.05) and decreased from (77.9 ±2.4)%to (66.7 ±2.9)%after treated with oxali-platin ( P<0.05 ) .The cell apoptotic rate was increased from ( 7.7 ±0.5 )% to ( 12.3 ±3.2 )% and from ( 14.7 ± 2.1)% to (34.0 ±2.1)% when RKO colon cancer cells were treated with 5-fluorouracil and oxaliplatin, respectively (P<0.05).CONCLUSION:Wip1 gene silencing enhances chemotherapy sensitivity of colon cancer cells.
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AIM:To investigate the role of side population ( SP) cells in multidrug resistance of colon cancer cells and microRNA biomarkers of SP cells in colon cancer cells .METHODS:SP cells in colon cancer cells were sorted by flow cytometry .The cell viability was measured by MTT method .MicroRNA expression profiles were detected by mi-croRNA chip.MicroRNA expression was verified by real-time PCR.RESULTS:The ratios of SP cells in HCT-15, HT-29 and LoVo colon cancer cell lines were 16.75%, 13.02%and 9.52%, respectively.In all 3 colon cancer cell lines, IC50 of the antitumor drugs including 5-fluorouracil , oxaliplatin and adriamycin for the SP cells were significantly higher than those for non-SP cells (P<0.05).MicroRNA profiling showed that miR-5000-3p, miR-5009-3p and miR-552 were all up-regulated in the SP cells of all 3 colon cancer cell lines .This result was verified by real-time PCR.CONCLUSION:miR-5000-3p, miR-5009-3p and miR-552 are all up-regulated in the SP cells of colon cancer cell lines , and may be the poten-tial microRNA biomarkers of SP cells in colon cancer .
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Objective To investigate the expression of CD44v3 and VEGF-C in human gastric cancers and the clinical significance. Methods The expression of CD44v3 and VEGF-C was detected by immunohistochemistry SP method in 92 gastric cancer tissues. Results Positive immunohistochemical stain for CD44v3 was identified in 27.2% of gastric cancer tissues. There was no correlation found among the expression of CD44v3 with sex, location,depth of invasion, WHO type, Lauren type as well as distant metastasis (all P ＞ 0. 05 ). The expression of CD44v3 was positively correlated with lymph node metastasis and tumor differentiation( all P ＜ 0. 05 ). Positive immunohistochemical stain for VEGF-C was identified in 48.9% of gastric cancer tissues. There was no correlation found among the expression of VEGF-C with sex,location, tumor differentiation, depth of invasion, WHO type, as well as distant metastasis ( all P ＞ 0. 05 ). The expression of VEGF-C was positively correlated with lymph node metastasis and Lauren type ( all P ＜0. 05). Cox-Regression reflected that only the lymphatic metastasis situation correlated with the life ratio( P =0. 015). Conclusion The expression of CD44v3 and VEGF-C could suggest the progression and metastatic potential value of gastric cancer,but didnt correlate with the life ratio. There was no cooperation between CD44v3 and VEGF-C in promoting gastric cancer metastasis.