Fast and Efficient Atmospheric NO2 Collection for Isotopic Analysis by a 3D-Printed Denuder System.

Being major species of atmospheric reactive nitrogen, nitrogen oxides (NOx = NO + NO2) have important implications for ozone and OH radical formation in addition to nitrogen cycles. Stable nitrogen isotopes (δ15N) of NOx have been sought to track NOx emissions and NOx chemical reactivities in the atmosphere. The current atmospheric NOx collection methods for isotopic analysis, however, largely suffer from unverified collection efficiency and/or low collection speed (<10 L/min). The latter makes it difficult to study δ15N(NOx) in pristine regions with low NOx concentrations. Here, we present a three-dimensional (3D)-printed honeycomb denuder (3DP-HCD) system, which can effectively collect atmospheric NO2 (a major part of NOx) under a variety of laboratory and field conditions. With a coating solution consisting of 10% potassium hydroxide (KOH) and 25% guaiacol in methanol, the denuder system can collect NO2 with nearly 100% efficiency at flow rates of up to 70 L/min, which is 7 times higher than that of the existing method and allows high-resolution (e.g., diurnal or finer resolution) NO2 collection even in pristine sites. Besides, the δ15N of NO2 collected by the 3DP-HCD system shows good reproducibility and consistency with the previously tested method. Preliminary results of online NO oxidation by a chrome trioxide (CrO3) oxidizer for simultaneous NO and NO2 collection are also presented and discussed.

An effective N6-methyladenosine-related long non-coding RNA prognostic signature for predicting the prognosis of patients with bladder cancer.

BACKGROUND: Bladder cancer (BLCA) typically has a poor prognosis due to high relapse and metastasis rates. A growing body of evidence indicates that N6-methyladenosine (m6A) and long non-coding RNAs (lncRNAs) play crucial roles in the progression of BLCA and the treatment response of patients with BLCA. Therefore, we conducted a comprehensive RNA-seq analysis of BLCA using data from The Cancer Genome Atlas (TCGA) to establish an m6A-related lncRNA prognostic signature (m6A-RLPS) for BLCA. METHODS: Consensus clustering analysis was used to investigate clusters of BLCA patients with varying prognoses. The least absolute shrinkage and selection operator Cox regression were used to develop the m6A-RLPS. The ESTIMATE and CIBERSORT algorithms were used to evaluate the immune composition. RESULTS: A total of 745 m6A-related lncRNAs were identified using Pearson correlation analysis (|R| > 0.4, p < 0.001). Fifty-one prognostic m6A-related lncRNAs were screened using univariate Cox regression analysis. Through consensus clustering analysis, patients were divided into two clusters (clusters 1 and 2) with different overall survival rates and tumor stages based on the differential expression of the lncRNAs. Enrichment analysis demonstrated that terms related to tumor biological processes and immune-related activities were increased in patient cluster 2, which was more likely to exhibit low survival rates. Nine m6A-related prognostic lncRNAs were finally determined and subsequently used to construct the m6A-RLPS, which was verified to be an independent predictor of prognosis using univariate and multivariate Cox regression analyses. Further, a nomogram based on age, tumor stage, and the m6A-RLPS was generated and showed high accuracy and reliability with respect to predicting the survival outcomes of BLCA patients. The prognostic signature was found to be strongly correlated to tumor-infiltrating immune cells and immune checkpoint expression. CONCLUSIONS: We established a novel m6A-RLPS with a favorable prognostic value for patients with BLCA. We believe that this prognostic signature can provide new insights into the tumorigenesis of BLCA and predict the treatment response in patients with BLCA.

Sexual Functions in Women With Stress Urinary Incontinence After Mid-Urethral Sling Surgery: A Systematic Review and Meta-Analysis of Prospective Randomized and Non-Randomized Studies.

