1.Changes of Intestinal Mucosal Barrier and Intestinal Flora in Rats with Severe Acute Pancreatitis.
Yan LI ; Hao WU ; Yiyun DENG ; Ruyi LIAO ; Lili XI ; Ping YAO
Journal of Biomedical Engineering 2015;32(2):412-417
This paper is to explore changes of intestinal mucosal barrier, intestinal flora, and bacterial translocation in rats with severe acute pancreatitis (SAP). Twenty four male SD rats were randomly divided into the control group (n = 10) and the experimental group (n = 14). The model of severe acute pancreatitis of rats was induced by the method of injecting adversely 5% sodium taurocholate into the common biliary-pancreatic duct. All of the rats were killed after 24 hours and the level of the serum amylase and the plasma endotoxin was determined after that. The pathological changes of pancreas and small intestine were observed through hematoxylin-eosin staining (HE staining) and the abdominal viscera bacterial translocation rates were tested. With the method of real-time polymerase chain reaction (RT-PCR) the quantity of the intestinal flora was analyzed. In the control group, the level of Escherichia coli, Lactobacillus and Bifidobacterium were 2.08 ± 1.29, 11.04 ± 7.55 and 12.21 ± 4.95, respectively. On the contrast, the level of Escherichia coli in the cecum contents was much higher (9.72 ± 3.58, P < 0.01), while the Lactobacillus number was decreased significantly (0.67 ± 0.34, P < 0.01), and the Bifidobacterium number was also decreased (4.59 ± 3.42, P < 0.05) in the experimental group, so the ratio of Bifidobacterium/Escherichia coli was reversed. Besides, in the experimental group, the plasma endotoxin positive rates and the bacterial translocation rates were much higher (P < 0.01 or P < 0.05) and the pathology scores of pancreas and small intestines were also significantly higher (P < 0.01) than those in the control group. These results indicated that in severe acute pancreatitis rats, the intestinal mucosal barrier was severely damaged and the dysbacteriosis occurs in the intestinal canal. And these might relate to the occurrence and development of multiple organ infection.
Animals
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Bacterial Translocation
;
Endotoxins
;
Intestinal Mucosa
;
pathology
;
Intestines
;
microbiology
;
Male
;
Pancreas
;
pathology
;
Pancreatitis
;
microbiology
;
pathology
;
Rats
;
Rats, Sprague-Dawley
2.Changes in the structure of intestinal mucosal flora in colorectal cancer patients.
Mei Mei HU ; Kai Yang CHEN ; Ning Yu WANG ; Yu Fan ZHAO ; Cheng Jin WEI ; Ling Xiang MENG ; Yong TANG ; Yu Ou TENG ; Hai Kuan WANG
Journal of Southern Medical University 2022;42(2):263-271
OBJECTIVE:
To investigate the changes in bacterial flora in fecal samples, at the tumor loci and in adjacent mucosa in patients with colorectal cancer (CRC).
METHODS:
We collected fecal samples from 13 patients with CRC and 20 healthy individuals and tumor and adjacent mucosa samples from 6 CRC patients. The differences in bacterial composition between the fecal and mucosa samples were analyzed with 16S rDNA sequencing and bioinformatics methods. We also detected the total number of bacteria in the feces using flow cytometry, isolated and identified the microorganisms in the fecal and mucosa samples using common bacterial culture media. We further tested the effects of 7 isolated bacterial strains on apoptosis of 3 CRC cell lines using lactate dehydrogenase detection kit.
RESULTS:
The bacterial α-diversity in the feces of healthy individuals and in adjacent mucosa of CRC patients was significantly higher than that in the feces and tumor mucosa in CRC patients (P < 0.05). Lactobacillaceae is a specific bacteria in the feces, while Escherichia, Enterococcus, and Fusobacterium are specific bacteria in tumor mucosa of CRC patients as compared with healthy individuals. Cell experiment with3 CRC cell lines showed that Bacteroides fragilis isolated from the tumor mucosa of CRC patients produced significant inhibitory effects on cell proliferation (P < 0.0001), while the isolated strain Fusobacterium nucleatum obviously promoted the proliferation of the cell lines (P < 0.001).
