[The association between Helicobacter pylori virulence factor genotypes and gastroduodenal diseases in children].

Objective: To investigate the association between Helicobacter pylori (Hp) virulence factor genotypes and the degree and activity of gastric mucosa pathological changes in pediatric gastroduodenal diseases. Methods: This retrospective cohort study was conducted from May 2020 to October 2020. The frozen strains of Hp, which were cultured with the gastric mucosa of 68 children with gastroscopy confirmed gastroduodenal diseases who visited the children's hospital of Zhejiang University School of Medicine from April 2012 to December 2014, were resuscitated. After extracting DNA from these Hp strains, PCR amplification and agarose gel electrophoresis were performed to determine the detection rate of cytotoxin-associated protein A (cagA),vacuolating cytotoxin A (vacA)(s1a、s1b/s2,m1/m2), outer inflammatory protein A (oipA),blood group antigen binding adhesin (babA),duodenal ulcer promoting protein A (dupA) genes; oipA genes were sequenced to determine the gene status. The patients were divided into different groups according to the findings of gastroscopy and gastric mucosa pathology. The detection rates of various virulence factor genotypes among different groups were compared using χ2 tests or Fisher's exact tests. Results: The 68 Hp strains all completed genetic testing. According to the diagnostic findings of gastroscopy, the 68 cases were divided into 47 cases of superficial gastritis and 21 cases of peptic ulcer. Regarding the pathological changes of gastric mucosa, 8 cases were mild, and 60 cases were moderate and severe according to the degree of inflammation; 61 cases were active and 7 cases inactive according to the activity of inflammation. The overall detection rates of cagA, vacA, vacA s1/m2, functional oipA, babA2, and dupA virulence factor genes were 100% (68/68), 100% (68/68), 94% (64/68), 99% (67/68), 82% (56/68), and 71% (48/68), respectively. In the superficial gastritis group, their detection rates were 100% (47/47), 100% (47/47), 96% (45/47), 98% (46/47), 81% (38/47), and 70% (33/47), respectively; in the peptic ulcer group, their detection rates were 100% (21/21), 100% (21/21), 90% (19/21), 100% (21/21), 86% (18/21), and 71% (15/21), respectively. There was no statistically significant difference between the two groups (all P>0.05). In the mild gastric mucosa inflammation group, the detection rates of the above six genotypes were 8/8, 8/8, 8/8, 7/8, 7/8, and 5/8, respectively; and in the moderate to severe inflammation groups, the detection rates were 100% (60/60), 100% (60/60), 93% (56/60), 100% (60/60), 82% (49/60), and 72% (43/60), respectively, with no statistically significant difference between the two groups (all P>0.05). In the active inflammation group, the detection rate of six genotypes were 100% (61/61), 100% (61/61), 93% (57/61), 98% (60/61), 82% (50/61), and 72% (44/61), respectively; and in the inactive inflammation group, they were 7/7, 7/7, 7/7, 7/7, 6/7, and 4/7, respectively. Again, there was no statistically significant difference between the two groups (all P>0.05). There was no statistically significant difference in the detection rate of combinations of 4 or 5 virulence factor genes among the different groups (all P>0.05). Conclusions: CagA, vacA, vacA s1/m2, functional oipA, babA2, and dupA genes are not associated with superficial gastritis and peptic ulcer in children, or with the degree and activity of gastric mucosa pathological inflammation. Different gene combinations of cagA, vacA, oipA, babA2, and dupA have no significant effects on predicting the clinical outcome of Hp infection in children.

[The relationship between genetic polymorphism of CYP2C19 and the efficacy of Helicobacter pylori eradication therapy in children].

