To demonstrate the fructose metabolism pathway in Bifidobacterium Longum NCC2705 and to construct its fermentation model, we explored the comparative proteome cultivating the strain on glucose or fructose, based on a proteomic reference map of B. longum NCC2705 constructed earlier. Then, we used matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry and electro-spray ionization tandem mass spectrometry (ESI-MS/MS) for differently expressed proteins identification. Furthermore, with semi-quantitative RT-PCR we determined the distinctively expressed proteins at the level of transcription. Proteomic comparison of glucose- and fructose-grown cells demonstrated much similarity. On the page of fructose there were all the enzymes and proteins that exist during the process of glucose degradation. We observed a greater variation of more than three-fold for the identified 9 spots representing 5 protein entries by MALDI-TOF MS. The sugar-binding protein specific to fructose (BL0033) and an ABC transporter ATP binding protein (BL0034) showed higher expression level from cells grown on fructose. It was also determined by semi-quantitative RT-PCR subsequently. BL0033 time course and concentration experiments showed that the induction time correlated to higher fructose concentration, and increased expression of BL0033. Fructose was catabolized via the same degradation pathway as glucose at the level of proteomics. BL0033 was induced by fructose. All results suggest that the uptake of fructose into the cell may be conducted by a specific ABC transport system, in which BL0033 and BL0034 as components might have played an important role.
Bifidobacterium ; chemistry ; genetics ; metabolism ; Culture Media ; Fermentation ; Fructose ; pharmacology ; Glucose ; pharmacology ; Proteome ; analysis ; genetics ; Proteomics ; methods

OBJECTIVES: To examine the changes of intestinal flora in children newly diagnosed with acute lymphoblastic leukemia (ALL) and the influence of chemotherapy on intestinal flora. METHODS: Fecal samples were collected from 40 children newly diagnosed with ALL before chemotherapy and at 2 weeks, 1 month, and 2 months after chemotherapy. Ten healthy children served as the control group. 16S rDNA sequencing and analysis were performed to compare the differences in intestinal flora between the ALL and control groups and children with ALL before and after chemotherapy. RESULTS: The ALL group had a significant reduction in the abundance of intestinal flora at 1 and 2 months after chemotherapy, with a significant reduction compared with the control group (P<0.05). Compared with the control group, the ALL group had a significant reduction in the diversity of intestinal flora before and after chemotherapy (P<0.05). At the phylum level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Actinobacteria at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05) and a significant increase in the relative abundance of Proteobacteria at 1 and 2 months after chemotherapy (P<0.05). At the genus level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Bifidobacterium at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05); the relative abundance of Klebsiella in the ALL group was significantly higher than that in the control group at 1 and 2 months after chemotherapy and showed a significant increase at 1 month after chemotherapy (P<0.05); the relative abundance of Faecalibacterium in the ALL group was significantly lower than that in the control group before and after chemotherapy and showed a significant reduction at 2 weeks and 1 month after chemotherapy (P<0.05). The relative abundance of Enterococcus increased significantly at 1 and 2 months after chemotherapy in the ALL group (P<0.05), and was significantly higher than that in the control group (P<0.05). CONCLUSIONS The diversity of intestinal flora in children with ALL is significantly lower than that in healthy children. Chemotherapy significantly reduces the abundance of intestinal flora and can reduce the abundance of some probiotic bacteria (Bifidobacterium and Faecalibacterium) and increase the abundance of pathogenic bacteria (Klebsiella and Enterococcus) in children with ALL.
Bacteria/genetics* ; Bifidobacterium ; Child ; Feces/microbiology* ; Gastrointestinal Microbiome ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy*

OBJECTIVES: To study the features of intestinal flora in children with food protein-induced proctocolitis (FPIP) by high-throughput sequencing. METHODS: A total of 31 children, aged <6 months, who experienced FPIP after exclusive breastfeeding and attended the outpatient service of the Third Affiliated Hospital of Zunyi Medical University from October 2018 to February 2021 were enrolled as the FPIP group. Thirty-one healthy infants were enrolled as the control group. Fecal samples were collected to extract DNA for PCR amplification. High-throughput sequencing was used to perform a bioinformatics analysis of 16S rDNA V3-V4 fragments in fecal samples. RESULTS: The diversity analysis of intestinal flora showed that compared with the control group, the FPIP group had a lower Shannon index for diversity (P>0.05) and a significantly higher Chao index for abundance (P<0.01). At the phylum level, the intestinal flora in both groups were composed of Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes. Compared with the control group, the FPIP group had a significant reduction in the composition ratio of Actinobacteria (P<0.001) and a significant increase in the composition ratio of Proteobacteria (P<0.05). At the genus level, the intestinal flora in the FPIP group were mainly composed of Escherichia, Clostridium, Enterococcus, Klebsiella, and Bifidobacterium, and the intestinal flora in the control group were mainly composed of Bifidobacterium and Streptococcus. Compared with the control group, the FPIP group had a significant reduction in the composition ratio of Bifidobacterium and Ruminococcus (P<0.05) and significant increases in the composition ratios of Clostridium and Shigella (P<0.05). CONCLUSIONS Compared with the control group, the FPIP group has a reduction in the diversity of intestinal flora and an increase in their abundance, and there are certain differences in several bacterial genera. These results suggest that changes in the composition of intestinal flora at genus level may play an important role in the development and progression of FPIP.
Bacteria/genetics* ; Bifidobacterium/genetics* ; Child ; Gastrointestinal Microbiome ; High-Throughput Nucleotide Sequencing/methods* ; Humans ; Infant ; Proctocolitis ; RNA, Ribosomal, 16S/genetics*

