BACKGROUND Hepatocellular carcinoma(HCC),the predominant form of primary liver cancer,is a key contributor to cancer-related deaths globally.However,HCC diagnosis solely based on blood biochemical markers lacks both s...BACKGROUND Hepatocellular carcinoma(HCC),the predominant form of primary liver cancer,is a key contributor to cancer-related deaths globally.However,HCC diagnosis solely based on blood biochemical markers lacks both sensitivity and specificity.AIM To investigate alterations of the fecal metabolome and intestinal bacteria and reveal the correlations among differential metabolites,distinct bacteria,and serum indicators.METHODS To uncover potentially effective therapeutic targets for HCC,we utilized nontargeted liquid chromatography-mass spectrometry and high-throughput DNA sequencing targeting the 16S rRNA gene.This comprehensive approach allowed us to investigate the metabolome and microbial community structure of feces samples obtained from patients with HCC.Furthermore,we conducted an analysis to assess the interplay between the fecal metabolome and intestinal bacterial population.RESULTS In comparison to healthy controls,a notable overlap of 161 differential metabolites and 3 enriched Kyoto Encyclopedia of Genes and Genomes pathways was observed in the HCC12(comprising patients with stage I and II HCC)and HCC34 groups(comprising patients with stage III and IV HCC).Lachnospira,Streptococcus,and Veillonella had significant differences in abundance in patients with HCC.Notably,Streptococcus and Veillonella exhibited significant correlations with serum indicators such as alpha-fetoprotein(AFP).Meanwhile,several differential metabolites[e.g.,4-keto-2-undecylpyrroline,dihydrojasmonic acid,1,8-heptadecadiene-4,6-diyne-3,10-diol,9(S)-HOTrE]also exhibited significant correlations with serum indicators such asγ-glutamyl transferase,total bilirubin,AFP,aspartate aminotransferase,and albumin.Additionally,these two genera also had significant associations with differential metabolites such as 1,2-Dipentadecanoyl-rac-glycerol(15:0/20:0/0:0),arachidoyl ethanolamide,and 4-keto-2-undecylpyrroline.CONCLUSION Our results suggest that the metabolome of fecal samples and the composition of intestinal bacteria hold promise as potential biomarkers for HCC diagnosis.展开更多
Probiotics are live microbial food supplements that have been shown to have beneficial effects on animal health.Endogenous probiotic bacteria have long been used for their proposed health promoting properties and have...Probiotics are live microbial food supplements that have been shown to have beneficial effects on animal health.Endogenous probiotic bacteria have long been used for their proposed health promoting properties and have become a hot research topic in growth improvement in aquaculture.The endogenous probiotic bacteria from intestines of Macrobrachium rosenbergii(giant river prawn)was explored for their probiotic potential,from which 367 bacterial strains were isolated from the intestine of M.rosenbergii.After 16 S rDNA sequence analysis,234 isolates were identified as Lactococcus garvieae,which accounted for 63.76%of the total number of culturable intestinal bacteria,suggesting that this bacterium was the main component of the microbiota.Furthermore,to reveal the probiotic properties of L.garvieae,this isolated bacterial strain was characterized morphologically,physiologically,and biochemically.Its enzyme production capacity,bacteriostatic activity,and resistance to acid,high temperature,and pH,were assessed.In vitro experiments showed that the L.garvieae(No.C 6 a 2)had a fast growth rate and entered the logarithmic phase rapidly.Besides,it had characteristics of acid-production and resistance,enzyme-producing capacity,and strong antibacterial activity against pathogenic Staphylococc us aureus,Aeromonas hydrophila,and Aeromonas veronii.However,it lacked the ability to tolerate high temperature.Our results provide novel data to deepen our understanding of the intestinal bacteria structure of M.rosenbergii and valuable information for probiotic screening and the application for M.rosenbergii.展开更多
AIM: To explore the relationship between small intestinalmotility and small intestinal bacteria overgrowth(SIBO) in Nonalcoholic steatohepatitis (NASH), andto investigate the effect of SIBO on the pathogenesisof NASH ...AIM: To explore the relationship between small intestinalmotility and small intestinal bacteria overgrowth(SIBO) in Nonalcoholic steatohepatitis (NASH), andto investigate the effect of SIBO on the pathogenesisof NASH in rats. The effect of cidomycin in alleviatingseverity of NASH is also studied. METHODS: Forty eight rats were randomly dividedinto NASH group (n = 16), cidomycin group (n = 16)and control group (n = 16). Then each group weresubdivided into small intestinal motility group (n = 8),bacteria group (n = 8) respectively. A semi-solid coloredmarker was used for monitoring small intestinal transit.The proximal small intestine was harvested under sterilecondition and processed for quantitation for aerobes(E. coli) and anaerobes (Lactobacilli). Liver pathologicscore was calculated to qualify the severity of hepatitis.Serum ALT, AST levels were detected to evaluate theseverity of hepatitis. RESULTS: Small intestinal transit was inhibited inNASH group (P < 0.01). Rats treated with cidomycinhad higher small intestine transit rate than rats in NASHgroup (P < 0.01). High fat diet resulted in quantitativealterations in the aerobes (E. coli ) but not in theanoerobics (Lactobacill). There was an increase in thenumber of E. coli in the proximal small intestinal florain NASH group than in control group (1.70 ± 0.12 log10(CFU/g) vs 1.28 ± 0.07 log10 (CFU/g), P < 0.01). TNF-αconcentration was significantly higher in NASH groupthan in control group (1.13 ± 0.15 mmol/L vs 0.57 ±0.09 mmol/L, P < 0.01). TNF-α concentration was lowerin cidomycin group than in NASH group (0.63 ± 0.09mmol/L vs 1.13 ± 0.15 mmol/L, P < 0.01). Treatmentwith cidomycin showed its effect by significantly loweringserum ALT, AST and TNF-α levels of NASH rats. CONCLUSION: SIBO may decrease small intestinalmovement in NASH rats. SIBO may be an importantpathogenesis of Nash. And treatment with cidomycin by mouth can alleviate the severity of NASH.展开更多
Poliumoside is representative of phenylethanoid glycosides, which are widely found in many plants. Poliumoside is also regarded as the main active component of Callicarpa kwangtungensis Chun(CK), though its oral bioav...Poliumoside is representative of phenylethanoid glycosides, which are widely found in many plants. Poliumoside is also regarded as the main active component of Callicarpa kwangtungensis Chun(CK), though its oral bioavailability in rat is extremely low(0.69%) and its in vivo and in vitro metabolism has not yet been systematically investigated. In the present study, an ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UPLC/Q-TOF-MS) method was employed to identify the metabolites and investigate the metabolic pathways of poliumoside in rat after oral administration 1.5 g·kg–1 of poliumoside. As a result, a total of 34 metabolites(30 from urine, 17 from plasma, and 4 from bile) and 9 possible metabolic pathways(rearrangment, reduction, hydration, hydrolyzation, dehydration, methylation, hydroxylation, acetylation, and sulfation) were proposed in vivo. The main metabolite, acteoside, was quantified after incubated with rat intestinal bacteria in vitro. In conclusion, the present study systematically explored the metabolites of poliumoside in vivo and in vitro, proposing metabolic pathways that may be significant for further metabolic studies of poliumoside.展开更多
