Objective:To investigate the association between intestinal colonization of segmented filamentous bacteria (SFB) and the risk of rotavirus infection, and the possible mechanisms by which SFB resist rotavirus infection.Methods:This case-control study enrolled 50 children aged 0 to 5 years who present to the outpatient Department of Children′s Hospital, Zhejiang University School of Medicine with diarrhea and positive stool tests for rotavirus. The children were divided into rotavirus enteritis group and control group consisting of 55 children with non-gastrointestinal and non-infectious surgical diseases.The age and sex composition of the two groups was matched. The DNA of the fecal flora was extracted and SFB was detected by real-time fluorescence quantitative PCR analysis. The children in the rotavirus enteritis group and the control group were subgrouped by age and sex to analyze the differences in SFB positivity rates between different groups, and further compare and analyze the differences in SFB positivity rates between these two groups of children in the ≤2 years old subgroup and the >2-5 years old subgroup. Neutralization test was performed with p3340 protein and rotavirus to determine the relationship between rotavirus infection rate and p3340 concentration in Vero cells. χ2 test or Fisher′s exact probability method was used for comparison between the two groups. Results:There were 50 children in the rotavirus enteritis group with an age of (1.7±0.9) years, and 55 children in the control group with an age of (1.8±1.1) years. The positive rate of SFB in children with rotavirus enteritis showed a declining trend across ages groups, with the highest rate of 10/14 in the ≤1 year old group, followed by 67% (14/21) in the >1-2 years old group, 9/15 in the >2-5 years old group, and there was no statistically significant difference ( P=0.867). The positive rate of SFB in the control group was 12/15 in the ≤1 year old group, 95% (19/20) in the >1-2 years old group, 50% (10/20) in the >2-5 years old group, with statistical significance ( P=0.004). The positive rate of SFB in children with rotavirus enteritis was 74% (20/27) in males and 56% (13/23) in females ( χ2=1.71, P=0.192). In the control group, it was 79% (22/28) in males and 70% (19/27) in females ( χ2=0.49, P=0.485). The positive rate of SFB was 66% (33/50) in the rotavirus enteritis group and 75% (41/55) in the control group, with no statistically significant ( χ2=0.56, P=0.454). In the children ≤2 years old, the SFB positivity rate was 69% (24/35) in the rotavirus enteritis group and 89% (31/35) in the control group, with a statistically significant difference ( χ2=4.16, P=0.041). However, in the children >2-5 years old, no statistically significant difference was observed, with the positive rate of SFB being 9/15 in the rotavirus enteritis group and 50% (10/20) in the control group ( P=0.734). Pearson correlation analysis revealed a negative correlation between rotavirus infection and SFB positivity ( r=-0.87, P<0.001). As the concentration of the p3340 specific protein increased, the luminescence intensity of the luciferase in the Vero cells, which were suitable for cultivating rotavirus, exhibited a decreasing trend ( F=4.17, P=0.001). Conclusions:SFB colonization in infants less than 2 years old is associated with a reduced risk of rotavirus infection. Cloning of specific SFB functional protein p3340 neutralizes rotavirus infection of Vero cells, and this mechanism of targeting rotavirus infection differs from the common antiviral mechanism.

Infant dyschezia is one of the most common functional gastrointestinal disorders in infants.The main clinical features include an emotionally state of strain, with scream, cry and red or purple in the face with each effort to defecate, which usually lasting 10-20 minutes and normal stool passes several times a day.The current diagnostic criteria of infant dyschezia is based on the Rome Ⅳ criteria.Pathogenesis of infant dyschezia is still unclear, which may be related to immature defecation pattern in infants, intestinal flora imbalance, brain-gut-microbiota axis disturbances and complementary feeding starting too early.Infant dyschezia is self-limited, and there is no need of specific treatment.The caregivers are recommended to avoid rectal stimulation and laxative intervention.

