Macrophage polarization in inflammatory bowel disease.

The growing prevalence of inflammatory bowel disease (IBD) has encouraged research efforts, which have contributed to gradual improvements in our understanding of IBD diagnosis and therapeutic approaches. The pathogenesis of IBD has not been fully elucidated; however, the combined actions of environmental, genetic, immune factors, and microbial organisms are believed to cause IBD. In the innate immune system, macrophages play important roles in maintaining intestinal health and in the development of IBD. Macrophages can be polarized from M0 into several phenotypes, among which M1 and M2 play critical roles in IBD development and the repair of intestinal homeostasis and damage. Certain macrophage-related IBD studies already exist; however, the functions of each phenotype have not been fully elucidated. As technology develops, understanding the link between macrophages and IBD has increased, including the growing knowledge of the developmental origins of intestinal macrophages and their performance of comprehensive functions. This review describes macrophage polarization in IBD from the perspectives of macrophage development and polarization, macrophage changes in homeostasis and IBD, metabolic changes, and the mechanisms of macrophage polarization in IBD. The discussion of these topics provides new insights into immunotherapy strategies for IBD. Video Abstract.

Case report: Diagnosis and treatment of DGAT1 deficiency-induced congenital diarrhea in two cases and literature review.

Introduction: Congenital diarrhea is a rare inherited intestinal disease characterized by persistent and severe diarrhea and malabsorption in the first few weeks after birth, which can be life-threatening. Some congenital diarrheal diseases are associated with mutations in the diacylglycerol acyltransferase 1 (DGAT1) gene. Case descriptions: This study delineated 2 cases of diarrhea and growth retardation, subsequently confirmed as congenital diarrhea via genetic testing, revealing that the etiology involved compound heterozygous mutations in the DGAT1 gene. Diagnostic assessments: High-MCT milk powder did not obtain an ideal outcome, whereas low-fat diets improved the symptoms of diarrhea and increased the body weigths. Disscussion: The two cases facilitated our understanding of the clinical features of, and treatments for, patients harboring a DGAT1 mutation and enriched the existing DGAT1 mutation database.

Evaluation of IgG/IgE-binding capacity and functional properties of enzymatic hydrolysis in skimmed cow milk system.

Cow's milk (CM) allergy is a common food allergy that seriously impacts the growth and development of infants and children. However, CM is an important source of nutrients, and few studies focus on the effects of enzymatic hydrolysis treatment on the whole skimmed CM system. In this study, the IgG/IgE-binding and functional properties of Alcalase-, Protamex-, and Flavourzyme-treated skimmed CM (AT, PT, and FT, respectively) were systematically evaluated. The results showed that the treatment groups were mainly composed of low molecular weight (MW) peptides (<3 kDa), accounting for 94.85%-97.90%. Additionally, the IgG reactivity of these peptides was significantly lower (p < 0.05) than those of higher MW peptides (10-30 kDa and >30 kDa). The IgE reactivity of FT with higher MW peptides was the lowest among these groups, with the OD value reaching 0.089. Moreover, the total amino acid content of hydrolysates of skimmed CM (HM) increased significantly (skimmed CM, 5.94 µg/mL; AT, 123.70 µg/mL; PT: 136.20 µg/mL; FT, 988.72 µg/mL) compared to that in skimmed CM. A total of 10, 10, and 7 flavor compounds were increased in AT, PT, and FT, respectively. Furthermore, the solubility, foamability, and emulsifying ability of HM were significantly improved, being 2.17-fold, 1.52-fold, and 1.96-fold higher in PT than in skimmed CM. These results lay a theoretical foundation for the development of hypoallergenic dairy products.

Therapeutic Potential of Curcumin in a Rat Model of Dextran Sulfate Sodium-Induced Ulcerative Colitis by Regulating the Balance of Treg/Th17 Cells.

