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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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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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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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Objective To investigate the infection and genotypes of Wolbachia in common mosquito species in Henan Province, so as to provide insights into management of mosquito-borne diseases. Methods Aedes, Culex and Anopheles samples were collected from cowsheds, sheepfolds and human houses in Puyang, Nanyang City and Xuchang cities of Henan Province from July to September, 2022, and the infection of Wolbachia was detected. The 16S rDNA and wsp genes of Wolbachia were amplified and sequenced. Sequence alignment was performed using the BLAST software, and the obtained 16S rDNA gene sequence was compared with the sequence of the 16S rDNA gene in GenBank database. In addition, the phylogenetic trees were created based on 16S rDNA and wsp gene sequences using the software MEGA 11.0. Results A total 506 female adult mosquitoes were collected from three sampling sites in Nanyang, Xuchang City and Puyang cities from July to September, 2022. The overall detection of Wolbachia was 45.1% (228/506) in mosquitoes, with a higher detection rate in A. albopictus than in Cx. pipiens pallens [97.9% (143/146) vs. 50.6% (85/168); χ2 = 88.064, P < 0.01]. The detection of Wolbachia in Cx. pipiens pallens was higher in Xuchang City (96.8%, 62/64) than in Nanyang (15.6%, 7/45) and Puyang cities (27.1%, 16/59) (χ2 = 89.950, P < 0.01). The homologies of obtained Wolbachia 16S rDNA and wsp gene sequences were 95.3% to 100.0% and 81.7% to 99.8%. Phylogenetic analysis based on wsp gene sequences showed Wolbachia supergroups A and B in mosquito samples, with wAlbA and wMors strains in supergroup A and wPip and wAlbB strains in supergroup B. Wolbachia strain wAlbB infection was detected in A. albopictus in Puyang and Nanyang Cities, while Wolbachia strain wPip infection was identified in A. albopictus in Xuchang City. Wolbachia strain wAlbA infection was detected in Cx. pipiens pallens sampled from three cities, and one Cx. pipiens pallens was found to be infected with Wolbachia strain wMors in Nanyang City. Conclusions Wolbachia infection is commonly prevalent in Ae. albopictus and Cx. pipiens pallens from Henan Province, and Wolbachia strains wAlbB and wAlbA are predominant in Ae. albopictus, while wPip strain is predominant in Cx. pipiens pallens. This is the first report to present Wolbachia wMors strain infection in Cx. pipiens pallens in Henan Province.
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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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@#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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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.
Subject(s)
Female , Humans , Vagina/microbiology , Uterine Cervical Neoplasms/genetics , Uterine Cervical Dysplasia , Cervix Uteri , Lactobacillus/genetics , Papillomavirus InfectionsABSTRACT
ObjectiveTo explore the relationship between the intestinal flora and the impairment of liver and kidney in HIV-infected men who have heterosexual sex with healthy women. MethodsFecal samples from 41 HIV-infected heterosexual men who have sex with women （PMSW） and 43 age- and BMI-matched healthy heterosexual men who have sex with women （NMSW） were collected and subjected to 16S rDNA sequencing. The blood levels of AST， ALT， TBIL， UREA， Cr， UA， β2-MG and other liver and kidney function indicators were measured. Bioinformatics methods were used to analyze the characteristics of the intestinal flora of the patients in these two groups， to compare the differential bacteria strains， and to analyze their correlation with liver and kidney function indicators. ResultsIn comparison with NMSW， the alpha diversity of intestinal flora was decreased in PMSW， and the beta diversity analysis showed significant differences in flora characteristics between the two groups （P<0.05）. The abundance of Clostridium， Phylum thick-walled， Trichosporon， and Clostridium tumefaciens decreased but Fusobacteriota increased （LDA score >4）. The comparison of liver and kidney function indexes revealed that AST， β2-MG levels were higher in PMSW than in NMSW， while TBIL was lower in PMSW than in NMSW. The number of patients with abnormal β2-MG was much higher in PMSW than in NMSW， and the difference was statistically significant （P<0.001）. It was also found that AST was negatively correlated with Clostridium （P<0.05）； TBIL was negatively correlated with Clostridium and positively correlated with Phylum thick-walled and Trichosporon （P<0.05）. β2-MG was negatively correlated with Phylum thick-walled， Clostridium， Trichosporon and Rumenococcus （P<0.05） and positively correlated with Clostridium （P<0.05）. ConclusionIn PMSW group， the alpha diversity of the flora is decreased. AST and β2-MG levels are increased， and TBIL level is decreased. These changes were significantly correlated with different strains of bacteria in the intestinal flora.