BACKGROUND: Stress urinary incontinence (SUI) is a common, distressing health issue which affects many women. Mid-urethral sling (MUS) surgeries are recommended as gold standard interventions, although evidence regarding their impact on female sexual function remains controversial. AIM: To provide high-quality evidence of the impact of MUS surgeries on sexual functions in women with SUI. METHODS: A systematic search of PubMed, Embase, and the Cochrane Library was conducted to identify studies assessing the effect of MUS treatments on sexual functions in women with SUI. The included studies were prospective randomized or non-randomized trials which assessed patients using 2 validated questionnaires, the Female Sexual Function Index (FSFI) and the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12). Pre- and postoperative data regarding sexual function were extracted. Meta-analysis of comparable data was performed using Review Manager (version 5.3) software. MAIN OUTCOME MEASURES: The relationship between MUS interventions and sexual functions was analyzed by pooling weighted mean differences (WMD) with 95% CI from studies which used either the FSFI or PISQ-12 questionnaires, before and after receiving surgical interventions. RESULTS: Of the 22 eligible studies, 13 utilized FSFI and 9 adopted the PISQ-12. Pooled analysis indicates that 6-month postoperative PISQ-12 scores were significantly higher than preoperative scores (WMD -3.31 points; 95% CI -5.32 to -1.30; P = .001). Similar results were found at the 12-month juncture (WMD -3.30 points; 95% CI -6.01 to -0.58; P = .02) and at 24 months (WMD -4.44 points; 95% CI -5.45 to -3.44; P < .00001). Likewise, pooled postoperative FSFI total scores were significantly higher than preoperative scores at 6 months (WMD -2.22 points; 95% CI -3.36 to -1.08; P = .00001) and 12 months (WMD -3.49 points; 95% CI -5.96 to -1.02; P = .006). Postoperative FSFI sub-scores also suggest that desire, arousal, orgasm, lubrication, satisfaction, and pain during sexual intercourse significantly improved postoperatively (all P < .05). Moreover, combined evidence highlighted a significant reduction in coital incontinence postoperatively (risk ratio 5.78; 95% CI 3.16-10.58; P < .00001). CLINICAL IMPLICATIONS: These assessment tools might be more appropriately used to create opportunities for counseling. STRENGTHS & LIMITATIONS: We encountered substantial heterogeneity and insufficient long term follow-up data. There is also a distinct lack of standards, in terms of data recording and reporting across this evidence base which adds to the problems with the PISQ-12 and FSFI which, even though validated, appear unsophisticated and not necessarily fit-for-purpose. CONCLUSION: This meta-analysis confirms that sexual functions do improve after MUS surgeries for women with SUI. Lai S, Diao T, Zhang W, et al. Sexual Functions in Women With Stress Urinary Incontinence After Mid-Urethral Sling Surgery: A Systematic Review and Meta-Analysis of Prospective Randomized and Non-Randomized Studies. J Sex Med 2020;17:1956-1970.

A visualization analysis of hotspots and global trends on pelvic floor dysfunction in cervical cancer.

BACKGROUND/OBJECTIVE: Cervical cancer is the major cause of cancer-related mortalities in women globally. It constitutes one of the life-threatening conditions for women in developing countries. The popularization of cervical cancer screening and the improvement of treatment levels has caused the mortality rate of cervical cancer to decrease gradually, but pelvic floor dysfunction before and after cervical cancer treatment has become prominent and attracted more and more attention. Bibliometric analysis has been carried out in this research. The main goal of this research is to provide a comprehensive insight into the knowledge structure and global research hotspots about pelvic floor dysfunction in cervical cancer. METHODS: Literature related to cervical cancer and pelvic floor dysfunction as of May 2023 was searched on the Web of Science Core Collection (WOSCC). The visualization and bibliometric analyses of the number and contents of publications were performed to analyze the temporal trends, spatial distribution, collaborative networks, influential references, keyword co-occurrence, and clustering. RESULTS: There were 870 publications from 74 countries or regions, with the U.S. publications in a leading position. Since 2020, the number of publications has rapidly increased with the emphasis on the quality of life of cervical cancer patients. Although pelvic floor dysfunction in cervical cancer mainly occurs in developing countries, developed countries have made great contributions to this disease. However, in developing countries such as China and India, the quality of publications needs to be improved. In this field, the studies focused on the sexual dysfunction or urinary incontinence of cervical cancer patients, and the most cited papers discussed the effect of cervical cancer treatment on the sexual activities of females. The frontier keywords were represented by pelvic radiotherapy and risk factors. CONCLUSION: This study provides an objective and comprehensive analysis of the literature available on pelvic floor dysfunction in cervical cancer and identifies future trends and current hotspots. It can provide a valuable reference for researchers in this field.

Efficacy of hematoporphyrin injection (HpD) photodynamic therapy in the treatment of widespread extramammary Paget's disease.

Photodynamic therapy (PDT) utilizing Hematoporphyrin Derivative (HpD) injection has been demonstrated as an efficacious treatment for various conditions, including Bowen's disease, subtypes of basal cell carcinomas, and actinic keratosis. While surgical resection is considered the primary treatment option for extramammary Paget's disease (EMPD), some patients may not be suitable candidates for surgical intervention. ALA-PDT may have some benefits in treating EMPD in select patients, while Hematoporphyrin Derivative-Photodynamic Therapy (HpD-PDT) has demonstrated promising potential as a cancer treatment. We present one case of vulvar extramammary Paget's disease (EMPD), that is a female patient with lesions in the vulva and involving the urethra. Due to advanced age, underlying diseases, the extensive affected area, and the specific location of the vulvar lesion, the patients were unable to undergo surgical treatment. Therefore, the patient declined traditional wide local excision and instead opted for hematoporphyrin photodynamic therapy. Treatment eliminated the tumor, but it recurred locally after 1.5 years of follow-up. Localized small-scale recurrence at the affected site can be treated with surgical resection or photodynamic therapy to achieve complete clearance of the lesion. However, the patient refuses further examination and treatment. EMPD has a high recurrence rate, but we propose that hematoporphyrin photodynamic therapy is an effective alternative to conventional surgery for treating this condition, even in case of recurrence.

m6A Methylation Patterns and Tumor Microenvironment Infiltration Characterization in Clear-Cell Renal Cell Carcinoma.

