Association between childhood obesity and gut microbiota:16S rRNA gene sequencing-based cohort study

BACKGROUND This study aimed to identify characteristic gut genera in obese and normal-weight children(8-12 years old)using 16S rDNA sequencing.The research aimed to provide insights for mechanistic studies and prevention strategies for childhood obesity.Thirty normal-weight and thirty age-and sex-matched obese children were included.Questionnaires and body measurements were collected,and fecal samples underwent 16S rDNA sequencing.Significant differences in body mass index(BMI)and body-fat percentage were observed between the groups.Analysis of gut microbiota diversity revealed lowerα-diversity in obese children.Differences in gut microbiota composition were found between the two groups.Prevotella and Firmicutes were more abundant in the obese group,while Bacteroides and Sanguibacteroides were more prevalent in the control group.AIM To identify the characteristic gut genera in obese and normal-weight children(8-12-year-old)using 16S rDNA sequencing,and provide a basis for subsequent mechanistic studies and prevention strategies for childhood obesity.METHODS Thirty each normal-weight,1:1 matched for age and sex,and obese children,with an obese status from 2020 to 2022,were included in the control and obese groups,respectively.Basic information was collected through questionnaires and body measurements were obtained from both obese and normal-weight children.Fecal samples were collected from both groups and subjected to 16S rDNA sequencing using an Illumina MiSeq sequencing platform for gut microbiota diversity analysis.RESULTS Significant differences in BMI and body-fat percentage were observed between the two groups.The Ace and Chao1 indices were significantly lower in the obese group than those in the control group,whereas differences were not significant in the Shannon and Simpson indices.Kruskal-Wallis tests indicated significant differences in unweighted and weighted UniFrac distances between the gut microbiota of normal-weight and obese children(P<0.01),suggesting substantial disparities in both the species and quantity of gut microbiota between the two groups.Prevotella,Firmicutes,Bacteroides,and Sanguibacteroides were more abundant in the obese and control groups,respectively.Heatmap results demonstrated significant differences in the gut microbiota composition between obese and normal-weight children.CONCLUSION Obese children exhibited lowerα-diversity in their gut microbiota than did the normal-weight children.Significant differences were observed in the composition of gut microbiota between obese and normal-weight children.

Seasonal changes in gut microbiota of sea cucumber over natural aestivation cycle

Sea cucumber Apostichopus japonicus is an ideal model organism for marine invertebrate aestivation;it annually enters a“sleeping state”for nearly 3 months when ambient water temperature range is 25–30℃.The natural fasting is accompanied by remodeling the intestinal biota and function,which is a part of host biology and could affect the gut microbiota.We investigatesd the impact of annual aestivation on gut microbiota using high-throughput sequencing of 16S rRNA amplicons.Results reveal a notable alteration in the composition of gut bacteria during aestivation during which various indigenous families and genera that exhibit a preference for dietary glycans(e.g.,family Rhodobacteraceae and Flavobacteriaceae)decreased,while the taxa capable of utilizing substrates derived from the host(e.g.,genus Akkermansia and Prevotella)increased,and so did certain opportunistic pathogenic bacteria.Moreover,the investigation delved into the gut morphology and immunity response of A.japonicus and reveal that the intestine of A.japonicus undergoes substantial atrophy and degeneration during aestivation.However,there was an observed augmentation in the levels of acid and neutral mucin within mucous cells,as well as an enhanced immune defense response(as evidenced by increased gene expression of AjTLR3,LITAF,Ajp105,and LYZ).These results imply that the composition of sea cucumber gut microbiota differed between aestivation and active periods,which potentially affects the intestinal functions of the host and the symbiotic relationship between host and its microbiota over the activeaestivation cycle.

SMRT sequencing and ddPCR reveal the complexity of developmental trajectories and temporal dynamics of gut bifidobacterial communities in infants