Objective:To analyze and summarize the clinical features in children with recurrent intussusception.Methods:This retrospective cohort study collected the clinical data of 126 children with recurrent intussusception who were admitted to the Children′s Hospital of Zhejiang University School of Medicine due to "abdominal pain, paroxysmal crying, vomiting, bloody stools" from January 1, 2015 to November 30, 2019. The clinical manifestations of recurrent intussusception between ≤3 years old group and >3 years old group were compared, the etiology and age characteristics of pathologic lead points (PLP) were analyzed, and the clinical characteristics of PLP group and non-PLP group were also compared. The χ 2 test and Mann-Whitney U test were used to compare the differences between groups. Results:A total of 126 children with recurrent intussusception were included, of whom 76 were males and 50 were females, with the age of 2.9 (1.7, 5.1) years. The proportion of children aged more than 1 year was 87.3% (110/126), and 48.4% (61/126) more than 3 years. Clinical manifestations mostly lacked the typical triad of symptoms. The percentage of paroxysmal crying in ≤ 3 years old group was significantly higher than that in >3 years old group (52.3% (34/65) vs. 24.6% (15/61), χ 2=10.17, P=0.001), while the percentage of abdominal pain was significantly lower than that in the >3 years old group (46.1% (30/65) vs. 75.4% (46/61), χ 2=11.25, P=0.001). The rate of positive ultrasound examination was 17.5% (22/126), and 63.6% (14/22) of them were diagnosed. The positive rate of CT examination was 4/13, of which 2 cases were diagnosed. In this study, 37 children were diagnosed with PLP by colonoscopy, laparoscopy or laparotomy, and 89 children were found without PLP. The positive rate of PLP in >3 years old group was significantly higher than that in ≤3 years old group (37.7% (23/61) vs. 21.5% (14/65), χ 2=3.96, P=0.046). Meckel′s diverticulum and juvenile polyp were the main contributors of PLP in ≤3 years old group, accounting for 7/14 and 3/14 respectively, while lymphoma and juvenile polyp accounted for 34.8% (8/23) and 26.1% (6/23), respectively in >3 years old group. Compared with non-PLP group, PLP group had higher age (5.2 (1.6, 6.7) vs. 2.7 (1.8, 4.2) years, Z=-2.26, P=0.01). However, there were no significant differences in gender and recurrence frequency between the two groups (both P>0.05). Conclusions:Recurrent intussusception is more common in children more than 1 year old, and has a wide spectrum of non-specific clinical presentations. Imaging examinations can be used to identify PLP. The most recurrent intussusception is idiopathic, but the presence of PLP should be alerted for, such as Meckel′s diverticulum, lymphoma and juvenile polyp. Colonoscopy sometimes is necessary, surgical exploration and treatment should be carried out in time.

Nowadays, more and more attention was attached to the problem of children′s growth and development.It is known that postnatal growth is mainly regulated by the conserved GH-IGF-1 axis that acts through endocrine and paracrine pathways.It is now well established that undernourished children harbor an altered microbiota, correlated with impaired growth.Moreover, many disorders of intestinal flora are always accompanied by growth retardation.These evidences show that there might be a certain relationship between intestinal flora and growth and development.Recent studies have demonstrated that intestinal flora may regulate the growth and development process through this axis, and GH-IGF-1 axis may also affect the composition and diversity of intestinal flora.This paper reviewes the bidirectional regulatory relationship between intestinal microbiota and the GH-IGF-1 axis to reveal the functional relationship between growth and development of children and GH-IGF-1 axis as well as the intestinal flora.By elucidating the influence of intestinal flora on growth and development, a new approach would be found for the application of therapeutic methods of microflora in the field of growth retardation in children.

Objective:To investigate the differences of gastric mucosa microbiota between children with chronic gastritis and duodenal ulcer under the condition of Helicobacter pylori (Hp) infection. Methods:This prospective cohort study involved 57 children with Hp infection diagnosed by gastric endoscopy who were admitted to the Children′s Hospital of Zhejiang University School of Medicine due to "abdominal pain, abdominal distension and vomiting" between January 2018 to August 2018. According to gastroscopy and pathological examination, the children were divided into chronic gastritis group and duodenal ulcer group. Gastric mucosa from Hp infected patients were sampled, and the flora DNA was analyzed by high-throughput sequencing. The statistical difference of α diversity, β diversity between two groups were analyzed. The relative abundance of the two groups in each taxonomic level was analyzed statistically. T test, Rank sum test or χ 2 test was used for comparison between the two groups. Results:A total of 57 children diagnosed with Hp infection were enrolled in this study, including 42 cases of chronic gastritis (the age was (9.3±2.8) years, 22 males and 20 females) and 15 cases of duodenal ulcer (the age was (11.1±3.3) years, 9 males and 6 females). Alpha diversity index Chao and ACE in Hp infected chronic gastritis group were significantly higher than those in Hp infected duodenal ulcer group (217±50 vs. 183±64, t=2.088, P=0.009;218±47 vs. 192±76, t=1.566, P=0.016, respectively). The Beta-diversity index such as nonmetric multidimensional scaling (NMDS) analysis were significantly different in the two groups (analysis of similarity R=0.304, P=0.028). Among the main bacteria genera, there were 6 genera with significant differences between the two groups, which were Prevotella (0.190% (0.008%-1.983%) vs. 0.021% (0.005%-2.398%), Z=-2.537, P=0.011), Alloprevotella (0.097% (0.010%-0.813%) vs. 0.015% (0.003%-0.576%), Z=-2.492, P=0.013), Haemophilus (0.109% (0.004%-0.985%) vs. 0.014% (0.004%-0.356%), Z=-2.900, P=0.004), Neisseria (0.074% (0.004%-0.999%) vs. 0.024% (0.003%-0.255%), Z=-2.718, P=0.007), Streptococcus (0.166% (0.008%-1.869%) vs. 0.045% (0.006%-0.879%), Z=-2.537, P=0.010), and an unclassified- Microbacteriaceae (0.214% (0.060%-1.762%) vs. 0.117% (0.010%-0.954%), Z=-2.120, P=0.034). Linear discriminant analysis (LDA) effect sized analysis showed that at the genus level, only Prevotella was significantly enriched in the duodenal ulcer group (LDA=2.90, P=0.010), while Streptococcus, Neisseria and Haemophilus were significantly enriched in the chronic gastritis group (LDA=2.83, 2.82, 2.69, P=0.011, 0.007, 0.004, respectively). Conclusions:The gastric mucosal microbiota in duodenal ulcer associated with Hp is significantly different from that in chronic gastritis. Hp may promote the occurrence of peptic ulcer together with gastric microbiota.