The pathogenesis of ulcerative colitis (UC) remains unclear, and it is believed that an imbalance of regulatory T (Treg) cells and T helper 17 (Th17) cells is related to the occurrence of UC. Curcumin has been confirmed to exert anti-inflammatory effects in bronchial asthma and osteoarthritis by regulating the balance of Treg/Th17 cells. This study aimed to explore the therapeutic potential of curcumin in dextran sulfate sodium (DSS)-induced UC rats by regulating the balance of Treg/Th17 cells. Disease activity index (DAI) scores were calculated. Changes in colon inflammation were observed using hematoxylin and eosin staining. Treg and Th17 cells in the spleen were detected by flow cytometry, and the levels of interleukin (IL)-10 and IL-17A were determined using enzyme-linked immunosorbent assay. In DSS-induced colitis, curcumin significantly ameliorated colitis symptoms by reducing the DAI and increasing colon length. Additionally, curcumin significantly increased the expression of Treg cells and decreased the expression of Th17 cells and the extent of histopathological damage. Furthermore, curcumin increased the expression of IL-10 and decreased the expression of IL-17A. Curcumin attenuates DSS-induced UC injury by regulating Treg/Th17 balance and related cytokine secretion. Thus, curcumin may be a promising therapeutic drug for treating UC.

LncRNA RP5-857K21.7 inhibits PDGF-BB-induced proliferation and migration of airway smooth muscle cells through the miR-508-3p/PI3K/AKT/mTOR axis.

The continuous increase in the prevalence of asthma poses a threat to human health. Despites numerous researches, the understanding of asthma development still remain elusive, hindering the development of effective treatment. Here, we explored the role of lncRNA RP5-857K21.7 (RP5-857K21.7) in the development of asthma and its potential molecular mechanism of regulation. Airway smooth muscle cells (ASMCs) were isolated and cultured after which some of the cells were induced with PDGF-BB to build an asthma cell model, and then, qRT-PCR analysis was used to measure the expression level of RP5-857K21.7 in the cell model. Result shows that the RP5-857K21.7 is significantly downregulated in PDGF-BB-induced ASMCs cells. Through CCK-8, transwell, and flow cytometry assay, we examined the functional impact of RP5-857K21.7 on the proliferation, migration, and apoptosis of the ASMCs, respectively, and found that the overexpression of RP5-857K21.7 markedly inhibit PDGF-BB-induced ASMCs cell proliferation, migration and induce apoptosis. Bioinformatics analysis predicted that the RP5-857K21.7 could sponge miR-508-3p and result was validated through a dual-luciferase reporter assay, biotinylated RNA pull-down assay, and RIP-qRT-PCR analysis. Mechanistically, RP5-857K21.7 regulates the PI3K/AKT/mTOR pathway by endogenously sponging miR-508-3p to inhibit PDGF-BB-induced ASMCs cell proliferation, migration and induce apoptosis. The current research suggests that the RP5-857K21.7 and its associated molecular pathway (miR-508-3p/PI3K/AKT/mTOR axis) might be a useful therapeutic target for the treatment of asthma disease.

The tumor-suppressive effects of alpha-ketoglutarate-dependent dioxygenase FTO via N6-methyladenosine RNA methylation on bladder cancer patients.

N6-methyladenosine (m6A) methylation participates in the progression of bladder cancer (BCa). Nevertheless, the regulatory mechanism of alpha-ketoglutarate-dependent dioxygenase FTO influencing the BCa progression has still remained elusive. In this study, to investigate the tumor-suppressive effects of FTO via m6A RNA methylation on BCa patients, a total of 15 cancer tissues and adjacent normal tissues (ANTs) were collected from BCa patients who received tumor resection in our hospital from September 2015 to December 2019. We found that the FTO expression was significantly reduced in cancer tissues compared with that in ANTs, which indicated a lower malignant potential and a higher overall survival rate. It was revealed that overexpression of FTO in two human urinary BCa cell lines (HT-1197 and HT-1376) significantly decreased the cell proliferation and invasion abilities compared with the negative controls, whereas the cell apoptosis was markedly enhanced. In addition, we noted that the changes in m6A methylation level mainly appeared at 5' untranslated region (5' UTR) of MALAT1 and NOTCH1 transcripts, and at 3' UTR of CSNK2A2 and ITGA6 transcripts, responding to the overexpression of FTO. Mechanistically, we found that the splicing factor, proline- and glutamine-rich (SFPQ) could influence the FTO-mediated m6A RNA demethylation, eventually affecting the gene expression. This study provided a new insight into the relationship between the FTO expression and the m6A RNA methylation, assisting scholars to better understand the pathogenesis of BCa.

