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Metal-free carbon-based materials are one of the most promising electrocatalysts toward 2-electron oxygen reduction reaction (2e-ORR) for on-site production of hydrogen peroxide (H2O2), which however suffer from uncontrollable carbonizations and inferior 2e-ORR selectivity. To this end, a polydopamine (PDA)-modified carbon catalyst with a dipole-dipole enhancement is developed via a calcination-free method. The H2O2 yield rate outstandingly reaches 1.8 mol gcat -1 h-1 with high faradaic efficiency of above 95% under a wide potential range of 0.4-0.7 VRHE, overwhelming most of carbon electrocatalysts. Meanwhile, within a lab-made flow cell, the synthesized ORR electrode features an exceptional stability for over 250 h, achieved a pure H2O2 production efficacy of 306 g kWh-1. By virtue of its industrial-level capabilities, the established flow cell manages to perform a rapid pulp bleaching within 30 min. The superior performance and enhanced selectivity of 2e-ORR is experimentally revealed and attributed to the electronic reconfiguration on defective carbon sites induced by non-covalent dipole-dipole influence between PDA and carbon, thereby prohibiting the cleavage of O-O in OOH intermediates. This proposed strategy of dipole-dipole effects is universally applicable over 1D carbon nanotubes and 2D graphene, providing a practical route to design 2e-ORR catalysts.
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BACKGROUND: To investigate the prevalence of common sexually transmitted infections (STIs) and the association of STI/human papillomavirus co-infection in young and middle-aged women with previous abnormal cervical findings referred for colposcopy. METHODS: 719 cervical-swab cytobrush specimens were obtained from women aged ≤ 50 years who were referred for colposcopy at Peking University First Hospital due to previous abnormal cervical findings. HPV 21 typing and a panel of pathogenic STIs were tested for using the 21 HPV GenoArray Diagnostic Kit (HBGA-21PKG; HybriBio, Ltd., Chaozhou, China) and a nucleic acid STI detection kit (HybriBio Ltd. Guangzhou, China), after which colposcopy with multipoint positioning biopsy was performed. RESULTS: The overall prevalence of STIs among HPV positive women with previous abnormal cervical cancer screening results was 63.7% (458/719), with Ureaplasma parvum serovar 3, Ureaplasma parvum serovar 6 and herpes simplex virus type 2 having significantly higher prevalence among high-risk HPV positive patients (19.3%, Χ2 = 5.725, P = 0.018; 21.5%, Χ2 = 4.439, P = 0.035; 5.7%, Χ2 = 4.184, P = 0.048). Among patients positive for the high-risk human papillomavirus, the prevalence of Neisseria gonorrhoeae infection in human papillomavirus 16/18 positive patients was significantly higher than that in other patients (2.5%, Χ2 = 4.675; P = 0.043). Histopathologically, Chlamydia trachomatis infection was more frequently detected in lower than or equal to low-grade squamous intraepithelial lesion infection status (13.0%, Χ2 = 3.368; P = 0.041). CONCLUSIONS: The high prevalence of HPV coinfection with other sexually transmitted pathogens, particularly Ureaplasma parvum serovar 3, Ureaplasma parvum serovar 6, and herpes simplex virus type 2, calls for routine STI screening and effective STI prevention and management in patients with abnormal cervical cancer screening results.
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Colposcopia , Infecções por Papillomavirus , Infecções Sexualmente Transmissíveis , Displasia do Colo do Útero , Neoplasias do Colo do Útero , Feminino , Humanos , Pessoa de Meia-Idade , Gravidez , Detecção Precoce de Câncer , Papillomavirus Humano , Papillomaviridae , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/epidemiologia , Infecções Sexualmente Transmissíveis/epidemiologia , Infecções Sexualmente Transmissíveis/diagnóstico , Ureaplasma , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/epidemiologiaRESUMO
BACKGROUND: The classification of benign and malignant microcalcification clusters (MCs) is an important task for computer-aided diagnosis (CAD) of digital breast tomosynthesis (DBT) images. Influenced by imaging method, DBT has the characteristic of anisotropic resolution, in which the resolution of intra-slice and inter-slice is quite different. In addition, the sharpness of MCs in different slices of DBT is quite different, among which the clearest slice is called focus slice. These characteristics limit the performance of CAD algorithms based on standard 3D convolution neural network (CNN). METHODS: To make full use of the characteristics of the DBT, we proposed a new ensemble CNN, which consists of the 2D ResNet34 and the anisotropic 3D ResNet to extract the 2D focus slice features and 3D contextual features of MCs, respectively. Moreover, the anisotropic 3D convolution is used to build 3D ResNet to avoid the influence of DBT anisotropy. RESULTS: The proposed method was evaluated on 495 MCs in DBT images of 275 patients, which are collected from our collaborative hospital. The area under the curve (AUC) of receiver operating characteristic (ROC) and accuracy of classifying benign and malignant MCs using decision-level ensemble strategy were 0.8837 and 82.00%, which were significantly higher than the experimental results of 2D ResNet34 (AUC: 0.8264, ACC: 76.00%) and anisotropic 3D ResNet (AUC: 0.8455, ACC: 76.00%). Compared with the results of 3D features classification in the radiomics, the AUC of the deep learning method with decision-level ensemble strategy was improved by 0.0435, and the F1 score was improved from 79.37 to 85.71%. More importantly, the sensitivity increased from 78.13 to 84.38%, and the specificity increased from 66.67 to 77.78%, which effectively reduced the false positives of diagnosis CONCLUSION: The results fully prove that the ensemble CNN can effectively integrate 2D features and 3D features, improve the classification performance of benign and malignant MCs in DBT, and reduce the false positives.
