207 ESOPHAGEAL BIOPSY AND SWAB MICROBIAL CHARACTERIZATION WITH HEALTH AND ESOPHAGEAL DISEASE IN CHINA

2020 ◽  
Vol 33 (Supplement_1) ◽  
Author(s):  
M Li ◽  
D Shao ◽  
G Song ◽  
F Meng ◽  
C Hao ◽  
...  

Abstract   Little is known about the esophageal microbiota and esophageal health or disease in China, especially esophageal squamous cell precancerous lesions. And there is currently no optimal sampling method for esophageal microbiota. Consequently, we compared microbial signature between and within esophageal biopsy and swab specimens from participants in healthy or with esophageal diseases. Methods A total of 236 subjects were recruited in the Linxian Cancer Hospital (Henan, China), including 5 groups as 70 healthy cases, 70 esophagitis cases, 70 low grade intraepithelial neoplasia (LGIN) cases, 19 high grade intraepithelial neoplasia (HGIN) cases and 7 esophageal squamous cell carcinoma (ESCC) cases. Sterile biopsies and swabs were used to collect paired samples from the same participants under the endoscopy examination. DNA was extracted with the MoBio PowerSoil Kit. Data from 16S rRNA gene sequencing were processed using QIIME2 and R to evaluate differences in alpha and beta diversity and taxonomic relative abundances. Results Alpha diversity was not significantly different between specimens (biopsy = 116.2, swab = 118.0, P > 0.05). In swabs, there were differences of observed OTUs among groups but not Shannon index, detailly, there was an inconspicuous decreasing from Health (125.0) to ESCC (114.0). About beta diversity, no distinct clustering by specimen was observed for weighted Unifrac distance and unweighted Unifrac distance. The 10 dominant genera were similar between specimens and among groups. Specially, Streptococcus was the most abundant except ESCCs; Fusobacterium was in the top ten only in ESCCs. And there was a decreasing of Prevotella 7 and an increasing of Haemophilus as progression. Conclusion Esophageal biopsy and swab specimens could yield similar bacterial composition. Both can be used to find clues of microbiota and disease. And esophageal microbial composition and progression characterizations could provide new idea and basis for large scale, prospective cohort studies in the future.

Author(s):  
Dehua Liu ◽  
Si Chen ◽  
Yawen Gou ◽  
Wenyong Yu ◽  
Hangcheng Zhou ◽  
...  

BackgroundGastric microbiota may be involved in gastric cancer. The relationship between gastrointestinal microbes and the risk of gastric cancer is unclear. This study aimed to explore the gastric and intestinal bacteria associated with gastritis and gastric precancerous lesions. We conducted a case-control study by performing 16S rRNA gene analysis of gastric biopsies, juices, and stool samples from 148 cases with gastritis or gastric precancerous lesions from Anhui and neighboring provinces, China. And we validated our findings in public datasets.ResultsAnalysis of microbial sequences revealed decreased bacterial alpha diversity in gastric bacteria during the progression of gastritis. Helicobacter pylori was the main contributor to the decreased microbial composition and diversity in the gastric mucosa and had little influence on the microbiota of gastric juice and feces. The gastric mucosal genera Gemella, Veillonella, Streptococcus, Actinobacillus, and Hemophilus had the higher degree of centrality across the progression of gastric precancerous lesions. And Acinetobacter may contribute to the occurrence of intraepithelial neoplasia. In addition, the microbial model of H. pylori-positive gastric biopsies and feces showed value in the prediction of gastric precancerous lesions.ConclusionsThis study identified associations between gastric precancerous lesions and gastric microbiota, as well as the changes in intestinal microbiota, and explored their values in the prediction of gastric precancerous lesions.


Author(s):  
Zhengqi Li ◽  
Lizhou Dou ◽  
Yueming Zhang ◽  
Shun He ◽  
Deli Zhao ◽  
...  