Objective:The differences of ovarian morphology,reproductive hormones,glucose and lipid metabolism and intestinal bacteria in rats with polycystic ovary syndrome(PCOS)induced by triazole were compared.Method:Eighteen ...Objective:The differences of ovarian morphology,reproductive hormones,glucose and lipid metabolism and intestinal bacteria in rats with polycystic ovary syndrome(PCOS)induced by triazole were compared.Method:Eighteen 21 SPF female SD rats were randomly divided into group A(3-week group),group B(5-week group)and group D(control group)by random number table.Group A received letrozole+CMC-Na mixture by gavage in the first 3 weeks and CMC-Na solution by gavage in the last 2 weeks,group B received letrozole+CMC-Na mixture by gavage for 5 weeks,and group D received CMC-Na solution by gavage for 5 weeks,and all three groups of rats were fed with normal diet.At the end of gavage,the body weight of rats in each group was observed,the histological changes of ovaries were observed by hematoxylin-eosin(HE)staining,the serum levels of estradiol(E2),follicle stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),total cholesterol(TC),triglyceride(TG),fasting blood glucose(Glu),fasting insulin(FINS)and lipopolysaccharide(LPS)were measured by enzyme-linked immunosorbent assay(ELISA),and the LH/FSH ratio and insulin resistance index(HOMA IR)were calculated;the intestinal bacteria of rats were detected by 16S rRNA technique.Result:1.Comparison of ovary histomorphology:Under light microscope,multiple luteum and oocytes were observed in mature follicles in group D,and granulosa cells were orderly arranged and multilayered,without cystic dilated follicles.There were no mature follicles in the ovarian tissues of group A and GROUP B.The follicles were irregular in structure and more cystic dilated follicles were visible.The number of granular cells in some follicles decreased or even disappeared.2.Comparison of sex hormone levels:compared with group D,T level in group B was significantly increased(P<0.001),and T level in group A had an upward trend(P>0.05);The LH/FSH levels in group A and B were significantly increased(P<0.001;P<0.001).Compared with group A,E2 in group B was significantly decreased(P<0.05)and T was significantly increased(P<0.01).3.Comparison of glucose and lipid metabolism levels:Compared with group D,TC levels in groups A and B were significantly increased(P<0.01;P<0.01).Compared with group A,TG in group B was significantly increased(P<0.05).There were no significant differences in Glu,FINS and HOMA-IR levels among all groups.4.Comparison of LPS levels:Compared with group D,the serum LPS levels of rats in groups A and B were significantly increased(P<0.001;P<0.01).5.Intestinal flora analysis and comparison:At the phylum level,compared with group D,the abundance of Firmicutes in group B increased(P<0.01),Firmicutes in group A showed an upward trend(P>0.05),and the abundance of Bacteroidetes in groups A and B decreased(P<0.05).At the genus level,compared with group D,Lactobacillus in group B increased(P<0.01).The results of LEfSe analysis showed that there were differences in the composition of various intestinal bacteria among the three groups(LDA>3).Conclusion:The phenotype of PCOS rats was related to the length of modeling,and the phenotypic characteristics of PCOS in rats at 5 weeks of modeling were more typical than those in rats at 3 weeks of modeling;PCOS can cause changes in intestinal flora,and the changes in the structure of intestinal flora between groups are related to different modeling duration.展开更多
AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. METH...AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. METHODS: Comparative quantification of 16S rDNAs from different intestinal bacterial components was used to quantify multiple intestinal bacteria. The 16S rDNAs of different bacteria were amplified simultaneously by cPCR. The LDR/LCR was examined to actualize the genotyping and quantification. Two beneficial (Bifidobacterium , Lactobacillus ) and three conditionally pathogenic bacteria (Enterococcus , Enterobacterium and Eubacterium ) were used in this detection. With cloned standard bacterial 16S rDNAs, standard curves were prepared to validate the quantitative relations between the ratio of original concentrations of two templates and the ratio ofthe fluorescence signals of their final ligation products. The internal controls were added to monitor the whole detection flow. The quantity ratio between two bacteria was tested. RESULTS: cPCR and LDR revealed obvious linear correlations with standard DNAs, but cPCR and LCR did not. In the sample test, the distributions of the quantity ratio between each two bacterial species were obtained. There were significant differences among these distributions in the total samples. But these distributions of quantity ratio of each two bacteria remained stable among groups divided by age or sex. CONCLUSION: The detection method in this study can be used to conduct multiple intestinal bacteria genotyping and quantification, and to monitor the human intestinal health status as well.展开更多
AIM To investigate the diversity of bacterial lactase genes in the intestinal contents of mice with antibiotics-induced diarrhea.METHODS Following 2 d of adaptive feeding, 12 specific pathogenfree Kunming mice were ra...AIM To investigate the diversity of bacterial lactase genes in the intestinal contents of mice with antibiotics-induced diarrhea.METHODS Following 2 d of adaptive feeding, 12 specific pathogenfree Kunming mice were randomly divided into the control group and model group. The mouse model of antibiotics-induced diarrhea was established by gastric perfusion with mixed antibiotics(23.33 m L·kg^(-1)·d^(-1)) composed of gentamicin sulfate and cephradine capsules administered for 5 days, and the control group was treated with an equal amount of sterile water. Contents of the jejunum and ileum were then collected and metagenomic DNA was extracted, after which analysis of bacterial lactase genes using operational taxonomic units(OTUs) was carried outafter amplification and sequencing.RESULTS OTUs were 871 and 963 in the model group and control group, respectively, and 690 of these were identical. There were significant differences in Chao1 and ACE indices between the two groups(P < 0.05). Principal component analysis, principal coordination analysis and nonmetric multidimensional scaling analyses showed that OTUs distribution in the control group was relatively intensive, and differences among individuals were small, while in the model group, they were widely dispersed and more diversified. Bacterial lactase genes from the intestinal contents of the control group were related to Proteobacteria, Actinobacteria, Firmicutes and unclassified bacteria. Of these, Proteobacteria was the most abundant phylum. In contrast, the bacterial population was less diverse and abundant in the model group, as the abundance of Bradyrhizobium sp. BTAi1, Agrobacterium sp. H13-3, Acidovorax sp. KKS102, Azoarcus sp. KH32 C and Aeromonas caviae was lower than that in the control group. In addition, of the known species, the control group and model group had their own unique genera, respectively.CONCLUSION Antibiotics reduce the diversity of bacterial lactase genes in the intestinal contents, decrease the abundance of lactase gene, change the lactase gene strains, and transform their structures.展开更多
AIM: To investigate the effects of psychological stress on small intestinal motility and bacteria and mucosa in mice, and to explore the relationship between small intestinal dysfunction and small intestinal motility ...AIM: To investigate the effects of psychological stress on small intestinal motility and bacteria and mucosa in mice, and to explore the relationship between small intestinal dysfunction and small intestinal motility and bacteria and mucosa under psychological stress. METHODS: Sixty mice were randomly divided into psychological stress group and control group. Each group were subdivided into small intestinal motility group (n= 10), bacteria group (n = 10), and D-xylose administered to stomach group (n= 10). An animal model with psychological stress was established housing the mice with a hungry cat in separate layers of a two-layer cage. A semi-solid colored marker (carbon-ink) was used for monitoring small intestinal transit. The proximal small intestine was harvested under sterile condition and processed for quantitation for aerobes (Escherichia coli) and anaerobes (Lactobacilli). The quantitation of bacteria was expressed as Iog10(colony forming units/g). D-xylose levels in plasma were measured for estimating trie damage of small intestinal mucosa. RESULTS: Small intestinal transit was inhibited (39.80±9.50% vs 58.79±11.47%,P<0.01) in mice after psychological stress, compared with the controls. Psychological stress resulted in quantitative alterations in the aerobes (E.coli). There was an increase in the number of E coli in the proximal small intestinal flora (1.78±0.30 log10(CFU/g) vs 1.37±0.21 log10(CFU/g), P<0.01), and there was decrease in relative proportion of Lactobacilli and E.coli of stressed mice (0.53±0.63 vs 1.14±1.07,P<0.05), while there was no significant difference in the anaerobes (Lactobacilli) between the two groups (2.31±0.70 log10 (CFU/g) vs 2.44±0.37 log10(CFU/g), P>0.05). D-xylose concentrations in plasma in psychological stress mice were significantly higher than those in the control group (2.90±0.89 mmol/L vs 0.97±0.33 mmol/L, P<0.01). CONCLUSION: Small intestinal dysfunction under psychological stress may be related to the small intestinal motility disorder and dysbacteriosis and the damage of mucosa probably caused by psychological stress.展开更多