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Neoplasias da Mama , Calcinose , Neoplasias da Mama/diagnóstico por imagem , Diagnóstico por Computador , Feminino , Humanos , Mamografia , Redes Neurais de Computação , Curva ROCRESUMO
This paper focuses on biofilms of Lactobacillus spp. - a type of normal flora isolated from healthy human vaginas of women of childbearing age; thereupon, it broadens the research scope of investigation of vaginal normal flora. The static slide culture method was adopted to foster biofilms, marked by specific fluorescence staining. Laser scanning confocal and scanning electron microscopy were used to observe the microstructure of the biofilms. Photographs taken from the microstructure were analysed to calculate the density of the biofilms. The body of Lactobacillus spp., though red, turned yellow when interacting with the green extracellular polysaccharides. The structure of the biofilm and aquaporin within the biofilm were imaged. Lactobacillus density increases over time. This study provides convincing evidence that Lactobacillus can form biofilms and grow over time in vitro. This finding establishes an important and necessary condition for selecting proper strains for the pharmaceutics of vaginal ecology.
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Biofilmes/crescimento & desenvolvimento , Lactobacillus/fisiologia , Vagina/microbiologia , Adulto , Feminino , Humanos , Microscopia Confocal , Microscopia Eletrônica de Varredura , Imagem ÓpticaRESUMO
OBJECTIVE: To evaluate the cure effectiveness and safety of sucrose gel in the treatment of bacterial vaginosis through a multi-center, randomized, double-blind, parallel controlled clinical study. METHODS: A clinical research method of multi-center, randomly double-blind, and dose group parallel comparison was adopted. In the study, 533 patients with bacterial vaginosis were randomly divided into two groups, which included 214 cases in the control group (5.0 g metronidazole gel) and 319 cases in the trial group (5.0 g sucrose gel ). The patients were treated with different medication according to the group where they were. All the cases in these two groups were treated with drugs vaginally twice in a day, morning and evening separately, for 5 days. The curative effect and safety evaluation were assessed from 7 to 10 days and 21 to 30 days after treatment respectively. RESULTS: The efficacy of the comprehensive clinical treatment showed that the cure rate of metronidazole gel group and sucrose gel group were 70.53% and 80.83% respectively 7 to 10 days after treatment. The recovery rate of Nugent score for vaginal smear were 71.50% and 81.15% respectively. The differences in the efficacy between these two groups were significant statistically (P<0.05). However, the cure rates of metronidazole gel group and sucrose gel group were 63.29% and 61.98% respectively 21 to 30 days after treatment. No statistically significant difference (P>0.05) could be found in the cure rates of the two groups. CONCLUSION: The clinical comprehensive efficacy and recovery of vaginal bacteria of sucrose gel group in the treatment of bacterial vaginosis were obviously superior to those of metronidazole gel 7 to 10 days after treatment. The susucrose gel could improve the clinical efficacy index and laboratory index of bacterial vaginosis. Other effects included the release of clinical symptoms, and the recovery of the normal micro-environment in the vagina according to the Nugent score. The curative efficacy of sucrose gel was equal to that of metronidazole gel 21 to 30 days after treatment. In the future, sucrose gel treatment can be a new strategy for the treatment of bacterial vaginosis. Various advantages can be taken to improve the cure rate of bacterial vaginosis and reduce the shortcomings produced by this disease.