OBJECTIVETo investigate the effects of bifidobacteria on malondialdehyde (MDA) content and intercellular adhesion molecule-1 (ICAM-1) expression in serum and ileum tissues of young rats with endotoxin-induced intestinal damage, and possible protective mechanisms of bifidobacteria on intestines.

METHODSEigteen-day-old Wistar rats were randomly administered with normal saline (NS), lipopolysaccharide (LPS, 5 mg/kg) or LPS + bifidobacteria. Bifidobacteria (0.5 mL, twice a day) was intragastrically administrated 7 days prior to LPS injection until the end of the experiment. MDA contents in serum and ileum were detected by the TBA method. Expression of ICAM-1 protein and mRNA were evaluated by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) after 2, 6, 24 and 72 hrs of LPS injection.

RESULTSThe serum and ileum MDA contents in the untreated LPS group increased significantly and reached a peak at 6 hrs of LPS injection when compared with the NS control group (ileum: 99.88 +/- 12.62 nmol/mg prot vs 84.25+/-12.96 nmol/mg prot, P < 0.05; serum: 1.67 +/- 0.30 nmol/mL vs 1.13 +/- 0.20 nmol/mL, P < 0.05). The MDA contents in ileum (92.75 +/- 9.28 nmol/mg prot) and serum (1.17 +/- 0.23 nmol/mL) in the bifidobacteria-treated group at 6 hrs of LPS injection were significantly lower than in the LPS group (P < 0.05). The expression of ICAM-1 protein in the untreated LPS group remarkably increased at 6, 12, 24 and 72 hrs of LPS injection when compared with the NS control group (P < 0.01). The bifidobacteria-treated group displayed lower ICAM-1 protein levels than the untreated LPS group at 72 hrs of LPS injection (P < 0.01). The ICAM-1 mRNA expression in the untreated LPS group significantly increased at 2 hrs of LPS injection when compared with the NS control group (P < 0.01). The ICAM-1 mRNA expression in the bifidobacteria-treated group began to decrease at 2 hrs of LPS injection and was reduced again at 24 hrs after experiencing increase at 6 and 12 hrs of LPS injection when compared with the untreated LPS group (P < 0.01).

CONCLUSIONSThe serum and ileum MDA contents and the expression of ICAM-1 protein and mRNA increased in young rats with endotoxin-induced intestinal damage. Bifidobacteria supplementation can decrease MDA contents and inhibit ICAM-1 expressions, thus providing protections for intestines.

Animals ; Bifidobacterium ; Ileum ; chemistry ; metabolism ; Intercellular Adhesion Molecule-1 ; analysis ; genetics ; Lipopolysaccharides ; toxicity ; Malondialdehyde ; analysis ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Time Factors

OBJECTIVETo observe the effect of pBBADs-OXM-transformed bifidobacteria on the body weight of obese mice.

METHODSB. longum was transformed with pBBADs-OXM by electroporation, and arabopyranose-induced oxyntomodulin expression by the bacterium was detected by ELISA. pBBADs-OXM-transformed bifidobacteria was administered orally obese mice on a daily basis with pBBADs-GFP-transformed bifidobacteria as the negative control, and the body weight changes of the mice were observed.

RESULTSOXM was detected by ELISA not only in the supernatant but also the precipitant of the transformed bacterial culture. The body weight of the obese mice fed with pBBADs-OXM-transformed bifidobacteria decreased significantly compared with that of the mice in the obese model group (P<0.05).

CONCLUSIONAdministration of pBBADs-OXM-transformed B.longum can reduce the body weight of obese mice.

Administration, Oral ; Animals ; Appetite Depressants ; administration & dosage ; metabolism ; Bifidobacterium ; genetics ; metabolism ; Body Weight ; drug effects ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Mice ; Obesity ; drug therapy ; Oxyntomodulin ; administration & dosage ; biosynthesis ; genetics ; Random Allocation ; Recombinant Proteins ; administration & dosage ; biosynthesis ; genetics

