Antibiotic pretreatment on the efficacy of Clostridium butyricum in the treatment of dextran sulfate sodium induced colitis and the influence of intestinal microbiota / 中华消化杂志

Objective:To investigate the effects of Clostridium butyricum on colitis and intestinal microbiota in mice with or without antibiotic pretreatment. Methods:Thirty specific pathogen free BALB/c mice were randomly divided into the blank control group, dextran sulfate sodium (DSS) group, antibiotic + DSS group, Clostridium butyricum + DSS group and antibiotic+ Clostridium butyricum + DSS group, with 6 mice in each group. After the mice were pretreated with quadruple antibiotics (ampicillin 1 g/L, neomycin 1 g/L, metronidazole 1 g/L, and vancomycin 0.5 g/L) in normal drinking water for 30 d, the mice colitis model was induced with DSS. At the same time, the mice in Clostridium butyricum + DSS group and antibiotics+ Clostridium butyricum + DSS group were given 1×10 6colony-forming unit (CFU) Clostridium butyricum by gavage. The effect of Clostridium butyricum on mice with colitis was evaluated by disease activity index (DAI), colon length and histopathological score. The level of serum inflammatory factors was detected by enxyme linked immunosorbent assay, and the effect of Clostridium butyricum on gut microbita in mice was determined by fecal 16S rRNA sequencing. Results:The general condition of mice of the blank control group were good, and their DAI scores fluctuated around 0. Since the fourth day after DSS drinking water was given, the mice of the DSS group showed signs of colitis such as weight loss, unformed stools and bloody stools. On the fourth day after intervention, the DAI score of Clostridium butyricum + DSS group was lower than that of DSS group (0.000±0.000 vs. 0.444±0.111), and the difference was statistically significant ( t=4.000, P=0.016 1). On the tenth and twelfth day after the intervention, the DAI scores of antibiotic+ Clostridium butyricum + DSS group were both lower than those of antibiotic+ DSS group (0.000±0.000 vs. 1.111±0.222, 0.667±0.000 vs. 1.889±0.222), and the differences were statistically significant ( t=5.000 and 5.500, both P<0.05). The histopathological score of mice colon tissue of Clostridium butyricum + DSS group was lower than that of DSS group (2.50±1.73 vs. 5.50±1.00), and the histopathological score of mice colon tissue of antibiotic+ Clostridium butyricum+ DSS group was lower than that of antibiotic+ DSS group (1.25±0.96 vs. 5.00±0.82), and the differences were statistically significant ( t=3.000 and 5.960, both P<0.05). The serum level of interleukin (IL)-1β Clostridium butyricum+ DSS group was higher than that of blank control group ((4.464±0.075) ng/L vs. (3.907±0.080) ng/L), the serum levels of tumor necrosis factor-α, IL-6 and IL-1β of Clostridium butyricum+ DSS group and antibiotic+ Clostridium butyricum + DSS group were all lower than those of DSS group ((2.402±0.383) ng/L , (1.845±0.345) ng/L vs. (6.958±1.084) ng/L, (1.752±0.146) ng/L, (1.307±0.048) ng/L vs. (3.537±0.608) ng/L, (4.464±0.075) ng/L, (4.066±0.190) ng/L vs. (7.477±0.339) ng/L), and the differences were statistically significant ( t=5.005, 3.964, 4.495, 4.693, 6.294, 8.674 and 8.774 , all P<0.05). The results of 16S rRNA sequencing showed that there were a significantly large number of anti-inflammatory or short-chain fatty acid producing bacteria in the gut microbiota of mice intervened by Clostridium butyricum, among which the dominant bacteria genus in Clostridium butyricum + DSS group and antibiotic+ Colstridium butyicum+ DSS group were Mucispirillum (linear discriminant analysis (LDA)=3.667 log10, P=0.004) and Stenotrophomonas (LDA=2.778 log10, P=0.044). In the antibiotic+ Clostridium butyricum+ DSS group, the dominant bacteria genus were Peptococcus (LDA=2.685 log10, P=0.018), Butyricimonas (LDA=2.712 log10, P=0.011), Bilophila (LDA=3.204 log10, P=0.014), Intestinimonas (LDA=3.346 log10, P=0.010), Candidatus- Saccharimonas (LDA=3.363 log10, P=0.029), Desulfovibrio (LDA=3.402 log10, P=0.025), Oscillibacter (LDA=2.870 log10, P=0.019) and Akkermansia (LDA=4.031 log10, P=0.005). Conclusions:Clostridium butyricum can effectively improve colitis in mice and regulate the intestinal microbial structure of mice, whlie antibiotic pretreatment can strengthen its regulation of intestinal microbiota to and enhance the efficacy of Clostridium butyricum.