Infant intestinal microbiome is closely linked with health and risk of disease. Bifidobacterium are important components of the infant gut and are known to confer various health effects on the host. However, few studies have described the precise composition and dynamics of early infant gut bifidobacterial communities. Thus, this was a pilot study aiming to describe the developmental trajectories and temporal dynamics of bifidobacterial communities in infants before 6 months of age. A total of 28 fecal samples from 4 infants(GF, ZZ, QM, TN, respectively)were collected and analyzed after 5, 15, 30, 60, 90, 120, 150, and 180 days of birth by a bifidobacteria-target method(based on single-molecule real-time sequencing of partial bifidobacterial rpsK genes)in conjunction with droplet digital polymerase chain reaction(ddPCR). The infant fecal microbiota comprised a total of 11 bifidobacterial species, including 4 major species, i.e., B. dentium(37.35%), B. catenulatum(32.04%), B. breve(22.24%), and B. animalis(8.02%). The infant microbiota showed highly individualized developmental trajectories. The leading species for GF was B. catenulatum, with a relatively stable developmental trajectory. In ZZ, B. breve was enriched, and the developmental trajectory was rather fluctuating. The most abundant species for QM and TN was B. dentium. The developmental trajectory of B. dentium in QM showed a trend of gradual decrease, whereas an opposite trend was seen in samples of TN. The results of ddPCR confirmed large variations in quantities of bifidobacteria between infants and suggested discordances in temporal dynamics of bifidobacterial communities during the first half year of infancy. In conclusion, our results suggested that the early infant gut bifidobacterial microbiota was highly complex and temporal dynamics, with individualized developmental trajectories, which should be considered in future research of infant gut microbiota.

High-throughput sequencing of 16S r RNA amplicons characterizes gut microbiota shift of juvenile sea cucumber Apostichopus japonicus feeding with three antibiotics

Sea cucumber Apostichopus japonicus is an important marine economic species in Asian countries due to its profound nutritional and medicinal value. So far, with the rapid development of intensifi ed artifi cial aquaculture of sea cucumbers, the use of antibiotics is still an inexpensive and dispensable way to treat pathogenic infections, especially during the nursery phase. However, there is little information on the eff ects of antibiotics on the intestinal microbiota of sea cucumber. Therefore an Illumina based sequencing method was used to examine the intestinal bacterial composition of juvenile A . japonicas following diets with three typical antibiotics (tetracycline, erythromycin, and norfl oxacin) under 15, 30, and 45 d. The fi ndings reveal that diff erent antibiotics have distinct eff ects on the growth performance of juvenile sea cucumbers. However, the richness and diversity of microbiota were barely aff ected by antibiotics but the community composition alterations indicated that the three antibiotics exhibited their respective patterns of reshaping the intestinal bacteria of juvenile sea cucumbers. In common, the abundance of some sensitive genera with helpful functions, such as Thalassotalea , Shewanella , Sulfi tobacter , and Halomonas decreased signifi cantly with exposure to antibiotics and the abundance of multiple potential pathogenic- and suspected antibiotic-resistant microorganisms like Arcobacter , Leucothrix , and Clostridium_sensu_stricto_1 was found increased signifi cantly in the antibiotic groups. These results suggest that low doses of antibiotics could aff ect the composition of the intestinal microbiota of sea cucumbers and might increase the risk of infection of the hosts. This study could help us to explore how antibacterial compounds modify the gut microbiota of sea cucumbers and provide theoretical guidance in hatchery management by scientifi c antibiotic use in sea cucumber mariculture.

Alterations in gut microbiota are related to metabolite profiles in spinal cord injury

Studies have shown that gut microbiota metabolites can enter the central nervous system via the blood-spinal cord barrier and cause neuroinflammation, thus constituting secondary injury after spinal cord injury. To investigate the correlation between gut microbiota and metabolites and the possible mechanism underlying the effects of gut microbiota on secondary injury after spinal cord injury, in this study, we established mouse models of T8–T10 traumatic spinal cord injury. We used 16 S rRNA gene amplicon sequencing and metabolomics to reveal the changes in gut microbiota and metabolites in fecal samples from the mouse model. Results showed a severe gut microbiota disturbance after spinal cord injury, which included marked increases in pro-inflammatory bacteria, such as Shigella, Bacteroides, Rikenella, Staphylococcus, and Mucispirillum and decreases in anti-inflammatory bacteria, such as Lactobacillus, Allobaculum, and Sutterella. Meanwhile, we identified 27 metabolites that decreased and 320 metabolites that increased in the injured spinal cord. Combined with pathway enrichment analysis, five markedly differential amino acids(L-leucine, L-methionine, L-phenylalanine, L-isoleucine and L-valine) were screened out, which play a pivotal role in activating oxidative stress and inflammatory responses following spinal cord injury. Integrated correlation analysis indicated that the alteration of gut microbiota was related to the differences in amino acids, which suggests that disturbances in gut microbiota might participate in the secondary injury through the accumulation of partial metabolites that activate oxidative stress and inflammatory responses. Findings from this study provide a new theoretical basis for improving the secondary injury after spinal cord injury through fecal microbial transplantation.

