OBJECTIVETo study the changes in gut microbiota and serum D-lactate level and their significance in children with recurrent pneumonia.

METHODSThe stool and blood samples were collected from 30 children with recurrent pneumonia (recurrent group), 30 children with acute pneumonia (acute group), and 15 children receiving surgical operation (surgery group). The 16S rRNA fluorescent quantitative polymerase chain reaction (FQ-PCR) was applied to determine the numbers of Bifidobacterium and Escherichia coli in stool samples, and the ratio between the logarithmic values of the numbers of Bifidobacterium and Escherichia coli (B/E value) was calculated. The serum D-lactate level was measured, and correlation analysis was performed.

RESULTSThe recurrent group had a significantly lower number of Bifidobacterium and a significantly lower B/E value than the acute group and the surgery group (P<0.05), as well as a significantly higher number of Escherichia coli than the surgery group (P<0.05). There was no significant difference in the number of Escherichia coli between the recurrent group and the acute group. The recurrent group had a significantly higher serum D-lactate level than the acute group and the surgery group (P<0.05). In the recurrent group, B/E value was negatively correlated with serum D-lactate level (r=-0.539, P<0.05).

CONCLUSIONSChildren with recurrent pneumonia may have biological and mechanical barrier damage in the intestinal mucosa.

Bacteria ; classification ; genetics ; isolation & purification ; Bifidobacterium ; genetics ; isolation & purification ; Child ; Child, Preschool ; Escherichia coli ; genetics ; isolation & purification ; Feces ; microbiology ; Female ; Gastrointestinal Microbiome ; Humans ; Lactates ; blood ; Pneumonia ; blood ; microbiology ; pathology ; Recurrence

OBJECTIVETo study the characteristics of the colonization of 8 species of bifidobacteria by systematically profiling fecal bifidobacterial community in the early life of infants.

METHODSFresh fecal samples including meconium samples were collected for culture and isolation of fecal bifidobacteria from 16 cases of full-term newborn infants born between March and April 2013 at their life of 2, 4, 7, 10, 14, 28, and 90 days. The isolated fecal bifidobacteria were taxonomically identified to genus and 8 species with PCR analysis.

RESULTSOne hundred and fifty-two predominant bifidobacteria strains were detected in the fecal samples, the detection rate of B. breve (22.4%) were the highest. Bifidobacteria were found in the feces of 8% infants 4 days after birth. The colonization rates increased to 54% and 60% at 28 days and 3 months respectively, significantly exceeding the colonization rate at 4 days after birth (P<0.05). Adult-type bifidobacteria B. catenulatum were found in the infants 10 days after birth, and infant-type bifidobacteria B. infantis were found at 14 days after birth, but infant-type bifidobacteria B. infantis were detected at a high level until 3 months after birth. The most tested infants had 2 species or less of bifidobacteria.

CONCLUSIONSIntestinal bifidobacteria in infants might have less diversity in early infancy. Infant-type bifidobacteria appear late, while adult-type bifidobacteria colonize earlier.

Bifidobacterium ; classification ; isolation & purification ; Breast Feeding ; Feces ; microbiology ; Female ; Humans ; Infant, Newborn ; Intestines ; microbiology ; Male

OBJECTIVETo investigate the changes in fecal flora and its correlation with the occurrence and progression of inflammatory bowel disease (IBD).

METHODSWe collected fresh fecal specimens from 167 IBD patients (including 113 with ulcerative colitis and 54 with Crohn's disease) and 54 healthy volunteers. The fecal flora was analyzed by gradient dilution method and the data of inflammatory markers including WBC, PLT, CRP and ESR were collected to assess the association between the fecal flora and the inflammatory markers.

RESULTSThe species Enterrococcus (6.60∓0.23, P<0.01), Saccharomyces (2.22∓0.27, P<0.05), Bacteriodes (5.57∓0.28, P<0.001), Bifidobacterium (5.08∓0.30, P<0.01), Peptococcus (6.22∓0.25, P<0.001), Lactobacillus (6.00∓0.26, P<0.001), and Clostridium (3.57∓0.30, P<0.05) all increased significantly, while Eubacterium (1.56∓0.24, P<0.01) reduced markedly in patients with ulcerative colitis compared with those in the control subjects. Enterrococcus (6.93∓0.28, P<0.01), Saccharomyces (2.73∓0.37, P<0.01), Bacteriodes (4.32∓0.52, P<0.05), Bifidobacterium (4.88∓0.42, P<0.05), Peptococcus (6.19∓0.32, P<0.01) and Lactobacillus (4.73∓0.47, P<0.001) all increased significantly and Eubacterium (1.01∓0.29, P<0.01) and Clostridium (0.87∓0.31, P<0.01) decreased in patients with Crohn's disease. The positivity rates of bacterial culture were consistent with the results of quantitative analysis of the fecal flora. The changes in fecal flora did not show a significant correlation with these inflammatory markers.