CONCLUSION
The bacterial flora in the feces, tumor mucosa and adjacent mucosa of CRC patients is significantly different from that in the feces of healthy individuals, and the fecal flora of CRC patients can not represent the specific flora of the tumor mucosa. Inhibition of F. nucleatum colonization in the tumor mucosa and promoting B. fragilis colonization may prove beneficial for CRC treatment.
Bacteria
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Colorectal Neoplasms/pathology*
;
Feces/microbiology*
;
Gastrointestinal Microbiome
;
Humans
;
Intestinal Mucosa
3.The study on bacteria invading the intestinal mucosa barrier in mice with fulminant hepatic failure.
Hong-Li SONG ; Sa LÜ ; Pei LIU
Chinese Journal of Hepatology 2011;19(3):214-217
OBJECTIVETo explore the mechanism of fulminate hepatic failure (FHF) complicated with spontaneous peritonitis (SBP) through the research of bacteria invading the intestinal mucosa barrier.
METHODS240 BalB/c male mice were divided into four groups as isotonic NS group (n = 40), lipopolysaccharide (LPS) group (n = 40), galactosamine (GalN) group (n = 40) and FHF model group (n = 120). Each mouse received same volume of NS, LPS (10 ug/kg), GalN (800 mg/kg) or LPS (10 ug/kg)/GalN (800 mg/kg) intraperitoneal injection according to its group. 8 mice were executed at 2, 6, 9, 12 and 24 hours after injection, respectively, and the liver and intestinal tissue samples were taken at the same time. ALT was measured by automatic biochemical analyzer and was compared between groups using Mann-Whitney U test. Liver and intestinal tissue received HE staining. The ultrastructure of intestinal mucosa and the method by which bacteria invaded the intestinal mucosa were observed by transmission electron microscopy. All data were analyzed by SPSS13.0 statistic software.
RESULTSALT level, results of hepatic pathology, mortality and clinical manifestations of mice in the FHF model group met the diagnostic criteria of FHF. Intestinal tissue was found with slight edema and little inflammatory cells infiltration through HE staining in all the 4 groups of mice 9 hours after injection. Microvilli were found broken, shed and shorten in the intestinal epithelial cells with incomplete tight junction (TJs) and obviously changed organelles in the FHF model group of mice observed by transmission electron microscope. Mass hemorrhagic necrosis of liver cells with remnant liver cells swelling and many inflammatory cells infiltration by HE staining in the FHF model group. But the changes in hepatic pathology and intestinal mucosa ultrastructure were not so obvious in the mice of NS, LPS and GalN groups. Bacteria penetrated the intestinal wall by pinocytosis 6 to 9 hours after injection in the FHF model group, the microvilli were broken off and TJs turned rupture in the areas that the bacteria penetrated. The bacteria were found in the form of cyst 12 hours after injection.
CONCLUSIONLPS (10 mg/kg)/GalN (800 mg/kg) combined injection was successful in establishing the FHF mice model. The rupture of TJs may provide conditions for intestinal bacteria to penetrate the intestinal mucosa in FHF. Rupture of TJs may be one of the reasons why FHF was complicated with SBP.
Animals ; Disease Models, Animal ; Intestinal Mucosa ; microbiology ; pathology ; Liver ; pathology ; Liver Failure, Acute ; microbiology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Tight Junctions ; microbiology ; pathology
4.Effect of Acupuncture in the Treatment of Young Pigs with Induced Escherichia coli Diarrhea.
Eun Sung PARK ; Seona JO ; Je Kyung SEONG ; Tchi Chou NAM ; Il Suk YANG ; Min Cheol CHOI ; Yeo Sung YOON
Journal of Veterinary Science 2003;4(2):125-128
The effect of acupuncture in the treatment of young pigs with induced enteropathogenic Escherichia coli diarrhea was histopathologically evaluated by routine hematoxylin and eosin stain. Thirty two pigs weighed 4-5kg and aged 21days old were used in this study. The animals with diarrhea were treated with traditional acupuncture, or enrofloxacin. In the group treated with traditional acupuncture, acupoint GV1 (Jiaochao) was used and in the group treated with antibiotics, enrofloxacin was injected intramuscularly. Ten pigs were inoculated with E. coli, but were not treated and served as nontreated control group. At postinoculation day 6, all pigs of the acupuncture and antibiotic treated groups recovered from diarrhea. In the ascending and descending colons of the nontreated control group, severe infiltration of inflammatory cells in the lamina propria was observed and in the fundic stomach, destruction of the fundic gland architecture and necrotic lesions were observed, however, in the same sites of the acupuncture and antibiotics treated groups, the mucosae of the colon and stomach were relatively similar to those of the normal group. These results indicate that acupuncture treatment is effective in controlling induced E. coli diarrhea in pigs at its early stage.