Uncontrolled preliminary study on the clinical efficacy of fecal microbiota transplantation in irritable bowel syndrome and its influence on gut microbiota / 中华消化杂志

Objective:To investigate the efficacy and safety of fecal microbiota transplantation (FMT) in the treatment of irritable bowel syndrome (IBS), and to explore the effects of FMT on the gut microbiota of IBS patients.Methods:From September 2016 to August 2017, at Guangzhou First People′s Hospital, 28 hospitalized IBS patients who underwent FMT treatment were enrolled. Before FMT, four and 12 weeks after FMT, all the IBS patients completed the irritable bowel syndrome quality of life scale (IBS-QOL), irritable bowel syndrome severity scoring system (IBS-SSS) and gastrointestinal symptom rating scale (GSRS). 16S rDNA sequencing was performed before FMT and four weeks after FMT. The effects of FMT on gut microbiota diversity and microbiota structure of IBS patients were analyzed respectively from the level of phylum, family and genus, and linear discriminant analysis effect size (LEfSe) was further used to screen the different bacteria. Paired t test and paired rank sum test were used for statistical analysis. Results:Twelve weeks after FMT, the scores of the six dimensions of IBS-QOL including dysthymia, behavioral disorder, auto imagery, health concerns, eating avoidance, and relationship expansion were all lower than those before FMT (43.750, 22.656 to 56.250 vs. 48.438, 32.031 to 60.938; 37.500, 18.750 to 56.250 vs. 46.429, 21.429 to 62.500; 31.250, 14.063 to 42.188 vs. 31.250, 18.750 to 50.000; 41.667, 27.083 to 56.250 vs. 50.000, 41.667 to 66.667; 54.167, 43.750 to 72.917 vs. 66.667, 58.333 to 83.333; 8.333, 0.000 to 33.333 vs. 16.667, 8.333 to 33.333, respectively), and the differences were statistically significant ( Z=-2.157, -3.429, -2.274, -3.197, -3.042 and -2.329, all P<0.05). Twelve weeks after FMT, the scores of the two dimensions of IBS-QOL including behavioral disorder and relationship expansion were both lower than those of four weeks after FMT (37.500, 18.750 to 56.250 vs. 39.286, 19.643 to 62.500 and 8.333, 0.000 to 33.333 vs. 16.670, 2.083 to 41.667, respectively), and the differences were statistically significant ( Z=-1.998 and -2.110, both P<0.05). Four and 12 weeks after FMT, the scores of IBS-SSS and GSRS were both lower than those before FMT ((190.32±106.51), (201.43±102.48) vs. (245.93±86.10) and 5.50, 4.00 to 9.00 and 5.50, 4.00 to 8.75 vs. 7.00, 6.00 to 9.75), and the differences were statistically significant ( t=4.402 and 3.848, Z=-3.081 and -3.609; all P<0.01). No serious adverse reactions occurred in the patients after FMT. At the phylum level, after FMT the abundance of Verrucomicrobia in the feces of IBS patients was richer than that before FMT (6.74% vs. 0.37%); at the family level, after FMT the abundance of Verrucomicrobiaceae in the feces of IBS patients was richer than that before FMT (6.74% vs. 0.37%); at the genus level, after FMT the abundance of Akkermansia was richer than that before FMT (6.74% vs. 0.37%); and the differences were statistically significant (all Z=-2.589, all P=0.010). The results of LEfSe method indicated that four weeks after FMT the abundance of Akkermansia in the gut microbiota of IBS patients was richer than that before FMT (6.74% vs. 0.37%), and the difference was statistically significant (linear discriminant analysis value=4.5, P=0.049). Conclusions:FMT is safe and effective in the treatment of IBS. The mechanism may be through upregulating the diversity of gut microbiota and changing the structure of gut microbiota of IBS patients.

Correlation between the severity of peptic ulcer bleeding and Helicobacter pylori type / 中华消化杂志

Objective@#To investigate the correlation between the severity of peptic ulcer bleeding (PUB) and the serum antibody typing of Helicobacter pylori (H.pylori).@*Methods@#From January 1, 2009 to December 31, 2018, at Guangzhou First People′s Hospital, 1 444 patients diagnosed with PUB and received H. pylori serum antibody test at the same time were enrolled and divided into high-risk group (324 cases) and low-risk group (1 120 cases) according to Forrest classification, and according to recurrent bleeding, the patients were divided into recurrent bleeding group (32 cases) and non-rebleeding group (1 412 cases). Serum H. pylori specific antibodies cytotoxin-associated gene A (CagA), vacuolating cytotoxin A (VacA) and urease were detected by protein array. The correlation between H. pylori positive rate, H. pylori type, PUB and rebleeding were analyzed. Chi-square test and logistic regression analysis were used for statistical analysis.@*Results@#Among 1 444 PUB patients, there were 709 patients with gastric ulcer bleeding (GUB) and 735 patients with duodenal ulcer bleeding (DUB). Previous history of peptic ulcer disease (odds ratio (OR)=1.501, P=0.006), the maximum diameter of ulcer over 2 cm (OR=2.484, P<0.01) and H. pylori infection (OR=1.508, P=0.005) were independent risk factors of the severity of PUB. The total H. pylori positive rate was 68.49% (989/1 444), H. pylori type Ⅰ was the main type. Of which, 61.34% (549/895) were CagA and VacA double positive strains, 31.73% (284/895) were VacA single positive bacteria and CagA single positive bacteria was only 6.93% (62/895). The positive rate of H. pylori of high-risk group was higher than that of low-risk group (75.31%, 244/324 vs. 66.52%, 745/1 120), and the difference was statistically significant (χ2=8.999, P=0.004). In addition, the more serious Forrest classification, the higher the detection rate of H. pylori, and the difference was statistically significant (χ2=11.840, P=0.037). There was no significant difference in the positive rate of H. pylori between recurrent bleeding group and non-rebleeding group (81.25%, 26/32 vs. 68.20%, 963/1 412; χ2=2.469, P>0.05). According to H. pylori antibody type, H. pylori type Ⅰ infection was mainly in both high-risk group and low-risk group. The positive rate of H. pylori type Ⅰ strain of high-risk group was higher than that of low-risk group (67.28%, 218/324 vs. 60.45%, 677/1 120), and the difference was statistically significant (χ2=4.986, P=0.026). There was no statistically significant difference in the positive rate of H. pylori between GUB group and DUB group (68.41%, 485/709 vs. 68.57%, 504/735; χ2=0.005, P>0.05).@*Conclusions@#The infection of H. pylori is positively correlated with the severity of PUB, but not correlated with early ulcer rebleeding. H. pylori type Ⅰ is the main pathogenic strain of GUB and DUB, and CagA and VacA double positive strain is the most common strain.