Effects of enzymatic hydrolysis on the allergenicity of natural cow milk based on a BALB/c mouse model.

Cow milk allergy is one of the most prevalent food allergies worldwide, particularly in infants and children. To the best of our knowledge, minimal research exists concerning the antigenicity of cow milk (CM). This study was performed to evaluate the allergenicity of enzymatically hydrolyzed cow milk (HM) in a BALB/c mouse model. The mice were randomly divided into 5 groups (n = 12/group), which were sensitized with phosphate-buffered saline, CM, and HM (Alcalase-, or Protamex-, or Flavorzyme-treated cow milk; Novo Nordisk; AT, PT, FT, respectively), respectively, using cholera toxin as adjuvant on d 0, 7, 14, 21. On d 28, the test mice were orally challenged with phosphate-buffered saline, CM, and HM (AT, PT, or FT) alone. Anaphylactic symptoms were monitored in the mice. Antibody, cytokine, histamine, and mouse mast cell protease-1 (mMCP-1) levels were measured using enzyme-linked immunosorbent assays. In addition, the numbers of T helper (Th)1 and Th2 cells, as well as the proportions of CD4+CD25+Foxp3+ Treg cells, in mouse spleens were detected using flow cytometry. Statistical significance was determined by one-way ANOVA. The results revealed significant differences between CM- and HM-challenged mice. Among these, the clinical scores of HM-challenged mice (AT, 1.50; PT, 2.00; FT, 1.92) were lower than those of CM-challenged mice (positive control, 2.83), but body weight and temperature of HM-challenged mice were higher than those of CM-challenged mice. In addition, significant reductions of allergen-specific IgE, IgG, histamine, and mMCP-1 were showed in HM-challenged mice, especially for histamine, ranging from 171.42 ng/mL to 214.94 ng/mL. Remarkable reductions of IL-4, IL-5, and IL-13 levels, as well as elevations of interferon-γ and IL-10 levels in the spleens of HM-challenged mice were also detected. Moreover, the number of Th2 cells decreased in the HM-challenged mice, to 2.36% (AT), 1.79% (PT), and 4.03% (FT), respectively, whereas the numbers of Th1 cells (AT, 6.30%; PT, 6.70%; FT, 6.56%) and the proportions of CD4+CD25+Foxp3+Tregs (AT, 8.86%; PT, 9.21%; FT, 9.16%) increased significantly. Our findings indicate that exposure to HM was sufficient to induce a shift toward a Th1 response, thereby reducing potential allergenicity. Importantly, these results will lay a theoretical foundation for the development of hypoallergenic CM products.

Deleterious Variants in ABCC12 are Detected in Idiopathic Chronic Cholestasis and Cause Intrahepatic Bile Duct Loss in Model Organisms.