In traditional Chinese medicine theory, Panax ginseng and Veratrum nigrum L. is an important incompatible herb pair. Studies on the content variation of main components and the influences on the metabolism in rat inte...In traditional Chinese medicine theory, Panax ginseng and Veratrum nigrum L. is an important incompatible herb pair. Studies on the content variation of main components and the influences on the metabolism in rat intestinal bacteria are useful to understand the mechanism of incompatibility of this herb pairs. In this study, the content variation of ginsenosides and their metaboltic profiles in the extracts of P. ginseng and compatibility of P. ginseng with V. nigrum L. （G-V） were investigated using relative quantitative method of electrospray ionization mass spec- trometry （ESI-MS） and UPLC-MSn, respectively. The relative contents of most ginsenosides were reduced in the extract of G-V. Furthermore, ginsenosides Rbt, Rb2, Rc and Rd could be metabolized to Rd, F2 and C-K in rat in- testinal bacteria. The metabolic speeds ofRbl, Rb2 and Re in the G-V extracts at ratios of 10 ： 5, 10 ： 7 and 10 ： 10 and the metabolic rates of ginsenosides Rbb Rb2 and Rc to Rd, Rd to F2 in all compatibility extracts were lower than that in the P ginseng extract. In conclusion, this study illustrated the mechanism of effect-reducing by comparison of the relative contents and metabolic profiles of ginsenosides after compatibility of P ginseng and V. nigrum L.展开更多
The extract of Radix aconiti was incubated with rat intestinal bacteria in vitro. Further, aconitine and hypaconitine standard were incubated at the same condition as the extract of Radix aconiti, respectively. Ultra ...The extract of Radix aconiti was incubated with rat intestinal bacteria in vitro. Further, aconitine and hypaconitine standard were incubated at the same condition as the extract of Radix aconiti, respectively. Ultra Performance Liquid Chromatography/Multi-Mass Spectrometry （UPLC/MSn） was used for detecting and identifying all the aco- nite alkaloids. Results showed that there were four metabolites which were identified as 8-butyryl-14-benzoylmesa- conine （m/z 660）, 8-propionyl-14-benzoylaconine （m/z 660）, 8-butyryl-14-benzoylaconine （m/z 674） and 8-valeryl-14-benzoylmesaconine （m/z 674） in the metabolized sample of the extract of Radix aconiti. The relative area ratio of them presented increasing trend during 6 d. On the basis of all results, we could concluded that substi- tution at N atom mainly influenced the metabolizing rate of diester-diterpenoid alkaloids （DDAs）, C-8 substitute was active metabolized site, intestinal bacterial metabolites of the aconite alkaloids in the extract of Radix aconiti were mainly the substitute with propionyl group, butyl group or valeryl group at C-8. This paper illustrated holistic metabolizing profile of the extract of Radix aconiti in vitro and possible metabolizing reaction type of main DDAs, which could provide reference for finding out potential bioactive components in the extract and the prescription of Chinese Medicine.展开更多
Objective To investigate the metabolic routes and metabolites of Rehmannia glutinosa and Cornus officinalis herb pair produced by gut microbiome from rats.Methods A rapid and sensitive ultra-performance liquid chromat...Objective To investigate the metabolic routes and metabolites of Rehmannia glutinosa and Cornus officinalis herb pair produced by gut microbiome from rats.Methods A rapid and sensitive ultra-performance liquid chromatography/quadrupole-time-offlight mass spectrometry（UPLC-Q-TOF/MS） technique combined with Metabolynx？software was established and successfully applied to identify the metabolites of the main bioactive components in the herb pair extract by rat intestinal bacteria.Results Four parent compounds（loganin,morroniside,catalpol,and acteoside） and their eight corresponding metabolites were detected and tentatively identified by the characteristics of their protonated ions.Hydrogenated and demethylated loganetin,dehydroxylated morronisid aglycone,caffeic acid,and its methylated product were the main metabolites.These metabolites suggested that the glycosides were firstly hydrolyzed to their aglycones by hydrolytic enzymes of the enteric microbial flora and subsequently to the other metabolites through hydrogenation,（de）-methylation,and de-hydroxylation.Conclusion The results may be helpful for the further investigation of the pharmacokinetic study of R.glutinosa and C.officinalis herb pair in vivo.展开更多
[ Objective ] The paper was to screen intestinal aerobic bacteria from 2nd instar larvae of Dendrolimu kikuchii and determine its toxicity. [ Method ] The intestinal samples of 2nd instar larvae of D. kikuchii were di...[ Objective ] The paper was to screen intestinal aerobic bacteria from 2nd instar larvae of Dendrolimu kikuchii and determine its toxicity. [ Method ] The intestinal samples of 2nd instar larvae of D. kikuchii were diluted in gredient and coated on plates to isolate strains, and a total of 5 strains of aerobic bacteria were isolated. With bacterial genomic DNA as template, the universal primers of bacterial 16S rDNA （27f and 1 492r） were used to amplify the template, 4 restriction enzymes Hae Ⅲ and Hind Ⅲ, Hinf Ⅰ and Taq Ⅰ were used to carry out ARDRA polymorphism analysis on PCR products. [ Result] Clustering map analysis showed that 5 strains of aerobic bacteria clustered into two different taxonomic operating units （OTU） in 95% similarity level, this indicated that the genetic diversity level of intestinal aerobic bacteria in 2nd instar larvae of D. kikuchii was relatively low. The indoor toxicity test showed that the death peaks of insects killed by intestinal bacteria were during 4 - 10 d, the insecticidal effect of strain 4 was the best with corrected mortality rate of 53.57% at 12 d. [ Conclusion] The paper provided reference for control of D. kikuchii.展开更多
Objective In order to use facultative gut bacteria as an alternate to animals for the initial gastrointestinal toxicity screening of heavy metals, a comparative study on rat intestinal epithelial cells and resident gu...Objective In order to use facultative gut bacteria as an alternate to animals for the initial gastrointestinal toxicity screening of heavy metals, a comparative study on rat intestinal epithelial cells and resident gut bacteria was undertaken. Methods in vitro growth rate of four gut bacteria, dehydrogenase （DHA） and esterase （EA） activity test, intestinal epithelial and bacterial cell membrane enzymes and in situ effect of arsenite were analysed. Results Growth profile of mixed resident population of gut bacteria and pure isolates of Escherichia coli, Pseudomonas sp., Lactobacillus sp., and Staphylococcus sp. revealed an arsenite （2-20 ppm） concentration-dependent inhibition. The viability pattern of epithelial cells also showed similar changes. DHA and EA tests revealed significant inhibition （40%-72%） with arsenite exposure of 5 and 10 ppm in isolated gut bacteria and epithelial cells. Decrease in membrane alkaline phosphatase and Ca^2＋-Mg^2＋-ATPase activities was in the range of 33%-55% in four bacteria at the arsenite exposure of 10 ppm, whereas it was 60%-65% in intestinal epithelial villus cells, in situ incubation of arsenite using intestinal loops also showed more or less similar changes in membrane enzymes of resident gut bacterial population and epithelial cells. Conclusion The results indicate that facultative gut bacteria can be used as suitable in vitro model for the preliminary screening of arsenical gastrointestinal cytotoxic effects.展开更多
Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the...Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear crylAh gene （800 ng mL^-1） and supercoiled plasmid DNA （800 ng mL^-1） under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon＇s index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic crylAh maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.展开更多
Intestinal bacteria play a role in the development of non-steroidal anti-inflammatory drugs （NSAID）-induced small intestinal injury. Agents such as probiotics, able to modi~ the gut ecology, might theoretically be u...Intestinal bacteria play a role in the development of non-steroidal anti-inflammatory drugs （NSAID）-induced small intestinal injury. Agents such as probiotics, able to modi~ the gut ecology, might theoretically be useful in preventing small intestinal damage induced by NSAIDs. The clinical studies available so far do suggest that some probiotic agents can be effective in this respect.展开更多
In vitro models of human colon carcinoma cell line(Caco-2 cell monolayer) and human intestinal bacteria were used to investigate the intestinal transport and biotransformation of resibufogenin and cinobufagin in Chan ...In vitro models of human colon carcinoma cell line(Caco-2 cell monolayer) and human intestinal bacteria were used to investigate the intestinal transport and biotransformation of resibufogenin and cinobufagin in Chan Su by HPLC/APCI-MSn. The experimental results of Caco-2 cell monolayer demonstrate that the apparent permeability coefficients(Papp) of resibufogenin and cinobufagin are higher than 10–6 cm/s, which indicates that both resibufogenin and cinobufagin have a good absorption in the small intestine. And the biotransformation result of human intestinal bacteria shows that resibufogenin has been transformed to 3-epiresibufogenin and cinobufagin has been transformed to 3-epicinobufagin, deacetylcinobufagin and 3-epideacetycinobufagin, respectively.展开更多