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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.
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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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ABSTRACT In this study, we investigated the chromosomes of three species of Sicarius spiders from the Brazilian Caatinga, using classical and molecular cytogenetic techniques. Based on the phylogenetic approach, we also discussed about the variation of diploid number, types of sex chromosome system and changes in the localization of ribosomal genes of Scytodoidea. Sicarius are Synspermiata spiders that together with the genera Loxosceles and Hexophthalma constitute the family Sicariidae. In this group, the available cytogenetic data showed a low diploid number range (2n♂=18 to 2n♂=23) and the presence of only multiple sex chromosome systems (X1X2Y and X1X20). Mitotic metaphase cells exhibited 2n♂=16+X1X2Y for Sicarius cariri and S. ornatus, and 2n♂=18+XY for S. tropicus. In these species, silver impregnation revealed nucleolar organizer region (Ag-NOR) on the terminal region of pair 1. In S. ornatus and S. tropicus, the results obtained with fluorescent in situ hybridization (FISH) using 18S rDNA probe were similar to Ag-NOR, however in S. cariri, the ribosomal sites were localized in the terminal region of the X1 sex chromosome. In this work, we presented the first description of a simple sex chromosome system for Sicariidae, helping to understand how the XY sex chromosome system evolved from the X1X2Y system. Additionally, FISH data incongruous with Ag-NOR indicate that the cytogenetic studies in Sicariidae allow investigating the relation between the karyotype evolution and the distribution and the activity of rDNA genes.
RESUMEN En este estudio, investigamos los cromosomas de tres especies de arañas Sicarius de la Caatinga brasileña, utilizando técnicas de citogenética clásica y molecular. Usando un enfoque filogenético, también discutimos la variación del número diploide, los tipos de sistema cromosómico sexual y los cambios en la localización de los genes ribosómicos en Scytodoidea. Los Sicarius son arañas Synspermiata que, junto con los géneros Loxosceles y Hexophthalma, constituyen a la familia Sicariidae. En este grupo, los datos citogenéticos disponibles mostraron un rango de número diploide bajo (2n♂=18 a 2n♂=23) y únicamente la presencia de sistemas de cromosomas sexuales múltiples (X1X2Y y X1X20). Las células mitóticas en metafase mostraron 2n♂=16+X1X2Y para Sicarius cariri y S. ornatus, y 2n♂=18+XY para S. tropicus. En estas especies, la impregnación de plata reveló la región organizadora nucleolar (Ag-NOR) en la región terminal del par 1. En S. ornatus y S. tropicus, los resultados obtenidos con la hibridación in situ fluorescente (FISH) utilizando la sonda de ADNr 18S fueron similares a los de Ag-NOR, sin embargo, en S. cariri los sitios ribosomales se localizaron en la región terminal del cromosoma sexual X1. En este trabajo, presentamos la primera descripción de un sistema cromosómico sexual simple para Sicariidae, ayudando a entender cómo el sistema cromosómico sexual XY evolucionó a partir del sistema X1X2Y. Además, los datos de FISH incongruentes con Ag-NOR indican que los estudios citogenéticos en Sicariidae permiten investigar la relación entre la evolución del cariotipo y la distribución y la actividad de los genes de ADNr.
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Background: Chronic HPV infection is a precursor of cervical cancer, which is largely caused by dysregulation of vaginal flora and other factors like abnormal H2O2, neuraminidase and insufficient vaginal hygiene. The relationship between HPV-induced cancer and vaginal microbiota is involved in the viral chronicity and also influences the disease prognosis. A meta-analysis system was used to evaluate the relationship between cervical lesions, HPV and vaginal microenvironment. Methods: PubMed, Web of Science, Cochrane and Embase databases were searched for relevant literature published from 2016 to December 2020. According to the inclusion and exclusion criteria, literature screening, data extraction and quality evaluation were carried out, and stata16 statistical software was used for Meta-analysis and systematic evaluation. Results: The overall relative risk of CST in 95% CI: 0.76-1.4, LSIL group compared with normal cytology group was 0.81. The overall relative risk of CST in the HSIL group and cervical cancer group was 0.77 and 1.26, respectively. It was found that there was publication bias in the HPV positive group (p-value of Begg and Egger were 0.067 and 0.247) and cervical cancer group (p-value of Begg and Egger were 0.677 and 0.457 respectively). There was a significant difference in CST III between HPV positive group and the LSIL group. Conclusion: Cervical lesions and HPV are related to the increase of vaginal microbial diversity, and HPV and LSIL groups are related to CST III, while HSIL and cervical cancer groups are related to CSTIV, which has a certain guiding significance for early clinical diagnosis, but further large-scale studies are needed to confirm our findings.