Correlation between the severity of peptic ulcer bleeding and Helicobacter pylori type / 中华消化杂志

Objective To investigate the correlation between the severity of peptic ulcer bleeding (PUB) and the serum antibody typing of Helicobacter pylori (H.pylori).Methods From January 1, 2009 to December 31, 2018, at Guangzhou First People's Hospital, 1 444 patients diagnosed with PUB and received H.pylori serum antibody test at the same time were enrolled and divided into high-risk group (324 cases) and low-risk group ( 1 120 cases ) according to Forrest classification , and according to recurrent bleeding , the patients were divided into recurrent bleeding group (32 cases) and non-rebleeding group (1 412 cases).Serum H.pylori specific antibodies cytotoxin-associated gene A (CagA), vacuolating cytotoxin A (VacA) and urease were detected by protein array .The correlation between H.pylori positive rate, H.pylori type, PUB and rebleeding were analyzed .Chi-square test and logistic regression analysis were used for statistical analysis . Results Among 1 444 PUB patients, there were 709 patients with gastric ulcer bleeding ( GUB) and 735 patients with duodenal ulcer bleeding ( DUB).Previous history of peptic ulcer disease ( odds ratio (OR)＝ 1.501, P＝0.006), the maximum diameter of ulcer over 2 cm (OR＝2.484, P?0.01) and H.pylori infection (OR＝1.508, P＝0.005) were independent risk factors of the severity of PUB .The total H.pylori positive rate was 68.49%(989/1 444), H.pylori type Ⅰwas the main type.Of which, 61.34%(549/895) were CagA and VacA double positive strains , 31.73%(284/895) were VacA single positive bacteria and CagA single positive bacteria was only 6.93%(62/895).The positive rate of H.pylori of high-risk group was higher than that of low-risk group (75.31%, 244/324 vs.66.52%, 745/1 120), and the difference was statistically significant (χ2 ＝8.999, P ＝0.004).In addition, the more serious Forrest classification , the higher the detection rate of H.pylori, and the difference was statistically significant (χ2 ＝11.840, P＝0.037).There was no significant difference in the positive rate of H.pylori between recurrent bleeding group and non-rebleeding group (81.25%, 26/32 vs.68.20%, 963/1 412; χ2 ＝2.469, P＞0.05).According to H.pylori antibody type, H.pylori typeⅠinfection was mainly in both high-risk group and low-risk group.The positive rate of H.pylori typeⅠstrain of high-risk group was higher than that of low-risk group (67.28%, 218/324 vs.60.45%, 677/1 120), and the difference was statistically significant ( χ2 ＝4.986, P ＝0.026).There was no statistically significant difference in the positive rate of H.pylori between GUB group and DUB group (68.41%, 485/709 vs. 68.57%, 504/735; χ2 ＝0.005, P＞0.05).Conclusions The infection of H.pylori is positively correlated with the severity of PUB, but not correlated with early ulcer rebleeding .H.pylori typeⅠis the main pathogenic strain of GUB and DUB, and CagA and VacA double positive strain is the most common strain .

Value of FibroScan combined with gamma-glutamyl transpeptidase-to-platelet ratio in predicting liver fibrosis in patients with chronic hepatitis B: A preliminary study / 临床肝胆病杂志

ObjectiveTo investigate the value of FibroScan combined with gamma-glutamyl transpeptidase-to-platelet ratio (GPR) in predicting liver fibrosis stage in patients with chronic hepatitis B (CHB). MethodsA total of 278 patients who were diagnosed with CHB by liver biopsy in Guangzhou Eighth People’s Hospital from January 2012 to December 2016 were enrolled. The value of GPR and FibroScan used alone or in combination in predicting liver fibrosis stage (F0-F4) was analyzed. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and the Mann-Whitney U test was used for further comparison between two groups. The Spearman’s correlation coefficient was used for correlation analysis. The area under the receiver operating characteristic curve (AUC) was used to evaluate liver fibrosis stage. ResultsWith liver biopsy as the gold standard, of all patients, 50 had stage F1 fibrosis, 104 had stage F2 fibrosis, 92 had stage F3 fibrosis, and 32 had stage F4 fibrosis. Liver stiffness measurement by FibroScan gradually increased with the increase in liver fibrosis stage (P＜0.05), and GPR also increased gradually in patients with stage F1, F2, and F3 liver fibrosis (P＜0.05). GPR and FibroScan were positively correlated with liver fibrosis (r=0.419 and 0.481, both P＜0.001), and GPR was positively correlated with FibroScan (r=0.436, P＜0.001). According to AUC, FibroScan combined with GPR had a better diagnostic efficiency than FibroScan (0.793 vs 0.739, Z=3.044, P=0.002) or GPR (0.793 vs 0.740, Z=2.389, P=0.037) alone in predicting progressive liver fibrosis (≥F3); FibroScan combined with GPR had a better diagnostic efficiency than GPR alone (0.782 vs 0714, Z=2.130, P=0.033) in predicting marked liver fibrosis (≥F2). ConclusionFibroScan combined with GPR has a certain advantage in predicting progressive liver fibrosis (≥F3) in CHB patients and can improve diagnostic efficiency.

