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ObjectiveTo explore the possible mechanism of the Yiqi Jiedu formula (YQ) in treating ischemic stroke (IS) from the perspective of the microbial-gut-brain axis (MGBA). MethodRats were randomly divided into five groups, with six in each group, including sham surgery group, model group, and low, medium, and high dose YQ groups (1, 5, and 25 mg·kg-1). Except for the sham surgery group, all other groups were established with a middle cerebral artery occlusion (MCAO) model using the thread occlusion method. The success of modeling was determined through neurobehavioral scoring, and the protective effect of YQ on IS was evaluated. Then, the changes in gut microbiota before and after MCAO modeling and YQ administration were compared using 16S rDNA sequencing technology, and the possible biological pathways related to the effect of this formula were analyzed. The expression of inflammatory factors such as interleukin-6 (IL-6), interleukin-17A (IL-17A), and interleukin-10 (IL-10) in serum was detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the expression of tight junction proteins ZO-1 and Occludin in brain and intestinal tissue, and hematoxylin-eosin staining (HE) was used to observe pathological changes in the cerebral cortex and colon, so as to validate the possible mechanism of action. ResultYQ significantly improved the neurobehavioral score of MCAO rats (P<0.01) and played a good regulatory role in intestinal microbial disorders caused by enriched pathogens and opportunistic pathogens during the acute phase. Among them, significantly changed microorganisms include Morgentia, Escherichia Shigella, Adlercreutzia, and Androbacter. Bioinformatics analysis found that these bacteria may be related to the regulation of inflammation in the brain. Compared with the blank group, the detection of inflammatory factors in the serum of IS model rats showed an increase in inflammatory factors IL-6 and IL-17A (P<0.01) and a decrease in the content of anti-inflammatory factor IL-10 (P<0.01). Compared with the model group, the content of inflammatory factors IL-6 and IL-17A in the serum of the treatment group decreased (P<0.05), and that of anti-inflammatory factor IL-10 increased (P<0.01). The expression results of barrier proteins ZO-1 and Occludin in brain and intestinal tissue showed that the expression levels of both decreased in IS model rats (P<0.05), while the expression levels of both increased in the treatment group (P<0.05). ConclusionAcute cerebral ischemia can lead to an imbalance of intestinal microbiota and damage to the intestinal barrier, and it can increase intestinal permeability. YQ can regulate intestinal microbiota imbalance caused by ischemia, inhibit systemic inflammatory response, and improve the disruption of the gut-blood brain barrier, preventing secondary cascade damage to brain tissue caused by inflammation. The MGBA may be an important mechanism against the IS.
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Objective:To study the changes of intestinal flora before and after dietary intervention in pregnant women with gestational diabetes mellitus (GDM) and explore its correlation with the results of oral glucose tolerance test (OGTT), using third generation high-throughput sequencing of full-length 16S rDNA.Methods:Thirty pregnant women who received regular antenatal care in the First Affiliated Hospital of Xinjiang Medical University between July 2022 and March 2023 were selected. Demographic data and laboratory examination results of these pregnant women were collected. The pregnant women with GDM (the GDM group) were given individualized dietary intervention, while the control group were given routine dietary guidance, for a total of 2 weeks. The stool was collected before and after the intervention in both groups, and a total of 60 stool samples were collected. Through the PacBio Sequencing platform, the third generation sequencing of full-length 16S rDNA was utilized to investigate the intestinal microbiome diversity. The composition, abundance and diversity of intestinal flora were compared between groups, both before and after the intervention.Results:There were 74 species showing significant differences in abundance between the two groups of pregnant women, 54 of which were enriched in the GDM group. The correlation analysis of blood glucose levels tested by OGTT as an environmental factor showed that Lachnospirales ( P=0.002) and unclassified Bacteria ( P=0.035) were positively correlated with OGTT-0h blood glucose ( P<0.05), while Christensenellales ( P=0.018)and Oscillospirales ( P=0.045) negatively. Lachnospirales ( P=0.027)and unclassified Bacteria ( P=0.028) were positively correlated with OGTT-1 h blood glucose ( P<0.05), while Oscillospirales ( P=0.025) negatively. There was a positive correlation between Lachnospirales ( P=0.027) and OGTT-2h blood glucose ( P<0.05). Conclusions:After the individualized diet intervention, the dominant bacteria of the intestinal flora changed and the fasting blood glucose of declined in pregnant women with GDM. It's a reasonable presumption that individualized diet intervention can contribute to the regulation of the disordered intestinal flora in pregnant women with GDM, which implies a role of dietary intervention in the treatment of GDM.