Acupuncture
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Animals
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Colon/cytology/microbiology/pathology
;
Diarrhea/therapy/*veterinary
;
Escherichia coli Infections/therapy/*veterinary
;
Gastric Mucosa/cytology/microbiology/pathology
;
Intestinal Mucosa/cytology/microbiology/pathology
;
Male
;
Stomach/cytology/microbiology/pathology
;
Swine
;
Swine Diseases/*microbiology/therapy
5.An experimental study on the prevention of enteral bacterial translocation in scalded rats by smectite powder.
Hai-tao SU ; Yi-shu LI ; Shu-liang LU ; Man SUN ; Chun QING ; Zong-yu LI ; Tie-bing SHAO ; Li-bing HUANG ; Bing QU ; Xin-bo YANG
Chinese Journal of Burns 2005;21(2):89-92
OBJECTIVETo explore the preventive and treatment effects of smectite powder on enteral bacterial translocation in scalded rats.
METHODSFifty-four Sprague-Dawley (SD) rats were randomly divided into three groups, i.e. normal control (A, n = 6), burn control (B, n = 24), and burn treatment (T, n = 24) groups. The rats in B and T groups were fed with tracing bacteria JM109, which was transfected with PUC19 plasmid in advance. The rats were subjected to 30% TBSA scald injury after the plasmid was shown to have colonized in the intestine. Smectite powder (0.6 g/day/kg) was fed to rats of T group immediately after the scalding, while those in B group received no smectite powder. Bacterial translocation in blood and mesenteric lymph nodes in all groups was observed and identified by enzyme digestion at 12 post scald hour (PSH) and on 1, 3 and 5 post-scald days (PSD). The contents of malondialdehyde (MDA) and superoxide dismutase (SOD) were determined in rat intestinal tissue. And the degree of injury to the entire small intestine was observed pathologically. The villus height of intestinal mucosa was measured, and the rate of epithelial nuclear splitting of mucosal crypts was calculated.
RESULTSThe number of rats with positive blood bacterial culture in B group was obviously higher than that in A and T groups (P < 0.05) on 1 and 5 PSD. The bacterial quantity in mesenteric lymph nodes (MLN) in T group on 1 PSD (38 +/- 16 CFU/g) and 5 PSD (68 +/- 20 CFU/g) were obviously lower than those in B group (228 +/- 67 vs 183 +/- 29 CFU/g, P < 0.05). There was significant difference in the intestinal contents of MDA and SOD between B and T groups at each time point (P < 0.05). The rat jejunum villus height and the epithelial nuclear splitting in the small intestine mucosa in T group were evidently higher than those in B group (P < 0.05 or 0.01).
CONCLUSIONSmectite powder is beneficial to the protection of the intestinal mucosa in scalded rats, and can effectively prevent postburn intestinal bacterial translocation in rats.
Animals ; Bacterial Translocation ; Burns ; drug therapy ; microbiology ; Intestinal Mucosa ; microbiology ; pathology ; Rats ; Rats, Sprague-Dawley ; Silicates ; therapeutic use
6.Helicobacter pylori and Telomerase Activity in Intestinal Metaplasia of the Stomach.