Analysis of clinicopathological characteristics and changing trend of 77 978 cases of colorectal polyps and colorectal cancer detected by colonoscopy in the past 27 years / 中华消化杂志

Objective To explore the detection rate of colorectal polyps and colorectal cancer under colonoscopy,and their clinicopathological characteristics and changing trend.Methods From January 1,1991 to December 31,2017,at Guangzhou First People's Hospital,the colorectal polyps and colorectal cancer diagnosed by colonoscopy were collected.The gender,age,location of colorectal polyps and colorectal cancer and pathological type were retrospectively analyzed.Chi-square test was performed for statistical analysis.Results Among 77 978 patients underwent routine colonoscope examination,male and female accounted for 50.18％ (39 132/77 978) and 49.82％ (38 846/77 978),respectively;and the age was (52.8 ± 15.7)years.A total of 23 240 patients with colorectal polyps were detected,and the total detection rate was 29.80％.The detection rate of colorectal polyps of male patients was higher than that of female patients (35.11％,13 741/39 132 vs.24.45％,9 499/38 846),and the difference was statistically significant (x2 =1 059.16,P ＜0.01),the age was (59.2 ± 14.6) years.The most common location of polyps was left colon,accounted for 48.56％ (11 286/23 240).The pathological type was adenoma (60.99％,13 726/22 504).Totally 5 204 patients with colorectal cancer were detected,the detection rate of 6.67％;the detection rate of male was higher than that of female (7.71％,3 018/39 132 vs.5.63％,2 186/38 846),and the difference was statistically significant (x2 =136.07,P ＜0.01),the age was (63.5 ± 12.5) years.The colorectal cancer was predominantly located in the left colon (73.71％,3 836/5 204).And the common pathological type was moderately differentiated adenocarcinoma (83.61％,4 351/5 204).Based on a nine-year as a time period,the detection rate of eolorectal polyps was 12.91％ (599/4 639) from 1991 to 1999,22.74％ (3 784/16 642) from 2000 to 2008,and 33.26％ (18 857/56 697) from 2009 to 2017,showing a significant upward trend (x2 =1 353.45,P ＜0.01).The detection rate of colorectal cancer was 9.03％ (419/4 639) from 1991 to 1999,8.59％ (1 430/ 16 642) from 2000 to 2008,and 5.92％ (3 355/56 697) from 2009 to 2017,showing a significant downward trend (x2 =191.89,P ＜ 0.01).There was no significant change in pathological types of colorectal polyps in 27 years (x2 =201.08,P ＞ 0.05);while moderately differentiated adenocarcinoma increased,however,the highly and poorly differentiated type decreased (x2 =249.27,P ＜ 0.01).No significant changes in lesion location was found (x2 =117.11 and 134.82,both P ＞ 0.05).Conclusions The detection rate of colorectal polyps increased in the past 27 years,while colorectal cancer decreased.The elderly men are susceptible populations.The most common location of colorectal polyps and cancer are left colon,which should be carefully examined during colonoscope examination.

Microbiota transplantation: concept, methodology and strategy for its modernization

Fecal microbiota transplantation (FMT) has become a research focus of biomedicine and clinical medicine in recent years. The clinical response from FMT for different diseases provided evidence for microbiota-host interactions associated with various disorders, including Clostridium difficile infection, inflammatory bowel disease, diabetes mellitus, cancer, liver cirrhosis, gut-brain disease and others. To discuss the experiences of using microbes to treat human diseases from ancient China to current era should be important in moving standardized FMT forward and achieving a better future. Here, we review the changing concept of microbiota transplantation from FMT to selective microbiota transplantation, methodology development of FMT and step-up FMT strategy based on literature and state experts' perspectives.

Clinical characteristics of clostridium difficile-associated diarrhea among patients in a tertiary care center in China

Objective: This study investigated the incidence, risk factors, and clinical characteristics of Clostridium difficile-associated diarrhea [CDAD] in Chinese patients

Methods: Fecal specimens of patients with antibiotic-associated diarrhea [AAD] were collected to test C. difficile toxin A and B using enzyme-linked fluorescent assay to identify CDAD. By adopting a nested case-control design, the matched people [ratio 1:3] without AAD were included as controls