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Abstract Potable water supply and sanitization in rural areas in developing countries are still inadequate. The main risk associated with unsafe drinking water is the infection with pathogenic microorganisms. Objective: In this study, we investigate the bacterial diversity and the potentially pathogenic bacteria in water samples from diffe rent points of distribution in three rural villages from the Andean region of Colombia. Methods: Illumina libraries for water samples were prepared and sequenced using 300 bp paired-end MiSeq protocol, the bioinformatic analyses were performed with Mothur pipeline and the phyloseq package in Rstudio. Results: The mi crobial community composition showed statistically significant differences according to the village and the sample origin. Alpha, Beta, and Gammaproteobacteria were the dominant class detected in all water samples. The most relevant pathogenic genera detected in the surface were Legionella, Mycobacterium, Yersinia, Burkholderia, and Rickettsia. In the tap water samples, potential pathogens like Streptococcus, Staphylococcus, Corynebacterium, Nocardia, and Escherichia/Shige lla were detected.
Resumen El suministro y potabilización del agua de consumo humano en las zonas rurales de los países en vías de desarrollo sigue siendo limitado. El principal riesgo asociado con el uso de agua no potable es la infección con microorganismos patógenos. Objetivo: En este estudio se investigó la diversidad bacteriana y la presencia de bacterias potencialmente patógenas en muestras de agua de diferentes puntos de distribución en tres asentamientos rurales de la región andina de Colombia. Métodos: Se prepararon y secuenciaron bibliotecas de amplicones (rDNA 16S) para muestras de agua utilizando la plataforma Illumina MiSeq con lecturas pareadas de 300 bases. Los análisis bioinformáticos se realizaron con el programa Mothur y el paquete estadístico Phyloseq en Rstudio. Resultados: La composición de la comunidad microbiana mostró diferencias estadísticamente significativas según el sitio y el origen de la muestra. Alfa, Beta y Gammaproteobac terias fueron las clase dominantes detectadas en todas las muestras de agua. Los géneros patógenos más relevantes detectados fueron Legionella, Mycobacterium, Yersinia, Burkholderia y Rickettsia. En las muestras de agua del grifo se detectaron patógenos potenciales como Streptococcus, Staphylococcus, Corynebacterium, Nocardia y Escherichia /Shigella.
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In aquaculture, microalgal-bacterial interaction has ecological significance, and thus demands better understanding for improvement of sustainability and productivity of large scale microalgal cultivation. Here, we assessed the bacterial diversity, including the uncultivable bacterial assemblage associated with the marine microalgae Isochrysis galbana using next generation sequencing approach. Isochrysis has been considered as one of the most favoured types of live feed in aquaculture and hence, chose Isochrysis galbana MBTDCMFRI S002. Total genomic DNA was extracted from I. galbana culture and 16S rDNA V3 region was sequenced with an Illumina MiSeq platform. A total of 30 different known bacterial genera were detected from 1190 identified operational taxonomic units. These bacterial phylotypes were affiliated to Alphaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteria, Acidimicrobiia, Bacteroidia, Flavobacteriia and Bacilli classes. These 30 bacterial genera comprise only 4.62% of the total OTUs obtained and remaining 95.38% of the sequences do not exhibit any similarity against known bacterial genera in the taxonomic database. The functional profile of bacterial communities was predicted using PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) analysis. The results indicated that these associated bacterial communities are mainly involved in environmental as well as genetic information processing, membrane transport and nutrient metabolism. These functions may mediate their interaction with the phytoplankton host, and thus improve bacterial survival in microalgal habitat. Overall, the present study enhances the understanding of microalgal-bacterial interaction in terms of diversity and functional role of associated microbial community.