The biosurfactants rhamnolipids and the “soil ecosystem engineers” earthworms are often used to remediate contaminated soils. However, the effects of rhamnolipids on earthworm intestinal flora and microbial communit...The biosurfactants rhamnolipids and the “soil ecosystem engineers” earthworms are often used to remediate contaminated soils. However, the effects of rhamnolipids on earthworm intestinal flora and microbial community in soil containing earthworms are not clearly understood. In our study, a 21-d microcosm experiment was carried out to reveal the effects of rhamnolipids on microbial abundance, composition, and metabolism, as well as contaminant degradation capacity. Both rhamnolipids and earthworms had positive effects on soil bacteria. Rhamnolipid-amended soil(RT) showed higher bacterial abundance and metabolic activity than earthworm-amended soil(ET), while the improvement in bacterial composition and contaminant degradation capacity by rhamnolipids was lower than that by earthworms. Notably, these effects were further amplified by the combined treatment of rhamnolipids and earthworms(RET). Specifically, the bacterial abundance(log-transferred) increased from 9.5 copies g-1in the control with no addition to 10.3, 10.6, and 11.1 copies g-1in ET, RT, and RET, respectively. Compared to ET, the relative abundance of the dominant phylum, Proteobacteria, increased from 41.66% to 51.67% in RET,and more pollutant-degrading bacteria were also enriched in RET. Therefore, the increases in bacterial abundance and contaminant-degrading bacteria led to the following ranking of soil dioxin removal rate: RET(77.28%) > ET(59.83%) > RT(24.65%) > control(4.71%). Moreover, the addition of rhamnolipids enhanced the abundance of bacterial functional genes involved in metabolism and environmental information processing. In addition, the composition and diversity of bacteria in the gut of earthworms were conspicuously affected by rhamnolipids, and the relative abundance of Microbacterium and Shewanella increased significantly(P < 0.05). Therefore, this study revealed that rhamnolipids remarkably influenced the abundance, composition, and metabolism of the microbial community in earthworm gut, further promoting the degradation rate of dioxin, providing theoretical support for optimizing the combined application of rhamnolipids and earthworms in soil bioremediation engineering and for the assessment of the ecological impact of rhamnolipids.展开更多
Lactic acid bacteria(LAB)are a representative probiotic.As the dominant flora in the human intestinal tract,LAB can regulate the balance of human intestinal flora and improve host health.The purpose of this study was ...Lactic acid bacteria(LAB)are a representative probiotic.As the dominant flora in the human intestinal tract,LAB can regulate the balance of human intestinal flora and improve host health.The purpose of this study was to isolate and screen LAB that are well suited to the intestinal characteristics of the Chinese population,with excellent probiotics and high antibacterial activity.After 16S ribosomal RNA(rRNA)homology and phylogenetic tree analysis,potential probiotics were tested for their antibacterial activity,resistance to artificial gastrointestinal fluid and drugs,surface hydrophobicity,and safety.Three strains of LAB with acid resistance,bile salt resistance,epithelial cell adhesion,and no multidrug resistance were selected:Lactobacillus salivarius,Leuconostoc lactis,and Lactobacillus paracasei.Analysis of the antibacterial active substances in the three strains and their fermentation broths revealed that the main antibacterial substances of L.lactis were organic acids,whereas those of L.salivarius and L.paracasei were organic acids and bacteriocins with broad-spectrum antibacterial activity.These three strains of probiotic LAB with high antibacterial activity were identified as bacterial resources that could potentially be used to develop probiotic preparations for the prevention and treatment of intestinal diseases caused by intestinal pathogens.展开更多
BACKGROUND While colorectal polyps are not cancerous,some types of polyps,known as adenomas,can develop into colorectal cancer over time.Polyps can often be found and removed by colonoscopy;however,this is an invasive...BACKGROUND While colorectal polyps are not cancerous,some types of polyps,known as adenomas,can develop into colorectal cancer over time.Polyps can often be found and removed by colonoscopy;however,this is an invasive and expensive test.Thus,there is a need for new methods of screening patients at high risk of developing polyps.AIM To identify a potential association between colorectal polyps and small intestine bacteria overgrowth(SIBO)or other relevant factors in a patient cohort with lactulose breath test(LBT)results.METHODS A total of 382 patients who had received an LBT were classified into polyp and non-polyp groups that were confirmed by colonoscopy and pathology.SIBO was diagnosed by measuring LBTderived hydrogen(H)and methane(M)levels according to 2017 North American Consensus recommendations.Logistic regression was used to assess the ability of LBT to predict colorectal polyps.Intestinal barrier function damage(IBFD)was determined by blood assays.RESULTS H and M levels revealed that the prevalence of SIBO was significantly higher in the polyp group than in the non-polyp group(41%vs 23%,P<0.01;71%vs 59%,P<0.05,respectively).Within 90 min of lactulose ingestion,the peak H values in the adenomatous and inflammatory/hyperplastic polyp patients were significantly higher than those in the non-polyp group(P<0.01,and P=0.03,respectively).In 227 patients with SIBO defined by combining H and M values,the rate of IBFD determined by blood lipopolysaccharide levels was significantly higher among patients with polyps than those without(15%vs 5%,P<0.05).In regression analysis with age and gender adjustment,colorectal polyps were most accurately predicted with models using M peak values or combined H and M values limited by North American Consensus recommendations for SIBO.These models had a sensitivity of≥0.67,a specificity of≥0.64,and an accuracy of≥0.66.CONCLUSION The current study made key associations among colorectal polyps,SIBO,and IBFD and demonstrated that LBT has moderate potential as an alternative noninvasive screening tool for colorectal polyps.展开更多
BACKGROUND The pathogenesis of gastroesophageal reflux disease(GERD)is closely associated with the intestinal bacteria composition and their metabolites.AIM To investigate whether washed microbiota transplantation(WMT...BACKGROUND The pathogenesis of gastroesophageal reflux disease(GERD)is closely associated with the intestinal bacteria composition and their metabolites.AIM To investigate whether washed microbiota transplantation(WMT)improves symptoms of nonerosive reflux disease(NERD)with proton pump inhibitor(PPI)dependency.METHODS Patients with recurrent NERD and PPI dependency at the First Affiliated Hospital of Guangdong Pharmaceutical University from 2017 to 2018 were included and divided into a WMT or PPI group treated with PPI with/without WMT.The endpoint was NERD symptom frequency evaluated 1 mo after WMT using reflux disease questionnaire(RDQ)and GERD questionnaire(GERDQ)scores,remission time,PPI dose,and the examination of intestinal mucosal barrier function.RESULTS In the WMT(n=15)and PPI(n=12)groups,the total remission rate at 1 mo after treatment was 93.3%vs 41.7%.Compared with the PPI group,the WMT group showed better results in GERDQ(P=0.004)and RDQ(P=0.003)and in remission months(8 vs 2,P=0.002).The PPI dose was reduced to some extent for 80%of patients in the WMT group and 33.3%in the PPI group.In 24 patients,intestinal mucosal barrier function was examined before treatment,and changes in the degree of damage were observed in 13 of these patients after treatment.Only one of the 15 patients had minor side effects,including a mushy stool two or three times a day,which resolved on their own after 1 wk.CONCLUSION This study is the first to demonstrate that WMT may be safe and effective for relieving NERD symptoms and reducing PPI dependency and recurrence.展开更多
基金Supported by the Department of Science and Technology of Shanxi Province,No.20210302124369the Health Commission of Shanxi Province,No.2021116Shanxi Administration of Traditional Chinese Medicine,No.2024ZYY2C054.