Important evidence indicates that the microbiota plays a key role in esophageal squamous cell carcinoma (ESCC). Here, paired saliva and brush specimens were obtained from 276 participants undergoing upper gastrointestinal endoscopic examination before or during screening for upper gastrointestinal (UGI) cancer. The esophageal microbiota was investigated by 16S rRNA gene profiling and next-generation sequencing. We observed that as the disease progressed, the α diversity in the saliva and cell brush samples decreased. Linear discriminant analysis effect size (LEfSe) results showed that in both the saliva and cell brush specimens, Granulicatella, Rothia, Streptococcus, Gemella, Leptotrichia and Schaalia were common biomarkers in patients with low-grade dysplasia, Lactobacillus was a common biomarker in patients with high-grade dysplasia, and Bosea, Solobacterium, Gemella, and Peptostreptococcus were common biomarkers in patients with esophageal cancer. The top 3 genera in the saliva and cell brush specimens had areas under the curve (AUCs) of 87.16 and 89.13%, respectively, to distinguish ESCC patients from normal people. The PICRUSt2 results identified in brush samples that patients with ESCC had decreased nitrate reductase functions. Our results suggest that future studies can focus on the function of the characteristic bacteria in ESCC.


2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1907.2-1907
Author(s):  
Y. Tsuji ◽  
M. Tamai ◽  
S. Morimoto ◽  
D. Sasaki ◽  
M. Nagayoshi ◽  
...  

Background:Anti-citrullinated protein antibody (ACPA) production is observed in several organs even prior to the onset of rheumatoid arthritis (RA), and oral mucosa is considered to be one of the important tissues. The presence of HLA-DRB1*SE closely associates with ACPA production. Saliva is considered to reflect the oral microbiota including periodontal disease. Alteration of oral microbiota of RA becomes to be normalized by DMARDs treatment, however, the interaction of HLA-DRB1*SE, ACPA and oral microbiota of RA patients remains to be elucidated.Objectives:The Nagasaki Island Study, which had started in 2014 collaborating with Goto City, is intended for research of the preclinical stage of RA, including ACPA/HLA genotype screening and ultrasound and magnetic resonance imaging examinations in high-risk subjects. Using the samples accumulated in this cohort, we have tried to investigate the difference of oral microbiota among RA patients and healthy subjects regarding to ACPA and HLA-DRB1*SE.Methods:Blood and salivary samples were obtained from 1422 subjects out of 4276 who have participated in the Nagasaki Island Study from 2016 to 2018. ACPA positivity was 1.7 % in total. Some of RA patients resided in Goto City participated in the Nagasaki Island Study. At this point, we selected 291 subjects, who were ACPA positive non-RA healthy subjects (n=22) and patients with RA (n=33, 11 subjects were ACPA positive and 22 ACPA negative respectively) as the case, age and gender matched ACPA negative non-RA healthy subjects (n=236) as the control. ACPA was measured by an enzyme-linked immunosorbent assay, and HLA genotyping was quantified by next-generation sequencing (Ref.1). The operational taxonomic unit (OUT) analysis using 16S rRNA gene sequencing were performed. The richness of microbial diversity within-subject (alpha diversity) was scaled via Shannon entropy. The dissimilarity between microbial community composition was calculated using Bray-Curtis distance as a scale, and differences between groups (beta diversity) were tested by permutational multivariate analysis of variance (PERMANOVA). In addition, UniFrac distance calculated in consideration of the distance on the phylogenetic tree were performed.Results:Median age 70 y.o., % Female 58.8 %. Among RA and non-RA subjects, not alpha diversity but beta diversity was statistically significance (p=0.022, small in RA). In RA subjects, both alpha and beta diversity is small (p<0.0001), especially significant in ACPA positive RA (Figure 1). Amongt RA subjects, presence of HLA-DRB1*SE did not show the difference but the tendency of being small of alpha diversity (p=0.29).Conclusion:Our study has suggested for the first time the association of oral microbiota alteration with the presence of ACPA and HLA-DRB1*SE. Oral dysbiosis may reflect the immunological status of patients with RA.References:[1]Kawaguchi S, et al. Methods Mol Biol 2018;1802: 22Disclosure of Interests:None declared