Il Kwun CHUNG ; Kyu Yoon HWANG ; In Ho KIM ; Hong Soo KIM ; Sang Heum PARK ; Moon Ho LEE ; Chang Jin KIM ; Sun Joo KIM
The Korean Journal of Internal Medicine 2002;17(4):227-233
BACKGROUND: Helicobacter pylori (H. pylori) has been considered a definitive carcinogen in gastric cancer. Telomerase is activated in gastric cancer and some premalignant gastric lesions, including intestinal metaplasia (IM). In this study, we evaluated the relationships of both H. pylori infection and telomerase activity with endoscopic and histologic features in IM. The effects of H. pylori eradication on endoscopic, histologic and biochemical changes were evaluated. METHODS: Endoscopic biopsies were obtained from 43 patients with IM for rapid urease, histologic and telomerase tests. The endoscopic and histologic features, H. pylori infection and telomerase were assessed. After H. pylori eradication, 15 patients were re-evaluated and compared after 4 months. RESULTS: Thirty-four (79.1%) patients were infected with H. pylori. The incidence of H. pylori infection was borderline correlated to the severity of IM (p=0.076). Telomerase was elevated in eight (18.6%) patients. Telomerase tends to be high in subtype III and endoscopic grade III of IM. After H. pylori eradication, endoscopic extent (p=0.039) and histologic severity (p=0.074) showed improvements, and telomerase decreased significantly (p=0.0001). CONCLUSION: Our data suggest that telomerase is associated with the severity and extent of IM and that H. pylori eradication improves the endoscopic and histologic features in IM, and decreases telomerase activity. H. pylori eradication can be considered one of the methods to prevent gastric cancer in patients with H. pylori-infected IM. Further long-term and large-scaled study will be needed.
Female
;
Helicobacter Infections/*enzymology
;
*Helicobacter pylori
;
Human
;
Intestinal Mucosa/enzymology/microbiology/*pathology
;
Male
;
Metaplasia/enzymology/microbiology
;
Middle Aged
;
Precancerous Conditions/enzymology/microbiology
;
Stomach Neoplasms/*enzymology/microbiology
;
Telomerase/*metabolism
7.Inflammatory Bowel Diseases and Enteric Microbiota.
The Korean Journal of Gastroenterology 2010;55(1):4-18
Intestinal mucosal layers are colonized by a complex microbiota that provides beneficial effects under normal physiological conditions, but is capable of contributing to chronic inflammatory disease such as inflammatory bowel disease (IBD) in susceptible individuals. Studies have shown that the enteric microbiota may drive the development of the gut immune system and can induce immune homeostasis as well as contribute to the development of IBD although the precise etiology is still unknown. Therefore, intestinal microbes seem to play a key role in the disease pathogenesis. Especially, dysbiosis, which is a shift in the composition of enteric microbiota to a nonphysiologic composition, is associated with one or more defects in mucosal immune functions, including microbe recognition, barrier function, intercellular communication, and anti-microbial effector mechanisms. This review focuses on the impact of enteric microbiota on the development and perpetuation of IBD. In addition, interactions with enteric bacteria and mucosal cells, including intestinal epithelial cells, dendritic cells, and T cells, to induce immune responses at mucosal surfaces have been discussed in the point of IBD pathogenesis. Further extension of the knowledge of enteric microbiota may lead to insights on the pathogenesis and new therapeutic strategies for IBD.
Bacterial Physiological Phenomena
;
Host-Pathogen Interactions
;
Humans
;
Inflammatory Bowel Diseases/*microbiology/pathology
;
Intestinal Mucosa/immunology/microbiology
;
Intestines/microbiology/pathology
;
T-Lymphocytes/immunology/metabolism
8.Ultrastructural study on route of gut bacterial translocation in a rat after spinal cord injury.
Chinese Journal of Applied Physiology 2015;31(6):561-566
OBJECTIVETo observe the ultrastructural change of the route of gut bacterial translocation in a rat with spinal cord injury (SCI).
METHODSForty Wistar rats were divided into the following groups: control group and 3 SCI groups (10 in each group). The rats in the SCI groups were established SCI model at 24 h, 48 h, and 72 h after SCI. Small intestine mucous membrane tissue was identified and assayed by transmission electron microscope, scanning electron microscope and immunofluorescence microscopy.
RESULTSSmall intestine mucous membrane tissue in control group was not damaged significantly, but those in SCI groups were damaged significantly. Proliferation bacteria in gut lumen attached on microvilli. The extracellular bacteria torn the intestinal barrier and perforated into the small intestinal mucosal epithelial cell. The bacteria and a lot of particles of the seriously damaged region penetrated into the lymphatic system and the blood system directly. Some bacteria were internalized into the goblet cell through the apical granule. Some bacteria and particles perforated into the submucosa of the M cell running the long axis of M cells through the tight junctions. In the microcirculation of mucosa, the bacteria that had already broken through the microvilli into blood circulation swim accompanying with erythrocytes.