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Sacarose/uso terapêutico , Vaginose Bacteriana/tratamento farmacológico , Administração Intravaginal , Método Duplo-Cego , Esquema de Medicação , Feminino , Géis/química , Humanos , Metronidazol/uso terapêuticoRESUMO
Bacterial vaginosis (BV) is an infection of the genital tract characterized by disturbance of the normally Lactobacilli-dominated vaginal flora due to the overgrowth of Gardnerella and other anaerobic bacteria. Gardnerella vaginalis, an anaerobic pathogen and the major pathogen of BV, produces sialidases that cleave terminal sialic acid residues off of human glycans. By desialylation, sialidases not only alter the function of sialic acid-containing glycoconjugates but also play a vital role in the attachment, colonization and spread of many other vaginal pathogens. With known pathogenic effects, excellent performance of sialidase-based diagnostic tests, and promising therapeutic potentials of sialidase inhibitors, sialidases could be used as a biomarker of BV. This review explores the sources of sialidases and their role in vaginal dysbiosis, in aims to better understand their participation in the pathogenesis of BV and their value in the diagnosis and treatment of BV.
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Vaginose Bacteriana , Feminino , Humanos , Vaginose Bacteriana/tratamento farmacológico , Vaginose Bacteriana/microbiologia , Neuraminidase/química , Ácido N-Acetilneuramínico , Gardnerella vaginalis , Vagina/microbiologiaRESUMO
Introduction: Human papillomavirus (HPV) infection, especially persistent high-risk HPV, is associated with cervical cancer. Female reproductive tract microecological disorders and lower genital tract infections have been increasingly correlated with HPV infection and cervical lesions. Due to their common risk factors and transmission routes, coinfection with other sexually transmitted infections (STIs) has become a concern. Additionally, the clinical significance of Mycoplasma subtypes appear to vary. This study aimed to assess the correlations between common STIs and HPV infection, and to investigate the clinical significance of Mycoplasma subtypes. Methods: We recruited 1,175 patients undergoing cervical cancer screening at the Peking University First Hospital gynecological clinic from March 2021 to February 2022 for vaginitis and cervicitis tests. They all received HPV genotyping and detection of STIs, and 749 of them underwent colposcopy and cervical biopsy. Results: Aerobic vaginitis/desquamative inflammatory vaginitis and STIs (mainly single STIs) were found significantly more often in the HPV-positive group than in the HPV-negative group. Among patients with a single STI, rates of infection with herpes simplex virus type 2 or UP6 in the HPV-positive group were significantly higher than in the HPV-negative group (ORadj: 1.810, 95%CI: 1.211-2.705, P=0.004; ORadj: 11.032, 95%CI: 1.465-83.056, P=0.020, respectively). Discussion: Through detailed Mycoplasma typing, a correlation was found between different Mycoplasma subtypes and HPV infection. These findings suggest that greater attention should be paid to detecting vaginal microecological disorders in those who are HPV-positive. Further, lower genital tract infections, including both vaginal infections and cervical STIs, are significantly more common among women who are HPV-positive and who thus require more thorough testing. Detailed typing and targeted treatment of Mycoplasma should become more routine in clinical practice.
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Mycoplasma , Infecções por Papillomavirus , Infecções do Sistema Genital , Infecções Sexualmente Transmissíveis , Neoplasias do Colo do Útero , Humanos , Feminino , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/epidemiologia , Papillomavirus Humano , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/epidemiologia , Detecção Precoce de Câncer , Relevância Clínica , Infecções Sexualmente Transmissíveis/diagnóstico , Infecções Sexualmente Transmissíveis/epidemiologiaRESUMO
Bacterial vaginosis (BV) is the most common infection of the lower reproductive tract among women of reproductive age. Recurrent infections and antibiotic resistance associated with biofilms remain significant challenges for BV treatment. Gardnerella species are commonly found in women with and without BV, indicating that genetic differences among Gardnerella isolates may distinguish pathogenic from commensal subgroups. This study isolated 11 Gardnerella strains from vaginal samples obtained from women with BV before or after treatment. The biofilm formation ability of each strain was examined by crystal violet staining. Eight strains were selected using phylogenetic analysis of the cpn60 sequences and classified as subgroups A (6/8), B (1/8), and D (1/8). The biofilm formation ability and antibiotic resistance profile of these strains was compared among the subgroups. Subgroup D had the strongest biofilm formation ability. Six of the planktonic strains exhibited resistance to the first-line BV drug, metronidazole, and one to clindamycin. Moreover, biofilm formation in vitro increased strain resistance to clindamycin. Two strains with strong biofilm ability, S20 and S23, and two with weak biofilm ability, S24 and S25, were selected for comparative genomic analysis. S20 and S23 were found to contain four key genes associated with biofilm formation and more genes involved in carbohydrate synthesis and metabolism than S24 and S25. Identifying differences in the expression of virulence factors between Gardnerella subgroups could inform the development of novel treatments for BV.