BACKGROUND & AIMS: The etiology of cholestasis remains unknown in many children. We surveyed the genome of children with chronic cholestasis for variants in genes not previously associated with liver disease and validated their biological relevance in zebrafish and murine models. METHOD: Whole-exome (n = 4) and candidate gene sequencing (n = 89) was completed on 93 children with cholestasis and normal serum γ-glutamyl transferase (GGT) levels without pathogenic variants in genes known to cause low GGT cholestasis such as ABCB11 or ATP8B1. CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 genome editing was used to induce frameshift pathogenic variants in the candidate gene in zebrafish and mice. RESULTS: In a 1-year-old female patient with normal GGT cholestasis and bile duct paucity, we identified a homozygous truncating pathogenic variant (c.198delA, p.Gly67Alafs∗6) in the ABCC12 gene (NM_033226). Five additional rare ABCC12 variants, including a pathogenic one, were detected in our cohort. ABCC12 encodes multidrug resistance-associated protein 9 (MRP9) that belongs to the adenosine 5'-triphosphate-binding cassette transporter C family with unknown function and no previous implication in liver disease. Immunohistochemistry and Western blotting revealed conserved MRP9 protein expression in the bile ducts in human, mouse, and zebrafish. Zebrafish abcc12-null mutants were prone to cholangiocyte apoptosis, which caused progressive bile duct loss during the juvenile stage. MRP9-deficient mice had fewer well-formed interlobular bile ducts and higher serum alkaline phosphatase levels compared with wild-type mice. They exhibited aggravated cholangiocyte apoptosis, hyperbilirubinemia, and liver fibrosis upon cholic acid challenge. CONCLUSIONS: Our work connects MRP9 with bile duct homeostasis and cholestatic liver disease for the first time. It identifies a potential therapeutic target to attenuate bile acid-induced cholangiocyte injury.

The JAK2 inhibitor AG490 regulates the Treg/Th17 balance and alleviates DSS-induced intestinal damage in IBD rats.

The pathogenesis of inflammatory bowel disease (IBD) remains unclear, and it is currently believed that an imbalance in regulatory T (Treg) cells/T helper 17 cells (Th17 cells) is related to the occurrence and development of IBD. Recently, the JAK2 inhibitor AG490 has been used in animal models such as rheumatoid arthritis and bronchial asthma models and shown to exert immunoregulatory functions that improve disorder in the Treg/Th17 cell balance. This study aimed to evaluate the effect of AG490 on the intestinal inflammatory process in an IBD rat model. A dextran sulfate sodium (DSS)-induced IBD rat model was established, and disease activity index (DAI) scores were calculated. The histopathological damage score was determined by haematoxylin-eosin (H&E) staining. Treg/Th17 cells in the spleen were detected by flow cytometry. The levels of interleukin (IL)-10, IL-6 and IL-17A were detected by enzyme-linked immunosorbent assay (ELISA). AG490 attenuated DSS-induced IBD injury by regulating the Treg/Th17 balance and related cytokine secretion to reduce the DAI and colonic tissue damage. Thus, AG490 may be a new method for effective treatment of IBD.

Evaluation of inflammatory bowel disease activity in children using serum trefoil factor peptide.

BACKGROUND: The diagnosis of IBD and evaluation of treatment require endoscopy, which is difficult in children. This study evaluated the use of TFF3 as a biomarker. METHODS: Permeability of the intestinal mucosa and serum TFF3 were assayed and colon tissue was harvested 7 days after inducing IBD in mice with TNBSA. TFF3 was monitored in 51 pediatric IBD patients stratified by active disease or remission and in 20 healthy children. Mucosal healing was assessed by the Simple Endoscopic Score for Crohn Disease and Baron scores in CD and UC patients. RESULTS: Histological evaluation revealed transmural inflammation of the colon in IBD model mice. Permeability of the intestinal mucosa and serum TFF3 were both higher in TNBSA-treated than in control mice (P < 0.05). TFF3 was higher in children with active IBD than in those in remission and in healthy children (P < 0.05). TFF3 was positively correlated with the SES-CD score (P < 0.05) but not with either the pediatric CDAI score or the serum CRP. The sensitivity of serum TFF3 for monitoring CD activity was 100% and the specificity was 76.2%. CONCLUSIONS: TFF3 level increased with CD activity, which is of significance for diagnosis and for evaluation of mucosal healing. TFF3 could also be a marker in pediatric UC, as TFF3 positively correlated with UCAI. IMPACT: The diagnosis and evaluation of IBD is difficult; endoscopy provides objective assessment; TFF3 can be a useful marker instead of endoscopy. TFF3 was increased in active CD of children; TFF3 can be used as a clinical marker of pediatric CD; TFF3 can diagnose and evaluate mucosal healing of CD. Pediatrician should pay attention to clinical marker; TFF3 level may be a key evaluation of mucosal healing of CD; the value of diagnosis of TFF3 in CD is important.