Objective: To investigate the relationship between genetic polymorphisms of cytochrome P450 2C19 (CYP2C19) and the efficacy of Helicobacter pylori (Hp) eradication therapy in children. Methods: The retrospective cohort study was conducted on 125 children with gastroscopy and positive rapid urease test (RUT) from September 2016 to December 2018 who presented to the Children's Hospital of Zhejiang University School of Medicine due to gastrointestinal symptoms including nausea, vomiting, abdominal pain, bloating, acid reflux, heartburn, chest pain, vomiting blood and melena. Hp culture and drug susceptibility test were carried out with gastric antrum mucosa before treatment. All the patients completed 2 weeks of standardized Hp eradication therapy and had 13C urea breath test 1 month after that, which was used to evaluate the curative effect. The DNA of gastric mucosa after RUT was analyzed and CYP2C19 gene polymorphism was detected. Children were grouped according to metabolic type. Combined with the results of Hp culture and drug susceptibility, the relationship between CYP2C19 gene polymorphism and the efficacy of Hp eradicative treatment was analyzed in children. Chi square test was used for row and column variables, and Fisher exact test was used for comparison between groups. Results: One hundred and twenty five children were enrolled in the study, of whom 76 were males and 49 females. The genetic polymorphism of CYP2C19 in these children found poor metabolizer (PM) of 30.4% (38/125), intermediate metabolizer (IM) of 20.8% (26/125), normal metabolizer (NM) of 47.2% (59/125), rapid metabolizer (RM) of 1.6% (2/125), and ultrarapid metabolizer (UM) of 0. There were statistically significant in positive rate of Hp culture among these groups (χ2=124.00, P<0.001). In addition, the successful rates of Hp eradication in PM, IM, NM and RM genotypes were 84.2% (32/38), 53.8% (14/26), 67.8% (40/59), and 0, respectively, with significant differences (χ2=11.35, P=0.010); those in IM genotype was significantly lower than that in PM genotype (P=0.011). With the same standard triple Hp eradicative regimen, the successful rate of Hp eradication for IM type was 8/19, which was lower than that of PM (80.0%, 24/30) and NM type (77.3%, 34/44) (P=0.007 and 0.007, respectively). There was a significant difference in the efficacy of Hp eradication treatment among different genotypes (χ2=9.72, P=0.008). According to the clarithromycin susceptibility result, the successful rate of Hp eradication treatment for IM genotype was 4/15 in the sensitive group and 4/4 in the drug-resistant group (χ2=6.97, P=0.018). Conclusions: The genetic polymorphism of CYP2C19 in children is closely related to the efficacy of Hp eradication treatment. PM has a higher successful rate of eradication treatment than the other genotypes.

[Eosinophilic esophagitis in children: analysis of 22 cases].

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.

The anti-cancer effects of cisplatin on hepatic cancer are associated with modulation of miRNA-21 and miRNA-122 expression.

OBJECTIVE: Cisplatin is an effective chemotherapeutic drug to treat hepatic cancer, but its efficacy is marred by extensive adverse effects. Micro (mi) RNAs are small regulatory RNAs that may be used as molecular targets to better fine-tune chemotherapy in hepatic cancer. In this study, we examined to what extent the anti-cancer effects of cisplatin are associated with expressions of miRNA (miR)-21 and miR-122. MATERIALS AND METHODS: The growth-inhibiting effects of cisplatin on the human hepatic cell line HepG2 were assessed by MTT assay, while cell apoptosis was documented using DAPI staining. Also, we tested the effects of cisplatin on tumour growth in a mouse tumour xenograft model. Finally, we quantified expression levels of miR-21 and miR-122 in cisplatin-treated HepG2 cells. RESULTS: We observed that cisplatin significantly decreased the growth of HepG2 cells (p < 0.05 vs control cells) at all tested concentration (5-80 µg/ml) after 24 or 48 hours of treatment. Microscopic studies demonstrated apoptotic signs in cisplatin-treated cells. In the mouse tumour xenograft model, tumour weights and volumes were significantly (p < 0.05 untreated animals) lower after treatment with cisplatin. Also, treatment of HepG2 cells for 48 hours with 20 µg/ml cisplatin was associated with significant decreases in miR-21 expression levels and up-regulation of miR-122. CONCLUSIONS: The anti-cancer effects of cisplatin are associated with down-regulation of miR-21 expression and up-regulation of miR-122.

Expression of transforming growth factor-ß1 and connective tissue growth factor in congenital biliary atresia and neonatal hepatitis liver tissue.