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Objective To investigate the effect of vitamin D(VD)on intestinal flora in spontaneously diabetic rats.Methods Zucker diabetic fatty rats(ZDF rats)were randomised to control(Con)group,VD control(VD)group,model(T2DM)group,and VD intervention(VD+T2DM)group.Fasting blood glucose profiles and oral glucose tolerance levels were determined in rats of each group.16S rDNA sequencing was used to assess changes in rat intestinal flora.OTU analysis(Venn diagram),α diversity analysis(chao1,observed species,PD whole tree,and shannon and simpson),βdiversity analysis(principal coordinate analysis(PCoA)),flora structure,and colony species variability analysis(linear discriminant analysis and influence factor(LEfSe)analysis)were also performed.Results VD intervention significantly improved fasting blood glucose levels and insulin resistance in T2DM rats(P<0.05).α diversity showed no significant differences in chao1,observed species,PD whole tree,and shannon and simpson indices between T2DM and VD+T2DM groups(P>0.05).β diversity analysis showed that the VD+T2DM group had more species similarity to the Con group than the T2DM group.The dominant bacteria of rat intestinal flora in each group were significantly different.In comparison to the T2DM group,the VD+T2DM group showed a decrease in abundance of Bacteroidetes and increases in abundances of Firmicutes and Clostridium XIVa.Conclusions VD improves fasting glucose elevation and insulin resistance in T2DM rats.VD improves the structure of intestinal flora,decreases Bacteroidetes,and elevates Firmicutes and Clostridium XIVa abundances in T2DM rats.
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ObjectiveThis study aims to explore the potential mechanism of Danggui Niantongtang in treating knee osteoarthritis (KOA) by regulating the intestinal flora through 16S rDNA analysis. MethodThirty-six C57BL/6J mice were subjected to anterior cruciate ligament transection (ACLT) to establish a KOA model and were randomly divided into the sham surgery group, model group, low-dose Danggui Niantongtang group (0.819 g·kg-1), medium-dose Danggui Niantongtang group (1.638 g·kg-1), high-dose Danggui Niantongtang group (3.276 g·kg-1), and Meloxicam group (0.975 mg·kg-1), with 6 mice in each group. Except for the treatment groups, the sham surgery group and model group were given normal saline by gavage. After 4 weeks of continuous intervention, feces and intact knee joints of the mice were collected. Hematoxylin-eosin (HE) staining and Safranin O-Fast Green staining were performed to observe the pathological changes in knee joint tissue morphology. The 16S rDNA sequencing was used to analyze changes in the abundance and diversity of intestinal microorganisms before and after treatment, along with corresponding functional predictions. ResultHigh-dose Danggui Niantongtang and Meloxicam significantly relieved pain symptoms in KOA mice, improved the disorder of joint structure, maintained the integrity of knee articular cartilage, increased the expression of type Ⅱ collagen alpha 1 (Col2a1) in articular cartilage, and decreased the expression of matrix metalloproteinase-13 (MMP-13). The results of 16S rDNA sequencing showed that high-dose Danggui Niantongtang could adjust the abundance and structure of intestinal microbial species. Compared with the sham surgery group, the abundance of Proteobacteria, Actinobacteria, Ruminococcus, and Bacteroides was significantly increased in the model group (P<0.05), while in the Danggui Niantongtang group, the abundance of these four flora was significantly reduced compared with the model group. Compared with the sham surgery group, the abundance of Verrucomicrobia, Oscillospira, and Akkermansia was significantly decreased in the model group (P<0.05), while in the Danggui Niantongtang groups, the abundance of these three flora was significantly increased compared with the model group (P<0.05). Functional pathway prediction of differential genera revealed that species differences among groups mainly involved metabolic pathways with high abundance associated with biosynthesis and precursors, as well as energy production, including amino acid biosynthesis, nucleotide and nucleoside biosynthesis, cofactors, prosthetic groups, electron carriers, and vitamin biosynthesis. ConclusionDanggui Niantongtang can effectively protect articular cartilage and delay the progression of KOA, possibly by regulating the structure of the intestinal flora, promoting probiotics, and inhibiting the growth of harmful pathogenic bacteria.
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ObjectiveTo observe the microbial changes in Wistar rats with liver injury caused by the Dictamni Cortex-Flavescens Sophora by high-throughput sequencing technology and investigate the potential mechanism of liver injury caused by the Dictamni Cortex-Flavescens Sophora. MethodFemale Wistar rats were randomly divided into four groups: normal group, as well as low-dose, medium-dose, and high-dose groups of traditional Chinese medicine (TCM). The rats were gavaged with the Dictamni Cortex-Flavescens Sophora in different doses (4.125, 8.25, 16.5 g·kg-1 of raw drug respectively) for 28 days, and the general condition was recorded. The liver-body weight ratio was calculated, and the biochemical indexes of serum were observed. The Hematoxylin-eosin (HE) staining was used to observe pathological changes in the liver, and 16S rDNA high-throughput sequencing was utilized to detect fecal microbial changes in rats. ResultCompared with the normal group, Dictamni Cortex-Flavescens Sophora increased the liver weights and liver-body weight ratios of Wistar rats. The difference in liver weight between the medium-dose and high-dose groups of TCM was statistically significant (P<0.05), and the liver-body weight ratios of the low-dose, medium-dose, and high-dose groups of TCM were all statistically significant (P<0.05). Compared with the normal group, serum albumin and cholesterol levels increased in the medium-dose and high-dose groups of TCM (P<0.05). The histopathology of the liver in the medium-dose and high-dose groups of TCM showed tiny vacuole-like changes. Compared with the normal group, there were obvious intestinal flora disorders after administration of Dictamni Cortex-Flavescens Sophora, and alpha diversity increased in the medium-dose and high-dose groups of TCM. The principal coordinates analysis showed that species increasingly deviated from the normal group as the administered dose increased. Compared with the normal group, the proportion of Firmicutes and Bacteroidota decreased after the drug administration, and the genus level of Parasutterella, Romboutsia, Turicibacter, Allobaculum, and Dubosiella increased. The genus level of Lachnospiraceae_NK4A136_group, Blautia, Erysipelatoclostridium, Muribaculum, and Ruminococcus_gnavus_group decreased. The correlation analysis showed that Parasutterella, Romboutsia, Turicibacter, Allobaculum, and Dubosiella were positively correlated with serum cholesterol and liver-body weight ratio, and lanchnospiraceae_NK4A136_group, Blautia, Muribaculum, Erysipelatoclostridium, and Ruminococcus_gnavus_group were negatively associated with serum cholesterol and liver-body weight ratio. ConclusionThe liver injury caused by Dictamni Cortex-Flavescens Sophora is manifested as a lipid metabolism disorder, and the mechanism is related to the increase in lipid metabolism-related microorganisms.