OBJECTIVE: To investigate the age distribution characteristics of intestinal segmented filamentous bacteria (SFB) in children and their relationship with intestinal mucosal immunity. METHODS: The fresh feces of 177 children and the ileocecal fluid of 47 children during colonoscopy were collected. The SFB was determined by real-time PCR. The concentration of secretory immunoglobulin A (sIgA) was determined by enzyme-linked immunosorbent assay. The numbers of interleukin 17A (IL-17A) cells and intraepithelial lymphocytes in the terminal ileum mucosa and the expression of transcription factors associated with the differentiation of T helper (Th) cells, T-box transcription factor (T-bet), forkhead box P3 (FOXP3), and retinoid-related orphan receptor gamma t (ROR-γt), were determined by immunohistochemistry. RESULTS: The positive rate of intestinal SFB in these children was 19.2% (34/177). Trend analysis showed that the positive rate of SFB was correlated with age: the rates for children aged 0-, 1-, 2-, 3-, 4-, 5-, 6-, and 7-15 years were 40%, 47%, 32%, 15%, 12%, 13%, 15% and 4% respectively (P<0.001). The concentration of sIgA in intestinal fluid was significantly higher in SFB-positive children (n=24) than in SFB-negative children (n=23) (P<0.01). The number of intraepithelial lymphocytes in the terminal ileum mucosa and the expression of T-bet, FOXP3, and ROR-γt were not significantly different between the SFB-positive group (n=12) and the SFB-negative group (n=11), but the number of IL-17A cells in the terminal ileum mucosa was significantly lower in the SFB-positive group than in the SFB-negative group (P<0.05). CONCLUSIONS Intestinal SFB colonization in children is age-related, and the colonization rate is relatively high in children under 3 years old. In SFB-positive children, the secretion of intestinal sIgA is increased, while the number of IL-17A cells in the terminal ileum is reduced.
Adolescent ; Age Distribution ; Bacteria ; Child ; Humans ; Immunity, Mucosal ; Intestinal Mucosa

Functional gastrointestinal disorders are a common disease in pediatrics.It's diagnosis and treatment are mainly based on Rome standard according to the symptom of complaint.The advances of diagnosis and treatment of functional gastrointestinal disorders in children based on the Rome Ⅳ standard were reviewed,and in order to improve the pediatrician's diagnostic levels for this kind of diseases.

Objective: Eosinophilic esophagitis (EoE) is a chronic immune-mediated esophageal disease.The current domestic reports of EoE in children is rare.The aim of this study was to analyze the clinical features, the diagnosis and treatment advance of EoE in children by case analysis and literature review. Method: Clinical data of 22 children with EoE from January, 2011 to December, 2015 in Children′s Hospital, Zhejiang University School of Medicine were recorded, retrospective analysis was performed on clinical presentation, gastroendoscopy and histopathological examination features and the treatment. Result: (1) Clinical data: EoE can occur at any age in children (5 months to 13 years). The most common clinical manifestations of EoE are vomiting and abdominal pain, 45% (10/22) and 41%(9/22) respectively. (2) Endoscopy and pathological features of esophageal mucosa: 11 cases with coarse mucous membrane (50%), 6 cases with congestion or erosion of esophageal membrane (27%), 5 cases with longitudinal crack (23%), 3 cases with ring uplift (14%), 3 cases with granular uplift (14%), 3 cases with normal mucosa(14%). Histopathologic manifestation is eosinophil infiltration and the eosinophil counts were all more than or equal to 15/HP. (3) Laboratory results: 13 cases had increasing eosinophil counts and eosinophils proportion (62%). (4)Allergy history: among 22 cases, 7 patients had allergy history (32%). (5) Situation of treatment and remission: 16 cases had clinical remission by oral omeprazole; 2 cases had clinical remission by oral Omeprazole and Montelukast sodium; 1 case acquired remission by elimination diet; 1 case acquired remission by elimination diet and oral prednisone. 2 cases dropped out; Only 2 patients received gastroendoscopy re-examination after 3 months and revealed esophageal mucosal histologic complete recovery. Conclusion The clinical symptoms of EoE in children varies.Esophageal mucosal features of gastroendoscopy examination in children with EoE were longitudinal crack, white exudates or plaques, paper mucosa, ring uplift and granular uplift.Most patients could achieve remission by using proton-pump inhibitors, only few children needed elimination diet and change formula, or even oral glucocorticoids.