BACKGROUND/AIMS: Several clinical trials have revealed various advantages for probiotics in inflammatory bowel disease (IBD). The aim of this study was to further investigate the effects of probiotic yogurt consumption on gut microbiota in patients with this disease. METHODS: A total of 305 participants were divided into three groups; group A (IBD patients receiving probiotic yogurt; n=105), group B (IBD patients receiving placebo; n=105), and control group (healthy individuals receiving probiotic yogurt; n=95). Stool samples were collected both before and after 8 weeks of intervention; and population of Lactobacillus, Bifidobacterium and Bacteroides in the stool specimens was measured by Taqman real-time PCR method. ': By the end of the intervention, no significant variations in the mean weight and body mass index were observed between three groups (p>0.05). However, the mean numbers of Lactobacillus, Bifidobacterium, and Bacteroides in group A were significantly increased compared to group B (p<0.001, p<0.001, and p<0.01, respectively). There were also significant differences in the mean numbers of either of three bacteria between group A and the healthy control group; however, these differences between two groups were observed both at baseline and the end of the intervention. CONCLUSIONS: Consumption of probiotic yogurt by patients with IBD may help to improve intestinal function by increasing the number of probiotic bacteria in the intestine and colon. However, many more studies are required in order to prove the concept.
Adult ; Bacteroides/genetics ; Bifidobacterium/genetics ; DNA, Bacterial/analysis ; Double-Blind Method ; Feces/microbiology ; Female ; Gastrointestinal Microbiome ; Humans ; Inflammatory Bowel Diseases/*drug therapy ; Intestines/microbiology ; Lactobacillus/genetics ; Male ; Middle Aged ; Placebo Effect ; Probiotics/*therapeutic use ; Real-Time Polymerase Chain Reaction

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

This study aims to analyze and compare the effect of cell wall-broken decoction pieces, conventional decoction pieces and conventional powder of Rhodiolae Crenulatae Radix et Rhizoma on the intestinal flora of normal mice. The conventional bacterial culture and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) were adopted for the mice after the oral administration for 14 days. According to the bacterial culture results, the 1/8 dose cell wall-broken decoction pieces group showed fewer Enterococcus and Escherichia coli bacillus but more Lactobacillus and Bifidobacterium than the conventional decoction pieces group and the traditional powder group (P <0.05). Meanwhile, on the basis of the PCR-DGGE results, the 1/8 dose cell wall-broken decoction pieces group revealed the highest Shannon-Wiener index (H) and species richness (S) among the seven groups, with extremely significant differences compared with the normal group (P <0.01), significant differences compared with the conventional decoction pieces group and the conventional powder group (P <0.05) and a high intra-group similarity. In conclusion, the long-term intake of 1/8 dose Rhodiolae Crenulatae Radix et Rhizoma cell wall-broken decoction pieces showed a certain effect in regulating intestinal tract by promoting the growth of Lactobacillus and Bifidobacterium. Furthermore, the intestinal flora community will become more stable.
Animals ; Bifidobacterium ; drug effects ; genetics ; growth & development ; Cell Wall ; Denaturing Gradient Gel Electrophoresis ; Intestines ; microbiology ; Lactobacillus ; drug effects ; genetics ; growth & development ; Mice ; Mice, Inbred C57BL ; Polymerase Chain Reaction ; Rhizome ; Rhodiola

OBJECTIVETo study the effect of Bifidobacterium on the expression of β-defensin-2 (BD-2) in intestinal tissue of neonatal rats with necrotizing enterocolitis (NEC).

METHODSA total of 40 rats were randomly divided into four groups: normal control, Bifidobacterium control, NEC model, and Bifidobacterium treatment, with 10 rats in each group. A rat model of NEC was induced by hypoxia, cold stimulation, and artificial feeding. The rats in the Bifidobacterium control and Bifidobacterium treatment groups were given Bifidobacterium via the gastric tube after cold stimulation once a day for three consecutive days. The morphological changes of the terminal ileum were observed under a light microscope and the intestinal injury score was determined. Immunohistochemistry and qRT-PCR were used to measure the protein and mRNA expression of BD-2 in the ileal mucosal tissue.

RESULTSThe NEC model group had a significantly higher intestinal injury score than the normal control, Bifidobacterium control, and Bifidobacterium treatment groups (P<0.05). The Bifidobacterium treatment group had a significantly higher intestinal injury score than the normal control and Bifidobacterium control groups (P<0.05). The mRNA and protein expression of BD-2 in the normal control group was significantly lower than in the Bifidobacterium control, NEC model, and Bifidobacterium treatment groups (P<0.05). The Bifidobacterium control group had significantly higher mRNA and protein expression of BD-2 than the NEC model and Bifidobacterium treatment groups (P<0.05). The Bifidobacterium treatment group had significantly higher mRNA and protein expression of BD-2 than the NEC model group (P<0.05).

CONCLUSIONSBifidobacterium can induce the expression of BD-2 in intestinal tissue of rats and reduce inflammatory response by increasing the expression of BD-2. This provides a protective effect on neonatal rats with NEC.

Animals ; Bifidobacterium ; Disease Models, Animal ; Enterocolitis, Necrotizing ; therapy ; Humans ; Infant, Newborn ; Intestinal Mucosa ; metabolism ; NF-kappa B ; physiology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology ; beta-Defensins ; analysis ; genetics ; physiology

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