Sarcopenia and gut microbiota alterations in patients with hematological diseases before and after hematopoietic stem cell transplantation

Objective: The aim of this study was to investigate the prevalence of sarcopenia(SP) and its relationship with gut microbiota alterations in patients with hematological diseases before and after hematopoietic stem cell transplantation(HSCT).Methods: A total of 108 patients with various hematological disorders were selected from Peking University People’s Hospital. SP was screened and diagnosed based on the 2019 Asian Sarcopenia Diagnosis Strategy. Physical measurements and fecal samples were collected, and 16S rRNA gene sequencing was conducted. Alpha and beta diversity analyses were performed to evaluate gut microbiota composition and diversity.Results: After HSCT, significant decreases in calf circumference and body mass index(BMI) were observed,accompanied by a decline in physical function. Gut microbiota analyses revealed significant differences in the relative abundance of Enterococcus, Bacteroides, Blautia and Dorea species before and after HSCT(P<0.05). Before HSCT, sarcopenic patients had lower Dorea levels and higher Phascolarctobacterium levels than non-sarcopenia patients(P<0.01). After HSCT, no significant differences in species abundance were observed. Alpha diversity analysis showed significant differences in species diversity among the groups, with the highest diversity in the postHSCT 90-day group and the lowest in the post-HSCT 30-day group. Beta diversity analysis revealed significant differences in species composition between pre-and post-HSCT time points but not between SP groups. Linear discriminant analysis effect size(LEfSe) identified Alistipes, Rikenellaceae, Alistipes putredinis, Prevotellaceae defectiva and Blautia coccoides as biomarkers for the pre-HSCT sarcopenia group. Functional predictions showed significant differences in anaerobic, biofilm-forming and oxidative stress-tolerant functions among the groups(P<0.05).Conclusions: This study demonstrated a significant decline in physical function after HSCT and identified potential gut microbiota biomarkers and functional alterations associated with SP in patients with hematological disorders. Further research is needed to explore the underlying mechanisms and potential therapeutic targets.

Molecular methods for colorectal cancer screening:Progress with next-generation sequencing evolution

Currently,colorectal cancer(CRC)represents the third most common malignancy and the second most deadly cancer worldwide,with a higher incidence in developed countries.Like other solid tumors,CRC is a heterogeneous genomic disease in which various alterations,such as point mutations,genomic rearrangements,gene fusions or chromosomal copy number alterations,can contribute to the disease development.However,because of its orderly natural history,easily accessible onset location and high lifetime incidence,CRC is ideally suited for preventive intervention,but the many screening efforts of the last decades have been compromised by performance limitations and low penetrance of the standard screening tools.The advent of next-generation sequencing(NGS)has both facilitated the identification of previously unrecognized CRC features such as its relationship with gut microbial pathogens and revolutionized the speed and throughput of cataloguing CRC-related genomic alterations.Hence,in this review,we summarized the several diagnostic tools used for CRC screening in the past and the present,focusing on recent NGS approaches and their revolutionary role in the identification of novel genomic CRC characteristics,the advancement of understanding the CRC carcinogenesis and the screening of clinically actionable targets for personalized medicine.

Relationship between hyperlipidemia and the gut microbiome of rats, characterized using high-throughput sequencing

Objective:To determine the effects of a high-fat diet(HFD)on the gut microbiome in rats,to explore the relationship between the intestinal flora and blood lipid profile.Methods:SpragueeDawley rats were fed an HFD for four weeks to induce hyperlipidemia,then 16S rRNA sequencing was used to compare the intestinal flora between hyperlipidemic and control diet-fed rats.Results:The microbiome of rats fed an HFD for four weeks differed from that of control diet-fed rats.Bacterial species that were less abundant were most affected by HFD feeding,among which were many pathogenic species,which became significantly more abundant.Eighteen genera were present in significantly different numbers in hyperlipidemic and control rats,more than half of which have been linked to infection and inflammation,or energy intake and obesity.The results indicated a type of stress response of the flora to a high-fat environment.In addition,the age of the rats tended to influence the gut microbial composition.Conclusion:These findings suggest that HFD may induce hyperlipidemia by affecting the gut microbial composition.Changes in the abundance of pro-inflammatory and pathogenic bacteria,and those that influence energy intake and obesity,may be important mediators of this.

Gut microbiota and non-alcoholic fatty liver disease

BACKGROUND： Non-alcoholic fatty liver disease （NAFLD） is a common disorder with poorly understood pathogenesis. Beyond environmental and genetic factors, cumulative data support the causative role of gut microbiota in disease development and progression.