CONCLUSIONIBD patients have fecal flora imbalance compared with the healthy controls, and this imbalance may contribute to the occurrence and progression of IBD. The decline of Eubacterium contributes to the occurrence and development of IBD.

Adult ; Bacteria ; isolation & purification ; Bacteroides ; isolation & purification ; Bifidobacterium ; isolation & purification ; Biomarkers ; analysis ; Clostridium ; isolation & purification ; Colitis, Ulcerative ; microbiology ; Crohn Disease ; microbiology ; Enterococcus ; isolation & purification ; Eubacterium ; isolation & purification ; Feces ; microbiology ; Female ; Humans ; Inflammatory Bowel Diseases ; etiology ; microbiology ; Lactobacillus ; isolation & purification ; Male ; Peptococcus ; isolation & purification ; Saccharomyces ; isolation & purification

OBJECTIVETo determine the changes in fecal Bifidobacterium species in patients with type 2 diabetes in comparison with healthy individuals.

METHODSThe bacterial DNA were extracted from the fecal samples from 50 type 2 diabetic patients and 30 healthy individuals. Real-time quantitative PCR was employed to determine the copy numbers of the bacteria in the fecal samples using 16S rRNA-targeted genus- and species-specific PCR primers for a selected group of fecal Bifidobacterium species including total Bifidobacterium, B.longum, B.breve, B.adolescent, and B. infantis.

RESULTSThe diabetic group had significantly lower copy numbers of total Bifidobacterium and B.adolescent compared to the healthy individuals (P<0.05).

CONCLUSIONType 2 diabetic patients have a lowered number of Bifidobacterium species in the gut microflora.

Aged ; Bifidobacterium ; classification ; isolation & purification ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; microbiology ; Feces ; microbiology ; Female ; Humans ; Intestines ; microbiology ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction

OBJECTIVETo examine the differential levels of fecal Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile between patients with hepatic cirrhosis and healthy controls. Fecal samples were collected from 29 patients with hepatic cirrhosis treated in the Department of Digestive Diseases at Zunyi Hospital between March and December of 2010.

METHODSFecal samples were collected from 13 healthy college students for use as controls. All samples were assessed by pH measurement, bacterial culture for turbidity, fluorescence in situ hybridization, and laser scanning confocal microscopy. The t-test and rank correlation test were used to determine statistical significance of intergroup differences in each tested parameter.

RESULTSThe feces of patients with hepatic cirrhosis had higher pH than that of healthy controls (6.79+/-0.64 vs. 6.18+/-0.74, P less than 0.05). The bacterial turbidity was not significantly different between the feces of hepatic cirrhosis patients and healthy controls (1.15+/-0.59 vs. 1.39+/-1.01, P more than 0.05). The numbers of Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile in feces of patients with hepatic cirrhosis were significantly lower than those of the controls (all P less than 0.01). No significant correlation was found between the number or ratio of bacteria species and the severity of hepatic cirrhosis (Child-Pugh scores; P more than 0.05).

CONCLUSIONThe total quantity of intestinal bacteria in patients with hepatic cirrhosis is not significantly different from that in healthy patients. However, the profile of intestinal bacteria is different, which may explain the increased pH of fecal samples from patients with hepatic cirrhosis, but the differential profile is not correlated to cirrhosis pathogenesis.

Adult ; Aged ; Aged, 80 and over ; Bacteroides ; isolation & purification ; Bifidobacterium ; isolation & purification ; Case-Control Studies ; Clostridium ; isolation & purification ; Enterobacteriaceae ; isolation & purification ; Feces ; microbiology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Liver Cirrhosis ; microbiology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo explore the intestinal mucosal barrier protective effect of herbal medicine Compound Tongfu Granule (CTG) in patients with liver cirrhosis of decompensation stage.

METHODSFifty patients enrolled were randomly assigned to the control group (26 cases) and the CTG group (24 cases), and 30 healthy adults were set up as normal control. After 2-week treatment, the intestinal permeability (IP, represented by urinary lactulose/mannitol excretion rate), plasma endotoxin (EDT) level, and change of enteric bacteria (EB) in patients were observed before and after treatment, and compared with those in the normal control.