Increasing evidence suggests the essential regulation of RNA N6-methyladenosine (m6A) modification in carcinogenesis and immune response. Nevertheless, the potential impacts of these modifications on the tumor microenvironment (TME) immune cell infiltration characteristics in clear-cell renal cell carcinoma (ccRCC) remain unclear. Utilizing a consensus clustering algorithm, we determined three m6A modification patterns and identified three m6A-related gene clusters among 569 ccRCC samples, which were associated with different biological functions and clinical outcomes. Thereafter, the m6A score was constructed using m6A-associated signature genes to accurately exploit the m6A modification patterns within individual tumors. The m6A score was further demonstrated to be noticeably related to ccRCC prognosis. In addition, the m6A score was found to be strongly correlated with tumor mutational burden (TMB), microsatellite instability, immune infiltration, immune checkpoint expression, and immunotherapy response, which was also validated in the pan-cancer analyses. Our findings thoroughly elucidated that m6A modification contributes to tumor microenvironment immune-infiltrating characteristics and prognosis in ccRCC. Assessing the m6A modification patterns of individual patients with ccRCC will offer novel insights into TME infiltration and help develop more effective treatment strategies.

Acute effect of sacral neuromodulation on video-urodynamic parameters of neurogenic lower urinary tract dysfunction in a patient with vesicoureteral reflux: a case report.

Neurogenic lower urinary tract dysfunction (NLUTD) is a common urological disease that causes long-term complications and severely reduces patient's quality of life. Sacral neuromodulation has proven to be an effective treatment for NLUTD. However, most previous studies have focused mainly on the efficacy and safety of sacral neuromodulation in the treatment of NLUTD and less on the changes in urodynamic parameters in patients before and after sacral neuromodulation. This study aimed to evaluate the effect of short-term sacral neuromodulation on the results of video-urodynamic parameters in a 63-year-old woman with NLUTD with vesicoureteral reflux. The patient was admitted to the Department of Urology of Beijing Hospital in January 2021 and examined using video-urodynamics. In the same month, the patient underwent the first stage of sacral neuromodulation, with an experience period of 2 weeks. After the experience period ended, video-urodynamics was performed again in February 2021. By comparing the two video-urodynamic results, the effect of short-term sacral neuromodulation on the anatomy and physiology of the lower urinary tract was determined. After 2 weeks of sacral neuromodulation treatment, video-urodynamic parameter analysis showed that while the urine storage period of the patient significantly improved, the voiding period was not significantly changed. This was specifically reflected in the improvement of bladder compliance, safe capacity of the bladder, and significant reduction in vesicoureteral reflux. The improvement of the safe capacity of the bladder effectively helped the patient to control the number of intermittent catheterizations within an acceptable range, which greatly improved her quality of life. Therefore, the patient underwent permanent sacral neuromodulation implantation in February 2021. This study suggests that short-term sacral neuromodulation can significantly improve lower urinary tract function and reduce vesicoureteral reflux in patients with NLUTD with vesicoureteral reflux. In short, we believe that sacral neuromodulation may be a good choice for patients with NLUTD.

The mechanism of oxymatrine on atopic dermatitis in mice based on SOCS1/JAK-STAT3 pathway.

Based on the suppressor of cytokine signaling 1 (SOCS1)/Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) pathway, the mechanism of oxymatrine in the treatment of atopic dermatitis (AD) was preliminarily explored in this study. C57BL/6 mice were induced to establish AD model by smearing carbotriol (MC903) on their back. The AD mice were randomly divided into model group, oxymatrine groups with three dosages (25, 50 and 100 mg/kg), (n = 10). Oxymatrine groups were intragastric administered once daily for 14 days. The same volume of saline was given in the normal control group and model group once daily for 14 days. Subsequently, HE staining was used to observe the pathological changes of skin tissue, ELISA was used to detect the levels of serum inflammatory factors including interleukin-4, 6 and 17 (IL-4, IL-6, and IL-17), tumor necrosis factor-α (TNF-α) and immunoglobulin E (IgE). Immunohistochemistry was used to detect the expression of suppressor of cytokine signaling 1 and CD3 in skin tissue, and Western blotting was used to detect the proteins in suppressor of cytokine signaling 1/JAK-STAT3 pathway. Compared with the normal control group, the pathological damage of mice in the model group, such as skin hyperplasia, edema, congestion and inflammatory infiltration, aggravated increased significantly. And the expression of serum inflammatory factors, CD3 positive expression and JAK-STAT3 pathway protein in the model group were increased (p < .05), and the expression of suppressor of cytokine signaling 1 protein (p < .05) was decreased. Compared with the model group, the above pathological damage of the mice was reduced, and the serum inflammatory factors, JAK-STAT3 pathway protein, and CD3 positive expression were decreased as a dose-dependant manner (p < .05), and the expression of suppressor of cytokine signaling 1 protein was increased as a dose-dependent manner (p < .05). Oxymatrine can improve the skin inflammation symptoms of AD mice by up regulating the expression of suppressor of cytokine signaling 1, inhibiting the activation of JAK-STAT3 pathway and blocking the activation of T lymphocytes.

N6-Methyladenosine-Related Long Non-coding RNA Signature Associated With Prognosis and Immunotherapeutic Efficacy of Clear-Cell Renal Cell Carcinoma.