文摘BACKGROUND Hepatocellular carcinoma(HCC),the predominant form of primary liver cancer,is a key contributor to cancer-related deaths globally.However,HCC diagnosis solely based on blood biochemical markers lacks both sensitivity and specificity.AIM To investigate alterations of the fecal metabolome and intestinal bacteria and reveal the correlations among differential metabolites,distinct bacteria,and serum indicators.METHODS To uncover potentially effective therapeutic targets for HCC,we utilized nontargeted liquid chromatography-mass spectrometry and high-throughput DNA sequencing targeting the 16S rRNA gene.This comprehensive approach allowed us to investigate the metabolome and microbial community structure of feces samples obtained from patients with HCC.Furthermore,we conducted an analysis to assess the interplay between the fecal metabolome and intestinal bacterial population.RESULTS In comparison to healthy controls,a notable overlap of 161 differential metabolites and 3 enriched Kyoto Encyclopedia of Genes and Genomes pathways was observed in the HCC12(comprising patients with stage I and II HCC)and HCC34 groups(comprising patients with stage III and IV HCC).Lachnospira,Streptococcus,and Veillonella had significant differences in abundance in patients with HCC.Notably,Streptococcus and Veillonella exhibited significant correlations with serum indicators such as alpha-fetoprotein(AFP).Meanwhile,several differential metabolites[e.g.,4-keto-2-undecylpyrroline,dihydrojasmonic acid,1,8-heptadecadiene-4,6-diyne-3,10-diol,9(S)-HOTrE]also exhibited significant correlations with serum indicators such asγ-glutamyl transferase,total bilirubin,AFP,aspartate aminotransferase,and albumin.Additionally,these two genera also had significant associations with differential metabolites such as 1,2-Dipentadecanoyl-rac-glycerol(15:0/20:0/0:0),arachidoyl ethanolamide,and 4-keto-2-undecylpyrroline.CONCLUSION Our results suggest that the metabolome of fecal samples and the composition of intestinal bacteria hold promise as potential biomarkers for HCC diagnosis.
基金Supported by the National Natural Science Foundation of China(No.32273121)the Natural Science Foundation of Zhejiang Province(No.LGN22C190019)+1 种基金the Huzhou Natural Science Foundation(No.2021YZ08)the earmarked fund for the China Agriculture Research System of MOF and MARA(No.CARS-48)。
文摘Probiotics are live microbial food supplements that have been shown to have beneficial effects on animal health.Endogenous probiotic bacteria have long been used for their proposed health promoting properties and have become a hot research topic in growth improvement in aquaculture.The endogenous probiotic bacteria from intestines of Macrobrachium rosenbergii(giant river prawn)was explored for their probiotic potential,from which 367 bacterial strains were isolated from the intestine of M.rosenbergii.After 16 S rDNA sequence analysis,234 isolates were identified as Lactococcus garvieae,which accounted for 63.76%of the total number of culturable intestinal bacteria,suggesting that this bacterium was the main component of the microbiota.Furthermore,to reveal the probiotic properties of L.garvieae,this isolated bacterial strain was characterized morphologically,physiologically,and biochemically.Its enzyme production capacity,bacteriostatic activity,and resistance to acid,high temperature,and pH,were assessed.In vitro experiments showed that the L.garvieae(No.C 6 a 2)had a fast growth rate and entered the logarithmic phase rapidly.Besides,it had characteristics of acid-production and resistance,enzyme-producing capacity,and strong antibacterial activity against pathogenic Staphylococc us aureus,Aeromonas hydrophila,and Aeromonas veronii.However,it lacked the ability to tolerate high temperature.Our results provide novel data to deepen our understanding of the intestinal bacteria structure of M.rosenbergii and valuable information for probiotic screening and the application for M.rosenbergii.
文摘AIM: To explore the relationship between small intestinalmotility and small intestinal bacteria overgrowth(SIBO) in Nonalcoholic steatohepatitis (NASH), andto investigate the effect of SIBO on the pathogenesisof NASH in rats. The effect of cidomycin in alleviatingseverity of NASH is also studied. METHODS: Forty eight rats were randomly dividedinto NASH group (n = 16), cidomycin group (n = 16)and control group (n = 16). Then each group weresubdivided into small intestinal motility group (n = 8),bacteria group (n = 8) respectively. A semi-solid coloredmarker was used for monitoring small intestinal transit.The proximal small intestine was harvested under sterilecondition and processed for quantitation for aerobes(E. coli) and anaerobes (Lactobacilli). Liver pathologicscore was calculated to qualify the severity of hepatitis.Serum ALT, AST levels were detected to evaluate theseverity of hepatitis. RESULTS: Small intestinal transit was inhibited inNASH group (P < 0.01). Rats treated with cidomycinhad higher small intestine transit rate than rats in NASHgroup (P < 0.01). High fat diet resulted in quantitativealterations in the aerobes (E. coli ) but not in theanoerobics (Lactobacill). There was an increase in thenumber of E. coli in the proximal small intestinal florain NASH group than in control group (1.70 ± 0.12 log10(CFU/g) vs 1.28 ± 0.07 log10 (CFU/g), P < 0.01). TNF-αconcentration was significantly higher in NASH groupthan in control group (1.13 ± 0.15 mmol/L vs 0.57 ±0.09 mmol/L, P < 0.01). TNF-α concentration was lowerin cidomycin group than in NASH group (0.63 ± 0.09mmol/L vs 1.13 ± 0.15 mmol/L, P < 0.01). Treatmentwith cidomycin showed its effect by significantly loweringserum ALT, AST and TNF-α levels of NASH rats. CONCLUSION: SIBO may decrease small intestinalmovement in NASH rats. SIBO may be an importantpathogenesis of Nash. And treatment with cidomycin by mouth can alleviate the severity of NASH.
文摘Poliumoside is representative of phenylethanoid glycosides, which are widely found in many plants. Poliumoside is also regarded as the main active component of Callicarpa kwangtungensis Chun(CK), though its oral bioavailability in rat is extremely low(0.69%) and its in vivo and in vitro metabolism has not yet been systematically investigated. In the present study, an ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UPLC/Q-TOF-MS) method was employed to identify the metabolites and investigate the metabolic pathways of poliumoside in rat after oral administration 1.5 g·kg–1 of poliumoside. As a result, a total of 34 metabolites(30 from urine, 17 from plasma, and 4 from bile) and 9 possible metabolic pathways(rearrangment, reduction, hydration, hydrolyzation, dehydration, methylation, hydroxylation, acetylation, and sulfation) were proposed in vivo. The main metabolite, acteoside, was quantified after incubated with rat intestinal bacteria in vitro. In conclusion, the present study systematically explored the metabolites of poliumoside in vivo and in vitro, proposing metabolic pathways that may be significant for further metabolic studies of poliumoside.