RESULTSBefore treatment, cirrhotic patients showed significantly higher levels of IP, EDT, and intestinal bacilli, but a lower amount of enteric bifidobacteria as compared with those the normal control. After 2-week treatment, levels of EDT and urinary excretion rate of lactulose in the CTG group were lowered more significantly than those in the control group (P < 0.05), while the amount of bifidobacteria in the CTG group increased accompanied with intestinal bacilli significantly lowered to near the levels in the normal control (P < 0.05, P < 0.01).

CONCLUSIONCTG can improve the intestinal barrier function, correct the intestinal bacteria disturbance, and significantly reduce the entero-derived endotoxemia in cirrhotic patients of decompensation stage.

Adult ; Aged ; Aged, 80 and over ; Bifidobacterium ; isolation & purification ; metabolism ; Cell Membrane Permeability ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Endotoxins ; metabolism ; Humans ; Intestinal Mucosa ; drug effects ; metabolism ; microbiology ; pathology ; Lactulose ; metabolism ; Liver Cirrhosis ; drug therapy ; metabolism ; microbiology ; pathology ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo study the effect of a low level of galacto-oligosaccharides (GOS) on intestinal bifidobacteria and lactobacilli, and fermentation characteristics in term infants by comparing with human milk and a standard infant formula without GOS.

METHODSA total of 371 term infants from four hospitals of China were enrolled. The infants started with breast feeding. After 1-2 weeks, some of the infants were changed to feeding with formula milk and then were randomly assigned to two formula-feeding groups: with or without GOS supplementation (2.4 g/L). Growth, stool characteristics, and side effects were recorded in a 3-month-follow-up. Faecal samples were collected for analysis of intestinal bacteria (culture technique), acetic acid (gas chromatography) and pH (indicator strip) at postnatal 3 months.

RESULTSCompared with the formula-feeding group without GOS, the contents of bifidobacteria, lactobacilli and acetic acid and stool frequency increased, and faecal pH decreased significantly in the GOS-formula-feeding and the human milk group. There were no significant differences between the GOS-formula-feeding and the human milk groups. Supplementation with GOS did not lead to an increase in the incidence of crying, regurgitation and vomiting.

CONCLUSIONSA supplementation of low levels of GOS in infant formula seemed to improve stool frequency, decrease faecal pH, and stimulate intestinal bifidobacteria and lactobacilli up to levels as found in breast-fed infants.

Bifidobacterium ; isolation & purification ; Dietary Supplements ; Galactose ; administration & dosage ; Humans ; Hydrogen-Ion Concentration ; Infant Formula ; Infant, Newborn ; Intestines ; microbiology ; Lactobacillus ; isolation & purification ; Oligosaccharides ; administration & dosage

OBJECTIVEThe aim of this study is designed to explore the anti-tumor effect of lipoteichoic acid (LTA) of Bifidobacterium on the expression of survivin in colon cancer LoVo cells and its possible regulatory mechanism.

METHODSThe changes of survivin mRNA and protein in LoVo cells treated with LTA of Bifidobacterium were detected by RT-PCR and Western blot. Meanwhile, the expressions of pAKT (the key protein kinase in P13K/AKT signal transduction pathway), p53 and PTEN were measured by Western blot.

RESULTSThere were overexpressions of survivin mRNA and protein in LoVo cells. After treated with different dose of LTA of Bifidobacterium, the expressions of survivin mRNA and protein were markedly decreased in a dose-dependent manner (P < 0.01). Besides, the activity of pAKT was decreased significantly (P < 0.01) and the expression of p53 and PTEN was increased (P < 0.01).

CONCLUSIONLTA of Bifidobacterium can down-regulate the expression of survivin in LoVo cells through inhibiting the activity of PI3K/AKT signal transduction pathway and up-regulate the expression of p53. Accordingly, the activity of caspases is increased, and apoptosis of LoVo cells occurs ultimately.

Apoptosis ; drug effects ; Bifidobacterium ; chemistry ; Blotting, Western ; Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Dose-Response Relationship, Drug ; Down-Regulation ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; Lipopolysaccharides ; isolation & purification ; pharmacology ; Microtubule-Associated Proteins ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; Teichoic Acids ; isolation & purification ; pharmacology ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVETo observe the changes of intestinal flora in diarrhea type irritable bowel syndrome with Pi-wei dampness-heat syndrome (IBS-PDS).

METHODSThe seven kinds of common intestinal bacteria in feces, including enteri bacillus, enterococci, saccharomycete, bifid bacteria, lactobacillus, bacteroides and peptococcus were studied in 21 patients suffered from IBS-PDS, and compared with those in 22 patients with IBS with deficiency of Pi syndrome (DPS) and 25 healthy subjects as control.