Results: Out of 56,172 inpatients, 39,882 [71.0%] used antibiotics, 470 suffered from AAD, and 93 were diagnosed with CDAD. The incidence of nosocomial CDAD was 166 per 100,000. The proportion of CDAD in AAD was 19.8%. CDAD patients presented with more severe clinical manifestations and exhibited more concurrent illness. Logistic regression analysis showed the risk factors of CDAD: advanced age, nasogastric tube-feeding, high APACHE II scores, high level of serum C-reaction protein, low level of serum albumin, severe underlining disease or comorbidity, and number of antibiotic intake. Twenty-nine patients [31.2%] were cured with vancomycin, 54 [58.1%] were cured after dual therapy of vancomycin plus metronidazole, 7 [7.5%] died of underlying diseases aggravated with CDAD, and 3 [3.2%] were transferred to other hospitals for personal reasons

Conclusion: The incidence of nosocomial CDAD in China was high. Some risk factors could predispose CDAD

Efficacy of Levofloxacin-based Triple Therapy Combined with Probiotics as A Rescue Therapy for Helicobacter pylori Re-eradication / 胃肠病学

Background:With the emergence of bacterial resistance,the efficacy of Helicobacter pylori(Hp)eradication therapy is decreasing in recent years. After the previous failed course of eradication therapy,the possibility of failure of re-eradication therapy is greater. Therefore,choosing the rescue therapy for Hp re-eradication is particularly important. Aims:To investigate the efficacy of levofloxacin-based triple therapy combined with probiotics as a rescue therapy for Hp re-eradication. Methods:A total of 192 patients with a previous failed course of standard triple therapy for Hp eradication were enrolled and randomly assigned into four groups. Group A:lansoprazole 30 mg + amoxicillin 1 000 mg + levofloxacin 200 mg bid for 14 days;group B:regimen of group A with the addition of bismuth potassium citrate 600 mg bid for 14 days;group C:bifidobacterium and lactobacillus triple live bacteria 2 000 mg tid for 14 days followed by regimen of group A for 14 days;group D:regimen of group A with the addition of bifidobacterium and lactobacillus triple live bacteria 2 000 mg tid for 14 days. At 4 weeks after end of treatment,Hp eradication was assessed by 13 C-urea breath test. Adverse effects during the course of treatment were recorded. Results:A total of 177(92. 2% )patients completed the study,the completion rates in group A,B,C and D were 87. 5%(42 / 48),83. 3%(40 / 48),97. 9%(47 / 48)and 100% , respectively;the completion rates in group C and D were significantly higher than those in group A and B(P 0. 05). Adverse effects included constipation,taste distortion,bloating and anorexia,the incidences of adverse effects in group A,B,C and D were 79. 2% ,95. 8% ,29. 2% and 22. 9% ,respectively;the incidences in group C and D were significantly lower than those in group A and B(P < 0. 05). Conclusions:Levofloxacin-based triple therapy combined with bifidobacterium and lactobacillus triple live bacteria can decrease the adverse effects of traditional triple and quadruple therapies and improve the patient compliance,thus increases the efficacy of rescue therapy for Hp re-eradication.

The expression of Wnt/beta-catenin signaling molecule in inflammatory bowel diseases treated with bone marrow mesenchymal stem cell transplantation / 中国组织工程研究

BACKGROUND:The Wnt/β-catenin signaling pathway is one of the most important signaling pathways in stem cel regulation, which is involved in regulation of cel proliferation and differentiation. OBJECTIVE:To investigate the expression of Wnt/β-catenin main signaling molecule in inflammatory bowel tissues treated with bone marrow mesenchymal stem cel transplantation. METHODS:2,4,6-Trinitrobenzene sulfonic acid was used for establishing inflammatory bowel diseases rat models. Bone marrow mesenchymal stem cels labeled with green fluorescent protein were transplanted into rat modelsviatail vein. Normal saline was injected as control. The expression of Wnt/β-catenin signaling molecule was detected in the large intestine tissue of inflammatory bowel disease rat models by quantitative RT-PCR at 14 and 28 days after transplantation. RESULTS AND CONCLUSION:Real-time quantitative PCR results showed that the expression of Wnt3a andβ-catenin in the inflammatory bowel tissue increased significantly (P 0.05). The expressions of Wnt3a, β-catenin and c-myc in the transplantation group were significantly lower than those in the control group after transplantation (P <0.05). These findings indicate that the Wnt/β-catenin signaling pathway plays important roles in inflammatory bowel disease and repair after bone marrow mesenchymal stem cel transplantation, while this pathway may promote stem cels differentiating into intestinal epithelium, promote recovery from inflammatory bowel disease, repair inflammatory area, and restore intestinal tissue homeostasis.

Screening main genes during mesenchymal stem cell transplantation in repair of inflammatory bowel tissue / 中国组织工程研究

BACKGROUND:Previous studies have verified that mesenchymal stem cells could be transplanted into inflammatory bowel mucosa to repair inflammatory bowel tissue. OBJECTIVE:To observe the differential gene expression in large intestine before and after mesenchymal stem celltransplantation in repair of inflammatory bowel tissue of rats using microarray technology, and to primarily discover the main genes during mesenchymal stem celltransplantation, differentiation, and reparation in inflammatory colorectal tissue region. METHODS:Healthy Sprague-Dawley rats were randomly divided into two groups. Experimental rat models of inflammatory bowel disease were established using trinitrobenzene sulfonic acid via enema. At 24 hours after model establishment, green fluorescent protein-labeled mesenchymal stem cells were infused via the caudal vein. The control group was treated with physiological saline by enema, instead of trinitrobenzene sulfonic acid. At 28 days, large intestine was obtained from the experimental group and control group. Differential y expressed genes were screened in the experimental and control groups using microarray technique. RESULTS AND CONCLUSION:The microarray analysis results showed that there were 388 differential genes in the control and experimental groups (P2), in which 191 were up-expressed, and 197 were down-expressed. Al of these genes were mainly involved in inflammatory reaction, immune reaction and celldifferentiation. In the top 10 up-regulation and down-regulation differential genes (total y 20 genes), 3 genes were involved in inflammation, 3 genes were involved in immune reaction, and 2 genes were related to stem celldifferentiation. In the 388 genes, 33 were related to signaling pathways (P<0.05), 6 related to inflammation, 8 related to immunity, and 5 related to stem celldifferentiation. Results suggested that the main genes involved in mesenchymal stem cells in repair of inflammatory bowel tissue were primarily screened using gene expression microarray technique.