基金National Natural Science Foundation of China(No.81674011)。
文摘Objective:The differences of ovarian morphology,reproductive hormones,glucose and lipid metabolism and intestinal bacteria in rats with polycystic ovary syndrome(PCOS)induced by triazole were compared.Method:Eighteen 21 SPF female SD rats were randomly divided into group A(3-week group),group B(5-week group)and group D(control group)by random number table.Group A received letrozole+CMC-Na mixture by gavage in the first 3 weeks and CMC-Na solution by gavage in the last 2 weeks,group B received letrozole+CMC-Na mixture by gavage for 5 weeks,and group D received CMC-Na solution by gavage for 5 weeks,and all three groups of rats were fed with normal diet.At the end of gavage,the body weight of rats in each group was observed,the histological changes of ovaries were observed by hematoxylin-eosin(HE)staining,the serum levels of estradiol(E2),follicle stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),total cholesterol(TC),triglyceride(TG),fasting blood glucose(Glu),fasting insulin(FINS)and lipopolysaccharide(LPS)were measured by enzyme-linked immunosorbent assay(ELISA),and the LH/FSH ratio and insulin resistance index(HOMA IR)were calculated;the intestinal bacteria of rats were detected by 16S rRNA technique.Result:1.Comparison of ovary histomorphology:Under light microscope,multiple luteum and oocytes were observed in mature follicles in group D,and granulosa cells were orderly arranged and multilayered,without cystic dilated follicles.There were no mature follicles in the ovarian tissues of group A and GROUP B.The follicles were irregular in structure and more cystic dilated follicles were visible.The number of granular cells in some follicles decreased or even disappeared.2.Comparison of sex hormone levels:compared with group D,T level in group B was significantly increased(P<0.001),and T level in group A had an upward trend(P>0.05);The LH/FSH levels in group A and B were significantly increased(P<0.001;P<0.001).Compared with group A,E2 in group B was significantly decreased(P<0.05)and T was significantly increased(P<0.01).3.Comparison of glucose and lipid metabolism levels:Compared with group D,TC levels in groups A and B were significantly increased(P<0.01;P<0.01).Compared with group A,TG in group B was significantly increased(P<0.05).There were no significant differences in Glu,FINS and HOMA-IR levels among all groups.4.Comparison of LPS levels:Compared with group D,the serum LPS levels of rats in groups A and B were significantly increased(P<0.001;P<0.01).5.Intestinal flora analysis and comparison:At the phylum level,compared with group D,the abundance of Firmicutes in group B increased(P<0.01),Firmicutes in group A showed an upward trend(P>0.05),and the abundance of Bacteroidetes in groups A and B decreased(P<0.05).At the genus level,compared with group D,Lactobacillus in group B increased(P<0.01).The results of LEfSe analysis showed that there were differences in the composition of various intestinal bacteria among the three groups(LDA>3).Conclusion:The phenotype of PCOS rats was related to the length of modeling,and the phenotypic characteristics of PCOS in rats at 5 weeks of modeling were more typical than those in rats at 3 weeks of modeling;PCOS can cause changes in intestinal flora,and the changes in the structure of intestinal flora between groups are related to different modeling duration.
文摘AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. METHODS: Comparative quantification of 16S rDNAs from different intestinal bacterial components was used to quantify multiple intestinal bacteria. The 16S rDNAs of different bacteria were amplified simultaneously by cPCR. The LDR/LCR was examined to actualize the genotyping and quantification. Two beneficial (Bifidobacterium , Lactobacillus ) and three conditionally pathogenic bacteria (Enterococcus , Enterobacterium and Eubacterium ) were used in this detection. With cloned standard bacterial 16S rDNAs, standard curves were prepared to validate the quantitative relations between the ratio of original concentrations of two templates and the ratio ofthe fluorescence signals of their final ligation products. The internal controls were added to monitor the whole detection flow. The quantity ratio between two bacteria was tested. RESULTS: cPCR and LDR revealed obvious linear correlations with standard DNAs, but cPCR and LCR did not. In the sample test, the distributions of the quantity ratio between each two bacterial species were obtained. There were significant differences among these distributions in the total samples. But these distributions of quantity ratio of each two bacteria remained stable among groups divided by age or sex. CONCLUSION: The detection method in this study can be used to conduct multiple intestinal bacteria genotyping and quantification, and to monitor the human intestinal health status as well.
基金Supported by the National Natural Science Foundation of China,No.81573951
文摘AIM To investigate the diversity of bacterial lactase genes in the intestinal contents of mice with antibiotics-induced diarrhea.METHODS Following 2 d of adaptive feeding, 12 specific pathogenfree Kunming mice were randomly divided into the control group and model group. The mouse model of antibiotics-induced diarrhea was established by gastric perfusion with mixed antibiotics(23.33 m L·kg^(-1)·d^(-1)) composed of gentamicin sulfate and cephradine capsules administered for 5 days, and the control group was treated with an equal amount of sterile water. Contents of the jejunum and ileum were then collected and metagenomic DNA was extracted, after which analysis of bacterial lactase genes using operational taxonomic units(OTUs) was carried outafter amplification and sequencing.RESULTS OTUs were 871 and 963 in the model group and control group, respectively, and 690 of these were identical. There were significant differences in Chao1 and ACE indices between the two groups(P < 0.05). Principal component analysis, principal coordination analysis and nonmetric multidimensional scaling analyses showed that OTUs distribution in the control group was relatively intensive, and differences among individuals were small, while in the model group, they were widely dispersed and more diversified. Bacterial lactase genes from the intestinal contents of the control group were related to Proteobacteria, Actinobacteria, Firmicutes and unclassified bacteria. Of these, Proteobacteria was the most abundant phylum. In contrast, the bacterial population was less diverse and abundant in the model group, as the abundance of Bradyrhizobium sp. BTAi1, Agrobacterium sp. H13-3, Acidovorax sp. KKS102, Azoarcus sp. KH32 C and Aeromonas caviae was lower than that in the control group. In addition, of the known species, the control group and model group had their own unique genera, respectively.CONCLUSION Antibiotics reduce the diversity of bacterial lactase genes in the intestinal contents, decrease the abundance of lactase gene, change the lactase gene strains, and transform their structures.
文摘AIM: To investigate the effects of psychological stress on small intestinal motility and bacteria and mucosa in mice, and to explore the relationship between small intestinal dysfunction and small intestinal motility and bacteria and mucosa under psychological stress. METHODS: Sixty mice were randomly divided into psychological stress group and control group. Each group were subdivided into small intestinal motility group (n= 10), bacteria group (n = 10), and D-xylose administered to stomach group (n= 10). An animal model with psychological stress was established housing the mice with a hungry cat in separate layers of a two-layer cage. A semi-solid colored marker (carbon-ink) was used for monitoring small intestinal transit. The proximal small intestine was harvested under sterile condition and processed for quantitation for aerobes (Escherichia coli) and anaerobes (Lactobacilli). The quantitation of bacteria was expressed as Iog10(colony forming units/g). D-xylose levels in plasma were measured for estimating trie damage of small intestinal mucosa. RESULTS: Small intestinal transit was inhibited (39.80±9.50% vs 58.79±11.47%,P<0.01) in mice after psychological stress, compared with the controls. Psychological stress resulted in quantitative alterations in the aerobes (E.coli). There was an increase in the number of E coli in the proximal small intestinal flora (1.78±0.30 log10(CFU/g) vs 1.37±0.21 log10(CFU/g), P<0.01), and there was decrease in relative proportion of Lactobacilli and E.coli of stressed mice (0.53±0.63 vs 1.14±1.07,P<0.05), while there was no significant difference in the anaerobes (Lactobacilli) between the two groups (2.31±0.70 log10 (CFU/g) vs 2.44±0.37 log10(CFU/g), P>0.05). D-xylose concentrations in plasma in psychological stress mice were significantly higher than those in the control group (2.90±0.89 mmol/L vs 0.97±0.33 mmol/L, P<0.01). CONCLUSION: Small intestinal dysfunction under psychological stress may be related to the small intestinal motility disorder and dysbacteriosis and the damage of mucosa probably caused by psychological stress.