Intestinal toxicity of deoxynivalenol is limited by supplementation with Lactobacillus plantarum JM113 and consequentially altered gut microbiota in broiler chickens

Background: Limited research has focused on the effect of Lactobacillus on the intestinal toxicity of deoxynivalenol(DON).The present study was conducted to investigate the role of Lactobacillus plantarum(L.plantarum) JM113 in protecting against the intestinal toxicity caused by DON.Methods: A total of 144 one-day-old healthy Arbor Acres broilers were randomly distributed into 3 treatments,including the CON(basal diet),the DON(extra 10 mg/kg deoxynivalenol),and the DL(extra 1 × 109 CFU/kg L.plantarum JM113 based on DON group) treatments.The growth performance,organ indexes,intestinal morphology,pancreatic digestive enzymes,intestinal secreted immunoglobulin A(sIgA),jejunal transcriptome,and intestinal microbiota were evaluated.Results: Compared with the CON and DL groups,the DON supplementation altered intestinal morphology,especially in duodenum and jejunum,where villi were shorter and crypts were deeper(P < 0.05).Meanwhile,the significantly decreased mRNA expression of jejunal claudin-1 and occludin(P < 0.05),ileal rBAT and jejunal GLUT1 of 21-day-old broilers(P < 0.05),as well as duodenal PepT1 and ileal rBAT of 42-day-old broilers were identified in the DON group.Moreover,supplementation with L.plantarum JM113 could increase duodenal expression of IL-10 and IL-12 of 21-dayold broilers,ileal s IgA of 42-day-old broilers,and the bursa of Fabricius index of 21-day-old broilers.Further jejunal transcriptome proved that the genes related to the intestinal absorption and metabolism were significantly reduced in the DON group but a significant increase when supplemented with extra L.plantarum JM113.Furthermore,the bacteria related to nutrient utilization,including the Proteobacteria,Escherichia,Cc-115(P < 0.05),Lactobacillus and Prevotella(P < 0.1) were all decreased in the DON group.By contrast,supplementation with L.plantarum JM113 increased the relative abundance of beneficial bacterium,including the Bacteroidetes,Roseburia,Anaerofustis,Anaerostipe,and Ruminococcus bromi(P < 0.05).Specifically,the increased abundance of bacteria in the DL group could be proved by the significantly increased caecal content of propionic acid,n-Butyric acid,and total short-chain fatty acid.Conclusions: L.plantarum JM113 enhanced the digestion,absorption,and metabolic functions of the gut when challenged with DON by reducing the injury to intestinal barriers and by increasing the abundance of beneficial bacterium.

Structural shift of gut microbiota during chemopreventive effects of epigallocatechin gallate on colorectal carcinogenesis in mice

AIM To investigate the effect of epigallocatechin gallate(EGCG) on structural changes of gut microbiota in colorectal carcinogenesis.METHODS An azoxymethane(AOM)/dextran sodium sulfate(DSS)-induced colitis mouse model was established. Fortytwo female FVB/N mice were randomly divided into the following three groups: group 1(10 mice, negative control) was treated with vehicle, group 2(16 mice, positive control) was treated with AOM plus vehicle, and group 3(16 mice, EG) was treated with AOM plus EGCG. For aberrant crypt foci(ACF) evaluation, the colons were rapidly took out after sacrifice, rinsed with saline, opened longitudinally, laid flat on a polystyrene board, and fixed with 10% buffered formaldehyde solution before being stained with 0.2% methylene blue in saline. For tumor evaluation, the colon was macroscopically inspected and photographed, then the total number of tumors was enumerated and tumor size measured. For histological examination, the fixed tissues were paraffin-embedded and sectioned at 5 mm thickness. Microbial genomic DNA was extracted from fecal and intestinal content samples using a commercial kit. The V4 hypervariable regions of 16 S r RNA were PCR-amplified with the barcoded fusion primers. Using the best hit classification option, the sequences from each sample were aligned to the RDP 16 S r RNA training set to classify the taxonomic abundance in QIIME. Statistical analyses were then performed.RESULTS Treatment of mice with 1% EGCG caused a significant decrease in the mean number of ACF per mouse, when compared with the model mice treated with AOM/DSS(5.38 ± 4.24 vs 13.13 ± 3.02, P < 0.01). Compared with the positive control group, 1% EGCG treatment dependently decreased tumor load per mouse by 85%(33.96 ± 6.10 vs 2.96 ± 2.86, respectively, P < 0.01). All revealed that EGCG could inhibit colon carcinogenesis by decreasing the number of precancerous lesions as well as solid tumors, with reduced tumor load and delayed histological progression of CRC. During the cancerization, the diversity of gut microbiota increased, potential carcinogenic bacteria such as Bacteroides were enriched, and the abundance of butyrate-producing bacteria(Clostridiaceae, Ruminococcus, etc.) decreased continuously. In contrast, the structure of gut microbiota was relatively stable during the intervention of EGCG on colon carcinogenesis. Enrichment of probiotics(Bifidobacterium, Lactobacillu, etc.) might be a potential mechanism for EGCG's effects on tumor suppression. Via bioinformatics analysis, principal coordinate analysis and cluster analysis of the tumor formation process, we found that the diversity of gut microbiota increased in the tumor model group while that in the EGCG interfered group(EG) remained relatively stable.CONCLUSION Gut microbiota imbalance might be a potential mechanism for the prevention of malignant transformation by EGCG, which is significant for diagnosis, treatment, prognosis evaluation, and prevention of colorectal cancer.

