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Objective:To explore the effect of probiotics Lactiplantibacillus plantarum(LP) WCFS1 by gavage on acute necrotizing pancreatitis (ANP) and associated ileum injury in mice. Methods:Twenty-four healthy male mice were gavaged with broad-spectrum antibiotics for 3 weeks to establish microbiota-depleted mice, and then randomly divided into control group (CON), ANP model group (ANP), LP gavage group (LP) and LP gavage and ANP induced group (LP+ ANP) , with 6 mice in each group. Mice in LP and LP+ ANP group were treated by gavage of LP (1×10 9 CFU/ml, 0.2 ml/day per mouse) for 1 week, while CON and ANP were gavaged with sterile phosphate buffered saline for 1 week instead. The ANP model was induced by intraperitoneal injection with caerulein (100 μg/kg) for 10 times with 1-hour interval between two injections and the 10th injection with lipopolysaccharide(LPS) 5 mg/kg intraperitoneally, and the mice were sacrificed 2 h later. Levels of LP in stool and ileal mucosa were detected by real-time PCR; the pancreas and ileum were collected for pathological examination to observe the extent of tissue inflammation and to score the pathology. Serum amylase activities were determined by enzymatic kinetic chemistry; serum inflammators levels and intestinal permeability were detected by ELISA; levels of inflammators in pancreatic and ileal tissues were detected by real-time PCR; ileal tight-junction proteins (occludin, claudin-1 and ZO-1) were measured by immunofluorescence staining. Results:LP levels in the stool and ileal mucosa of mice were significantly increased after LP gavage, and the differences were statistically significant (913.30±39.12 vs 2.39±1.39, 23.11±0.50 vs 1.38±0.28, all P value <0.05). The pathological scores of pancreatic tissue of CON, LP, ANP and LP+ ANP group were (0.26±0.41), (0.17±0.26), (8.55±0.46) and (6.30±0.45); the serum amylase activities were (219.70±19.73), (217.60±11.30), (2896.24±98.32) and (1837.13±131.60)U/L, IL-1β were (0.87±0.28), (1.4±0.85), (67.41±6.45) and (36.33±5.65)pg/ml, IL-6 were (0.74±0.27), (0.16±0.16), (280586.12±39163.92) and (107912.75±31283.47)pg/ml, IL-10 were (35.52±5.27), (50.99±15.34), (2008.45±184.83) and (3070.35±403.71)pg/ml; the expression level of pancreatic IL-1β mRNA was 1.42±0.39, 0.95±25, 20.53±0.50 and 10.69±1.01, IL-6 mRNA was 1.31±0.44, 0.93±0.023, 21.97±1.71 and 11.54±1.75, IL-10 mRNA was 0.93±0.14, 0.75±0.15, 0.99±0.21 and 1.76±0.19; there was no significant difference between LP and CON group, and pancreatic pathological scores, serum amylase、IL-1β and IL-6 levels, and the expression level of pancreatic IL-1β and IL-6 mRNA were significantly decreased in LP+ ANP group compared with those in ANP group, while serum IL-10 levels and the expression level of pancreatic IL-10 mRNA were significantly increased compared with ANP group, and all the differences were statistically significant (all P values <0.05). The pathological scores of ileal tissue of CON, LP, ANP and LP+ ANP group were 0, 0, (3.17±0.41) and (1.67±0.52); the levels of serum DAO of CON, LP, ANP and LP+ ANP group were (0.03±0.03), (0.02±0.02), (0.50±0.05) and (0.49±0.06)ng/ml; LPS levels were (2.75±0.35), (3.74±0.28), (7.19±0.92) and (5.88±0.38)ng/ml; the expression level of ileal IL-1β mRNA was 1.21±0.20, 1.17±0.09, 1.81±0.25 and 1.63±0.21; IL-6 mRNA was 1.01±0.29, 2.83±0.42, 54.45±8.50 and 16.87±4.42; IL-10 mRNA was 1.12±0.41, 6.09±2.51, 11.65±1.47 and 29.86±2.93. There was no significant difference between LP and CON group, except that the ileal IL-10 mRNA expression was significantly higher than that of CON group. Ileal pathological scores, serum LPS levels and the expression level of ileal IL-6 mRNA were significantly lower in LP+ ANP group than those in ANP group, while the expression level of ileal IL-10 mRNA was significantly higher than that of ANP group; the expression of ileal tight junction proteins (ocludin, claudin-1, ZO-1) was significantly higher than those in ANP group, and all the differences were statistically significant (all P values <0.05). Conclusions:LP WCFS1 gavage could ameliorate the injury of pancreatic and ileal barrier in caerulein-induced ANP mice.