RESULTSAs compared with the healthy subjects, the levels of enteri bacillus and enterococci were significantly increased (P<0.01), the levels of bifid bacteria, Lactobacillus and Peptococcus were significantly decreased (P < 0.01), and saccharomycete and Bacteroides were insignificantly different in patients with PDS. As compared with patients with DPS, the levels of enteri bacillus, enterococci, bifid bacteria, Lactobacillus, Peptococcus and Bacteroidaceae were significantly increased except the level of saccharomycete.

CONCLUSIONThere may be alteration of intestinal flora in patients with IBS-PDS.

Adult ; Bifidobacterium ; isolation & purification ; Diagnosis, Differential ; Diarrhea ; etiology ; microbiology ; Enterobacteriaceae ; isolation & purification ; Female ; Humans ; Intestines ; microbiology ; Irritable Bowel Syndrome ; complications ; microbiology ; Lactobacillus ; isolation & purification ; Male ; Medicine, Chinese Traditional ; Middle Aged

OBJECTIVETo study the effect of bifidobacterium genomic DNA on umbilical cord blood mononuclear cell (CBMC), and investigate the immunoregulation of bifidobacterium DNA and explore possible mechanisms by which bifidobacterium acts against allergic reaction.

METHODSBifidobacterium genomic DNA (bDNA) and human DNA (hDNA) were extracted with phenol/chloroform/isoamyl alcohol and stored at -20 degrees C for later use. Parts of bDNA were completely digested with DNaseI (d-bDNA) at 37 degrees C. CBMCs were isolated with Ficoll from umbilical cord blood and incubated at 37 degrees C in a 5% CO2 humidified incubator. These cells were divided into four groups, control group: without any stimulant; bDNA group: stimulated with 25 microg/ml bDNA; d-bDNA group: stimulated with 25 microg/ml d-bDNA; hDNA group: stimulated with 25 microg/ml hDNA. The cells were stimulated with different stimulants in vitro, at the end of incubation culture supernatant and cells were collected. IL-12 and IL-10 levels in the culture supernatant were measured by enzyme linked immuno sorbent assay (ELISA); cells secreting IL-4 and IFN-gamma were counted by enzyme linked immunospot (ELISPOT) assay; and total RNA was isolated from the cells to assay T-bet and GATA3 mRNA expression levels by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSSix hours after stimulation there was no significant difference in IL-12 level in supernatant among the four groups; 12 hours after stimulation, IL-12 level in supernatant of bDNA treated group was significantly higher than that of each of the other groups, so were the results obtained at 24 hours and 48 hours after stimulation (P < 0.05). No significant difference could be detected in IL-12 level in supernatant among the other 3 groups. On the other hand, 6 hours after stimulation there was no significant difference in IL-10 level in supernatant among the four groups. But 12 and 24 hours after stimulation IL-10 level in supernatant of bDNA treated group was lower than that of each of the other groups, but the difference was not statistically significant. The count of IFN-gamma secreting cells of bDNA treated group was higher than that of the other groups, while IL-4 secteting cells of bDNA treated group were lower than that of the other groups. After bDNA stimulation, nuclear factor T-box expressed in T cells (T-bet) mRNA expression was conspicuously enhanced as compared to the other three groups (P < 0.05). GATA3 mRNA transcription in CBMC had no significant change after bDNA stimulation.

CONCLUSIONbDNA could promote secretion of Th1 type cytokine IL-12, while Th2 type cytokine IL-10 level of cell supernatant was decreased. bDNA could stimulate secretion of IFN-gamma by CBMC and inhibit secretion of IL-4. T-bet mRNA expression was highly enhanced after bDNA stimulation. bDNA could upregulate Th1 type response, which may be one of important mechanisms by which bifidobacterium inhibit allergic response.

Bifidobacterium ; cytology ; genetics ; Cell Culture Techniques ; DNA, Bacterial ; biosynthesis ; metabolism ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; immunology ; GATA3 Transcription Factor ; genetics ; Humans ; Infant, Newborn ; Interferon-gamma ; immunology ; secretion ; Interleukin-10 ; immunology ; secretion ; Interleukin-12 ; immunology ; secretion ; Interleukin-4 ; immunology ; secretion ; Leukocytes, Mononuclear ; immunology ; secretion ; RNA, Messenger ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; T-Box Domain Proteins ; genetics ; Th1 Cells ; drug effects ; immunology ; secretion