Fecal microbiota transplantation ameliorates experimental colitis via gut microbiota and T-cell modulation

BACKGROUND Emerging evidence has demonstrated that fecal microbiota transplantation(FMT)has a promising therapeutic effect on mice with experimental colitis and patients with ulcerative colitis(UC),although the mechanism of FMT is unclear.AIM To evaluate the protective effect of FMT on UC and clarify its potential dependence on the gut microbiota,through association analysis of gut microbiota with colon transcriptome in mice.METHODS Dextran sodium sulfate(DSS)-induced experimental colitis was established and fecal microbiota was transplanted by gavage.Severity of colon inflammation was measured by body weight,disease activity index,colon length and histological score.Gut microbiota alteration was analyzed through 16S ribosomal ribonucleic acid sequencing.The differentially expressed genes(DEGs)in the colon were obtained by transcriptome sequencing.The activation status of colonic T lymphocytes in the lamina propria was evaluated by flow cytometry.RESULTS Compared with the DSS group,the weight loss,colon length shortening and inflammation were significantly alleviated in the FMT group.The scores of disease activity index and colon histology decreased obviously after FMT.FMT restored the balance of gut microbiota,especially by upregulating the relative abundance of Lactobacillus and downregulating the relative abundance of Clostridium_sensu_stricto_1 and Turicibacter.In the transcriptomic analysis,128 DEGs intersected after DSS treatment and FMT.Functional annotation analysis suggested that these DEGs were mainly involved in T-lymphocyte activation.In the DSS group,there was an increase in colonic T helper CD4^(+)and T cytotoxic CD8^(+)cells by flow cytometry.FMT selectively downregulated the ratio of colonic CD4^(+)and CD8^(+)T cells to maintain intestinal homeostasis.Furthermore,Clostri dium_sensu_stricto_1 was significantly related to inflammation-related genes including REG3G,CCL8 and IDO1.CONCLUSION FMT ameliorated DSS-induced colitis in mice via regulating the gut microbiota and T-cell modulation.

Characteristics of mucosa-associated gut microbiota during treatment in Crohn's disease

BACKGROUND The dysbiosis of the gut microbiome is evident in Crohn's disease(CD) compared with healthy controls(HC), although the alterations from active CD to remission after treatment are unclear.AIM To characterize the mucosa-associated gut microbiota in patients with CD before and after the induction therapy.METHODS The basic information was collected from the subjects and the CD activity index(CDAI) was calculated in patients. A 16S rRNA sequencing approach was applied to determine the structures of microbial communities in mucosal samples including the terminal ileal, ascending colon, descending colon and rectum. The composition and function of mucosa-associated gut microbiota were compared between samples from the same cohort of patients before and after treatment.Differential taxa were identified to calculate the microbial dysbiosis index(MDI)and the correlation between MDI and CDAI was analyzed using Pearson correlation test. Predictive functional profiling of microbial communities was obtained with PICRUSt.RESULTS There were no significant differences in microbial richness among the four anatomical sites in individuals. Compared to active disease, the alpha diversity of CD in remission was increased towards the level of HC compared to the active stage. The principal coordinate analysis revealed that samples of active CD were clearly separated from those in remission, which clustered close to HC. Sixty-five genera were identified as differentially abundant between active and quiescent CD, with a loss of Fusobacterium and a gain of potential beneficial bacteria including Lactobacillus, Akkermansia, Roseburia, Ruminococcus and Lachnospira after the induction of remission. The combination of these taxa into a MDI showed a positive correlation with clinical disease severity and a negative correlation with species richness. The increased capacity for the inferred pathways including Lipopolysaccharide biosynthesis and Lipopolysaccharide biosynthesis proteins in patients before treatment negatively correlated with the abundance of Roseburia,Ruminococcus and Lachnospira.CONCLUSION The dysbiosis of mucosa-associated microbiota was associated with the disease phenotype and may become a potential diagnostic tool for the recurrence of disease.