Inhibiting microRNA-7 Expression Exhibited a Protective Effect on Intestinal Mucosal Injury in TNBS-Induced Inflammatory Bowel Disease Animal Model.

This study aimed to explore the expression and correlation of microRNA-7 (miR-7) and trefoil factor 3 (TFF3) genes and proteins in inflammatory bowel disease (IBD) mouse models and to elucidate the effect of miR-7 inhibition in the intestinal mucosa in IBD models. A TNBS-induced IBD mouse model was established. Changes in intestinal inflammation were observed by HE staining, and the expression levels of miR-7 and TFF3 were detected by RT-PCR. After miRNA-antagomir injection, the degree of colonic tissue damage and the expression levels of miR-7 and TFF3 in intestinal tissues were compared. TNBS-induced IBD mice showed significant weight loss, significantly decreased disease activity index (DAI), and a significantly increased pathological damage score. miR-7 was highly expressed in the colon tissue of IBD mice, and TFF3 was downregulated. Inhibition of the expression of miR-7 improved the stool characteristics and fecal occult blood (OB) of IBD mice, significantly increased the expression of TFF3 protein, and decreased the pathological damage scores. In the IBD mouse model, miR-7 posttranscriptionally regulates TFF3. The inhibition of miR-7 expression improves some clinical manifestations of IBD mice, reduces the pathological damage of the intestinal mucosa, and shows a protective effect in IBD.

Phenotypic characteristics and clinical manifestations of inflammatory bowel disease in infants and children under 2 years of age in Liaoning Province, China: five of six infants with IL-10R mutations.

A retrospective analysis of the phenotypic characteristics, clinical manifestations, investigations and interleukin-10 receptor (IL-10R) status of inflammatory bowel disease (IBD) in infants and children under 2 years of age in Liaoning Province, China between January 2015 and October 2016 is described. Six patients without a family history of IBD were diagnosed with Crohn disease, and IL-10R mutations were detected in five of them. Compound heterozygous patients with IL-10R mutations had severe clinical manifestations, were resistant to standard medication for IBD and had a poor prognosis as haematopoietic stem cell transplantation (HSCT) was not undertaken. Two of these patients died of suspected septicaemia. IBD in infants and children under 2 years of age is life-threatening when patients with IL-10R mutations do not receive allogeneic HSCT.

MicroRNA-7-5p regulates the proliferation and migration of intestinal epithelial cells by targeting trefoil factor 3 via inhibiting the phosphoinositide 3-kinase/Akt signalling pathway.

Trefoil factor 3 (TFF3) reconstructs the epithelial barrier by stimulating epithelial cell migration and proliferation, and significantly contributes to intestinal mucosal damage and healing. In a previous study, TFF3 was identified as a novel target of microRNA-7-5p (miR-7-5p). The aim of the present study was to investigate the roles and mechanisms of miR-7-5p in the proliferation and migration of intestinal epithelial cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to determine the expression level of miR-7-5p in the experimental groups. In addition, western blot analysis was performed to examine the expression levels of TFF3, phosphoinositide 3-kinase (PI3K), Akt and phosphorylated (p)-AKT when miR-7-5p or TFF3 was overexpressed, and the effects of miR-7-5p and TFF3 on LS174T cell proliferation and migration were simultaneously investigated. miR-7-5p was demonstrated to decrease the expression level of TFF3, and inhibit LS174T cell proliferation and migration, which was accompanied by decreased expression levels of PI3K and p-Akt. miR-7-5p was decreased following combined treatment with the TFF3 plasmid and miR­7-5p mimics, compared with treatment with miR-7-5p mimics alone, which was accompanied by increased expression levels of TFF3, PI3K and p-Akt, and enhanced LS174T cell proliferation and migration effects. The expression levels of miR-7-5p in the miRNA negative control (NC) + LY294002 group, the miR­7-5p mimic + LY294002 group, and the miR-7-5p mimic + TFF3 plasmid + LY294002 group were higher than those in the NC group, the miR-7-5p mimic group and the miR-7-5p mimic + TFF3 plasmid group, respectively. Accordingly, the expression level of TFF3 was downregulated and the proliferation and migration ability of the cells was downregulated. The present study demonstrates that overexpressed miR-7-5p may inhibit the proliferation and migration of LS174T cells by targeting the expression of TFF3 via inhibiting the PI3K/Akt signalling pathway. The PI3K/Akt signalling pathway may exert a feedback regulation effect on miR-7-5p, inhibiting the activity of this signalling pathway, which increases the miR-7-5p expression levels and further enhances the effects of miR-7-5p on cell proliferation and migration.