Increasing evidence suggests that N6-methyladenosine (m6A) and long non-coding RNAs (lncRNAs) play important roles in cancer progression and immunotherapeutic efficacy in clear-cell renal cell carcinoma (ccRCC). In this study, we conducted a comprehensive ccRCC RNA-seq analysis using The Cancer Genome Atlas data to establish an m6A-related lncRNA prognostic signature (m6A-RLPS) for ccRCC. Forty-four prognostic m6A-related lncRNAs (m6A-RLs) were screened using Pearson correlation analysis (|R| > 0.7, p < 0.001) and univariable Cox regression analysis (p < 0.01). Using consensus clustering, the patients were divided into two clusters with different overall survival (OS) rates and immune status according to the differential expression of the lncRNAs. Gene set enrichment analysis corroborated that the clusters were enriched in immune-related activities. Twelve prognostic m6A-RLs were selected and used to construct the m6A-RLPS through least absolute shrinkage and selection operator Cox regression. We validated the differential expression of the 12 lncRNAs between tumor and non-cancerous samples, and the expression levels of four m6A-RLs were further validated using Gene Expression Omnibus data and Lnc2Cancer 3.0 database. The m6A-RLPS was verified to be an independent and robust predictor of ccRCC prognosis using univariable and multivariable Cox regression analyses. A nomogram based on age, tumor grade, clinical stage, and m6A-RLPS was generated and showed high accuracy and reliability at predicting the OS of patients with ccRCC. The prognostic signature was found to be strongly correlated to tumor-infiltrating immune cells and immune checkpoint expression. In conclusion, we established a novel m6A-RLPS with a favorable prognostic value for patients with ccRCC. The 12 m6A-RLs included in the signature may provide new insights into the tumorigenesis and allow the prediction of the treatment response of ccRCC.

TNFSF13B and PPARGC1A expression is associated with tumor-infiltrating immune cell abundance and prognosis in clear cell renal cell carcinoma.

Growing evidence suggests that the tumor microenvironment (TME) plays crucial roles in tumor progression and treatment efficacy in clear cell renal cell carcinoma (ccRCC), which typically has a poor prognosis due to high relapse and metastasis rates. We comprehensively analyzed ccRCC RNA-sequencing data from The Cancer Genome Atlas (TCGA) database to identify candidate prognostic TME-related genes involved in ccRCC. We used the ESTIMATE and CIBERSORT algorithms to estimate the proportions of immune cells, stromal cells, and tumor-infiltrating immune cells (TICs) in the TME in ccRCC samples from 539 patients. By examining the intersection of the differentially expressed genes (DEGs) obtained by Cox regression analysis and protein-protein interaction network, we identified five overlapping DEGs (IGLL5, MZB1, HSD11B1, TNFSF13B, and PPARGC1A). Further analysis revealed that TNFSF13B expression was elevated in ccRCC tumor tissues and negatively associated with overall survival. PPARGC1A expression exhibited the opposite patterns. Immunohistochemical analysis of 35 paired ccRCC and adjacent normal tissues confirmed the in-silico results. Gene set enrichment analysis revealed that genes in the groups with high TNFSF13B and PPARGC1A expression were enriched mainly in immune-related activities. In the group with low PPARGC1A expression, genes were enriched in metabolic pathways. CIBERSORT analysis of TIC proportions revealed that Tregs and CD8 T-cell abundance correlated positively with TNFSF13B expression, but negatively with PPARGC1A expression. These findings demonstrate that TNFSF13B and PPARGC1A are prognostic predictors and possible therapeutic targets in ccRCC.

Survival in Patients With Metastatic Prostate Cancer Undergoing Radiotherapy: The Importance of Prostate-Specific Antigen-Based Stratification.

OBJECTIVES: To explore the effectiveness of radiotherapy in mPCa patients with different PSA stratifications based on the cancer database of a large population. BACKGROUND: Screening criteria for patients with metastatic prostate cancer, who are candidates for radiotherapy, are rarely reported. PATIENTS AND METHODS: We identified 22,604 patients with metastatic prostate cancer in the Surveillance, Epidemiology, and End Results database and divided them into a radiotherapy group and a control group. Patients with metastatic prostate cancer were divided into subgroups according to their levels of prostate-specific antigen to evaluate the efficacy of radiotherapy. They were also divided into six subgroups according to their prostate-specific antigen levels. We used multivariate Cox analysis to evaluate overall survival and cancer-specific survival. After 1:1 propensity score matching, Kaplan-Meier analysis was used to explore the difference in overall survival and cancer-specific survival in the radiotherapy and control group. RESULTS: In all, 5,505 patients received radiotherapy, compared to 17,099 in the control group. In the multivariate Cox analysis, radiotherapy improved overall survival (hazard ratio [HR]: 0.730, 95% confidence interval [CI]: 0.636-0.838; P<0.001) and cancer-specific survival (HR: 0.764, 95% CI: 0.647-0.903; P=0.002) in patients with a PSA level of 4-10 ng/mL. Similar results were obtained by Kaplan-Meier analysis after 1:1 propensity score matching. In patients with prostate-specific antigen levels between 4-10 ng/mL, the overall survival (P<0.001) and cancer-specific survival (P<0.05) in the radiotherapy group was significantly better than those in the control group. CONCLUSION: The result of this large population-based study shows that rigorous selection of appropriate metastatic prostate cancer patients for radiotherapy can benefit prognosis significantly. This can be the basis for future prospective trials.