2020 ◽  
Vol 4 (6) ◽  
Author(s):  
Yinyin Zhang ◽  
Hong Lin ◽  
Huizhen Fan

As a drug-mechanical combination technology, photodynamic (PDT) can achieve accurate and targeted therapy for malignant tumors and benign diseases through the production of reactive oxygen species, oxygen free radicals or singlet oxygen by photosensitizers at specific wavelengths. Compared with traditional surgery, it has the advantages of selective killing, repeatable treatment, preserving target organ function and so on. The purpose of this study was to explore the clinical value of photodynamic therapy in cervical precancerous lesions by taking the patients with low-grade cervical intraepithelial neoplasia (CIN1) with high-risk human papillomavirus (HR-HPV) persistent infection diagnosed by "three-step diagnosis and treatment procedure" as an example. Using HiPorfin as a photosensitizer, photodynamic therapy was performed 48 hours after intravenous drip. Set laser wavelength 630nm, light dose density 137.58J/cm2, transmission efficiency 1.42, output power 2w. 3cm columnar optical fiber was placed around the 2cm in the cervical canal to cover all the lesions, and the irradiation time was 900s (600s in the cervical canal and 300s outside the cervical canal). The patients were given oxygen inhalation for 6 hours after operation, and the patients were observed for itching and other discomfort, and paid attention to avoid light. Photodynamic therapy was performed again in the same way on the second day. After two months of treatment, pathological biopsy showed chronic cervicitis, indicating that the disease had been effectively controlled. Theoretically, although the patient is not the absolute indication of photodynamic therapy (that is, meeting CIN ? or CIN ?, having fertility requirements and not undergoing surgery), this therapy can remove not only the superficial lesions inside and outside the cervix, but also the potential lesions not found under colposcopy. It can also block the persistent infection of HPV by inhibiting the expression of HPV18, E6 and E7mRNA in Hela cells. In combination with Baofukang suppository, it can block HPV infection. Increase the negative conversion rate of cervical HPV and reduce the probability of recurrence after CIN1 cure. For young female patients with persistent HR-HPV infection and fertility requirements, photodynamic therapy is an effective choice for clinical treatment of CIN1.


2020 ◽  
Author(s):  
Shintaro Sukegawa ◽  
Sawako Ono ◽  
Keisuke Nakano ◽  
Kiyofumi Takabatake ◽  
Hotaka Kawai ◽  
...  

Abstract Background This study was conducted to compare histological diagnostic accuracy between conventional method and liquid-based cytology (LBC) method and to clarify the effectiveness of cytology. Methods Histological diagnoses of 251 cases were classified as negative (no malignancy lesion, inflammation, and mild/moderate dysplasia) and positive [oral intraepithelial neoplasia/carcinoma in situ (OIN/CIS) and squamous cell carcinoma (SCC)]. Cytological diagnoses were classified as negative for intraepithelial lesion or malignancy (NILM), oral low-grade squamous intraepithelial lesion (OLSIL), oral high-grade squamous intraepithelial lesion (OHSIL), or SCC. Cytological diagnostic results were compared with histology results. Results Of NILM cytology cases, the most frequent case was negative [LBC n = 50 (90.9%), conventional n = 22 (95.7%)]. Among OLSIL cytodiagnoses, the most common was negative (LBC n = 34; 75.6%, conventional n = 14; 70.0%). Among OHSIL cytodiagnoses (LBC n = 51, conventional n = 23), SCC was the most frequent (LBC n = 31; 60.8%, conventional n = 7; 30.4%). Negative cases were common (LBC n = 13; 25.5%, conventional n = 14; 60.9%). Among SCC cytodiagnoses SCC was the most common (LBC n = 16; 88.9%, conventional n = 14; 87.5%). Regarding the diagnostic results of cytology, assuming HSIL and SCC as cytologically positive, the LBC method/conventional method showed a sensitivity of 79.4%/76.7%, specificity of 85.1%/69.2%, false positive rate of 14.9%/30.7%, and false negative rate of 20.6%/23.3%. Conclusions LBC method was superior to conventional method in cytodiagnosis. It was especially superior for LSIL and HSIL. Because of the false positive and false negative cytodiagnoses, it is necessary to make a comprehensive diagnosis considering the clinical findings.