Gastroesophageal reflux (GER) and cow's milk allergy (CMA) are common disorders in infants. In recent years, more and more research has investigated the relationship between these two diseases. Some studies reported that about half of the cases of GER in infants younger than 1 year may be an association with CMA. Therefore, overall understanding the role of CMA on the pathogenesis of GER has a great importance on improving clinical level of diagnosis and therapy. This review article tried to elaborate advances in research on the relationship between CMA and GER in infants, including epidemiology, pathogenesis, clinical manifestations, diagnosis and treatment.
Gastroesophageal Reflux ; diagnosis ; etiology ; Humans ; Infant ; Milk Hypersensitivity ; complications

OBJECTIVETo understand the junction protein expression of gastric mucosa including occlusal proteins (occludin), closed protein-4 (claudin-4), zonula occluden-1(ZO-1), epithelial cadherin (E-cadherin), and β ring protein (β-catenin) and the clinical significance in children with Helicobacter pylori (Hp) infection.

METHODSeventy patients in whom gastric endoscopy was performed because of nausea, vomiting, abdominal pain, bloating, acid reflux, melena, and other gastrointestinal symptoms were enrolled in this study from Dec. 2010 to Apr. 2013 in our hospital. Informed consent was signed by their parents, and the study was in accordance with the principles of medical ethics. Hp positivity was confirmed if both respiratory urea test (RUT) and Hp were positive by gastric mucosal pathology. Gastric mucosal samples from 70 patients were enrolled in this study, 23 of them were Hp negative, 47 of them were Hp positive (24 cases without peptic ulcer, 23 cases with peptic ulcer). The mRNA levels and protein expression of tight junction protein of gastric mucosa were measured by RT-PCR and Western blot respectively. The location and semi quantitative content of E-cadherin and β-catenin in gastric mucosa were detected by immunohistochemical staining method.

RESULTThe mRNA level of E-cadherin, β-catenin, ZO-1 in the Hp positive group regardless of peptic ulcer was significantly lower than that in the Hp negative group. Hp positive without peptic ulcer group were 0.0008, 0.0040, 0.0014, respectively; Hp positive with peptic ulcer group were 0.0010, 0.0090, 0.0013, respectively; Hp negative group were 0.0137, 0.0423, 0.0198, respectively (F values were 36.956, 39.893, 38.962, respectively, all P<0.05). The expression of claudin-4 mRNA in Hp positive group with peptic ulcer increased significantly, the difference among Hp positive group with peptic ulcer, Hp positive group without peptic ulcer and Hp negative group was statistically significant (0.1438 vs. 0.0926 vs. 0.0789) (F value was 11.964, P<0.05), while the difference of occludin mRNA levels among the three groups was not statistically significant.Immunohistochemistry results showed that the score of E-cadherin, β-catenin positive cell in the Hp positive patients were also significantly lower than that in the Hp negative group (t values were 3.981 and 2.340, all P<0.05, respectively). Western blot results showed that the protein levels of β-catenin in Hp positive group with peptic ulcer were significantly lower than that in Hp negative group, while the protein levels of E-cadherin in Hp positive patients regardless of peptic ulcer were decreased significantly in Hp negative group.

CONCLUSIONOur results revealed that the tight junction protein E-cadherin, β-catenin, ZO-1 expression of gastric mucosa were decreased in children with Hp infection, while claudin-4 expression was increased in Hp positive patients with peptic ulcer, suggesting that damage to gastric epithelial barrier function may be the main pathogenesis of Hp associated gastric diseases in children.

Blotting, Western ; Cadherins ; metabolism ; Child ; Claudin-4 ; metabolism ; Gastric Mucosa ; metabolism ; pathology ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Immunohistochemistry ; Occludin ; metabolism ; Peptic Ulcer ; metabolism ; microbiology ; RNA, Messenger ; Tight Junction Proteins ; metabolism ; Tight Junctions ; metabolism ; Zonula Occludens-1 Protein ; metabolism ; beta Catenin ; metabolism