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In aquaculture, microalgal-bacterial interaction has ecological significance, and thus demands better understanding for improvement of sustainability and productivity of large scale microalgal cultivation. Here, we assessed the bacterial diversity, including the uncultivable bacterial assemblage associated with the marine microalgae Isochrysis galbana using next generation sequencing approach. Isochrysis has been considered as one of the most favoured types of live feed in aquaculture and hence, chose Isochrysis galbana MBTDCMFRI S002. Total genomic DNA was extracted from I. galbana culture and 16S rDNA V3 region was sequenced with an Illumina MiSeq platform. A total of 30 different known bacterial genera were detected from 1190 identified operational taxonomic units. These bacterial phylotypes were affiliated to Alphaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteria, Acidimicrobiia, Bacteroidia, Flavobacteriia and Bacilli classes. These 30 bacterial genera comprise only 4.62% of the total OTUs obtained and remaining 95.38% of the sequences do not exhibit any similarity against known bacterial genera in the taxonomic database. The functional profile of bacterial communities was predicted using PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) analysis. The results indicated that these associated bacterial communities are mainly involved in environmental as well as genetic information processing, membrane transport and nutrient metabolism. These functions may mediate their interaction with the phytoplankton host, and thus improve bacterial survival in microalgal habitat. Overall, the present study enhances the understanding of microalgal-bacterial interaction in terms of diversity and functional role of associated microbial community.
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The diversity of yeast species on grape berries changes depending on various factors. Determination of indigenous yeast intensity and diversity on grape berries is important for increasing the sensory characteristics of wine as well as fermentation efficiency. The natural yeast biota of grape berries affects wine flavour and quality by producing some secondary metabolites and hydrolytic enzymes. Despite the application of different nonconventional yeasts in food and fermentation industries, many significant researches are conducted in finding and improving the new strains having industrially important enzyme activities. Invertase enzyme has a vital role in the food industry in which it increases the sweetness of food without crystallizing them. Here, we studied yeast diversity in grape berries from selected localities and also their invertase activity. We collected grape berries from Alphonse, K?nal? Yap?ncak, Çavu?, Efes Karas?, Cinsaut, Atasar?s? and Isabella grape varieties cultivated in Bozcaada island and Gelibolu peninsula. Twenty-one yeast species belonging to seven genera were identified. The yeast strains having high invertase activities were identified with 5.8S-ITS rDNA sequencing technique. The diversity of yeast biota on berries collected from Gelibolu was greater than that of Bozcaada. Metschnikowia pulcherrima, Cryptococcus laurentii and Rhodotorula glutinis yeast species were dominant yeast species on grape berries. A total of 294 sucrose grown yeast strains showed growth on sucrose, and 19 of them exhibit the highest invertase activity that is not glucose repressible. These 19 yeast strains were identified as Starmerella bacillaris using 5.8S ITS rDNA region and the phylogenetic analysis was inferred with the Maximum Parsimony method.
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Objective:To investigate the regulation of intestinal microbiota by Roux-en-Y gastric bypass (RYGB) on patients with obesity or obesity combined with diabetes.Methods:The retrospective and descriptive study was conducted. The stool samples before and after surgery and clinical data of 20 patients with obesity, including 9 simple obesity cases and 11 obesity combined with diabetes cases, who underwent RYGB in the First Affiliated Hospital of Ji′nan University from July 2016 to August 2017 were collected. There were 11 males and 9 females, aged (33±11)years. Observation indicators: (1) changes in composition and structure of intestinal microflora; (2) changes of intestinal microflora in simple obesity patients after operation; (3) changes of intestinal microflora in obesity combined with diabetes patients after operation. Follow up was conducted using telephone interview or outpatient examinations to detect the body mass, the application of antimicrobial agent and the blood glucose control of patients. According to the unified training points, the stool samples were collected and stored into the DNA stabilizer, and then conducted to laboratory analysis within 45 hours. The follow up was