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BACKGROUND:Exercise has a regulatory effect on intestinal flora dysbiosis,which can effectively protect the beneficial flora and improve the intestinal environment.However,the effect of treadmill exercise on the structure and diversity of intestinal microbial community in Parkinson's disease and the specific mechanism are not clear. OBJECTIVE:Using 16S rDNA technique to analyze the effect of treadmill exercise on the structure and diversity of the intestinal flora of rats with Parkinson's disease,and to investigate the mechanism of non-pharmacological treadmill exercise to improve Parkinson's disease. METHODS:Twelve of the 18 Sprague-Dawley rats were randomly selected to make animal models of Parkinson's disease using unilateral 2-point nigrostriatal injection of 6-hydroxydopamine.The remaining six rats were used as sham-operation group,which were injected with the same dose of saline containing 0.2%ascorbic acid using the same positioning and injection method.After successful modeling,12 rats with Parkinson's disease were randomly divided into model group and treadmill exercise group(n=6 per group).The treadmill exercise group was subjected to a middle and low intensity tread mill exercise,10 m/min,30 minutes per day,5 days per week for 4 weeks.Fresh feces were collected and stored in liquid nitrogen 24 hour after the last exercise session,and the changes in fecal flora were analyzed by 16S rDNA sequencing technique. RESULTS AND CONCLUSION:Treadmill exercise significantly improved behavior and nigrostriatal tyrosine hydroxylase-positive cell expression in rats with Parkinson's disease model and alleviated changes in the structure and diversity of the gut microbial community caused by Parkinson's disease,increased the number of operational taxonomic units and modulated Alpha and Beta diversity in rats.At the phylum and genus levels,the abundance ratio of Firmicutes/Bacteroidetes in the model group decreased compared with the sham-operated group,while beneficial bacteria such as Prevotella,Bacteroides,and Clostridium_XlV increased significantly after treadmill exercise.To conclude,treadmill exercise has a significant modulating effect on behavioral abnormalities,toxic damage to dopaminergic neurons and gut microbial imbalance caused by Parkinson's disease,alleviates the symptoms of flora-related diseases,and has a positive effect on the improvement of Parkinson's disease.
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BACKGROUND:Osteonecrosis of the femoral head is a common and disabling disease,which is mainly characterized by microcirculation disorders and bone cell metabolism disorders.Luzhongjiangu decoction was developed by Shandong Academy of Chinese Medicine and used in the form of soup in the clinic,which has good efficacy in the treatment of osteonecrosis of the femoral head.However,its mechanism of action has not been clarified. OBJECTIVE:To study the effect mechanism of Luzhongjiangu decoction on intestinal flora in rats with osteonecrosis of the femoral head based on 16S rDNA sequencing technique. METHODS:The model of osteonecrosis of the femoral head was established in Wistar rats by intragastric administration of retinoic acid.The therapeutic effect of Luzhongjiangu decoction was evaluated by serum hormone,bone histopathology and serum hormone levels.16s rDNA sequencing technique was used to detect the intestinal flora of rats in the blank control group,model group and middle-dose Luzhongjiangu decoction group.The corresponding library was constructed and OTU clustering and microbial community diversity and abundance analysis were carried out to determine the composition of intestinal flora and the changes of species and diversity among groups. RESULTS AND CONCLUSION:Luzhongjiangu decoction could significantly increase the expression of osteocalcin,osteopontin and other osteogenic related factors,alleviate the destruction of bone trabeculae,increase bone mineral density,and had a significant therapeutic effect on osteonecrosis of the femoral head,of which the middle dose group showed the most significant effect.The results of intestinal flora sequencing showed that Luzhongjiangu decoction improved the flora disorder of rats with osteonecrosis of the femoral head to some extent,and screened out different colonies such as Bacillus,Desulfurizans,Desulfurization,Isobacteria,Bifidobacterium and so on;it could up-regulate the abundance of beneficial bacteria such as Bifidobacterium,down-regulate the abundance of harmful bacteria such as Desulfovibrio,and improve the structure of intestinal flora.Functional prediction analysis indicated that Luzhongjiangu decoction could mainly affect amino acid metabolism and energy metabolism.Correlation analysis showed that the differential bacteria of Bifidobacterium and Intestinimonas in the middle dose group of Luzhongjiangu decoction were positively correlated with vitamin D3,estradiol and calcitonin,and negatively correlated with prostaglandin E2.In the model group,Escherichia-Shigella,Desulfovibrio,Globicatella and Streptococcus were positively correlated with prostaglandin E2 and negatively correlated with vitamin D3,estradiol and calcitonin.To conclude,Luzhongjiangu decoction may play a role in the treatment of osteonecrosis of the femoral head by regulating the structure of intestinal flora,up-regulating the abundance of beneficial bacteria and affecting the secretion of vitamin D3,estradiol,calcitonin and prostaglandin E2.