MicroRNA­7­5p regulates the expression of TFF3 in inflammatory bowel disease.

Trefoil factor 3 (TFF3) serves an important role in intestinal mucosal damage and healing, and contributes to the pathogenesis and treatment of inflammatory bowel disease (IBD). The aim of the present study was to determine the association between TFF3 and microRNA­7­5p (miR­7­5p) in IBD. Tissue immunohistochemistry was applied to evaluate the relative expression of TFF3, and reverse transcription­quantitative polymerase chain reaction was performed to determine the expression of miR­7­5p in lesional tissue obtained from patients with IBD and healthy control tissues. A dual­luciferase reporter assay was used to investigate whether TFF3 was a target of miR­7­5p, and western blotting was performed to determine the expression of TFF3 when miR­7­5p was overexpressed or suppressed. The protein expression levels of TFF3 were decreased and miR­7­5p was overexpressed in the lesional tissue of patients with IBD compared with in healthy control tissues. TFF3 was identified as a target of miR­7­5p, and TFF3 protein expression was negatively regulated by miR­7­5p in human colonic epithelial LS174T cells. The present study demonstrated a negative association between the expression of miR­7­5p and TFF3 in IBD lesional tissues and normal tissues. In conclusion, TFF3 was identified as a novel target of miR­7­5p and miR­7­5p may serve as a promising therapeutic target for IBD.

A double-blinded randomized trial on growth and feeding tolerance with Saccharomyces boulardii CNCM I-745 in formula-fed preterm infants / Ensaio duplo-cego randomizado sobre crescimento e tolerância de alimentação com a Saccharomyces boulardii CNCM I-745 em neonatos prematuros alimentados com fórmula

Abstract Objective: The use of probiotics is increasingly popular in preterm neonates, as they may prevent necrotizing enterocolitis sepsis and improve growth and feeding tolerance. There is only limited literature on Saccharomyces boulardii CNCM I-745 (S. boulardii) in preterm infants. Method: A prospective, randomized, case-controlled trial with the probiotic S. boulardii (50 mg/kg twice daily) was conducted in newborns with a gestational age of 30-37 weeks and a birth weight between 1500 and 2500 g. Results: 125 neonates were enrolled; 63 in the treatment and 62 in the control group. Weight gain (16.14 ± 1.96 vs. 10.73 ± 1.77 g/kg/day, p < 0.05) and formula intake at maximal enteral feeding (128.4 ± 6.7 vs. 112.3 ± 7.2 mL/kg/day, p < 0.05) were significantly higher in the intervention group. Once enteral feeding was started, the time needed to reach full enteral feeding was significantly shorter in the probiotic group (0.4 ± 0.1 vs. 1.7 ± 0.5 days, p < 0.05). There was no significant difference in sepsis. Necrotizing enterocolitis did not occur. No adverse effects related to S. boulardii were observed. Conclusion: Prophylactic supplementation of S. boulardii at a dose of 50 mg/kg twice a day improved weight gain, improved feeding tolerance, and had no adverse effects in preterm infants >30 weeks old.