2020 ◽  
Vol 98 (6) ◽  
Author(s):  
Riley D Messman ◽  
Zully E Contreras-Correa ◽  
Henry A Paz ◽  
George Perry ◽  
Caleb O Lemley

Abstract The knowledge surrounding the bovine vaginal microbiota and its implications on fertility and reproductive traits remains incomplete. The objective of the current study was to characterize the bovine vaginal bacterial community and estradiol concentrations at the time of artificial insemination (AI). Brangus heifers (n = 78) underwent a 7-d Co-Synch + controlled internal drug release estrus synchronization protocol. At AI, a double-guarded uterine culture swab was used to sample the anterior vaginal tract. Immediately after swabbing the vaginal tract, blood samples were collected by coccygeal venipuncture to determine concentrations of estradiol. Heifers were retrospectively classified as pregnant (n = 29) vs. nonpregnant (n = 49) between 41 and 57 d post-AI. Additionally, heifers were classified into low (1.1 to 2.5 pg/mL; n = 21), medium (2.6 to 6.7 pg/mL; n = 30), and high (7.2 to 17.6 pg/mL; n = 27) concentration of estradiol. The vaginal bacterial community composition was determined through sequencing of the V4 region from the 16S rRNA gene using the Illumina Miseq platform. Alpha diversity was compared via ANOVA and beta diversity was compared via PERMANOVA. There were no differences in the Shannon diversity index (alpha diversity; P = 0.336) or Bray–Curtis dissimilarity (beta diversity; P = 0.744) of pregnant vs. nonpregnant heifers. Overall, bacterial community composition in heifers with high, medium, or low concentrations of estradiol did not differ (P = 0.512). While no overall compositional differences were observed, species-level differences were present within pregnancy status and estradiol concentration groups. The implications of these species-level differences are unknown, but these differences could alter the vaginal environment thereby influencing fertility and vaginal health. Therefore, species-level changes could provide better insight rather than overall microbial composition in relation to an animal’s reproductive health.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Lisa Karstens ◽  
Nazema Y. Siddiqui ◽  
Tamara Zaza ◽  
Alecsander Barstad ◽  
Cindy L. Amundsen ◽  
...  

AbstractThe urinary microbiome has been increasingly characterized using next-generation sequencing. However, many of the technical methods have not yet been specifically optimized for urine. We sought to compare the performance of several DNA isolation kits used in urinary microbiome studies. A total of 11 voided urine samples and one buffer control were divided into 5 equal aliquots and processed in parallel using five commercial DNA isolation kits. DNA was quantified and the V4 segment of the 16S rRNA gene was sequenced. Data were processed to identify the microbial composition and to assess alpha and beta diversity of the samples. Tested DNA isolation kits result in significantly different DNA yields from urine samples. DNA extracted with the Qiagen Biostic Bacteremia and DNeasy Blood & Tissue kits showed the fewest technical issues in downstream analyses, with the DNeasy Blood & Tissue kit also demonstrating the highest DNA yield. Nevertheless, all five kits provided good quality DNA for high throughput sequencing with non-significant differences in the number of reads recovered, alpha, or beta diversity.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Lang Yang ◽  
Hua Jin ◽  
Xiao-li Xie ◽  
Yang-tian Cao ◽  
Zhen-hua Liu ◽  
...  

Abstract Background Endoscopic resection has been used for high-grade intraepithelial neoplasia (HGIN) and superficial esophageal squamous cell carcinoma (ESCC) with limited risk of lymph node metastasis. However, some of these lesions cannot be accurately diagnosed based on forceps biopsy prior to treatment. In this study we aimed to investigate how to solve this histological discrepancy and avoid over- and under-treatment. Methods The medical records of patients with superficial esophageal squamous cell neoplasia who underwent endoscopic resection at our hospital from January 2012 to December 2019 were reviewed retrospectively. The histological discrepancy between the biopsy and resected specimens was calculated and its association with clinicopathological parameters was analyzed. Results A total of 137 lesions from 129 patients were included. The discrepancy rate between forceps biopsy and resected specimens was 45.3% (62/137). Histological discrepancy was associated with the histological category of the biopsy (p < 0.001). In addition, 17 of the 30 (56.7%) biopsies that was diagnosed as indefinite/negative for neoplasia or low-grade intraepithelial neoplasia were upgraded to HGIN or ESCC after resection. The upgrade was due to lesion size ≥ 10 mm (p = 0.002) and type B intrapapillary capillary loops (p < 0.001). Moreover, 34 of the 83 biopsies that were diagnosed with HGIN were upgraded to ESCC after resection, which was related to lesion size (p = 0.001), location (p = 0.018), and pink color sign (p = 0.002). Conclusions Histological discrepancy between forceps biopsy and resected specimens is common in clinical practice. Recognizing the risk factors for each histological category of biopsy may reduce these discrepancies and improve clinical management.


PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e8168
Author(s):  
Diana H. Taft ◽  
Samir Akre ◽  
Nicolas Madrid ◽  
Andre Knoesen ◽  
David A. Mills ◽  
...  

Dedicated lactation rooms are a modern development as mothers return to work while still providing breastmilk to their absent infants. This study describes the built environment microbiome of lactation rooms and daycares, and explores the influence of temperature and humidity on the microbiome of lactation rooms. Sterile swabs were used to collect samples from five different sites in lactation rooms at University of California, Davis and from five different sites in daycares located in Davis, California. DNA from the swabs was extracted and the V4 region of the 16S rRNA gene was sequenced using Illumina MiSeq. Temperature and relative humidity data were collected on a subset of the lactation rooms. Sampled lactation rooms could be either dedicated lactation rooms or could also serve other functions (e.g., combined lactation room and restroom lounge). The majority of sequence reads were identified as belonging to family Moraxellaceae, with 73% of all reads included in analysis identified as an unknown species of Acinetobacter. Alpha diversity was analyzed using the Shannon index, while beta diversity was analyzed using unweighted and weighted UniFrac distance. The Jaccard distance was used to measure amount of change at sampling locations between time points for analysis of the impact of temperature and humidity on the microbiome. There were significant differences in the beta diversity of the microbiome of lactation rooms by room type. There were also significant differences in the beta diversity of the microbiome by sample collection location. There were no significant differences in either alpha or beta diversity associated with room temperature or humidity. Additional studies are needed to understand if the differences in lactation room type may result in differences in the breastmilk microbiome of milk collected in those rooms, and to what extent any such differences may influence the infant microbiome.


2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Qianqian Zhang ◽  
Liju Zong ◽  
Hui Zhang ◽  
Wei Xie ◽  
Fan Yang ◽  
...  

Over 10% of patients diagnosed with cervical intraepithelial neoplasia (CIN) have no lesions detected in their cervical conization specimens. The purpose of this study was to determine the factors related to the absence of such lesions. We particularly sought to investigate whether the expression of B7-H4 in precancerous lesions and cancer of the uterine cervix plays a role in the presence or absence of residual lesions in conization specimens and whether this protein is associated with T cells (i.e., Foxp3+ regulatory T cells, CD4+, and CD8+) and interferon-γ production. Of the 807 patients with CIN treated by conization, 104 (12.9%) had no lesions in their conization specimens. Seventy-five of these patients were deemed the study group and were matched with 75 patients who did have CIN detected in their conization specimens (the control group). Immunohistochemistry and immunofluorescence staining were used to detect B7-H4, Foxp3, CD4, CD8, and interferon-γ in the 75 pairs of specimens obtained via biopsy; 20 samples were found to have chronic cervicitis, and another 20 had squamous cell carcinoma of the cervix. Menopause, the absence of human papillomavirus, low-grade histological findings, and a diagnosis of CIN1 and CIN2 on biopsy correlated with a low probability of lesions on conization specimens. B7-H4 expression was detected in 11.1% of CIN2, 46.6% of CIN3, and 70% of cervical cancer samples, but not in tissues representing chronic cervicitis or CIN1. B7-H4 expression was associated with the presence of lesions on conization specimens, increased regulatory T cells, decreased CD8+ T cells, and lower interferon-γ production. These data suggest that close follow-up and thorough reevaluation should be considered for patients diagnosed with CIN2 who are negative for B7-H4 expression on biopsy before proceeding with cervical conization.


Sign in / Sign up

Export Citation Format

Share Document