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Objective:To study the effects of group B streptococcus (GBS) colonization during late pregnancy on vaginal microbiota and neonatal outcomes.Methods:From September 2020 to September 2021, pregnant women receiving prenatal care and delivered in our hospital were prospectively enrolled. They were assigned into GBS(+) group and GBS(-) group based on the results of GBS culture and/or PCR tests of vaginal secretions. The mothers were also assigned into early-onset infection(EO) group and non-early-onset infection(non-EO) group based on the presence or absence of early-onset infection of their neonates. The vaginal microbiota and neonatal outcomes were compared between these groups.Results:A total of 125 cases were enrolled, including 65(52.0%) in GBS(+) group and 60(48.0%) in GBS(-) group. 24 cases (19.2%) were in EO group and 101 cases (80.8%) in non-EO group. The incidences of premature rupture of membranes (PROM), amniotic fluid contamination, chorioamnionitis and early-onset neonatal infection in GBS(+) group were significantly higher than GBS(-) group(all P<0.05).The abundances of Streptococcus and Ureaplasma in vaginal flora of GBS(+) group were higher than GBS(-) group ( P<0.01), whereas Rhodococcus, Phyllobacterium and Bifidobacterium were lower than GBS(-) group ( P<0.05).The EO group had significantly higher abundance of enterococcus than the non-EO group ( P<0.05). Mothers with GBS colonization and neonates with early-onset infection had the highest abundance of Escherichia/Shigella ( P=0.04). Mothers with GBS colonization and neonates without early-onset infection showed the highest abundance of Gardnerella ( P=0.04). Conclusions:GBS colonization during late pregnancy increases the incidences of PROM, amniotic fluid contamination, chorioamnionitis and early-onset neonatal infection. GBS colonization causes abnormal vaginal flora in pregnant women. The increases of Ureaplasma, Streptococcus, Escherichia/Shigella and Enterococcus in vaginal microbiota maybe associated with early-onset neonatal infection.
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Objective To compare the differences in the composition and diversity of intestinal flora between knee osteoarthritis(KOA)patients with dampness-heat obstruction syndrome,cold-dampness blockage syndrome and healthy people;To explore the characteristics of intestinal flora of KOA patients with the above two TCM syndrome types.Methods According to the inclusion and exclusion criteria,dampness-heat obstruction syndrome group,cold-dampness blockage syndrome group,and healthy control group were screened,with 10 cases for each group,and then their fecal samples were collected.16S rDNA sequencing technology was used to compare the differences of intestinal flora among groups by Alpha and Beta diversity analysis.Results There was no significant difference in species richness of intestinal flora among the three groups,but there was statistical significance in species diversity between cold-dampness blockage syndrome group and dampness-heat obstruction syndrome group,and healthy control group(P<0.05).There were statistical significance among the three groups in intestinal flora composition(P= 0.001).At the phylum level,Bacteroidota and Firmicutes were significantly dominant,and at genus level,the abundance of Prevotella in dampness-heat obstruction syndrome group and cold-dampness blockage syndrome group increased.The abundance of Enterobacteriaceae,Lachnospira and Klebsiella was relatively large in dampness-heat obstruction syndrome group.The abundance of Prevotella and Pseudomonas in cold-dampness blockage syndrome group was large.Conclusion There are differences in the intestinal flora structure between KOA patients with dampness-heat obstruction syndrome and cold-dampness blockage syndrome.