up to November 2018. Measurement data with normal distribution were represented as Mean± SD, and independent-samples t test was used for inter-group comparison and paired-samples t test was used for intra-group comparison. Measurement data with skewed distribution were represented as M( Q1, Q3), and Wilcoxon signed rank test of two independent samples was used for inter-group comparison. Count data were described as absolute numbers, and the chi-square test, ANOSIM analysis, linear discriminant (LEfSe) analysis and the Metastats analysis were used for inter-group comparison. Results:(1) Changes in composition and structure of intestinal microflora. The Shannon index of α diversity of preoperative intestinal microflora in simple obesity patients and obesity combined with diabetes patients was 4.37±0.69 and 4.47±0.85, respectively, showing no significant difference between them ( t=0.28, P>0.05). Results of preoperative LEfSe analysis showed that there were differences in the bacterial abundance of Firmicutes and Bacteroidea between simple obesity patients and obesity combined with diabetes patients. The abundances of Parasutterella in simple obesity patients and obesity combined with diabetes patients was 0.000 113 0(0, 0.004 378 2) and 0.008 464 0(0.001 325 7, 0.034 983 1), respectively, showing a significant difference between them ( Z=2.12, P<0.05). Results of preoperative PCoA analysis showed that the contribution rates of principal component 1, principal component 2 and principal component 3 were 24.98%, 22.24% and 16.33% in simple obesity patients and obesity combined with diabetes patients and results of ANOSIM comparison showed that there was no significant difference in preoperative intestinal microflora between them ( r=?0.11, P>0.05). The Shannon index of α diversity of postoperative intestinal microflora in simple obesity patients and obesity combined with diabetes patients was 4.60±0.65 and 4.66±0.40, respectively, showing no significant difference between them ( t=0.24, P>0.05). Results of postoperative LEfSe analysis showed that there were differences in the bacterial abundance of Bacteroidea, Proteus and Firmicutes between simple obesity patients and obesity combined with diabetes patients. The abundances of Morganella and Coprococcus_2 in simple obesity patients and obesity combined with diabetes patients were 0.000 192 0(0.000 011 9,0.001 569 0), 0(0,0) and 0(0,0), 0.000 054 1(0,0.000 419 0), showing significant differences between them ( Z=2.70, 2.29, P<0.05). (2) Changes of intestinal microflora in simple obesity patients after operation. There were 10 genera of bacteria of intestinal bacteria changing after surgery, including 7 species of bacteria increasing in the Firmicutes and the Proteobacteria as Veillonella, Morganella, Granulicatella, Aeromonas, Streptococcus, Rothia and Megasphaera and the bacteria decreasing in the Firmicutes and the Actinobacteria as Ruminococcus_torques_group, Romboutsia and Erysipelo-trichaceae_UCG-003. Results of LEfSe analysis showed that the bacteria significantly enriched in simple obesity patients before surgery were Ruminococcus_torques_group, Romboutsia and Erysipelotri-chaceae_UCG-003, belonging to Firmicutes, and the bacteria significantly enriched in simple obesity patients after surgery were Rothia, Granulicatella, Enterococcus, Streptococcus, Megasphaera, Veillonella, A eromonas and Morganella, belonging to Actinobacteria, Firmicutes and Proteobacteria. (3) Changes of intestinal microflora in obesity combined with diabetes patients after operation. There were 16 bacteria of intestinal bacteria increasing after surgery, including Streptococcus, Veillonella, Haemophilus, Pluralibacter, Gemella, Lachnospiraceae_NC2004_group, Granulicatella,Aeromonas, uncultured_ bacterium_f_ Saccharimonadaceae, R uminiclostridium_9, Butyricicoccus, Fusobacterium, Anaerotruncus, Fusicateni-bacter, Klebsiella and E ubacterium_eligens_group, which belonged to the Firmicutes and the Proteo-bacteria. Results of LEfSe analysis showed that the bacteria significantly enriched in obesity combined with diabetes patients before surgery were Fusicatenibacter, Tyzzerella_3 and Butyricicoccus, belonging to the Firmicutes, and the bacteria significantly enriched in obesity combined with diabetes patients after surgery were Gemella, Granulicatella, Enterococcus, Streptococcus, Lachnospiraceae_NC2004_group, Eubacterium_eligens_group, Anaerotruncus, Ruminiclostridium_9, Anaeroglobus, Veillonella, Fusobacterium, uncultured_bacterium_f_Saccharimonadaceae, Aeromonas, Klebsiella, Pluralibacter, Proteus and Haemophilus, belonging to the Firmicutes and the Proteobacteria. Conclusions:RYGB can significantly increases the intestinal microflora abundance in simple obesity patients and obesity combined with diabetes patients. The two types of patients have specific changes in intestinal microflora at the genus level.