Salmonella Typhimurium infection disrupts but continuous feeding of Bacillus based probiotic restores gut microbiota in infected hens

Background:The gut microbiota plays an important role in the colonisation resistance and invasion of pathogens.Salmonella Typhimurium has the potential to establish a niche by displacing the microbiota in the chicken gut causing continuous faecal shedding that can result in contaminated eggs or egg products.In the current study,we investigated the dynamics of gut microbiota in laying chickens during Salmonella Typhimurium infection.The optimisation of the use of an infeed probiotic supplement for restoration of gut microbial balance and reduction of Salmonella Typhimurium load was also investigated.Results:Salmonella infection caused dysbiosis by decreasing(FDR<0.05)the abundance of microbial genera,such as Blautia,Enorma,Faecalibacterium,Shuttleworthia,Sellimonas,Intestinimonas and Subdoligranulum and increasing the abundance of genera such as Butyricicoccus,Erysipelatoclostridium,Oscillibacter and Flavonifractor.The higher Salmonella Typhimurium load resulted in lower(P<0.05)abundance of genera such as Lactobacillus,Alistipes,Bifidobacterium,Butyricimonas,Faecalibacterium and Romboutsia suggesting Salmonella driven gut microbiota dysbiosis.Higher Salmonella load led to increased abundance of genera such as Caproiciproducens,Acetanaerobacterium,Akkermansia,Erysipelatoclostridium,Eisenbergiella,EscherichiaShigella and Flavonifractor suggesting a positive interaction of these genera with Salmonella in the displaced gut microbiota.Probiotic supplementation improved the gut microbiota by balancing the abundance of most of the genera displaced by the Salmonella challenge with clearer effects observed with continuous supplementation of the probiotic.The levels of acetate and butyrate in the faeces were not affected(P>0.05)by Salmonella challenge and the butyrate level was increased by the continuous feeding of the probiotic.Probiotic supplementation in Salmonella challenged chickens resulted in higher level of propionate.Continuous probiotic supplementation decreased(P<0.05)the overall mean load of Salmonella in faeces and had a significant effect on Salmonella load reduction in internal organs.Conclusions:Salmonella challenge negatively impacts the diversity and abundance of many gut microbial genera involved in important functions such as organic acid and vitamin production.Strategic feeding of a Bacillus based probiotic helps in restoring many of the microbial genera displaced by Salmonella Typhimurium challenge.

Gut mucosal microbiota profiles linked to colorectal cancer recurrence

BACKGROUND Emerging evidence links gut microbiota to various human diseases including colorectal cancer(CRC)initiation and development.However,gut microbiota profiles associated with CRC recurrence and patient prognosis are not completely understood yet,especially in a Chinese cohort.AIM To investigate the relationship between gut mucosal microbiota profiles and CRC recurrence and patient prognosis.METHODS We obtained the composition and structure of gut microbiota collected from 75 patients diagnosed with CRC and 26 healthy controls.The patients were followed up by regular examination to determine whether tumors recurred.Triplet-paired samples from on-tumor,adjacent-tumor and off-tumor sites of patients diagnosed with/without CRC recurrence were analyzed to assess spatial-specific patterns of gut mucosal microbiota by 16S ribosomal RNA sequencing.Next,we carried out bioinformatic analyses,Kaplan-Meier survival analyses and Cox regression analyses to determine the relationship between gut mucosal microbiota profiles and CRC recurrence and patient prognosis.RESULTS We observed spatial-specific patterns of gut mucosal microbiota profiles linked to CRC recurrence and patient prognosis.A total of 17 bacterial genera/families were identified as potential biomarkers for CRC recurrence and patient prognosis,including Anaerotruncus,Bacteroidales,Coriobacteriaceae,Dialister,Eubacterium,Fusobacterium,Filifactor,Gemella,Haemophilus,Mogibacteriazeae,Pyramidobacter,Parvimonas,Porphyromonadaceae,Slackia,Schwartzia,TG5 and Treponema.CONCLUSION Our work suggests that intestinal microbiota can serve as biomarkers to predict the risk of CRC recurrence and patient death.