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Background Multiple studies have shown a close relationship between changes in gut microbiota composition and obesity, and research results are influenced by factors such as race and geographical location, but there are few studies on children. Objective To analyze the diversity of gut microbiota related to obesity in a population of 2-6 years old, observe the distribution characteristics and species differences of gut microbiota between obese/overweight and normal weight groups, and explore the association betweenobese/overweight and gut microbiota diversity. Methods Fecal samples were collected from 74 children aged 2-6 years in Shanghai, including 18 obese/overweight individuals, 6 males and 12 females (male to female ratio of 1∶2), and 56 normal weight individuals, 18 males and 38 females (male to female ratio is nearly 1∶2). The 16S rDNA was extracted from bacteria in fecal samples, followed by PCR amplification, cDNA construction, and high-throughput sequencing. Naive Bayes algorithm was used to perform taxonomic analysis (phylum, class, order, family, genus, species) and community diversity analysis (Sobs index, Shannon index, Shannoneven index, Coverage index, PD index, and principal co-ordinates analysis) on representative sequences and abundance of amplicon sequence variants (ASV). Wilcoxon rank sum test, P-value multiple test correction, and analysis of similarities were used to test differences between the two groups to obtain information on the distribution characteristics and species differences of intestinal microbiota in children. Results Seventy-four fecal samples were sequenced, and the sequencing results were subjected to quality control and filtering. A total of 4905306 optimized sequences were obtained, resulting in 1860 ASVs. The diversity data analysis of ASVs generated 889 species annotation results at 8 taxonomic levels. The alpha diversity analysis showed that the richness (Sobs index), diversity (Shannon index), evenness (Shannoneven index), and phylogenetic diversity (PD index) of fecal community of the obese/overweight children were increased compared to those of the normal weight children, but there were no statistical differences between the two groups (P>0.05). The beta diversity analysis showed that there was little difference in the composition of microbial species between the two groups, and no significant clustering separation was observed. The results of species composition analysis at phylum, order, family, and genus levels of 74 samples showed a consistent core microbiota structure in the two groups of gut microbiota, but there were differences in microbiota composition. The differences in microbial community composition between the two groups were manifested at the taxonomic levels of order, family, and genus, among which phylum Firmicutes, order Erysipelotrichales, family Erysipelatocyclostridiaceae, genus Erysipelotrichaceae_ UCG-003 and genus Catenibacterium were significantly enriched in the obese/overweight group and contributed significantly to the phenotypic difference of obese/overweight [linear discriminant analysis (LDA)=3.72, P<0.01; LDA=3.29, P<0.05). Phylum Proteobacteria, order Enterobacterales, family Enterobacteriaceae, genus unclassified was significantly enriched in the normal weight group and contributed significantly to the phenotypic difference of normal body weight (LDA=3.93, P<0.05). Conclusion The richness and diversity of gut microbiota in obese/overweight children aged 2-6 years in Shanghai are increased, but there is no difference compared to normal weight children. There is a difference in the composition of gut microbiota between the obese/overweight group and the normal weight group.
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Aim To evaluate the hypolipidemic effect of the total phenylpropanoid glycosides extracted from Ligustrum robustum (Roxb.) Blume (LRTPG) on hyperlipidemic golden hamsters and explore its regulatory effect on intestinal flora. Methods Sixty hamsters were randomly divided into a control group, a model group, a positive drug group, LRTPG-L group, LRTPG-M group, and LRTPG-H group. After the successful induction of the model by high-fat diet, the animals were continuously administered for four weeks, and their blood lipids and liver lipids were detected. The formed feces from the colorectal region of the hamsters in the control group, model group and LRTPG-H group were collected for 16S rDNA sequencing. Results LRTPG reduced serum TG, TC, LDL-C and liver TG, TC concentrations significantly in hyperlipidemic hamsters. The results of the intestinal microbiota sequencing showed that compared to the control group, LRTPG significantly decreased the relative abundance of the phylum Firmicutes and increased the relative abundance of the phylum Bacteroidetes and Verrucomicrobia (P < 0.01) at the phylum level. At the family level, LRTPG significantly increased the relative abundance of Christensenellaceae, Peptococcaceae, and Verrucomicrobiaceae (P < 0.05 or P < 0.01). At the genus level, LRTPG significantly increased the relative abundance of Oscillospira, Oscillibacter, Flavonifractor and Akkermansiaceae (P < 0.05 or P < 0.01). These changes in the flora were beneficial to the hypolipidemic effect of LRTPG. Conclusion LRTPG may exert its hypolipidemic effect by improving the intestinal flora disorder caused by a high-fat diet in golden hamsters.