文摘In traditional Chinese medicine theory, Panax ginseng and Veratrum nigrum L. is an important incompatible herb pair. Studies on the content variation of main components and the influences on the metabolism in rat intestinal bacteria are useful to understand the mechanism of incompatibility of this herb pairs. In this study, the content variation of ginsenosides and their metaboltic profiles in the extracts of P. ginseng and compatibility of P. ginseng with V. nigrum L. （G-V） were investigated using relative quantitative method of electrospray ionization mass spec- trometry （ESI-MS） and UPLC-MSn, respectively. The relative contents of most ginsenosides were reduced in the extract of G-V. Furthermore, ginsenosides Rbt, Rb2, Rc and Rd could be metabolized to Rd, F2 and C-K in rat in- testinal bacteria. The metabolic speeds ofRbl, Rb2 and Re in the G-V extracts at ratios of 10 ： 5, 10 ： 7 and 10 ： 10 and the metabolic rates of ginsenosides Rbb Rb2 and Rc to Rd, Rd to F2 in all compatibility extracts were lower than that in the P ginseng extract. In conclusion, this study illustrated the mechanism of effect-reducing by comparison of the relative contents and metabolic profiles of ginsenosides after compatibility of P ginseng and V. nigrum L.
文摘The extract of Radix aconiti was incubated with rat intestinal bacteria in vitro. Further, aconitine and hypaconitine standard were incubated at the same condition as the extract of Radix aconiti, respectively. Ultra Performance Liquid Chromatography/Multi-Mass Spectrometry （UPLC/MSn） was used for detecting and identifying all the aco- nite alkaloids. Results showed that there were four metabolites which were identified as 8-butyryl-14-benzoylmesa- conine （m/z 660）, 8-propionyl-14-benzoylaconine （m/z 660）, 8-butyryl-14-benzoylaconine （m/z 674） and 8-valeryl-14-benzoylmesaconine （m/z 674） in the metabolized sample of the extract of Radix aconiti. The relative area ratio of them presented increasing trend during 6 d. On the basis of all results, we could concluded that substi- tution at N atom mainly influenced the metabolizing rate of diester-diterpenoid alkaloids （DDAs）, C-8 substitute was active metabolized site, intestinal bacterial metabolites of the aconite alkaloids in the extract of Radix aconiti were mainly the substitute with propionyl group, butyl group or valeryl group at C-8. This paper illustrated holistic metabolizing profile of the extract of Radix aconiti in vitro and possible metabolizing reaction type of main DDAs, which could provide reference for finding out potential bioactive components in the extract and the prescription of Chinese Medicine.
基金National Natural Science Foundation of China(No.81072996,81102743)Priority Academic Programs Development of Jiangsu Higher Education Institutions(PAPD)
文摘Objective To investigate the metabolic routes and metabolites of Rehmannia glutinosa and Cornus officinalis herb pair produced by gut microbiome from rats.Methods A rapid and sensitive ultra-performance liquid chromatography/quadrupole-time-offlight mass spectrometry（UPLC-Q-TOF/MS） technique combined with Metabolynx？software was established and successfully applied to identify the metabolites of the main bioactive components in the herb pair extract by rat intestinal bacteria.Results Four parent compounds（loganin,morroniside,catalpol,and acteoside） and their eight corresponding metabolites were detected and tentatively identified by the characteristics of their protonated ions.Hydrogenated and demethylated loganetin,dehydroxylated morronisid aglycone,caffeic acid,and its methylated product were the main metabolites.These metabolites suggested that the glycosides were firstly hydrolyzed to their aglycones by hydrolytic enzymes of the enteric microbial flora and subsequently to the other metabolites through hydrogenation,（de）-methylation,and de-hydroxylation.Conclusion The results may be helpful for the further investigation of the pharmacokinetic study of R.glutinosa and C.officinalis herb pair in vivo.
基金Supported by Opening Fund for Key Laboratory of Forest Disaster Warning and Control in Yunnan Province(ZK09A102)National Natural Science Founda-tion of China(31100007)~~
文摘[ Objective ] The paper was to screen intestinal aerobic bacteria from 2nd instar larvae of Dendrolimu kikuchii and determine its toxicity. [ Method ] The intestinal samples of 2nd instar larvae of D. kikuchii were diluted in gredient and coated on plates to isolate strains, and a total of 5 strains of aerobic bacteria were isolated. With bacterial genomic DNA as template, the universal primers of bacterial 16S rDNA （27f and 1 492r） were used to amplify the template, 4 restriction enzymes Hae Ⅲ and Hind Ⅲ, Hinf Ⅰ and Taq Ⅰ were used to carry out ARDRA polymorphism analysis on PCR products. [ Result] Clustering map analysis showed that 5 strains of aerobic bacteria clustered into two different taxonomic operating units （OTU） in 95% similarity level, this indicated that the genetic diversity level of intestinal aerobic bacteria in 2nd instar larvae of D. kikuchii was relatively low. The indoor toxicity test showed that the death peaks of insects killed by intestinal bacteria were during 4 - 10 d, the insecticidal effect of strain 4 was the best with corrected mortality rate of 53.57% at 12 d. [ Conclusion] The paper provided reference for control of D. kikuchii.
文摘Objective In order to use facultative gut bacteria as an alternate to animals for the initial gastrointestinal toxicity screening of heavy metals, a comparative study on rat intestinal epithelial cells and resident gut bacteria was undertaken. Methods in vitro growth rate of four gut bacteria, dehydrogenase （DHA） and esterase （EA） activity test, intestinal epithelial and bacterial cell membrane enzymes and in situ effect of arsenite were analysed. Results Growth profile of mixed resident population of gut bacteria and pure isolates of Escherichia coli, Pseudomonas sp., Lactobacillus sp., and Staphylococcus sp. revealed an arsenite （2-20 ppm） concentration-dependent inhibition. The viability pattern of epithelial cells also showed similar changes. DHA and EA tests revealed significant inhibition （40%-72%） with arsenite exposure of 5 and 10 ppm in isolated gut bacteria and epithelial cells. Decrease in membrane alkaline phosphatase and Ca^2＋-Mg^2＋-ATPase activities was in the range of 33%-55% in four bacteria at the arsenite exposure of 10 ppm, whereas it was 60%-65% in intestinal epithelial villus cells, in situ incubation of arsenite using intestinal loops also showed more or less similar changes in membrane enzymes of resident gut bacterial population and epithelial cells. Conclusion The results indicate that facultative gut bacteria can be used as suitable in vitro model for the preliminary screening of arsenical gastrointestinal cytotoxic effects.
基金supported by the National Basic Research Program of China (2007CB109203 and 2009CB118902)
文摘Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear crylAh gene （800 ng mL^-1） and supercoiled plasmid DNA （800 ng mL^-1） under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon＇s index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic crylAh maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.
文摘Intestinal bacteria play a role in the development of non-steroidal anti-inflammatory drugs （NSAID）-induced small intestinal injury. Agents such as probiotics, able to modi~ the gut ecology, might theoretically be useful in preventing small intestinal damage induced by NSAIDs. The clinical studies available so far do suggest that some probiotic agents can be effective in this respect.