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Bacterial vaginosis (BV) is the most common infection of the lower reproductive tract among women of reproductive age, characterized by a depletion of health-associated Lactobacillus and an overgrowth of anaerobes. Metronidazole has been recommended as a first-line therapy for treating BV for decades. Although most cases are cured by the treatment, recurrent infections of BV seriously affect women's reproductive health. Until now, limited information on the vaginal microbiota has been explored at the species level. Here, we adopted a single molecular sequencing approach for the 16S rRNA gene, named FLAST (full-length assembly sequencing technology), to analyze the human vaginal microbiota that improved species-level resolution for taxonomy and identified microbiota alterations in the vaginal tract in response to treatment with metronidazole. Appling high-throughput sequencing, we identified 96 and 189 novel full-length 16S rRNA gene sequences in Lactobacillus and Prevotella, respectively, which had not previously been reported in vaginal samples. Moreover, we found that Lactobacillus iners was significantly enriched in the cured group before metronidazole treatment, and that was maintained in a high frequency after the treatment, suggesting an important role for this species in response to metronidazole treatment. Our research also highlights the importance of the single-molecule paradigm for progressing the field of microbiology and applying these insights to better understand the dynamic microbiota during BV treatment. Subsequent novel treatment approaches should be proposed to improve BV treatment outcomes, optimize the vaginal microbiome, and reduce gynecological and obstetric sequelae. IMPORTANCE Bacterial vaginosis (BV) is a common infectious disease of the reproductive tract. Metronidazole treatment, as the first line of treatment, frequently fails at recovery of the microbiome. However, the precise types of Lactobacillus and other bacteria involved in BV remain unclear, and this has resulted in a failure to identify potential markers to predict clinic outcomes. In this study, we adopted a 16S rRNA gene full-length assembly sequencing technology for the taxonomy analysis and evaluation of vaginal microbiota before and after treatment with metronidazole. We additionally identified 96 and 189 novel 16S rRNA gene sequences in Lactobacillus and Prevotella species, respectively, in vaginal samples, which improves our understanding of the vaginal microbiota. Moreover, we found that the abundance of Lactobacillus iners and Prevotella bivia before treatment was associated with a lack of cure. These potential biomarkers will help to facilitate future studies aimed at improving BV treatment outcomes, optimize the vaginal microbiome, and reduce adverse sexual and reproductive outcomes.
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Microbiota , Vaginose Bacteriana , Feminino , Humanos , Metronidazol/farmacologia , Metronidazol/uso terapêutico , Vaginose Bacteriana/tratamento farmacológico , Vaginose Bacteriana/complicações , Vaginose Bacteriana/microbiologia , RNA Ribossômico 16S/genética , Vagina/microbiologia , Microbiota/genética , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
OBJECTIVE: To analyze the diversity of healthy women's vaginal dominant flora in different physiological states with PCR-denaturing gradient gel electrophoresis (PCR-DGGE), to establish the basis for the future study of lower genital tract infections, and to provide a theoretical basis for the development of vaginal probiotics adapted to Chinese females. METHODS: Women who underwent routine gynecologic examinations in Peking University First Hospital from October 2009 to January 2010 were enrolled, including 30 at reproductive age and 30 at post menopause age. Vaginal samples were collected and then total bacterial DNA was extracted. Universal bacterial primers were used to amplify the V3 region of 16S rDNA gene. PCR products were analyzed by denatured gradient gel electrophoresis (DGGE). Featured bands on DGGE were recovered, cloned and sequenced. Alignment with known sequence was made by using Blast on GenBank to identify bacterial strains so as to analyze the diversity of healthy women's vaginal dominant flora in different physiological states. RESULTS: (1) Vaginal flora in healthy Chinese women at their reproductive age were relatively simple, Lactobacillus were the dominant bacteria. Common bacteria included: Lactobacillus crispatus, Lactobacillus iners,and Lactobacillus gasseri. While L. iners were the dominant vaginal bacteria that could not be recognized by traditional method. (2) Vaginal bacteria in the women at post menopause age were more complex than in those at reproductive age. Common bacteria included: Lactobacillus iners, Lactobacillus crispatus, Escherichia coli, Streptococcus agalactiae, Streptococcus gallolyticus, Veillonella sp., Streptococcus intermedius, Streptococcus anginosus, Prevotella sp., Anaerococcus lactolyticus and Bacteroides fragilis. CONCLUSION: Based on the DGGE analysis, the most common vaginal bacteria in Chinese healthy women at reproductive age are Lactobacillus crispatus, Lactobacillus iners,and Lactobacillus gasseri. the most common vaginal bacteria in healthy post-menopausal women are Lactobacillus iners, Lactobacillus crispatus, Escherichia coli, Streptococcus sp., Prevotella sp.,Bacteroides fragilis and lactic acid producing Veillonella sp. and Anaerococcus lactolyticus. Meanwhile Lactobacillus iners was firstly found by DGGE to be the dominant vaginal bacteria in different physiological states.