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@#Abstract: Objective To investigate the composition and diversity of midgut microbial community of Haemaphysalis longicornis infected with severe fever with thrombocytopenia syndrome virus (SFTSV). Methods The midgut DNA of three group Haemaphysalis longicornis infected with SFTSV was extracted, and the 16S rDNA gene of the sample was sequenced by HiSeq platform. The composition and diversity of endosymbiotic microbial community were clarified by OTU cluster analysis and alpha diversity analysis. Results The midgut microbial clusters of the three groups infected with SFTSV were 143, 113, 163 OTUs respectively; the sparsity curve and abundance grade curve showed that the data had sufficient sequencing depth, and the midgut of Haemaphysalis longicornis infected with SFTSV was rich in microbial composition, but the species distribution was uneven. The analysis of microbial community composition showed that Proteobacteria, Firmicutes and Actinobacteria were the main dominant bacteria at the phyla level. At the class level, Gammaproteobacteria, Bacilli, Betaproteobacteria and Actinomycetia were the main dominant bacteria. At the order level, Legionellales, Bacillales, Burkholderiales and Actinomycetales were the main dominant orders. At the family level, Coxiellaceae, Bacillaceae, Moraxellaceae and Rhodococcaceae were the main dominant families. At the genus level, the relative abundance of Coxiella was the highest, followed by Aeribaillus and Azonexus. Alpha diversity analysis showed that the average Shannon index was 139.67, the average Simpson index was 0.48, the average Chao index was 145.06, and the average ACE index was 147.11. Conclusions The species diversity of intestinal microorganisms in Haemaphysalis longicornis infected with SFTSV is rich. The results provide a basis for further exploring the interaction between intestinal microbes of Haemaphysalis longicornis and SFTSV and developing new ideas for the prevention and control of ticks and tick-borne diseases.
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Objective To investigate the effects of Osteoking on hyperglycemia and regulating gut microbiota in db/db mice.Methods Wildtype mice were used as the control group,and db/db mice were randomly divided into model and Osteoking groups.After intragastric administration for 12 weeks,fasting blood glucose and serum glycosylated hemoglobin and insulin levels were measured,changes in intestinal microflora were determined,and functional pathways related to intestinal microflora in mice were predicted by 16S rDNA sequencing.Results Compared with the model group,Osteoking decreased fasting blood glucose(P<0.01),serum glycosylated hemoglobin(P<0.01),and the insulin resistance index(P<0.01),and increased insulin content(P<0.01)in db/db mice.Osteoking increased the abundance of beneficial intestinal microflora and decreased the abundance of harmful bacteria.Moreover,the abundance of Marvinbryantia was increased.Osteoking alleviated the decrease in metabolism of D-arginine and D-ornithine,sphingolipid,and galactose metabolism(P<0.05)and inhibited lysine degradation,the sulfur relay system,and propanoate metabolism(P<0.05).Conclusions Osteoking has hypoglycemic properties and improves the intestinal microflora imbalance in db/db mice.
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Objective: To explore the characteristics and correlations of vaginal flora in women with cervical lesions. Methods: A total of 132 women, including 41 women diagnosed with normal cervical (NC), 39 patients with low-grade cervical intraepithelial neoplasia (CIN 1), 37 patients with high-grade cervical intraepithelial neoplasia (CIN 2/3) and 15 patients with cervical squamous cell carcinoma (SCC), who came from the gynecological clinic of Second Hospital of Shanxi Medical University during January 2018 to June 2018, were enrolled in this study according to the inclusive and exclusive criteria strictly. The vaginal flora was detected by 16S rDNA sequencing technology. Co-occurrence network analysis was used to investigate the Spearman correlations between different genera of bacteria. Results: The dominant bacteria in NC, CIN 1 and CIN 2/3 groups were Lactobacillus [constituent ratios 79.4% (1 869 598/2 354 098), 63.6% (1 536 466/2 415 100) and 58.3% (1 342 896/2 301 536), respectively], while Peptophilus [20.4% (246 072/1 205 154) ] was the dominant bacteria in SCC group. With the aggravation of cervical lesions, the diversity of vaginal flora gradually increased (Shannon index: F=6.39, P=0.001; Simpson index: F=3.95, P=0.012). During the cervical lesion progress, the ratio of Lactobacillus gradually decreased, the ratio of other anaerobes such as Peptophilus, Sneathia, Prevotella and etc. gradually increased, and the differential bacteria (LDA score >3.5) gradually evolved from Lactobacillus to other anaerobes. The top 10 relative abundance bacteria, spearman correlation coefficient>0.4 and P<0.05 were selected. Co-occurrence network analysis showed that Prevotella, Peptophilus, Porphyrinomonas, Anaerococcus, Sneathia, Atopobium, Gardnerella and Streptococcus were positively correlated in different stages of cervical lesions, while Lactobacillus was negatively correlated with the above anaerobes. It was found that the relationship between vaginal floras in CIN 1 group was the most complex and only Peptophilus was significantly negatively correlated with Lactobacillus in SCC group. Conclusions: The increased diversity and changed correlations between vaginal floras are closely related to cervical lesions. Peptophilus is of great significance in the diagnosis, prediction and early warning of cervical carcinogenesis.