Resumo Objetivo: O uso de probióticos está cada vez mais popular em neonatos prematuros, já que podem prevenir a enterocolite necrosante (ECN) e a sepse e aumentar o crescimento e a tolerância de alimentação. Há apenas uma literatura limitada sobre a Saccharomyces boulardii CNCM I-745 (S. boulardii) em neonatos prematuros. Método: Um ensaio de caso-controle prospectivo randomizado com o probiótico S. boulardii (50 mg/kg duas vezes por dia) foi feito com recém-nascidos com idade gestacional de 30 a 37 semanas e peso ao nascer entre 1.500 e 2.500 g. Resultados: Foram incluídos 125 neonatos, 63 no grupo de tratamento e 62 no de controle. O ganho de peso (16,14 ± 1,96 em comparação com 10,73 ± 1,77 g/kg/dia, p < 0,05) e a ingestão de fórmula com nutrição enteral máxima (128,4 ± 6,7 em comparação com 112,3 ± 7,2 mL/kg/dia, p < 0,05) foram significativamente maiores no grupo de intervenção. Assim que a nutrição enteral foi iniciada, o tempo necessário para atingir a nutrição enteral completa foi significativamente menor no grupo probiótico (0,4 ± 0,1 em comparação com 1,7 ± 0,5 dia, p < 0,05). Não houve diferença significativa em sepse. Não ocorreu ECN. Não foi observado efeito colateral relacionado à S. boulardii. Conclusão: A suplementação profilática de S. boulardii em uma dose de 50 mg/kg duas vezes por dia melhorou o ganho de peso, aumentou a tolerância de alimentação e não teve efeito colateral em neonatos prematuros > 30 semanas de idade.

A double-blinded randomized trial on growth and feeding tolerance with Saccharomyces boulardii CNCM I-745 in formula-fed preterm infants.

OBJECTIVE: The use of probiotics is increasingly popular in preterm neonates, as they may prevent necrotizing enterocolitis sepsis and improve growth and feeding tolerance. There is only limited literature on Saccharomyces boulardii CNCM I-745 (S. boulardii) in preterm infants. METHOD: A prospective, randomized, case-controlled trial with the probiotic S. boulardii (50mg/kg twice daily) was conducted in newborns with a gestational age of 30-37 weeks and a birth weight between 1500 and 2500g. RESULTS: 125 neonates were enrolled; 63 in the treatment and 62 in the control group. Weight gain (16.14±1.96 vs. 10.73±1.77g/kg/day, p<0.05) and formula intake at maximal enteral feeding (128.4±6.7 vs. 112.3±7.2mL/kg/day, p<0.05) were significantly higher in the intervention group. Once enteral feeding was started, the time needed to reach full enteral feeding was significantly shorter in the probiotic group (0.4±0.1 vs. 1.7±0.5 days, p<0.05). There was no significant difference in sepsis. Necrotizing enterocolitis did not occur. No adverse effects related to S. boulardii were observed. CONCLUSION: Prophylactic supplementation of S. boulardii at a dose of 50mg/kg twice a day improved weight gain, improved feeding tolerance, and had no adverse effects in preterm infants >30 weeks old.

[Efficacy of infliximab in the treatment of Crohn's disease in children].

OBJECTIVE: To evaluate the efficacy and safety of infliximab in the treatment of Crohn's disease in children. METHODS: Thirteen children who were diagnosed with Crohn's disease and received routine comprehensive treatment and infliximab (5 mg/kg) between January 2011 and December 2014 were enrolled. The changes in their clinical manifestations, laboratory indices, and Pediatric Crohn's Disease Activity Index (PCDAI) after the 30-week treatment were analyzed retrospectively. Meanwhile, endoscopy was performed to evaluate therapeutic effects. RESULTS: The symptoms such as abdominal pain, diarrhea, and bloody stool were relieved soon after infliximab treatment, with no recurrence observed; after the 30-week treatment, the white blood cell count, erythrocyte sedimentation rate, C-reactive protein, and the PCDAI decreased, while the hemoglobin increased significantly compared with those before treatment (P<0.05). After infliximab treatment, two children underwent endoscopy. The endoscopy showed that one child was cured, and the other child failed to respond to the treatment. No adverse drug reactions were seen in all patients. CONCLUSIONS: Infliximab treatment has significant clinical effects in children with Crohn's disease, with no obvious adverse reactions, and therefore, it can be applied as one of the preferred alternatives for treatment of Crohn's disease in children.