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Impressões Digitais de DNA , Escherichia coli/isolamento & purificação , Lactobacillus/isolamento & purificação , Vagina/microbiologia , Adulto , Idoso , Biodiversidade , China , Eletroforese em Gel de Gradiente Desnaturante , Feminino , Humanos , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Streptococcus/isolamento & purificaçãoRESUMO
Bacterial vaginosis (BV) is the most common infectious disease of the reproductive tract in women of childbearing age. It often manifests as an imbalance in the vaginal microbiome, including a decrease in Lactobacillus and an increase in anaerobic bacteria. While Gardnerella spp. are considered a major cause of BV, they are also detected in the vaginal microbiome of healthy women. G. vaginalis was the only recognized species of Gardnerella until a recent study characterized three new species, G. leopoldii, G. piotii, and G. swidsinskii. This review describes the different types and genetic diversity of Gardnerella, as well as new findings on the correlation between different Gardnerella spp. and BV.
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Microbiota , Vaginose Bacteriana , Feminino , Gardnerella , Gardnerella vaginalis/genética , Humanos , Microbiota/genética , Vagina/microbiologia , Vaginose Bacteriana/microbiologiaRESUMO
Bacterial vaginosis is characterized as a polymicrobial dysbiosis with the loss of Lactobacillus spp. and growth of multiple anerobic bacteria, including Gardnerella, Prevotella and Atopobium ranked as the top three most abundant. A total of nine Gardnerella genomospecies have been identified, yet the association between their distribution or any exact Lactobacillus species with BV occurrence or prognosis remains controversial. A total of 308 patients and 62 healthy women who sought annual examinations were recruited, with 130 BV patients and 41 healthy women who met our inclusion criteria finally included. Vaginal samples were used for microscopic examination, 16S rRNA sequencing, bacterial culture and isolation. Isolates of Gardnerella vaginalis, Fannyhessae vaginae (used to be called Atopobium vaginae) and Lactobacillus iners were used for competition tests. We found that the relative abundances of Gardnerella, Prevotella and Atopobium were elevated in BV patients compared to healthy people (p<0.0001), yet no significant differences were found among patients with different clinical outcomes (p>0.05). Seven out of nine Gardnerella genomospecies were present in both BV patients and healthy women, and the relative abundances of all detected genomospecies were higher in BV patients (p<0.05). Cured patients possessed higher GS03 than intermediate and failed patients (p=0.005, 0.0337). L. iners was significantly higher in cured patients than in the other two groups (p=0.0021, p<0.0001), and its ability to inhibit the growth of G. vaginalis and F. vaginae was validated. In summary, seven Gardnerella genomospecies were detected in Chinese BV patients, but no association of its distribution and BV occurrence or prognosis was found. The relative abundance of L. iners was higher in cured patients, and its antimicrobial activity against G. vaginalis and F. vaginae was validated through in vitro inhibition experiment. L. iners could become a predictive indicator of clinical outcomes of BV patients, and its antimicrobial function might be beneficial to BV patients.
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Anti-Infecciosos , Vaginose Bacteriana , Humanos , Feminino , Gardnerella , Vaginose Bacteriana/tratamento farmacológico , RNA Ribossômico 16S/genética , Lactobacillus , Antibacterianos/farmacologia , Antibacterianos/uso terapêuticoRESUMO
Mixed vaginitis is the result of the simultaneous presence of different pathogenic processes mediated by at least two types of vaginal pathogens. Among the various types of mixed vaginitis presentations, bacterial vaginosis (BV) plus vulvovaginal candidiasis (VVC) presents to be the most prevalent form. Mixed vaginitis affects the health of women of all ages worldwide. However, few studies have focused on clinical manifestations, pathogenesis, diagnostic criteria, or therapy of mixed vaginitis. We recruited 48 symptomatic patients with clinical diagnoses of VVC complicated with BV, they were treated with oral metronidazole combined with local clotrimazole and followed to assess the drug efficacy and vaginal microbiome alterations before and after treatment. The vaginal microbiome in BV+VVC mixed vaginitis patients was altered significantly after the combined drug treatment within a unique form different from a simple overlay mode of BV and VVC, the key bacteria including Gardnerella and Atopobium, Lactobacillus. The combined drug therapy for the mixed vaginitis in this study was effective and enhanced treatment for BV may be more favorable because of more difficulty in dealing with BV according to the treatment outcome. The abundance of Lactobacillus in patients with mixed vaginitis affects the recovery of the vaginal microbiome as well as the prognosis, and the abundance should be actively restored. This is the first study to investigate the composition, diversity, and other characteristics of the vaginal microbiome in patients with BV+VVC mixed vaginitis before and after drug treatment, our results provide clues to improving the cure rate and reducing recurrences.