CONCLUSIONThe routes of bacterial translocation interact and format a vicious circle. At early step, the transcellular pathway of bacterial translocation is major. Following with the destroyed small intestine mucous, the routes of bacterial translocation through the lymphatic system and the blood system become direct pathways. The goblet cell-dendritic cell and M cell pathway also play an important role in the bacterial translocation.
Animals ; Bacteria ; Bacterial Translocation ; Epithelial Cells ; microbiology ; Goblet Cells ; microbiology ; Intestinal Mucosa ; microbiology ; pathology ; ultrastructure ; Intestine, Small ; microbiology ; pathology ; ultrastructure ; Microvilli ; microbiology ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; microbiology
9.Treatment with total hepatic vascular exclusion and reperfusion for the intestinal barrier in rats.
De-quan CAO ; Yan-ping CHEN ; Yong-guo LI ; Ye-tian CHANG
Journal of Central South University(Medical Sciences) 2005;30(4):433-436
OBJECTIVE:
To investigate the influence of treatment with total hepatic vascular exclusion and reperfusion on the intestinal barrier in rats.
METHODS:
The total hepatic vascular exclusion and reperfusion model was built after the block of hepatic portal, suprahepatic and infraheptic vena cava for 20 minutes. Sixty Sprague-Dawley rats were divided randomly into 2 groups: sham operation group (Group A, n=30) and total hepatic vascular exclusion and reperfusion treatment group (Group B, n=30). Each group was subdivided randomly into 3 subgroups (n=10) according to different experiment time points as follows: at the end of the total hepatic vascular exclusion (T0), 4 reperfusion after total hepatic vascular exclusion (T1) and the 48 h survival. Portal vein blood gas was analysed at T0. At T0 and T1 the following items were detected: the level of portal vein blood D-lactate, tumor necrosis factor-alpha (TNF-alpha), the MDA concentration and pathologic morphology change of intestinal mucosa.
RESULTS:
Compared with Group A, the PCO2 at T0 in Group B increased while pH, P02, and HCO3- decreased significantly (P < 0.05). The level of portal blood D-lactate, TNF-alpha and intestinal mucosa MDA at T0 and T1 was significantly higher (P < 0.05, or P < 0.01). The histologic damage in the intestinal mucosa was observed in Group B, and the rat survival in Group B was lower than that in Group A (P < 0.05).
CONCLUSION
The treatment with total hepatic vascular exclusion and reperfusion can damage the intestinal barrier in rats.
Animals
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Bacterial Translocation
;
Female
;
Intestinal Mucosa
;
microbiology
;
pathology
;
Ischemia
;
pathology
;
physiopathology
;
Liver
;
blood supply
;
Male
;
Random Allocation
;
Rats
;
Rats, Sprague-Dawley
;
Reperfusion Injury
;
pathology
;
physiopathology
10.Detection of Lawsonia intracellularis in diagnostic specimens by one-step PCR.
Dong Kyun SUH ; Suk Kyung LYM ; You Chan BAE ; Keun Woo LEE ; Won Pil CHOI ; Jae Chan SONG
Journal of Veterinary Science 2000;1(1):33-37
Lawsonia intracellularis is not culturable with a standard bacteriologic culture. One step PCR assay as a clinical diagnostic method was developed for the rapid detection of porcine proliferative enteritis (PPE) caused by L. intracellularis. Primers were designed based on the p78 DNA clone of L. intracellularis. The one step PCR resulted in the formation of a specific 210-bp DNA product derived from L. intracellularis. The nonspecific amplification product was not detected with swine genomic DNA or other bacterial strains causing similar symptoms to L. intracellularis infection. The one step PCR was as sensitive as 100 pg of L. intracellularis genomic DNA. We applied this method to field specimens diagnosed as PPE by macroscopic observation. Of 17 mucosal scraping specimens, 16(94%) were identified as positive to PPE and 15(88%) of 17 feces specimens. These results suggest that the one step PCR can be used as a rapid diagnostic method for L. intracellularis infection.
Animals
;
Base Sequence
;
DNA Primers
;
Desulfovibrionaceae Infections/diagnosis/*veterinary
;
Ileum/microbiology/pathology
;
Intestinal Mucosa/microbiology/pathology
;
Lawsonia Bacteria/genetics/*isolation & purification
;
Polymerase Chain Reaction/*methods
;
Reproducibility of Results
;
Sensitivity and Specificity
;
Swine
;
Swine Diseases/*diagnosis/microbiology