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Objective:To establish culture system for mouse pancreatic ductal organoids and investigate the morphology and physiological functions of the organoids.Methods:Pancreatic tissues were taken from C57BL/6 mice (6-8 weeks) and digested by collagenase Ⅳ. The pancreatic ducts were separated and collected and then the pancreatic organoids were cultured in the complete medium after Matrix gel embedding. Morphological evaluation of the organoids was performed after hematoxylin-eosin staining. The expression and localization of markers for organoids were identified by Western Blot and immunofluorescence staining; and the expression and localization of ion channels and antimicrobial peptides of the organoids were detected by agarose gel electrophoresis and immunofluorescence staining.Results:Mouse pancreatic organoids were successfully established, which could be stably passaged for 10 generations. The organoids grew spherically and formed a duct-like structure. The internal cavity corresponded to the lumen of pancreatic duct tissue. The pancreatic organoids stably expressed stem progenitor cell marker gene SOX9 and ductal epithelial cell-specific gene KRT19, which were both localized in the epithelium. The organoids did not express amylase. The organoids maintained stable expression of epithelial ion channels Clcn1, Kcnma1, CFTR, Slc12a5, Slc26a3, Slc26a6 and Scnn1a, low expression of Ano1 and no expression of Clcn3, Kcna1, Kcna2, Kcnd3, Kcnh1, Atp12a, Slc4a4, Slc9a1, Slc12a2 and Slc26a11; and CFTR highly expressed in epithelial cells. The organoids maintained high expression of antimicrobial peptides Reg3a, CRAMP and glycoprotein 2, low expression of Defb1, Defb2, and Defb3 and no expression of Defa1 and Defa4; and both CRAMP and Reg3a were expressed in the epithelial cells and secreted into the lumen of the organoids.Conclusions:Mouse pancreatic organoids are successfully established, which can be stably passaged. The organoids maintain the characteristics of ductal epithelial cells and can be used as an in vitro model to study the physiology of pancreatic ducts.
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Background: Studies have indicated that dysfunction of intestinal barrier is closely related to the occurrence and development of acute necrotizing pancreatitis (ANP). Aims: To investigate the intestinal barrier injury in mice with ANP induced by L-arginine and its mechanism on aggravating ANP. Methods: Thirty C57BL/6 mice were randomly divided into sham-operation (SO) group and ANP group. Mice in ANP group were intraperitoneally injected with 8% L-arginine (4.5 g/kg) twice at an interval of 1 hour, while mice in SO group were intraperitoneally injected with an equal volume of 0.9% NaCl solution. Mice were sacrificed 24, 48, 72 hours after the establishment of ANP. HE staining was used to evaluate the pathological score of pancreas and intestine. Serum amylase, lipase were determined. Inflammatory factors (IL-1β, IL-6, TNF-α) and intestinal permeability (DAO, D-lactic acid, LPS) were measured by ELISA assay. Western blotting was used to detect protein expressions of TLR4, MyD88 in pancreatic tissue and ileal tissue. Results: Compared with SO group, histopathological score of pancreatic tissue and ileal tissue were significantly increased at corresponding-time in ANP group (P<0.05), serum levels of amylase, lipase, inflammatory factors (IL-1β, IL-6, TNF-α) and intestinal permeability (DAO, D-lactic acid, LPS) were significantly increased (P<0.05), protein expressions of TLR4, MyD88 in pancreatic tissue and ileal tissue were significantly increased (P<0.05). Conclusions: Intestinal permeability is increased in ANP mice induced by L-arginine, and the secretion of LPS is increased, which may enhance the intestinal inflammation and systematic inflammation via activating TLR4-MyD88 signaling pathway, thereby aggravating ANP.
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Mucin 2 (MUC2) is a kind of heavily glycosylated protein secreted by intestinal goblet cells. As the main component of intestinal mucus, MUC2 plays an important role in preventing the pathogenic microorganism from invasion as well as in assisting the colonization of intestinal probiotics. Intestinal flora is involved in many physiological functions of the human body. It modulates the synthesis, secretion and degradation of MUC2 through different mechanisms to affect the quality and quantity of MUC2. This article reviewed the effect and mechanism of intestinal flora on MUC2.
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It aimed to analyze the price disclosure system of genetic drugs in Australia.Based on the analysis of background and historical evolution,it tracked each step of price disclosure on the basis of descriptive statistics and analvsis so as to provide reterences for constructing the price disclosure system of genetic drugs led by market factor under the effective conduct of future medical reform policy in China.
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@# Objective To explore the effect of Botulinum toxin type A (BTX-A) injection combined with rehabilitation training and medicated bath on spasm for children with cerebral palsy. Methods 80 children with spastic and mixed cerebral palsy were randomly divided into two groups with 40 cases in each group. The control group received physical therapy, and the observation group received BTX-A injection and rehabilitation training and medicated bath. They were assessed with modified Ashworth scale (MAS) and Gross Motor Function Measure (GMFM-88) before and 3 months after treatment. Results The scores of MAS and GMFM were better in 2 groups after treatment (P<0.05) and the observation group was better than that of the control group (P<0.05). Conclusion BTX-A injection combined with medicated bath can reduce muscle tension, improve gross motor function of children with spastic and mixed cerebral palsy.
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Shenjincao injection was prepared from the extract of Lycopodium cernuum L. by ultrafiltration or wateralcohol methods,respectively,and then injected intra-peritoneally in groups of rats contaminated with quartz powder to assess their prophylactic effects. After 5 weeks of treatment,the rats were dissected and the fresh and dried weights,collagen content and the pathologic grading of the lungs were examined. Results showed that Shenjincao injection prepared with ultrafiltration was effective for the prophylactic treatment of experimental silicnsis of rats,while that prepared with water-alcohol method was devoid of such effect. The antisilicosis active principle of Shenjincao was explored and discussed.