Gut Microbiota Dysbiosis in Cafeteria Diet Fed Sprague Dawley Rats

Diet plays a major role in the body physiology and metabolism. The quantity, nature and stability of the macronutrients present in the diet have a major impact on the composition of gut microbiota. Gut microbiota plays a major role in the body metabolism and leads to obese or lean phenotype. Bacteriodetes, Firmicutes, Proteobacteria and Actinobacteria are the major microbes that inhabit in the region of the gut. We made an attempt to study the effects of Cafeteria (CAF) diets and normal chow diets on diet consumption, weight gain, metabolism and composition of gut microbiota in fecal and cecum samples from three weeks old Sprague Dawley (SD) rats (n = 18/group) using 16S rDNA high throughput sequencing. Results revealed that distinctive diet based phenotypical changes were observed in some of the Cafeteria diet fed rats. Interestingly, some weight gain resistant (WGR) animals in Cafeteria diet fed groups show similar trend like that of control normal chow fed rats. Fecal microbiome analysis indicates that the ratio of Bacteriodetes is higher than the Firmicutes in cecum samples of Cafeteria diet fed rats whereas no significant difference is found in fecal samples of Cafeteria diet fed rats and as well as in control rats. Further analysis of other taxa at the level of family and genus of microbial abundance are also discussed. Our study suggests that contribution of gut microbiota towards obesity is not at the phylum level, and microbiome composition even at the level of species or strain may exert impact on the metabolism of the Cafeteria diet.

Koumiss Consumption Alleviates Symptoms of Patients with Chronic Atrophic Gastritis:A Possible Link to Modulation of Gut Microbiota

Intestinal dysbiosis is closely related to a variety of medical conditions,especially gastrointestinal diseases.The present study aimed to investigate the effects of koumiss on chronic atrophic gastritis(CAG)in an outpatient clinical trial(n=10;all female subjects aged 41-55;body mass index ranging from 19.5 to 25.8).Each patient consumed three servings of koumiss per day(i.e.250 ml daily before each of 3 meals)for a 60-day period.The improvement of patients'symptoms was monitored by comparing the total scores of symptoms before and after the treatment.Meanwhile,the changes in the patients’fecal microbiota composition and specific blood parameters were determined.After the 60-day koumiss administration,significant symptom improvements were observed,as evidenced by the reduction of the total symptoms score,and changes in blood platelet and cholesterol levels.The changes in patients’fecal microbiota composition were found.The patients’fecal microbiota fell into two distinct enterotypes,Bacteroides dorei/Bacteroides uniformis(BB-enterotype)and Prevotella copri(P-enterotype).Significant less Bacteroides uniformis was found in the BB-enterotype patient group,while significant more butyrate-producing bacteria(e.g.Eubacterium rectale and Faecalibacterium prausnitzii)were found in the P-enterotype patient group,following koumiss administration.After stopping koumiss consumption,the relative abundance of some biomarker taxa returned to the original level,suggesting that the gut microbiota modulatory effect was not permanent and that continuous koumiss administration was required to maintain the therapeutic effect.In conclusion,koumiss consumption could alleviate the symptoms of CAG patients.Our results may help understand the mechanism of koumiss in alleviating CAG disease symptoms,facilitating the development of such products with desired therapeutic functions.

Associations of gut microbiota with dyslipidemia based on sex differences in subjects from Northwestern China

BACKGROUND The gut microbiota(GM)has been proven to play a role in the regulation of host lipid metabolism,which provides a new theory about the pathogenesis of dyslipidemia.However,the associations of GM with dyslipidemia based on sex differences remain unclear and warrant elucidation.AIM To investigate the associations of GM features with serum lipid profiles based on sex differences in a Chinese population.METHODS This study ultimately recruited 142 participants(73 females and 69 males)at Honghui Hospital,Xi’an Jiaotong University.The anthropometric and blood metabolic parameters of all participants were measured.According to their serum lipid levels,female and male participants were classified into a high triglyceride(H_TG)group,a high total cholesterol(H_CHO)group,a low high-density lipoprotein cholesterol(L_HDL-C)group,and a control(CON)group with normal serum lipid levels.Fresh fecal samples were collected for 16S rRNA gene sequencing.UPARSE software,QIIME software,the RDP classifier and the FAPROTAX database were used for sequencing analyses.RESULTS The GM composition at the phylum level included Firmicutes and Bacteroidetes as the core GM.Different GM features were identified between females and males,and the associations between GM and serum lipid profiles were different in females and males.The GM features in different dyslipidemia subgroups changed in both female patients and male patients.Proteobacteria,Lactobacillaceae,Lactobacillus and Lactobacillus_salivarius were enriched in H_CHO females compared with CON females,while Coriobacteriia were enriched in L_HDL-C females.In the comparison among the three dyslipidemia subgroups in females,Lactobacillus_salivarius were enriched in H_CHO females,and Prevotellaceae were enriched in L_HDL-C females.Compared with CON or H_TG males,Prevotellaceae,unidentified_Ruminococcaceae,Roseburia and Roseburia_inulinivorans were decreased in L_HDL-C males(P value<0.05),and linear discriminant analysis effect size analysis indicated an enrichment of the above GM taxa in H_TG males compared with other male subgroups.Additionally,Roseburia_inulinivorans abundance was positively correlated with serum TG and total cholesterol levels,and Roseburia were positively correlated with serum TG level.Furthermore,Proteobacteria(0.724,95%CI:0.567-0.849),Lactobacillaceae(0.703,95%CI:0.544-0.832),Lactobacillus(0.705,95%CI:0.547-0.834)and Lactobacillus_salivarius(0.706,95%CI:0.548-0.835)could distinguish H_CHO females from CON females,while Coriobacteriia(0.710,95%CI:0.547-0.841),Coriobacteriales(0.710,95%CI:0.547-0.841),Prevotellaceae(0.697,95%CI:0.534-0.830),Roseburia(0.697,95%CI:0.534-0.830)and Roseburia_inulinivorans(0.684,95%CI:0.520-0.820)could discriminate H_TG males from CON males.Based on the predictions of GM metabolic capabilities with the FAPROTAX database,a total of 51 functional assignments were obtained in females,while 38 were obtained in males.This functional prediction suggested that cellulolysis increased in L_HDL-C females compared with CON females,but decreased in L_HDL-C males compared with CON males.CONCLUSION This study indicates associations of GM with serum lipid profiles,supporting the notion that GM dysbiosis may participate in the pathogenesis of dyslipidemia,and sex differences should be considered.