We investigated the expression of transforming growth factor-ß1 (TGF-ß1) and connective tissue growth factor (CTGF) in the liver tissue of infants with congenital biliary atresia and neonatal hepatitis, as well as the relationship between the expression of the two factors and liver fibrosis. Thirty-six infants who met the cholestasis criteria were classified into congenital biliary atresia and neonatal hepatitis groups. All specimens were stained with hematoxylin and eosin and Masson's trichrome, and the degree of liver fibrosis was assessed. The scope and level of CTGF and TGF-ß1 expression in the different specimens was evaluated by immunohistochemistry and observation. Liver fibrosis in the congenital biliary atresia group was more advanced than that in the neonatal hepatitis group, and the difference was significant (P < 0.01). In the neonatal hepatitis patients, CTGF and TGF-ß1 were mainly expressed in the hepatocytes, while they were expressed in both hepatocytes and biliary epithelial cells in the congenital biliary atresia patients, and in these patients the expression was significantly stronger than in the neonatal hepatitis patients (P < 0.01). With the aggravation of hepatic fibrosis, CTGF and TGF-ß1 expression levels in liver tissue gradually increased, and their expression levels were significantly correlated (P < 0.01). Liver fibrosis is present in both congenital biliary atresia and neonatal hepatitis patients. The gradual increase of CTGF and TGF-ß1 expression levels in liver tissue is associated with liver fibrosis. Early expression of CTGF and TGF-ß1 in biliary epithelial cells may be involved in the pathogenesis of congenital biliary atresia.

Effect of glutamine on intestinal barrier function following liver transplantation in rats.

OBJECTIVES: Glutamine is an important fuel for intestinal mucosal epithelial cells, and it promotes intestinal mucosal cell differentiation and proliferation. Most liver transplantation (LT) patients suffer from intestinal barrier dysfunction. Whether enteral glutamine supplementation has beneficial effects on intestinal barrier function following LT is not known. We investigated the effect of glutamine (Gln) supplementation on NF-κB and on the intestinal barrier in rats after an allogenic LT with concomitant immunosuppressive therapy. MATERIALS AND METHODS: Inbred Sprague-Dawley rats (n=40) receiving allogenic LT were randomly divided into Gln and control groups (n=20, each). Gln group rats were administered Gln (0.4 g/kg·day) by gastric infusion for 6 days, while control rats received saline. Ten rats from each group were sampled for basal parameters on the 3rd day, prior to LT. The remaining 10 from each group were sampled after receiving LT. Twenty inbred Sprague-Dawley rats were selected as donors. The 20 recipients underwent orthotopic LT after 3 days of treatment and were given immunosuppressive therapy for 6 days post-operation. They were euthanized for sample collection on the 7th day. NF-κB protein in the intestinal mucosa, portal plasma Gln, endotoxin and TNF-α levels, ileocecal sIgA content, bacterial translocation and mucosal ultrastructure were assessed. RESULTS: On the postoperative day 6, the Gln group had increased plasma Gln and ileocecal sIgA (secretory IgA). Gln group also showed improvement in mucosal microvilli structure and had reduced levels of intestinal mucosal NF-κB, portal endotoxin and TNF-α and decreased bacterial translocation as compared to the control group. CONCLUSIONS: Parenteral supplementation of glutamine ameliorated mucosal injury during allogenic LT, and improved intestinal barrier function. These findings suggest that glutamine supplementation may be an effective therapy to ensure successful recovery from liver transplantation.

L-arginine enhances arginine deiminase induced human lymphoma cell growth inhibition through NF-kBp65 and p53 expression in vitro.