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Objective:To investigate the effects of long-term high-fat diet on bone mineral density and intestinal flora in mice.Methods:Sixteen male C57BL/6 mice were randomly divided into control group (NC group) and high-fat group (HF group). After 24 weeks of high-fat feeding, biochemical indicators such as blood glucose and blood lipids were detected, bilateral femurs were taken and bone microstructure was analyzed with micro-computered tomography (micro-CT), and changes of intestinal microbial composition and proportion were revealed using 16S rDNA sequencing technology.Results:Compared with the control group, the serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in HF group were significantly increased. Micro-CT uncovered that the bone mineral density (Tb.BMD), bone volume fraction (BV/TV) and the number of trabecular bone (Tb.N) decreased, yet structural model index (SMI) and the trabecular fraction (Tb.Sp) increased in the HF group mice. The gut microbiota 16S rDNA sequence analysis showed that the proportion of Proteobacter was significantly increased and the proportions of pachycete, warty microbacterius, and actinomycete were reduced in HF group at the phyla level. The proportion of Bacteroidetes S24-7_norank in the NC group was significantly higher than that in the HF group, and the multilevel discriminant analysis of species differences (LEfSe) identified that the difference was significant, yet the proportion of Bacteroides, Pseudo-Prevotella, Desulfovibrio, Altobacter, and Helicobacter in the HF group were higher than those in the NC group, which were significant differences in Altobacter and Helicobacter at genus level.Conclusion:Long-term high-fat feeding can cause the destruction of femoral trabecular structure, decrease in the number of trabeculus bones, and bone mineral density in C57BL/6 mice. It also leads to significant changes in the composition and proportion of the intestinal flora.
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Objective:To investigate the changes of intestinal microecology in the early stage of sepsis rat model by 16S rDNA sequencing.Methods:Sixty male Sprague-Dawley (SD) rats were randomly divided into cecal ligation and puncture (CLP) group and sham operation group (Sham group), with 30 rats in each group. In the CLP group, sepsis rat model was reproduced by CLP method; the rats in the Sham group only underwent laparotomy without CLP. At 24 hours after the operation, the intestinal feces and serum samples of 8 rats in each group were collected. The survival rate of the rest rats was observed until the 7th day. The level of serum tumor necrosis factor-α (TNF-α) was detected by enzyme-linked immunosorbent assay (ELISA). Intestinal feces were sequenced by 16S rDNA sequencing technology. The operational taxonomic unit (OTU) data obtained after sequence comparison and clustering was used for α diversity and β diversity analysis, principal coordinate analysis and linear discriminant analysis effect size analysis (LEfSe) to observe the changes of intestinal microecology in early sepsis rats and excavate the marker flora.Results:At 24 hours after the reproduction of the model, the rats in the CLP group showed shortness of breath, scattered hair and other manifestations, and the level of serum TNF-α increased significantly as compared with that in the Sham group (ng/L: 43.95±9.05 vs. 11.08±3.27, P < 0.01). On the 7th day after modeling, the cumulative survival rate of the Sham group was 100%, while that of the CLP group was 31.82%. Diversity analysis showed that there was no significant difference in α diversity parameter between the Sham group and the CLP group (number of species: 520.00±52.15 vs. 492.25±86.61, Chao1 richness estimator: 707.25±65.69 vs. 668.93±96.50, Shannon index: 5.74±0.42 vs. 5.79±0.91, Simpson index: 0.93±0.03 vs. 0.94±0.05, all P > 0.05). However, the β diversity analysis showed that the difference between groups was greater than that within groups whether weighted according to OTU or not (abundance weighted matrix: R = 0.23, P = 0.04; abundance unweighted matrix: R = 0.32, P = 0.01). At the phylum level, the abundance of Proteobacteria and Candidatus_sacchari in the CLP group increased significantly as compared with the Sham group [18.100% (15.271%, 26.665%) vs. 6.974% (2.854%, 9.764%), 0.125% (0.027%, 0.159%)% vs. 0.018% (0.008%, 0.021%), both P < 0.05]. At the genus level, the abundance of opportunistic pathogen including Helicobacter, Ruthenium, Streptococcus, Clostridium ⅩⅧ in the CLP group was significantly higher than that in the Sham group [5.090% (1.812%, 6.598%) vs. 0.083% (0.034%, 0.198%), 0.244% (0.116%, 0.330%) vs. 0.016% (0.008%, 0.029%), 0.006% (0.003%, 0.010%) vs. 0.001% (0%, 0.003%), 0.094% (0.035%, 0.430%) vs. 0.007% (0.003%, 0.030%), all P < 0.05], and the abundance of probiotics such as Alloprevotella and Romboustia was significantly lower than that in the Sham group [7.345% (3.662%, 11.546%) vs. 22.504% (14.403%, 26.928%), 0.113% (0.047%, 0.196%) vs. 1.229% (0.809%, 2.29%), both P < 0.01]. LEfSe analysis showed that the probiotics belonging to Firmicutes were significantly enriched in the Sham group, and Romboustia was the most significantly enriched species. Opportunistic pathogens such as Helicobacter, Streptococcus and Clostridium ⅩⅧ were significantly enriched in the CLP group, Helicobacter_NGSU_ 2015 was the most significantly enriched species. Conclusion:In the early stage of sepsis, the intestinal microbiota structure of rats is significantly changed, which mainly shows that the abundance of Alloprevotella and other probiotics is significantly reduced, while that of Helicobacter and other opportunistic pathogens is significantly increased.