Trends in incidence and clinical feature of hypertriglyceridemic acute pancreatitis: clinical review of 20 years / 中华胰腺病杂志

Objective To retrospectively investigate the clinical features and incidence trends of hypertriglyceridemic acute pancreatitis (HLAP) in Guangzhou First People's Hospital for 20 years.Methods The medical records of 1 362 patients,who were admitted to Guangzhou First People's Hospital with acute pancreatitis during January 1991 to December 2010,were reviewed and 99 patients met the HLAP criterion.These patients were divided into four groups according to years (1991 ～ 1995,1996 ～2000,2001 ～2005,and 2006 ～2010).The incidence and clinical features were compared among the 4 groups.Results Among the 99 patients,there were 61 males and 38 females,the mean age was (44 ± 12) years old,and young male was predominant in HLAP.HLAP accounted for 7.3％ of all AP patients,and the incidence was increased 1.5 times during the 20 years period (5.4％,5.7％,6.7％,and 8.3％ respectively).The serum amylase level was (513.3 ±462.7) mmol/L,TG level was (12.7 ± 7.0) mmol/L,Ranson score was (1.2 ± 1.1),CTSI score was 2.2 ± 1.1,incidence of SIRS was 36.4％,and the incidence of MODS was 18.2％,and the incidence of pancreatic pseudocyst was 5.1％,but the incidence and mortality of SAP was not changed with time.The incidence and mortality of SAP of HLAP were significantly higher than those of acute biliary pancreatitis (20.2％ vs 8.5％,6.1％ vs 2.8％).The rate of blood purification increased from 0 to 10.7％,and the hospital stay was decreased from 26 d to 14 d,and the difference between the two groups was statistically significant (P ＜ 0.05).Conclusions Incidence of HLAP is increased gradually,with a trend towards more severe disease,but the prognosis is improved.

Analysis of clinical pathological characteristics and pattern changes of 2643 gastric polyps in the past 15 years / 中华消化杂志

Objective To investigate the clinical pathological characteristics and pattern changes of gastric polyps in the past 15 years.Methods From 1998 to 2012,the cases of gastric polyps diagnosed by gastroscopy were collected.Data of gender,age,location,number,size,Yamata type,pathological type,Helicobacter pylori (H.pylori) infection were retrospectively analyzed.The data were analyzed by Chi square test.Results Among 62 975 cases received gastroscopy,gastric polyps were detected in 2 643 patients and the gastric polyps detection rate was 4.20％,which of female (5.91％,1 790/30 269) was higher than that of male (2.61％,853/32 706).And patients aged ≥60 years old has a high prevalence of gastric polyps; the detection rate was 6.39％(1 124/17 590).The positive rate of H.pylori infection in patients detected with gastric polyp was 12.70％ (321/2 527).Gastric body (40.41％,1 068/2 643) and fundus (26.79％,708/2 643) were two predilection sites of gastric polyps.About 69.05％(1 825/2 643) patients' polyps were solitary,74.76％(1 976/2 643) patients' polyps were less than 0.5 cm in diameter and mainly were Yamata type Ⅰ (30.68％,811/2 643) or type Ⅱ (54.41％,1 438/2 643).The common pathological types were hyperplastic polyps (43.80％,1 109/2 532),inflammatory polyps (34.24％,867/2 532),fundic-gland polyps (19.55％,495/2 532) and adenomatous polyps (2.41％,61/2 532).The detection rate of polyps gradually increased in the past 15 years.Between 1998 to 2002,2003 to 2007,2008 to 2012,the average of 5-year detection rates were 1.11％,2.50％ and 6.49％,respectively.However,the proportion of pathological types didn't change significantly.Conclusions The detection rate of gastric polyps gradually increased year by year,and elderly woman is at high risk.Hyperplastic and inflammatory polyps ware the most common types.