OBJECTIVE: Arginine deiminase (ADI) and L-arginine (L-Arg) can act as anti-tumor agents in-vitro and in-vivo. However, the mechanism of ADI and L-Arg as anti-tumor agents has not been clearly shown. MATERIALS AND METHODS: With the goal of understanding the role of ADI and L-Arg in inhibition of cell growth, we used the Ramos human lymphoma cell line, which is known to be ADI-sensitive, and observed the p53 and NF-κBp65 protein expression after ADI and arginine treatment. After determining an optimal experimental ADI concentration (0.01 U/ml), we studied the effects of ADI treatment, when combined with different concentrations of L-arginine (control, ADI only, ADI with 10 mM/ml Arg, ADI with 30 mM/ml Arg, and ADI with 50 mM/ml Arg). An MTT assay was used to assess cell survival after treatment, Western blot analysis to determine the levels of the NF-κBp65, p53 and NO mediators and nitric oxide assays were used to determine nitrite levels. RESULTS AND CONCLUSIONS: L-arginine enhanced ADI-induced inhibited cell growth through expression of NF-κBp65 and p53 in a dose-dependent manner.

Regulation of apoptosis in human gastric cancer cell line SGC-7901 by L-arginine.

Aim: L-arginine (LArg) is an amino acid that has immunomodulating and anti-tumor effects. It is possible that anti-tumor effects of L-Arg are due to induction of apoptosis in tumor cells. The present study assessed anti-proliferating and pro-apoptotic effects of L-Arg in human gastric cancer cell line SGC7901. Methods: Cell proliferation was quantified by MTT assay. Apoptosis was assessed using flow cytometry and FITCAnnexinV/ propidium iodide staining. Expression and activation of proteins pertinent to apoptosis (Bcl2, surviving, p53, and XIAP) were studied using PCR, Western blot, and activity assays. Results: L-Arg significantly inhibited growth of SCG-7901 gastric cancer cells and down-regulated expression of anti-apoptotic gene Bcl-2 and survivin. By contrast, expression of p53 was upregulated by L-Arg. Conclusion: Regulation of apoptosis by L-Arg via down-regulation of anti-apoptotic proteins Bcl-2 and surviving, and up-regulation of pro-apoptotic protein p53 may represent the mechanism behind antitumor effects of L-Arg.

Regulation of apoptosis in human gastric cancer cell line SGC-7901 by L-arginine.

AIM: L-arginine (L-Arg) is an aminoacid that has immunomodulating and antitumor effects. It is possible that antitumor effects of L-Arg are due to induction of apoptosis in tumor cells. The present study assessed antiproliferating and proapoptotic effects of L-Arg in human gastric cancer cell line SGC-7901. METHODS: Cell proliferation was quantified by MTT assay. Apoptosis was assessed using flow cytometry and FITC-Annexin-V/propidium iodide staining. Expression and activation of proteins pertinent to apoptosis (Bcl-2, surviving, p53, and XIAP) were studied using PCR, Western blot, and activity assays. RESULTS: L-Arg significantly inhibited growth of SCG-7901 gastric cancer cells and downregulated expression of antiapoptotic gene Bcl-2 and survivin. By contrast, expression of p53 was upregulated by L-Arg. CONCLUSION: Regulation of apoptosis by L-Arg via downregulation of antiapoptotic proteins Bcl-2 and surviving, and upregulation of proapoptotic protein p53 may represent the mechanism behind antitumor effects of L-Arg.

Starch properties of mutant rice high in resistant starch.

As the staple food of over half the world's population, hot cooked rice high in resistant starch (RS) is of particular interest, which will have greater impact in the dietary prevention of diabetes and hyperlipidemia. A mutant rice high in RS in hot cooked rice, described as RS111, was comparatively studied with the wild type and common rice. Despite obviously low RS content in the raw milled rice, the RS content in the hot cooked rice of mutant RS111 was significantly higher than that of the wild type and common rice and, correspondingly, in vitro starch hydrolysis by porcine pancreatic alpha-amylase tends to be incomplete with low hydrolysis extent for the cooked mutant rice high in RS. Obvious differences in physicochemical properties, starch granule morphology, pasting properties, thermal properties, and X-ray diffraction pattern were observed among the mutant RS111, wild type, and common indica rice. The high-RS mutant was characterized by significantly higher apparent amylose content and crude lipid content, higher percentage of oval-shaped granules and bigger oval size, reduced paste viscosity, and low onset temperature, peak temperature, final temperature, enthalpy of gelatinization, and crystallinity.