Young age increases the risk of lymph node positivity but improves prognosis in patients with bladder cancer treated via cystectomy: a population-based study.

BACKGROUND: Age and lymph node positivity are significant prognostic indicators in patients with bladder cancer. This study aimed to investigate the impact of age on lymph node positivity and bladder cancer outcomes. METHODS: Patients with bladder cancer who underwent cystectomy with at least one lymph node examined between 2004 and 2015 were identified from the Surveillance, Epidemiology, and End Results database. Cochran-Armitage trend tests and logistic regression analyses were used to evaluate the association between age and lymph node positivity in all T stages. Multivariate Cox regression analysis was used to analyze the effect of age on overall survival (OS) and cancer-specific survival (CSS). RESULTS: Overall, 13,251 patients were identified, 648 of whom were under 50 years of age (4.89%). Lymph node positivity was negatively associated with increasing age in each stage except in non-invasive-muscular bladder cancer. In the multivariable analysis, age was an independent prognostic factor for OS and CSS in both the overall cohort and the lymph node positivity group. CONCLUSIONS: In patients with bladder cancer undergoing cystectomy, young age at diagnosis is associated with a higher risk of lymph node positivity and superior outcomes. These findings may guide clinicians in selecting suitable treatments, determining the aggressiveness of lymph node involvement, and predicting survival outcomes in patients of different ages.

Nomograms to predict individual prognosis of patients with primary signet ring cell carcinoma of the urinary bladder.

BACKGROUND: Signet ring cell carcinoma (SRCC) is a rare but highly malignant variant of bladder carcinoma. Nomograms have demonstrated good accuracy in predicting the prognosis and guiding the management of pure urothelial carcinoma (UC). However, no accurate and applicable nomogram has been formulated for primary SRCC cases. This study aimed to determine significant prognostic factors and to construct nomograms for predicting the survival outcomes of patients with primary SRCCs of the urinary bladder. METHODS: A total of 317 eligible patients diagnosed with SRCC were analyzed using the 2004-2016 data from the Surveillance, Epidemiology, and End Results database. Univariate and multivariate analyses were performed to explore the prognostic values. Nomograms were established to estimate the overall survival (OS) and cancer-specific survival (CSS) based on the Cox regression results. The performance of SRCC nomograms was evaluated using the concordance index and calibration curves. Survival curves were applied according to various surgical methods, lymph node status, and risk groups distinguished by nomograms. RESULTS: Two nomograms included common indicators that were significantly associated with OS and CSS, including T stage, M stage, tumor size, surgery, and the lymph node ratio (LNR). The nomograms indicated appreciable accuracy in predicting the OS and CSS, with concordance index of 0.723 [95% confidence interval (95% CI: 0.692-0.754] and 0.740 (95% CI: 0.701-0.779), respectively. The calibration curves revealed satisfactory consistency between the prediction of deviation correction and ideal reference line. CONCLUSIONS: The two nomograms developed in this study showed high accuracy and reliability in predicting the survival outcomes of patients with SRCC and could be used to comprehensively assess the risk of SRCC. Moreover, they could assist in the optimal treatment selection for such patients.

Prevalence, genetic diversity and implications for public health of Enterocytozoon bieneusi in various rodents from Hainan Province, China.

BACKGROUND: Rodents, globally overpopulated, are an important source for zoonotic disease transmission to humans, including Enterocytozoon bieneusi (one of the most prevalent zoonotic pathogens). Here, we studied the prevalence and performed genetic analyses of E. bieneusi in rodents from the Hainan Province of China. METHODS: A total of 603 fresh fecal samples were gathered from 369 wild rats, 117 bamboo rats, 93 Asiatic brush-tailed porcupine and 24 red-bellied squirrels. The wild rats were identified to the species level by amplification of a 421-bp region of the cytb gene from fecal DNA using PCR. Genotype analysis was performed by amplification of the internal transcribed spacer (ITS) region of rDNA of E. bieneusi using PCR. RESULTS: Seven wild rat species were identified. The average rate of infection with E. bieneusi was 15.8% (95/603) with 18.7% (69/369) in wild rats, 11.9% (25/210) in farmed rodents and 4.2% (1/24) in red-bellied squirrels. Sixteen E. bieneusi genotypes were identified, including 9 known genotypes (D, Type IV, PigEBITS7, Peru8, Peru11, ESH02, S7, EbpA and CHG5), and 7 novel genotypes (HNR-I to HNR-VII). Genotype D (44.2%, 42/95) predominated, followed by PigEBITS7 (20.0%, 19/95), HNR-VII (15.8%, 15/95), Type IV (5.3%, 5/95), HNR-III (2.1%, 2/95), HNR-VI (2.1%, 2/95) and each of the remaining 10 genotypes (1.1%, 1/95). The phylogenetic analysis of the ITS region of E. bieneusi divided the identified genotypes into the following four groups: Group 1 (n = 13), Group 2 (n = 1), Group 12 (n = 1), and the novel Group 13 (n = 1). CONCLUSIONS: To our knowledge, this is the first report on the identification of E. bieneusi in rodents from Hainan, China. The zoonotic potential of the identified E. bieneusi genotypes suggested that the rodents poses a serious threat to the local inhabitants. Thus, measures need to be taken to control the population of wild rats in the areas investigated in this study, along with identification of safe methods for disposal of farmed rodent feces. Additionally, the local people should be made aware of the risk of disease transmission from rodents to humans.