Effect of gastrointestinal heat retention syndrome on gut microbiota in children with upper respiratory tract infection and lung-heat syndrome

Objective:Gastrointestinal heat retention syndrome(GHRS)is associated with lung-heat syndrome and is related to recurrent respiratory infection.Upper respiratory tract infection(URTI)lung heat syndrome is common in children.The study will explore the effect of GHRS on the structure and function of gut microbiota in children with URTI lung-heat syndrome.Methods:Participants were divided into both groups using the self-developed URTI scale and the“GHRS Diagnostic Scale$Pediatric Part”:GHRS-positive children(LS group)and GHRS-negative children(L group).General information,clinical symptoms,and stool were collected.We used 16S rRNA amplicon sequencing technology to determine the gene sequence of the V3eV4 region in feces and measure the gut microbiota of the both groups at the genus level.Results:A total of 23 children were included in the both groups.There were 12 cases in the LS group and 11 cases in the L group.There was no statistical difference between the both groups in age,gender,height,weight,and body mass index.The effective sequences shared by the both groups accounted for 85.66%of the total.In the gut microbiota,there was no difference in the a diversity and the b diversity between the both groups.Compared with the L group,the LS group had a significant increase in the relative abundance of the Ruminococcus gnavus group,Prevotella-9,Staphylococcus,and Actinomyces(P<.05).The functions of the both groups of microbiota primarily concentrate on metabolism,genetic information processing,and environmental information processing.The relative abundance of signaling molecules and interactions in the LS group were higher than that in the L group(P<.05).The redundancy analysis(RDA)showed that the URTI score had the greatest impact on the distribution of microbiota.Conclusion:GHRS may affect the development of URTI lung-heat syndrome by changing the relative abundances of gut microbiota.

极早产儿生后1个月肠道菌群的动态变化及分布特征

目的了解极早产儿生后1个月肠道菌群的动态变化及分布特征,为益生菌早期干预提供理论依据。方法采取前瞻性研究方法,选取2022年9月至2023年3月本院收治的极早产儿为研究对象,收集生后第7、14、21、28天的粪便标本行16 S rRNA高通量测序及生物信息分析。结果共纳入极早产儿35例,男22例,女13例,出生胎龄210±11天,出生体重1419±339 g,收集粪便样本140份。在门水平上检测到的优势菌群包括厚壁菌门、变形菌门,占80%以上;属水平上检测到的优势菌群主要以条件致病菌为主,包括埃希菌属、梭菌属、葡萄球菌属、不动杆菌属和克雷伯杆菌属,而双歧杆菌属相对丰度均<5%。关键菌群差异分析发现,门及属水平的差异菌群主要为拟杆菌门(P=0.029)、蓝藻菌门(P=0.011)及葡萄球菌属(P=0.010)、罗氏菌属(P=0.040)。肠道菌群的多样性分析发现,四个时间点Alpla多样性指数Ace值及Shannon值比较差异无统计学差异(P>0.05),而Chao值呈逐渐下降趋势(P=0.001);四个时间点的Beta多样性分析Weighted-unifrac值分别为0.412(0.281~0.493)、0.498(0.214~0.526)、0.428(0.289~0.490)、0.143(0.077~0.423),差异有统计学意义(P<0.001)。结论极早产儿生后7~28天肠道优势菌群从厚壁菌门逐渐转变为变形菌门,并以条件致病菌为主,而双歧杆菌属定植数量少,肠道菌群多样性呈下降趋势。