The protective effect of trefoil factor 3 on the intestinal tight junction barrier is mediated by toll-like receptor 2 via a PI3K/Akt dependent mechanism.

Trefoil factor peptides are highly conserved secreted molecules characterized by heat and enzymatic digestion resistance. Intestinal trefoil factor 3 (TFF3) protects and repairs the gastrointestinal mucosa and restores normal intestinal permeability, which is dependent on the integrity of the tight junction (TJ) barrier and the TJ associated proteins claudin-1, zona occludens-1 (ZO-1) and occludin. Despite the important role of intestinal barrier dysfunction in the pathogenesis of inflammatory bowel diseases, the underlying mechanisms and associated molecules remain unclear. In the present study, we show that TFF3 and toll-like receptor 2 (TLR2) are functionally linked and modulate intestinal epithelial permeability via a mechanism that involves the PI3K/Akt pathway. We used the Caco-2 cell model to show that TLR2 and TFF3 inhibit the IL-1ß induced increase in permeability and release of proinflammatory cytokines, and that this effect is mediated by activation of PI3K/Akt signaling. TLR2 silencing downregulated the expression of TFF3 and overexpression of TLR2 and TFF3 increased the levels of phospho-Akt. TFF3 overexpression significantly upregulated the expression of ZO-1, occludin and claudin-1 and this effect was abrogated by inhibition of the PI3K/Akt pathway. Taken together, our results indicate that TLR2 signaling selectively enhances intestinal TJ barrier integrity through a mechanism involving TFF3 and the activation of the PI3K/Akt pathway.

[Screening and identification of high-yield poly(ß-malic acid) bacterial strain].

OBJECTIVE: To isolate and identify the high-yield poly-malic acid (PMLA) bacterial strains from the nature. METHODS: Samples were collected and cultured. The high-yield PMLA bacterial strains were screened through morphological observation, qualitative PMLA tests by HPLC and ITS sequence analysis on the isolated bacterial strains. RESULTS: A high-yield PMLA strain II 04 was isolated, the yield of PMLA of the strain reached to 26.23g/L in the rotary shaker at 25 degree for 7d. From morphological observation and ITS sequences analysis, the strain belonged to Aureobasidium pullulans, and named as Aureobasidium pullulans ZUCC-41. CONCLUSION: A high-yield bacterial strain has been isolated from the nature and identified to be Aureobasidium pullulans.

Protective effect of intestinal trefoil factor on injury of intestinal epithelial tight junction induced by platelet activating factor.

Intestinal barrier dysfunction plays an important role in the pathogenesis of inflammatory bowel disease (IBD). To evaluate the effect of intestinal trefoil factor (ITF) on increased intestinal permeability and its association with tight junction proteins, an in vitro intestinal epithelia barrier model was established with Caco-2 cells and treated with platelet-activating factor (PAF). We found that exposing cells to 0.3 M ITF (30 min before or 30 min after PAF treatment) attenuated the PAF-induced changes in transepithelial electrical resistance and Lucifer yellow flux. A quantitative RT-PCR and western blot analysis revealed that ITF suppressed PAF-induced downregulation of tight junction proteins claudin-1 and ZO-1 expression; furthermore, an abnormal localization and distribution of these proteins was inhibited, as assessed by immunofluorescence staining. These results suggest that ITF decreases mucosal permeability and shows potential as a therapy for treating IBD.