Lentivirus-mediated over-expression of beta-catenin accelerates proliferation and migration of mesenchymal stem cells / 中国组织工程研究

BACKGROUND:β-catenin is the most critical signaling molecule in the Wnt/β-catenin signaling pathway, which is involved in the regulation of cellproliferation, differentiation and tissue self-healing balance. OBJECTIVE:To construct a stableβ-catenin over-expression lentivirus-mediated vector and to transfect mesenchymal stem cells line for investigating its effects on proliferation and migration of mesenchymal stem cells. METHODS:Over-expression vector, PLV-EF1A-catenin-RFP, was constructed and transfected the 293T cellto infect mesenchymal stem cells, and positive cells were selected with puromycin. The up-regulated efficiency of targetingβ-catenin gene at mRNA level was detected by real-time quantitative PCR, the effect on proliferation of mesenchmal stem cellwas assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve, and the migration ability was detected by Transwel motility assay. RESULTS AND CONCLUSION:The lentiviral vector targetingβ-catenin gene was constructed successful y, and a stable mesenchymal stem cellline that up-regulatedβ-catenin was established. Real-time quantitative PCR results showed that the expression ofβ-catenin gene was efficiently up-regulated by infecting PLV-EF1A-catenin-RFP (P<0.05). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve showed that celldoubling time was shortened after infected with pLV-EF1A-catenin-RFP (P<0.05), indicating that the over-expression of theβ-catenin gene successful y increased the proliferative capability of mesenchymal stem cells. The Transwel assay also showed similar increasing results on the migration ability (P<0.01). The lenvivirus-mediated over-expression of theβ-catenin gene can be used to increase the proliferation and migration abilities of the mesenchymal stem cells.

Infliximab on intestinal barrier injure in rats with acute necrotizing pancreatits / 中华胰腺病杂志

Objective To evaluate the effects of infliximab (TNF-α monoclonal antibody ) on intestinal barrier injury in ANP complicated with MODS in a rat model.MethodsThirty SD rats were randomly divided into sham operation group (SO),ANP group and infliximab treatment group.Sodium taurocholate (4.5％) was injected into the pancreatic duct to induce ANP complicated with MODS model.Infliximab (8 mg/kg) was injected via tail vein in 6h after modeling in infliximab group.Same amount of 0.9％ NS was injected into the pancreatic duct in SO group.After 24 h of modeling,all rats were sacrificed,intestine and pancreas samples were collected for pathologic examination.The blood samples were harvested.The serum levels of amylase,TNF-α,diamine oxidase( DAO),D-lactate,and the rate of carbon propelling in ileum were measured.ResultsThe serum levels of amylase were ( 1125 ± 331 ),( 11024 ± 2203 ),( 545 ±30) U/L in SO group,ANP group and infliximab group; the serum levels of TNF-α were (12.1 ± 4.0),(107.6 ± 18.5),(75.8 ±5.9) U/L; the pathological scores of pancreas were 2.25 ±0.38,14.10 ±0.22,3.93 ± 0.67,the difference among the 3 groups was statistically significant ( P ＜ 0.05 ).The pathological scores of intestine were 2.29 ± 0.32,6.61 ± 0.58,3.91 ± 0.41 ; the DAO levels were ( 87.88 ± 34.51 ),(146.30 ±12.99),(115.00 ± 18.58) ng/ml; the D-lactate levels were (1.50 ±0.49),(2.32 ± 0.35),(2.02 ± 0.25 )mmol/L; and the rates of carbon propelling in ileum were (0.64 ± 0.04 )％,(0.28 ±0.08)％,(0.52 ±0.09)％,the difference among the 3 groups was statistically significant (P ＜0.05).ConclusionsInfliximab can effectively prevent dysfunction of intestinal barrier and improve motility in ANP rats.

The relationship between colorectal neoplasia and nonalcoholic fatty liver disease / 中华消化内镜杂志

ObjectiveTo investigate the relationship between colorectal neoplasia and nonalcoholic fatty liver disease (NAFLD).MethodsData of 809 patients who had undergone colonoscopy and epigastric CT scan in the same period were reviewed for gender,age,history of diabetes,hypertension,hyperlipidemia,NAFLD,colonoscopy and pathology results.Correlation of colorectal adenoma (CRA),colorectal carcinoma (CRC) and NAFLD were studied with multivariate Logistic analysis.Results NAFLD (P ＜0.0001,OR 2.06,95％ CI 1.53-2.76) was the independent risk factor for CRA,where the incidence of CRA was higher than that in the control group.ConclusionPatients with NAFLD is the risk population of colorectal adenomas.

The relation between interleukin-10 promoter-1082 and -819 sites gene polymorphism with irritable bowel syndrome / 中华消化杂志

Objective To explore the relation between IL-10 promoter region gene polymorphism and irritable bowel syndrome (IBS).Methods By polymerase chain reaction combined with restrition fragment length polymophism (PCR-RFLP),gene type of IL-10 promoter -1082 and -819 sites in 313 IBS patients and 281 controls was analyzed.Results The distribution of IL-10-1082 and-819 allele frequencies in IBS group,control group and total was in accordance with Hardy-Weinberg equilibrium law.The frequency of IL-10-819 T allele in diarrhea subtype (79.8％) and mixed IBS subgroup (77.1％) was significantly higher than that in control group (65.7％).There were no significant differences in IL-10-1082 A/G allele frequency between each subtypes and control group (P＞0.05),however there was statistically difference between diarrhea subtype and mixed IBS subgroup (P＜0.05).The frequency of-819 T/T genotype in IBS group (51.1 ％ )was significantly higher than that of control group (40.2％),the frequency of C/T genotype was significantly lower than that of control group,and the difference was statistically significant (all P＜0.05).The IL-10-819 T/T allele frequency of all IBS subtypes was significantly higher than that of control group; however C/T allele genotype frequency of all IBS subtypes was significantly lower than that of control group,and the difference was statistically significant (all P ＜ 0.05). There was no significant difference of C/C allele genotype between subtypes (P＜0.05).There was no significant difference of -1082 allele genotype between IBS group and control group (P＞0.05).The frequency of -1082 A/A genotype in diarrhea subtype of IBS patients (93.3％) was significantly higher than that of mixed IBS subtype (82.4％),while the frequency of A/G genotype was lower than that of mixed IBS subtype,and the difference was statistically significant (all P ＜ 0.05 ); there was no significant difference between other IBS subtypes and control group (P＞0.05).Conclusion IL-10-819 promoter T/Tgenotype may be related to IBS pathogenesis.