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Background Esophageal cancer is a common gastrointestinal tumor with a high incidence in China. Some studies suggest that intestinal flora is significantly related to the occurrence and development of tumors and other diseases. Traditional 16S rDNA sequencing technology only provides taxonomic resolution at genus level. Objective Based on PacBio single molecule real time (SMRT) sequencing technology to identify characteristic microbial biomarkers associated with esophageal cancer at the species level. Methods A total of 120 newly diagnosed cases of esophageal cancer were recruited and 60 healthy patients with matched sex and age were recruited as the control group. Fresh stool samples were collected from all subjects. Full-length 16S rDNA sequencing was performed on samples from 4 patients with esophageal cancer and 1:1 matched healthy controls using the third-generation sequencing PacBio SMRT technology, and the structural differences of intestinal flora were analyzed based on the sequencing results. Function prediction was performed by PICRUSt software. Large population samples were validated by screening different gut microbes by linear discriminant analysis and linear discriminant analysis effect size to identify esophageal cancer-associated gut microbes. Results Based on sequencing samples, the results of α diversity analysis showed that the Ace, Chao1, Simpson Diversity, and Shannon Wiener indices of the esophageal cancer group were higher than those of the healthy control group (P<0.05), and the results of β diversity showed that the scattered clusters of the esophageal cancer group and the healthy control group were separated, which meant that there were differences in the structure of intestinal flora between the two groups. It was found at the phylum level that the abundances of Proteobacteria, Bacteroidetes, and Firmicutes in the intestinal flora of the esophageal cancer group were increased. At the genus level, the relative abundances of Spirospira, Pasteurella, Roxella, and Bacteroides in the esophageal cancer group were increased. At the species level, there were 11 microbial species with increased relative abundances in the esophageal cancer group, including Enterobacter sp. E.20, Bacteroides ovatus V975, and Faecalibacterium prausnitzii, and the microbial species with decreased relative abundances in the esophageal cancer group were Ralstonia pickettii, Enterobacter unclassified, and Streptococcus salivarius JIM8777. The PICRUSt functional annotation found differences in alanine, aspartate and glutamate metabolism (map00250), peptidoglycan (map00550), one carbon pool by folate (map00670), thiamine metabolism (map00730), and biosynthesis of amino acids (map01230) between the two groups. The results of the population validation study showed that the abundances of Enterobacter sp E.20 and Bacteroides massilience in the esophageal cancer group were increased, the abundance of Streptococcus salivarius JIM8777 was decreased, and the differences between the two groups were statistically significant (P<0.05). By establishing receiver operating characteristic analysis for representative species level biomarkers, the area under curve (AUC) of combining Enterobacter sp E.20, Streptococcus salivarius JIM8777, and Bacteroides massilience was 0.779, higher than single diagnosis (AUC=0.610, 0.608, and 0.659, respectively). Conclusion There are significant differences in gut microbiota between the esophageal cancer group and the healthy control group. The combination of Enterobacter sp E.20, Streptococcus salivarius JIM8777, and Bacteroides Massilience has potential application value for the diagnosis of esophageal cancer.