Molecular detection of Enterocytozoon bieneusi in farm-raised pigs in Hainan Province, China: infection rates, genotype distributions, and zoonotic potential.

Enterocytozoon bieneusi is a zoonotic fungal pathogen with a high degree of host diversity that can parasitize many animals, including humans. Pigs may play an important role in the epidemiology of E. bieneusi as reservoir hosts. Nevertheless, the genotypes of E. bieneusi in pigs in China remain poorly understood. The aim of this study was to determine the prevalence of E. bieneusi infection amongst pigs raised on farms from four cities of Hainan Province, using nested polymerase chain reaction (PCR) of the partial small subunit of the ribosomal RNA gene, and to identify genotypes of E. bieneusi isolates based on sequence analysis of the ribosomal internal transcribed spacer (ITS) region. Among 188 stool samples, E. bieneusi was detected in 46.8% (88/188). Eight genotypes including four known (EbpA, CS-4, MJ14, and CHG19) and four novel (HNP-I - HNP-IV) genotypes were identified. Using phylogenetic analysis, genotypes EbpA, CS4, CHG19, HNP-III, and HNP-IV were clustered into zoonotic Group 1, while the remaining three genotypes (MJ14, HNP-I, and HNP-II) clustered into Group 10. The high prevalence of zoonotic genotypes of E. bieneusi among pigs suggests that pig farming is a potential source of human infection. Additionally, this is the first identification of genotypes in Group 10 in pigs indicating unique epidemic features of E. bieneusi in pigs in Hainan Province, the southernmost part of China.

TITLE: Détection moléculaire d'Enterocytozoon bieneusi chez les porcs d'élevage dans la province de Hainan en Chine : taux d'infection, répartition des génotypes et potentiel zoonotique. ABSTRACT: Enterocytozoon bieneusi est un pathogène fongique zoonotique avec une grande diversité d'hôte qui peut parasiter de nombreux animaux, y compris les humains. Les porcs peuvent jouer un rôle important dans l'épidémiologie d'E. bieneusi en tant qu'hôtes réservoirs. Néanmoins, les génotypes d'E. bieneusi chez le porc en Chine restent mal connus. Le but de cette étude était de déterminer la prévalence de l'infection par E. bieneusi chez les porcs élevés dans des fermes de quatre villes de la province de Hainan, en utilisant la réaction en chaîne par polymérase emboîtée (PCR) de la petite sous-unité partielle du gène de l'ARN ribosomal et de identifier les génotypes des isolats d'E. bieneusi sur la base d'une analyse de séquence de la région des espaceurs internes transcrits ribosomiques (ITS). Sur 188 échantillons de selles, E. bieneusi a été détecté dans 46,8 % (88/188). Huit génotypes, dont quatre génotypes connus (EbpA, CS-4, MJ14 et CHG19) et quatre génotypes nouveaux (HNP-I à IV), ont été identifiés. Dans une analyse phylogénétique, les génotypes EbpA, CS4, CHG19, HNP-III et HNP-IV étaient regroupés dans le groupe zoonotique 1, tandis que les trois génotypes restants (MJ14, HNP-I et HNP-II) étaient regroupés dans le groupe 10. La prévalence élevée des génotypes zoonotiques d'E. bieneusi chez les porcs suggère que l'élevage porcin est une source potentielle d'infection humaine. De plus, il s'agit de la première identification de génotypes du groupe 10 chez les porcs, indiquant des caractéristiques épidémiques uniques d'E. bieneusi chez les porcs dans la province de Hainan, la partie la plus méridionale de la Chine.

Genotyping and zoonotic potential of Enterocytozoon bieneusi in cattle farmed in Hainan Province, the southernmost region of China.

Enterocytozoon bieneusi is an intestinal pathogen that infects a wide range of species, including humans. Cattle constitute an important host for E. bieneusi; however, there is a scarcity of information on the prevalence and genotyping of E. bieneusi in cattle in the Hainan Province of China. In this study, PCR analysis of 314 fecal samples from cattle in six cities of Hainan was performed for genotype identification. The average prevalence of E. bieneusi in these animals was 9.9% (31/314), and ranged from 0.0% (0/12) to 20.5% (8/39). Five known genotypes - EbpC (n = 14), BEB4 (n = 12), J (n = 2), I (n = 1), and CHG5 (n = 1) - and a novel genotype: HNC-I (n = 1) - were identified. Genotypes EbpC and HNC-I were placed in zoonotic Group 1, and the remaining four genotypes (BEB4, J, I, and CHG5) were placed in Group 2. Since 93.5% of the genotypes found in the cattle (29/31) (EbpC, BEB4, J, and I) have previously been found in humans, these genotypes are probably involved in the transmission of microsporidiosis to humans.