Epidemiology of nonalcoholic fatty liver disease: a follow-up study for 4 years / 中华消化杂志

Objective To investigate the prevalence,incidence,death causes and natural course of nonalcoholic fatty liver disease (NAFLD) as well as the risk factors of NAFLD development in population of Guangdong province.Methods On the basis of previous epidemiological survey in 2005,3545 subjects were followed up (medium 4 years) for death causes.Of which,624 subjects (re-survey group) received interview,physical examinations,biochemical tests and ultrasonography again.The natural course and the risk factors of the NAFLD were analyzed.Results Of 3543 subjects,219subjects were failed to followed up and 4 out of 467 NAFLD patients (8.6‰) died of cardiovascular and cerebrovascular diseases during follow-up period.In rest 2857 subjects,22 (7.7‰) died of cardiovascular/cerebrovascular diseases and cancers.However,there were 117 NAFLD patients in resurvey group at the baseline.The NAFLD remained in 51 (43.6%) patients,aggravated in 26 (22.2%),alleviated in 21 (18.0%) and regressed in 19 (16.2%) at the endpoint.Meanwhile 185(36.5 %) out of 507 normal subjects developed NAFLD at the endpoint.The standard prevalence of NAFLD increased from 10.5% (2005) to 22.6% (2009,P<0.05).The standard incidence was 4.9% per year with male (5.1%) being higher than female (4.7%,P<0.05).The cumulative incidence was significantly higher in urban cities (43.1%) than in rural areas (27.4%,P<0.05).Cox proportional hazards regression showed that the age (OR= 1.893),waist circumference (OR=2.512),waist-to-hip ratio (WHR,OR= 1.400),body mass index (BMI,OR=2.581) and serum cholesterol levels (OR = 1.587) were associated with the development of NAFLD (P < 0.05).Multivariate regression logistic analysis revealed that age and BMI were associated factors for NAFLD.Decreased waist circumference was prone to remission.Conclusions The higher prevalence and incidence of NAFLD were noted in population of Guangdong province.The NAFLD patients mainly died of cardiovascular/cerebrovascular diseases,but not liver disease.Obesity,aging and dyslipidemia are high risk factors for NAFLD development.

A nested case-control study on genetic polymorphisms for patients with non-alcoholic fatty liver disease / 中华消化杂志

Objective To investigate the relationship between the genetic polymorphisms, which played roles in the pathogenesis of metabolic syndrome (MS), and susceptibility of non-alcoholic fatty liver disease (NAFLD) in Han people in Guangdong province. Methods The subjects were selected from an epidemiologie survey in Guangdong province. Fifty to 117 adult NAFLD patients, who met the criteria of Chinese guideline for diagnosis of NAFLD and had typically clinical, biochemical signs and abdominal ultrasonography, were recruited in the study. By using 1 : 1 matched method of nested case-control study, same numbers of people without NAFLD were included as controls. The genetic analyses was performed by using genomic DNA extracted from peripheral blood. Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) was applied to detect the single nucleotide polymorphisms (SNPs) at 9 sites in 7 candidate genes. Results Most SNPs of the genes were related to the susceptibility of NAFLD. Some of them had positive relation (increasing the risk) such as tumor necrosis factor (TNF)-α-238, adiponectin-45, leptin-2548, peroxisome proliferator-activated receptors (PPAR) γ-161 and phosphatidylethanolamine N-methyltransferase (PEMT)-175. Some had negative relation (decreasing the risk) including adiponectin-276 and hepatic lipase-514. And some had no relation (TNF-α-380 and PPAR g coactivator-1α-482). Conclusion Most cytokines' SNPs of candidate genes discovered in MS patients are related to the susceptibility of NAFLD.

Endoscopic pyloromyotomy in congenital hypertrophic pyioric stenosis / 中华消化内镜杂志

Objective To evaluate the short-term effect and safety of endoscopic pyloromyotomy for congenital hypertrophic pyioric stenosis(CHPS).Methods Twelve infants,including 8 boys and 4 girls,mean age(57.5±14.4)days,who were diagnosed as CHPS,underwent endoscopic pyloromyotomy by using an ultra-thin gastroscope.Incisions were made from the duodenal to the antral side of the pylorus by an endoscopic electrosurgical sphincterome or a needle knife.Results These 12 patients underwent 13 endoscopic operations.Vomiting ceased in 1-10 days in 11 cases,of which one patient's vomiting was recurrent 35 days after the first procedure,which was improved 10 days later after another endoscopic operations.Another one did not stop vomiting because of cytomegalovirus hepatitis,whose symptoms were relieved after secondary treatment.No complications were observed except for minor melena in one case after the therapy.Conclusion Endoscopicall pyloromyotomy in CHPS is effective and safe in short-term follow-up.