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Candidíase Vulvovaginal , Microbiota , Vaginose Bacteriana , Candidíase Vulvovaginal/diagnóstico , Candidíase Vulvovaginal/tratamento farmacológico , Feminino , Humanos , Metronidazol/uso terapêutico , Vagina/microbiologia , Vaginose Bacteriana/diagnóstico , Vaginose Bacteriana/tratamento farmacológico , Vaginose Bacteriana/microbiologiaRESUMO
Background: The relationship between pelvic organ prolapse (POP), an aging-related disease, and the senescence-related protein mitofusin 2 (Mfn2) has rarely been studied. The aim of the present study was to explore the therapeutic effects of the downregulation of Mfn2 expression by stem cells on POP through animal experiments. Methods: First, a rat POP model was constructed by ovariectomy and traction. The rats in the non-pelvic organ prolapse (NPOP) and POP groups were divided into four groups for negative controls (N1−N4, N1: NPOP-normal saline; N2: NPOP-untransfected stem cells; N3: NPOP-short hairpin negative control (NPOP-sh-NC); N4: NPOP-short hairpin-Mfn2 (NPOP-sh-Mfn2)), and four groups for prolapse (P1−P4, P1: POP-normal saline; P2: POP-untransfected stem cells; P3: POP-sh-NC; P4: POP-sh-Mfn2), respectively. Stem cells were then cultured and isolated. The expression of Mfn2 was inhibited by lentivirus transfection, and the stem cells were injected into the uterosacral ligament of the rats in each group. The expression levels of Mfn2 and procollagen 1A1/1A2/3A1 in the uterosacral ligaments of the rats were observed at 0, 7, 14, and 21 days after injection. Results: Compared to the rats in the NPOP group, the POP rats had significant prolapse. The Mfn2 expression in the uterosacral ligaments of the POP rats was significantly increased (p < 0.05, all), and the expression of procollagen 1A1/1A2/3A1 was significantly decreased (p < 0.001, all). The POP rat model maintained the same trend after 21 days (without stem cell injection). At day 14, compared to the rats in the N1 group, the Mfn2 expression in the uterosacral ligament of the rats in the N4 group was significantly decreased (p < 0.05, all), and the expression of procollagens was significantly increased (p < 0.05, all). Similarly, compared to the rats in the P1 group, the Mfn2 expression in the uterosacral ligament of the rats in the P4 group was significantly decreased (p < 0.05, all), and the expression of procollagens was significantly increased (p < 0.05, all). Similarly, on day 21, the Mfn2 mRNA and protein expression in the uterosacral ligament of the POP and NPOP rats was significantly decreased (p < 0.05, all), and the expression of procollagens was significantly increased (p < 0.05, all) in the rats in the sh-Mfn2 group (N4, P4) compared to the rats in the saline group (N1, P1). Conclusions: The downregulation of Mfn2 expression by stem cells decreased the expression of Mfn2 and increased the expression of procollagen1A1/1A2/3A1 in the uterosacral ligament of the POP rats; this effect was significant 14−21 days after the injection. Thus, Mfn2 may be a new target for POP control.
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GTP Fosfo-Hidrolases/metabolismo , Células-Tronco Mesenquimais , Proteínas Mitocondriais/metabolismo , Prolapso de Órgão Pélvico , Animais , Regulação para Baixo , Feminino , Hidrolases/genética , Ligamentos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Prolapso de Órgão Pélvico/genética , Prolapso de Órgão Pélvico/metabolismo , Prolapso de Órgão Pélvico/terapia , Pós-Menopausa , Pró-Colágeno/genética , Pró-Colágeno/metabolismo , Ratos , Solução Salina/metabolismoRESUMO
The mechanism underlying unwanted structural variations induced by CRISPR-Cas9 remains poorly understood, and no effective strategy is available to inhibit the generation of these byproducts. Here we find that the generation of a high level of translocations is dependent on repeated cleavage at the Cas9-targeting sites. Therefore, we employ a strategy in which Cas9 is fused with optimized TREX2 to generate Cas9TX, a Cas9 exo-endonuclease, which prevents perfect DNA repair and thereby avoids repeated cleavage. In comparison with CRISPR-Cas9, CRISPR-Cas9TX greatly suppressed translocation levels and enhanced the editing efficiency of single-site editing. The number of large deletions associated with Cas9TX was also reduced to very low level. The application of CRISPR-Cas9TX for multiplex gene editing in chimeric antigen receptor T cells nearly eliminated deleterious chromosomal translocations. We report the mechanism underlying translocations induced by Cas9, and propose a general strategy for reducing chromosomal abnormalities induced by CRISPR-RNA-guided endonucleases.