TITLE: Génotypage et potentiel zoonotique d'Enterocytozoon bieneusi chez les bovins élevés dans la province de Hainan, la région la plus au sud de la Chine. ABSTRACT: Enterocytozoon bieneusi est un pathogène intestinal qui infecte un large éventail d'espèces, y compris les humains. Le bétail constitue un hôte important pour E. bieneusi, mais les informations sur la prévalence et le génotypage d'E. bieneusi chez les bovins de la province de Hainan en Chine sont rares. Dans cette étude, une analyse PCR de 314 échantillons fécaux provenant de bovins dans six villes de Hainan a été réalisée pour l'identification du génotype. La prévalence moyenne d'E. bieneusi chez ces animaux était de 9,9 % (31/314), et variait de 0,0 % (0/12) à 20,5 % (8/39). Cinq génotypes connus, EbpC (n = 14), BEB4 (n = 12), J (n = 2), I (n = 1) et CHG5 (n = 1), et un nouveau génotype, HNC-I (n = 1), ont été identifiés. Les génotypes EbpC et HNC-I sont placés dans le groupe zoonotique 1, et les quatre génotypes restants (BEB4, J, I et CHG5) sont placés dans le groupe 2. Puisque 93,5 % (29/31) (EbpC, BEB4, J et I) des génotypes trouvés chez les bovins ont déjà été trouvés chez l'homme, ces génotypes sont probablement impliqués dans la transmission de la microsporidiose à l'homme.

Genotyping of Enterocytozoon bieneusi among captive long-tailed macaques (Macaca fascicularis) in Hainan Province: High genetic diversity and zoonotic potential.

Enterocytozoon bieneusi is a potentially important zoonotic pathogen. However, there is no information on E. bieneusi infection of captive long-tailed macaques (Macaca fascicularis) in Hainan Province, China. Here 193 fecal specimens of M. fascicularis were collected from a breeding base in Hainan Province, China, housing non-human primates for experimental use. E. bieneusi was identified and genotyped by nested PCR analysis of the internal transcribed spacer (ITS) region of the rRNA gene. A total of 59 (30.6%) specimens were PCR-positive for E. bieneusi and 16 ITS genotypes were identified including nine known genotypes: Type IV (n = 19), D (n = 11), CM1 (n = 8), PigEBITS7 (n = 4), Pongo2 (n = 4), Peru8 (n = 3), Peru11 (n = 1), WL21 (n = 1) and CM2 (n = 1) and seven novel genotypes HNM-I to HNM-VII (one each). Importantly, genotypes D, Type IV, Peru8, PigEBITS7, and Peru11, which were the predominant (38/59, 64.4%) genotypes identified among captive M. fascicularis in this study, are also well-known human-pathogenic genotypes. All the genotypes of E. bieneusi identified here, including the seven novel ones, belonged to zoonotic Group 1. This is the first report of the identification of E. bieneusi in M. fascicularis in Hainan Province, China. The finding that the numerous known human-pathogenic types and seven novel genotypes of E. bieneusi all belong to zoonotic Group 1 indicates the possibility of transmission of this important pathogenic parasite between M. fascicularis and humans.

PCR-Based Detection of Cryptosporidium spp. and Enterocytozoon bieneusi in Farm-Raised and Free-Ranging Geese (Anser anser f. domestica) From Hainan Province of China: Natural Infection Rate and the Species or Genotype Distribution.

Cryptosporidium spp. and Enterocytozoon bieneusi are two important zoonotic pathogens that can infect humans and a broad range of animal hosts. However, few studies have been conducted to study infection of the two pathogens in domestic geese until now. The aims of the present study were to determine the prevalence of natural infection, and the species or genotype distribution of Cryptosporidium and E. bieneusi in farm-raised and free-ranging geese from Hainan Province of China. In total, 266 fecal samples of geese were collected (142 farm-raised and 124 free-ranging geese). Cryptosporidium spp. and E. bieneusi were identified by nested PCR and sequencing analysis of the SSU rRNA and the ITS region of the rRNA genes. A total of 4.1% (12/226) of the geese were positive for Cryptosporidium spp., with 0.7% identified in the farm-raised geese and 7.0% in the free-ranging geese. Two bird-adapted species/genotypes were identified: C. baileyi (n = 1) and Cryptosporidium goose genotype I (n = 11). Meanwhile, E. bieneusi was found in 13.9% (37/266) of geese, with 8.9% identified in the farm-raised and 21.8% in the free-ranging geese. Eleven genotypes of E. bieneusi were identified constituted with six known genotypes: D (n = 13), I (n = 5), CHG2 (n = 1), CHG3 (n = 5), and CHG5 (n = 1), and five novel genotypes named HNE-I to V (one each). All of the genotypes identified in the geese here belonged to zoonotic Groups 1 or 2. This study is the first to demonstrate the presence of Cryptosporidium spp. and E. bieneusi in domestic geese from Hainan, China, and provides baseline data that will be useful for controlling and preventing these pathogens in goose farms. The geese infected with E. bieneusi, but not with Cryptosporidium, should be considered potential public health threats.