الموضوعات
Female , Humans , Vagina/microbiology , Uterine Cervical Neoplasms/genetics , Uterine Cervical Dysplasia , Cervix Uteri , Lactobacillus/genetics , Papillomavirus Infectionsالملخص
@#The HACEK organisms consist of the non-influenzae Haemophilus sp., Aggregatibacter sp., Cardiobacterium sp., Eikenella corrodens and Kingella sp. are responsible for a sizable percentage of infective endocarditis cases worldwide with the mortality rate of 18%. Amongst them, Aggregatibacter actinomycetemcomitans is the most common pathogen strongly associated with infective endocarditis. A. actinomycetemcomitans forms part of the oral microbiota and is also the etiological agent of periodontitis. Here, we present a case of a 37-year-old man with underlying obstructive uropathy, that sought treatment for postural hypotension and symptomatic anaemia with fever. Later, he had developed decompensated congestive cardiac failure with aortic regurgitation. A cardiac echocardiogram revealed the presence of vegetation on the aortic valve. Blood culture grew A. actinomycetemcomitans, and he was treated with furosemide and ceftriaxone. A further dental examination showed the patient is having chronic periodontitis, which could be the possible source of A. actinomycetemcomitans causing infective endocarditis. The patient was then transferred to the National Heart Centre for the first time for further management after completion of 4 weeks of intravenous antibiotics. As the pathogen is fastidious, rapid and newer technology like MALDI-TOF mass spectrometry provides rapid and accurate identification for appropriate patient clinical management.
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ObjectiveTo investigate the effect and mechanism of Dahuang Zhechongwan (DHZCW) on adenine-induced renal fibrosis in rats from the perspective of intestinal flora. MethodThirty-six SD rats were randomly divided into a blank group, a model group, and high-, medium- and low-dose DHZCW groups (0.168, 0.084, 0.042 g·kg-1), and a pirfenidone group (200 mg·kg-1), with 6 rats in each group. Except for those in the blank group, rats in other groups were treated with adenine suspension (250 mg·kg-1) by gavage for 28 days for renal fibrosis model induction. Subsequently, they received drug intervention for 4 weeks. Urine samples were collected from rats in metabolic cages, and renal function indicators including blood urea nitrogen (BUN), urea, creatinine (Crea), cystatin C (Cys C), and 24-hour urine protein (24 h TP) were measured. Kidney samples were collected and subjected to hematoxylin-eosin (HE) staining and Masson's trichrome staining to observe the pathological changes in rat renal tissues. Western blot was used to detect the expression levels of key effector proteins α-smooth muscle actin (α-SMA), type Ⅰ collagen (ColⅠ), and type Ⅲ collagen (ColⅢ) in the kidneys. High-throughput sequencing of 16S rDNA was used to analyze the species diversity of rat intestinal flora. ResultCompared with the blank group, the model group showed increased BUN, urea, Crea, Cys C, and 24 h TP levels (P<0.01). Compared with the model group, the high-, medium-, and low-dose DHZCW groups, as well as the pirfenidone group, showed significant reductions in BUN, urea, Crea, Cys C, and 24 h TP levels (P<0.01), indicating that DHZCW intervention significantly improved renal function. In the model group, renal tissues exhibited significant fibrotic changes, and the protein levels of α-SMA, ColⅠ, and ColⅢ were significantly increased (P<0.01) compared to those in the blank group. Compared with the model group, the high-dose DHZCW group and the pirfenidone group had relatively normal tissue structure, with no significant pathological damage observed. However, fibrotic changes were observed in the medium- and low-dose DHZCW groups, with the changes being more significant in the low-dose group. The protein levels of α-SMA, ColⅠ, and ColⅢ were significantly decreased in the high-, medium-, and low-dose DHZCW groups, as well as the pirfenidone group (P<0.01), indicating that DHZCW effectively reduced abnormal collagen deposition and inhibited renal fibrosis. From the perspective of intestinal flora, at the phylum level, compared with the blank group, the model group showed a significant increase in the abundance of Firmicutes and a decrease in Bacteroidetes, leading to a significant imbalance in their ratio. At the family level, the model group decreased the abundance of Lachnospiraceae, Prevotellaceae, and Bacteroidota_unclassified, and increased the abundance of Ruminococcaceae, Lactobacillaceae, and Oscillospiraceae. At the genus level, the model group showed significantly reduced abundance of Firmicutes_unclassified, Bacteroidota_unclassified, and Prevotellaceae_UCG-001, etc., and increased abundance of UCG-005, Clostridia_UCG-014_unclassified, etc. Compared with the model group, DHZCW effectively reduced the abundance of potential pathogenic bacteria and increased the abundance of beneficial bacteria, regulating the intestinal flora. ConclusionDHZCW can effectively improve renal function and inhibit renal fibrosis, and its mechanism of action may be related to the regulation of intestinal flora.
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Aims@#This study was aimed to analyze the antagonistic ability of Pantoea sp. against Ralstonia solanacearum, which causes bacterial wilt in potato plants and to analyze the antimicrobial compounds produced by the antagonistic bacteria. @*Methodology and results @#Pantoea sp. was isolated from the rhizosphere of healthy potato plants in Taman Tanda village, Baturiti, Tabanan, Bali. The diffusion well method tested the isolates' antagonistic ability against R. solanacearum. The Pantoea sp., which showed the best inhibition zone was then identified morphologically, biochemically and molecularly using 16S rDNA gene sequencing and its analysis. Five isolates of Pantoea sp. were isolated from the rhizosphere of potato plants. Pantoea sp. 4 showed the best inhibition against R. solanacearum and was identified as Pantoea agglomerans. Filtrate of P. agglomerans was tested against R. solanacearum to analyze the inhibition ability. The minimum inhibitory concentration (MIC) value of the P. agglomerans filtrate was 6%. Fraction 10 of P. agglomerans filtrate showed the highest inhibition zone (17 mm). The active compounds of fraction 10 characterized as antimicrobial were decamethyl tetrasiloxane, hexamethylcyclotrisiloxane, 2,4-dimethyl Benzo[h]quinoline and arsenous acid. @*Conclusion, significance and impact of study@#Pantoea agglomerans isolated from potato rhizosphere in Baturiti, Bali, have the potential as an antagonist agent against R. solanacearum that causes bacterial wilt. The active compound produced by P. agglomerans can act as an antibacterial against the pathogen R. solanacearum. Pantoea agglomerans culture can be used as a biocontrol agent against R. solanacearum.