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Proteína 9 Associada à CRISPR , Edição de Genes , Proteína 9 Associada à CRISPR/genética , Sistemas CRISPR-Cas/genética , Endonucleases/genética , Endonucleases/metabolismo , Humanos , RNA Guia de Cinetoplastídeos/química , RNA Guia de Cinetoplastídeos/genética , Translocação GenéticaRESUMO
Bacterial vaginosis (BV) and its recurrence are most commonly associated with the formation of Gardnerella species biofilm. Probiotics are typically used to treat BV; however, the optimal period of Lactobacillus probiotic application in BV treatment remains uncertain. The present study aimed to explore the effects of Lactobacillus rhamnosus and Lactobacillus casei on various stages of biofilm formation in Gardnerella species. The biofilm-forming ability of seven strains, including one Gardnerella vaginalis ATCC 14018 and six clinically isolated Gardnerella species, was determined via gentian violet staining assay. Moreover, the sensitivity of the planktonic and biofilm forms toward metronidazole and clindamycin was assessed via microdilution broth method. L. rhamnosus Xbb-LR-1 and L. casei Xbb-LC-1 were added during various stages of biofilm formation in Gardnerella species and were cocultured for 24 h. The biofilm thickness of each sample was determined via confocal laser scanning microscopy (CLSM). The absolute quantities of Gardnerella species in each sample was obtained via real time polymerase chain reaction method, and the pH value was obtained using a pH indicator paper. Biofilm formation by Gardnerella species in a medium with distinct pH values was observed via gentian violet staining, CLSM, and scanning electron microscopy (SEM). The biofilm increased the resistance of Gardnerella species toward metronidazole and clindamycin. L. rhamnosus added at the initial biofilm formation stage in Gardnerella species exhibited highest inhibitory effect, with a percentage inhibition of 38.17% ± 1.35%. When the pH value of the culture medium was <4.5 or >6.5, ATCC 14018 could hardly form a biofilm; however, at pH ≥4.5 and ≤6.5, it was able to form a stronger biofilm. The amount of biofilm attained maximum value at optical density of 3.29 ± 0.28 (595 nm), pH 5.5, and at 36 h. Biofilm formation increases the resistance of Gardnerella species toward antibiotics. Maintaining an acidic vaginal environment with pH <4.5 and a vaginal microbiota dominated by Lactobacillus remarkably prevents the formation of Gardnerella species biofilm at the initial stage, which further has a significant impact on the treatment and prevention of biofilm-related infections.
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Lacticaseibacillus casei , Lacticaseibacillus rhamnosus , Probióticos , Biofilmes , Feminino , Gardnerella , Gardnerella vaginalis , Humanos , VaginaRESUMO
OBJECTIVE: To evaluate the effective dose and safety of S-Gel in the treatment of bacterial vaginosis. METHODS: Clinical research method of multi- center, randomly double-blind, and dose group parallel comparison was adopted. In the study, 96 bacterial vaginosis patients were randomized into three groups: Group A, S-Gel 5.0 g, 32 patients; Group B, S-Gel 7.5 g, 32 patients; Group C, placebo, 32 patients. The patients were treated with different methods. Safety and efficacy were analyzed 3 to 4 days and 8 days after the treatment, respectively. RESULTS: The efficacy of comprehensive clinical treatment showed that: 8 days after the treatment, the cure rates of group A (5.0 g), group B (7.5 g), and group C (placebo) were 84.38%, 86.67%, and 3.13% respectively. No difference of statistic significance was found in groups A and B, difference of statistical significance was found in group A and group C, group B and group C respectively (P<0.001). CONCLUSION: As compared with placebo, S-Gel 7.5 g and 5.0 g bid (in the morning and evening) could obviously improve the clinical efficacy index and laboratory index of bacterial vaginosis. Other effects included the release of clinical symptoms, and the recovery of the normal micro-environment in the vagina. No significant difference was found in the cure rates of the two dose groups.
