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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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[ 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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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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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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Objective To investigate the expression of intestinal epithelial stem cell markers Msil and Hesl in mouse acute radiation enteritis (RE) model. Methods Forty female BALB/c mice were undergone one-time whole abdominal irradiation by 300 cGy/min β-ray. Thirty mice were selected before irradiation (Day 0) and every six mice were euthanized at day 3,5,7 and 14 after irradiation. The small intestinal were detached and the expression of Msil and Hesl were detected in middle part of small intestinal by quantitative RT-PCR, Western blots, and immunohistochemistry. Subsequently,the difference of Msil and Hesl expression was compared. Results The mouse acute RE model was successfully established. At the 5th day after radiation, the results of quantitative RT-PCR indicated that the relative ΔCt value of Msil and Hesl expression were 15. 17 ± 0. 47 and 15. 83 ± 0. 24respectively, the results of Western blot showed that the integrated intensity were 0. 443±0. 055 and 0. 301 + 0. 047 for Msil and Hesl. The expressions were significantly higher than other time points at RNA and protein levels (P<0. 05). The results of immunohistochemistry revealed that Msil and Hesl were highly expressed in intestinal crypts at 5th day and 7th day. Conclusions The expression of Msil and Hesl, upregulated before wound repair in RE model, indicates that intestinal epithelial repair is related to intestinal stem cell proliferation.
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AIM: To investigate the short-term efficacy and safety of sulfasalazine (SASP) 3 g per day in the treatment of patients with mild and moderate ulcerative colitis (UC). METHODS: 122 patients were treated with SASP ( 1 g, t.i.d.) for 6 weeks. The data of clinical manifestations, colonoscopic and histological involvements were compared before and after the treatment of UC. The short-period efficacy and adverse reactions were evaluated in 110 patients. RESULTS: The therapeutic project was carried out in the 110 out of 122 patients. After 110 patients were treated for 6 weeks, the clinical, colonoscopic and histological remission were 71.8%, 21.8% and 16.4%, respectively. Among the 79 patients with clinical remission, 58.2% and 67.1% of them remained grade 1 in colonoscopic and histological findings, respectively. The curative rates and the effective rates were 63.9% and 82.0%, respectively. Among the 122 patients treated with SASP, 21 of them ( 17.2%) had adverse reactions. Except 4 patients suffered urticaria and leukopenia, no patients quitted the treatment because of obvious adverse reaction. CONCLUSION: SASP ( 3 g per day) can be an effective and safe medicine in treatment of patients with mild and moderate UC, but more than half of the patients in clinical remission still have light inflammation in colonoscopy and histology.
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0.05) before and after treatment. The rates of eosinophil infiltration: 98.2% vs. 80.4% in the mild UC (P
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AIM:To investigate the preparation techniques and anti-tumor effects both in vitro and in vivo of a novel nanoparticles control-releasing preparation of 5-fluorouracil(5-FU)by intravenous injection.METHODS:With polylactic acid(PLA)as marix materials,we adopted ultrasound emulsification method to prepare PLA enveloped 5-FU nanoparticles(5-FU-NPs).Scanning electricity microscopy was used to observe the morphology of 5-FU-NPs and laser optical scattering experiment was conducted to determine its diameter distribution.The drug-carrying capacity(ratio)of the nanoparticles was determined by means of high-power liquid chromatography(HPLC)and MTT test was used to observe cytotoxicity in vitro.The anti-tumor effects were determined at different dosages,frequencies of taking drugs in vivo.RESULTS:Scanning electron microscopy showed that the 5-FU-NPs were globular particles with smooth surface in an average particle diameter of 191.9 nm with a normal distribution,and the drug-carrying capacity of 5-FU-NPs was 15.2%.5-FU-NPs had the same anti-cancer effect as unenveloped drug in vitro and showed typical dose-effect relationship.Compared to naked 5-FU,5-FU-NPs presented significant difference(P
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Protective effect of Panax notoginseng saponins(PNS) on cerebral cortex cells and pial microcirculation in acute hypoxic animals and their possible mechanisms were studied. The results showed that: PNS prevented reduction of PaO_2, (hypoxic group 27.15?5.67, PNS group 39.63?6.38, P
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AIM: To observe the effects of aspirin and prostaglandin E_2 (PGE_2) on the cell viability and cell cycle in SW1990 human pancreatic carcinoma cell lines, and to investigate the mechanisms of aspirin-induced growth inhibition and cell cycle arrest. METHODS: After incubated with aspirin or PGE_2 and their combination, the viability of SW1990 cells was measured by MTT assay. The levels of intracellular PGE_2 were determined by ELISA. The effects of aspirin or PGE_2 on cell cycle were investigated by flow cytometry (FCM). The expression of p21~ Wafl/cipl and p27~ Kipl/pic2 (the cyclin-dependent kinase inhibitors) were analyzed by Western blotting. RESULTS: Aspirin could inhibit the growth of cells and level of intracellular PGE_2 in a dose-dependent manner. Aspirin enhanced the expression of p21~ Wafl/cipl and p27~ Kipl/pic2 and induced cell cycle arrest at G_0/G_1 phase. PGE_2 increased the cell viability of SW1990 cells. However, it couldn't antagonize the changes of cell viability and cell cycle that induced by aspirin. CONCLUSIONS: The inhibitory effects of aspirin on growth and cell cycle of pancreatic carcinoma cells might not be mediated by a COX-dependent pathway completely. Cell cycle arrest induced by aspirin might be associated with up-regulation of p21~ Wafl/cipl and p27~ Kipl/pic2 .