基金Supported by the National Natural Science Foundation of China(Nos.30772721 and 30873360)
文摘In vitro models of human colon carcinoma cell line(Caco-2 cell monolayer) and human intestinal bacteria were used to investigate the intestinal transport and biotransformation of resibufogenin and cinobufagin in Chan Su by HPLC/APCI-MSn. The experimental results of Caco-2 cell monolayer demonstrate that the apparent permeability coefficients(Papp) of resibufogenin and cinobufagin are higher than 10–6 cm/s, which indicates that both resibufogenin and cinobufagin have a good absorption in the small intestine. And the biotransformation result of human intestinal bacteria shows that resibufogenin has been transformed to 3-epiresibufogenin and cinobufagin has been transformed to 3-epicinobufagin, deacetylcinobufagin and 3-epideacetycinobufagin, respectively.
基金financially supported by the National Key Research and Development Program of China (No. 2018YFC1803100)the Youth Innovation Promotion Association of Chinese Academy of Sciences (No. 2018350)。
文摘The biosurfactants rhamnolipids and the “soil ecosystem engineers” earthworms are often used to remediate contaminated soils. However, the effects of rhamnolipids on earthworm intestinal flora and microbial community in soil containing earthworms are not clearly understood. In our study, a 21-d microcosm experiment was carried out to reveal the effects of rhamnolipids on microbial abundance, composition, and metabolism, as well as contaminant degradation capacity. Both rhamnolipids and earthworms had positive effects on soil bacteria. Rhamnolipid-amended soil(RT) showed higher bacterial abundance and metabolic activity than earthworm-amended soil(ET), while the improvement in bacterial composition and contaminant degradation capacity by rhamnolipids was lower than that by earthworms. Notably, these effects were further amplified by the combined treatment of rhamnolipids and earthworms(RET). Specifically, the bacterial abundance(log-transferred) increased from 9.5 copies g-1in the control with no addition to 10.3, 10.6, and 11.1 copies g-1in ET, RT, and RET, respectively. Compared to ET, the relative abundance of the dominant phylum, Proteobacteria, increased from 41.66% to 51.67% in RET,and more pollutant-degrading bacteria were also enriched in RET. Therefore, the increases in bacterial abundance and contaminant-degrading bacteria led to the following ranking of soil dioxin removal rate: RET(77.28%) > ET(59.83%) > RT(24.65%) > control(4.71%). Moreover, the addition of rhamnolipids enhanced the abundance of bacterial functional genes involved in metabolism and environmental information processing. In addition, the composition and diversity of bacteria in the gut of earthworms were conspicuously affected by rhamnolipids, and the relative abundance of Microbacterium and Shewanella increased significantly(P < 0.05). Therefore, this study revealed that rhamnolipids remarkably influenced the abundance, composition, and metabolism of the microbial community in earthworm gut, further promoting the degradation rate of dioxin, providing theoretical support for optimizing the combined application of rhamnolipids and earthworms in soil bioremediation engineering and for the assessment of the ecological impact of rhamnolipids.
基金the National Key Research and Development Program of China(No.2016YFD0400605)。
文摘Lactic acid bacteria(LAB)are a representative probiotic.As the dominant flora in the human intestinal tract,LAB can regulate the balance of human intestinal flora and improve host health.The purpose of this study was to isolate and screen LAB that are well suited to the intestinal characteristics of the Chinese population,with excellent probiotics and high antibacterial activity.After 16S ribosomal RNA(rRNA)homology and phylogenetic tree analysis,potential probiotics were tested for their antibacterial activity,resistance to artificial gastrointestinal fluid and drugs,surface hydrophobicity,and safety.Three strains of LAB with acid resistance,bile salt resistance,epithelial cell adhesion,and no multidrug resistance were selected:Lactobacillus salivarius,Leuconostoc lactis,and Lactobacillus paracasei.Analysis of the antibacterial active substances in the three strains and their fermentation broths revealed that the main antibacterial substances of L.lactis were organic acids,whereas those of L.salivarius and L.paracasei were organic acids and bacteriocins with broad-spectrum antibacterial activity.These three strains of probiotic LAB with high antibacterial activity were identified as bacterial resources that could potentially be used to develop probiotic preparations for the prevention and treatment of intestinal diseases caused by intestinal pathogens.
基金Supported by the Key-Area Research and Development Program of Guangdong Province,No.2022B1111070006the Guangdong Innovation Research Team for Higher Education,No.2021KCXTD025.
文摘BACKGROUND While colorectal polyps are not cancerous,some types of polyps,known as adenomas,can develop into colorectal cancer over time.Polyps can often be found and removed by colonoscopy;however,this is an invasive and expensive test.Thus,there is a need for new methods of screening patients at high risk of developing polyps.AIM To identify a potential association between colorectal polyps and small intestine bacteria overgrowth(SIBO)or other relevant factors in a patient cohort with lactulose breath test(LBT)results.METHODS A total of 382 patients who had received an LBT were classified into polyp and non-polyp groups that were confirmed by colonoscopy and pathology.SIBO was diagnosed by measuring LBTderived hydrogen(H)and methane(M)levels according to 2017 North American Consensus recommendations.Logistic regression was used to assess the ability of LBT to predict colorectal polyps.Intestinal barrier function damage(IBFD)was determined by blood assays.RESULTS H and M levels revealed that the prevalence of SIBO was significantly higher in the polyp group than in the non-polyp group(41%vs 23%,P<0.01;71%vs 59%,P<0.05,respectively).Within 90 min of lactulose ingestion,the peak H values in the adenomatous and inflammatory/hyperplastic polyp patients were significantly higher than those in the non-polyp group(P<0.01,and P=0.03,respectively).In 227 patients with SIBO defined by combining H and M values,the rate of IBFD determined by blood lipopolysaccharide levels was significantly higher among patients with polyps than those without(15%vs 5%,P<0.05).In regression analysis with age and gender adjustment,colorectal polyps were most accurately predicted with models using M peak values or combined H and M values limited by North American Consensus recommendations for SIBO.These models had a sensitivity of≥0.67,a specificity of≥0.64,and an accuracy of≥0.66.CONCLUSION The current study made key associations among colorectal polyps,SIBO,and IBFD and demonstrated that LBT has moderate potential as an alternative noninvasive screening tool for colorectal polyps.
文摘BACKGROUND The pathogenesis of gastroesophageal reflux disease(GERD)is closely associated with the intestinal bacteria composition and their metabolites.AIM To investigate whether washed microbiota transplantation(WMT)improves symptoms of nonerosive reflux disease(NERD)with proton pump inhibitor(PPI)dependency.METHODS Patients with recurrent NERD and PPI dependency at the First Affiliated Hospital of Guangdong Pharmaceutical University from 2017 to 2018 were included and divided into a WMT or PPI group treated with PPI with/without WMT.The endpoint was NERD symptom frequency evaluated 1 mo after WMT using reflux disease questionnaire(RDQ)and GERD questionnaire(GERDQ)scores,remission time,PPI dose,and the examination of intestinal mucosal barrier function.RESULTS In the WMT(n=15)and PPI(n=12)groups,the total remission rate at 1 mo after treatment was 93.3%vs 41.7%.Compared with the PPI group,the WMT group showed better results in GERDQ(P=0.004)and RDQ(P=0.003)and in remission months(8 vs 2,P=0.002).The PPI dose was reduced to some extent for 80%of patients in the WMT group and 33.3%in the PPI group.In 24 patients,intestinal mucosal barrier function was examined before treatment,and changes in the degree of damage were observed in 13 of these patients after treatment.Only one of the 15 patients had minor side effects,including a mushy stool two or three times a day,which resolved on their own after 1 wk.CONCLUSION This study is the first to demonstrate that WMT may be safe and effective for relieving NERD symptoms and reducing PPI dependency and recurrence.