Assuntos
Sacarose/administração & dosagem , Cremes, Espumas e Géis Vaginais/administração & dosagem , Vaginose Bacteriana/tratamento farmacológico , Adolescente , Adulto , Método Duplo-Cego , Feminino , Humanos , Pessoa de Meia-Idade , Adulto JovemRESUMO
Adhesion to the gastrointestinal tract is considered to be important for bifidobacteria to colonize the human gut and exert their probiotic effects. Some cell surface proteins of bifidobacteria, known as adhesins, play critical roles in the binding to host cells or the extracellular matrix (ECM). To elucidate the mechanisms associated with the adhesion of Bifidobacterium longum BBMN68, a centenarian originated potential probiotic, PSORTdb was employed to identify putative extracellular localized proteins in the B. longum BBMN68. Of the 560 predicted extracellular proteins, 21 were further identified as putative adhesion proteins using the conserved domain database of NCBI, and four were successfully overexpressed in the heterologous host, Lactococcus lactis NZ9000. Notably, a recombinant strain expressing FimM showed a significantly increased adhesive affinity for both HT-29 and mucus-secreting LS174T goblet cells (2.2- and 5.4-fold higher than that of the control strain, respectively). Amino acid sequence alignment showed that FimM is a major pilin subunit protein containing a Cna-B type domain and a C-terminal LPKTG sequence. However, in silico analysis of the fimM-coding cluster revealed that BBMN68_RS10200, encoding a pilus-specific class C sortase, was a pseudogene, indicating that FimM may function as a surface adhesin that cannot polymerize into a pili-like structure. Immunogold electron microscopy results further confirmed that FimM localized to the surface of L. lactis NZfimM and B. longum BBMN68 but did not assemble into pilus filaments. Moreover, the adhesive affinity of L. lactis NZfimM to fibronectin, fibrinogen, and mucin were 3.8-, 2.1-, and 3.1-fold higher than that of the control. The affinity of FimM for its attachment receptors was further verified through an inhibition assay using anti-FimM antibodies. In addition, homologs of FimM were found in Bifidobacterium bifidum 85B, Bifidobacterium gallinarum CACC 514, and 23 other B. longum strains by sequence similarity analysis using BLASTP. Our results suggested that FimM is a novel surface adhesin that is mainly present in B. longum strains.
RESUMO
Background/Purpose: Lactobacillus colonization is important to maintain urogenital flora stability and prevent pathogenic infection. Different Lactobacillus species have distinct properties and effects on the urogenital flora. To select probiotics that colonize the vagina and provide protection against pathogenic infection, we evaluated the adhesion of five Lactobacillus strains and their inhibitory effects on the adhesion of pathogens to vaginal epithelial cells (VECs). Methods and Materials: (1) Lactobacillus adhesion experiments: VK2/E6E7 and primary VECs were used to evaluate the adhesion of two Lactobacillus gasseri and three Lactobacillus crispatus strains. The adhesion of these five Lactobacillus strains was compared. (2) Adhesion inhibition experiments: The inhibitory effects of the five Lactobacillus strains on the adhesion of pathogens (Gardnerella, Mobiluncus, Candida albicans, Streptococcus agalactiae, Staphylococcus aureus, Escherichia coli, and Enterococcus faecalis) were evaluated by adhesion exclusion, displacement, and competition experiments. Results: (1) Lactobacillus adhesion was stronger in the primary VECs than in the VK2/E6E7 VECs (P < 0.05). The adhesion of the three L. crispatus strains was stronger than that of the two L. gasseri strains (P < 0.05). L. crispatus 4# showed the strongest adhesion. (2) The exclusion, displacement, and competition experiments showed that all five Lactobacillus strains significantly inhibited the adhesion of the seven pathogenic strains to the VECs (P < 0.05). The displacement effect was stronger than the exclusion and competition effects of each Lactobacillus strain. (3) The results of the exclusion, displacement, and competition experiments indicated that L. gasseri 1# showed the strongest adhesion inhibition of C. albicans and S. agalactiae. L. crispatus 3# showed the strongest adhesion inhibition of S. aureus, whereas L. crispatus 4# showed the strongest adhesion inhibition of Gardnerella, Mobiluncus, E. coli, and E. faecalis. Conclusion: The source of the VECs might not affect the selection of the most adhesive Lactobacillus strain. L. crispatus showed stronger VEC adhesion than L. gasseri. The degree of antagonism of the Lactobacillus strains toward the different pathogens varied. This result provides incentives for personalized clinical treatment.