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Aims@#The study aims to investigate the bacterial community profiles on the surface of red algae (Kappaphycus alvarezii) and persistent bacteria that can adapt to long-term cultivation in a closed circulation system.@*Methodology and results@#Kappaphycus alvarezii explants were cultured in a controlled laboratory setting for 30 days to investigate related bacterial adaptability to controlled culture conditions. Bacterial isolates associated with seedlings were subjected to 16S rDNA amplification and sequencing, followed by the construction of a phylogenetic tree using MEGA X software. The results show distinct microbial composition between the first and 30th days. The derived phylogenetic tree features three dominant phyla: Proteobacteria (Vibrio and Thalassospira), Pseudomonadota (Pseudoalteromonas, Alteromonas, Grimontia, Ruegeria, Phaebacter and Bacterioplanes) and Firmicutes (Bacillus). A comparative examination of these two bacterial groups (day 1 and day 30) reveals evidence of persistent marine bacteria, such as the genera Vibrio, Pseudoalteromonas, Alteromonas, Phaaebacter and Bacterioplanes, that successfully adapt to long-term cultivation within closed circulation systems.@*Conclusion, significance and impact of study@#The findings of this study contribute to the understanding of bacterial ecology in the controlled red algae cultivation environment and also provide valuable insights into the optimization of an ideal closed cultivation system for sustainable K. alvarezii production, benefiting the seaweed industry.
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Objective @#To investigate the differences in gut microbiota and metabolites between urban and rural middle school students and explore their significance in gut homeostasis , so as to establish a healthy lifestyle and diet for children.@*Methods @#Fecal samples were collected from middle school students in Hefei ( n = 14) and Jixi county ( n = 18 , Southern Anhui) , aged 13. 0 - 13. 5 years. Stool samples were sequenced by 16S ribosomal DNA (LC⁃MS) , followed by bioinformatic analysis. @*Results @# Lachnoclostridium and Anaerostipes were dominant in the urban students that had been reported to be associated with colorectal cancer, atherosclerosis , depression and other disorders. In the village children , Ruminococcaceae UCG⁃002 , Barnesiella and Eubacterium dominated. An increased proportion of these microbes were related to metabolism of bile acids , short⁃chain fatty acids , lipid and carbohydrate decomposition , and play an important role in maintaining immune balance and physiological function. Additionally , significant differences in gut metabolites of the two groups were noted , mainly in arachidonic acid metabolism , platelet activation , serotonin metabolism , vitamin absorption , primary bile acid metabolism and other pathways.@*Conclusion @#Adolescent students of urban and mountainous areas differ in gut microbiota and metabo⁃ lites. Rural children have a healthy bacterial flora and metabolites in guts due to a reasonable lifestyle and diet in comparison with the city children.
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Multiple organs are physiologically and pathologically interconnected during aging, and the brain plays a central role in this process. There is a direct two-way communication between the brain and the gut called “brain-gut interaction”, which is of great significance for the study of aging, and the molecular mechanism remains to be further studied. The aim of this study is to explore the mechanism of aging in the context of brain-gut interaction. The results of general physical signs of mice showed that the amount of exercise decreased, body weight and food intake decreased significantly in aged mice (P < 0. 001, P<0. 05). The thymus index of aged mice was significantly lower than that of normal mice (P< 0. 05), and the thymic pathological results showed that the thymic cortex of aging mice was thinner, the boundary between medulla and cortex was blurred, and the cells were loosely arranged. Metabolomics analysis revealed 317 differential metabolites in feces and 100 differential metabolites in hippocampus. The results of microbiome showed that Bacteroidetes and Firmicutes were the dominant phyla of gut microbiota. Bacteroidetes showed an upward trend and Firmicutes showed a downward trend after aging. KEGG pathway results showed that 26 metabolic pathways were related to the study of aging, among which galactose metabolism, ABC transporter and purine metabolism were of great significance for the brain-gut interaction. The results of Spearman correlation analysis of the three groups showed that the types of metabolites involved were mainly lipids and lipid-like molecules and organic acids and derivatives, and the gut microbiota involved were mainly Bacteroidetes and Firmicutes. In conclusion, the present study demonstrated that the synergistic changes between brain and gut in aging mice were related to the mechanism of aging, which provided new insights into the mechanism of aging process.