Genotype identification and phylogenetic analysis of Enterocytozoon bieneusi in farmed black goats (Capra hircus) from China's Hainan Province.

Enterocytozoon bieneusi is an important pathogen commonly found in humans and animals. Farmed animals with close contact to humans are important hosts of E. bieneusi. The role of goats in the transmission of E. bieneusi, however, remains unclear. In this study, 341 fresh fecal samples of black goats were collected from five locations in Hainan Province, China. Enterocytozoon bieneusi was identified and genotyped by sequences of the internal transcribed spacer (ITS) region. Phylogenetic analysis was performed by constructing a neighbor-joining tree of the ITS gene sequences. The average prevalence of E. bieneusi in black goats was 24.0% (82/341) with rates ranging from 6.3% (4/63) to 37.2% (32/86) across the locations (χ2 = 17.252, p < 0.01). Eight genotypes of E. bieneusi were identified, including six known genotypes: CHG5 (n = 47); CHG3 (n = 23); CHG2 (n = 4); CM21 (n = 3); D (n = 2); and AHG1 (n = 1), and two novel genotypes termed HNG-I (n = 1) and HNG-II (n = 1). In the phylogenetic tree, genotype D was clustered into Group 1 and the other identified genotypes were included in Group 2. This represents the first report identifying E. bieneusi in black goats from Hainan Province, with a high prevalence and wide occurrence demonstrated. The two new genotypes identified provide additional insights into the genotypic variations in E. bieneusi. Due to the small percentage of zoonotic genotypes in these animals, there is minimal risk of zoonotic transmission of E. bieneusi.

TITLE: Identification du génotype et analyse phylogénétique d'Enterocytozoon bieneusi chez des chèvres noires (Capra hircus) de la province de Hainan, en Chine. ABSTRACT: Enterocytozoon bieneusi est un agent pathogène important que l'on trouve couramment chez l'homme et les animaux. Les animaux d'élevage, en contact étroit avec l'homme, sont des hôtes importants d'E. bieneusi. Le rôle des chèvres dans la transmission d'E. bieneusi reste toutefois incertain. Dans cette étude, 341 échantillons de fèces fraîches de chèvres noires ont été prélevés dans cinq sites de la province de Hainan, en Chine. Enterocytozoon bieneusi a été identifié et génotypé par des séquences de la région de l'espaceur interne transcrit (ITS). L'analyse phylogénétique a été réalisée en construisant un arbre de jonction voisine des séquences du gène ITS. La prévalence moyenne d'E. bieneusi chez les chèvres noires était de 24,0 % (82/341), avec des taux allant de 6,3 % (4/63) à 37,2 % (32/86) dans tous les sites (χ2 = 17,252, p < 0,01). Huit génotypes d'E. bieneusi ont été identifiés, dont six génotypes connus: CHG5 (n = 47) ; CHG3 (n = 23) ; CHG2 (n = 4) ; CM21 (n = 3) ; D (n = 2) ; AHG1 (n = 1) et deux nouveaux génotypes appelés HNG-I (n = 1) et HNG-II (n = 1). Dans l'arbre phylogénétique, le génotype D appartenait au groupe 1 et les autres génotypes identifiés étaient inclus dans le groupe 2. Il s'agit du premier rapport identifiant E. bieneusi chez des chèvres noires de la province de Hainan, avec une prévalence élevée et une occurrence étendue. Les deux nouveaux génotypes identifiés fournissent des informations supplémentaires sur les variations génotypiques chez E. bieneusi. En raison du faible pourcentage de génotypes zoonotiques chez ces animaux, le risque de transmission zoonotique d'E. bieneusi est minime.

Cryptosporidium spp. in wild rats (Rattus spp.) from the Hainan Province, China: Molecular detection, species/genotype identification and implications for public health.

Wild rats (Rattus spp.) carry many zoonotic pathogens including Cryptosporidium. Due to the close proximity of rats to humans in urban environments, the potential for disease transmission is high. Cryptosporidium is a protozoan parasite which when ingested causes serious human illness. Despite its importance, genetic characterization of Cryptosporidium in wild rats in the Hainan province of China has not been performed. In this study, we analyzed the occurrence and genetics of Cryptosporidium in wild rats from Hainan, China. From December 2017 to October 2018, 150 wild rats were captured and fresh fecal material was collected from intestinal sections. Rat species were identified by PCR-based amplification and analysis of the vertebrate cytochrome b (cytb) gene. Cryptosporidium was examined by PCR amplification of the partial small subunit of ribosomal DNA (SSU rDNA). C. viatorum were subtyped by PCR analysis of the gp60 gene. A total of four rat species were identified including Asian house rats (Rattus tanezumi) (n = 46), brown rats (Rattus norvegicus) (n = 56), Edward's long-tailed rats (Leopoldamys edwardsi) (n = 38) and muridae (Niviventer fulvescens) (n = 10), with Cryptosporidium positive rates of 73.9%, 28.6%, 55.3% and 40.0%, respectively (average infection rate: 50.0%, 75/150. Sequence analysis confirmed the presence of four Cryptosporidium species and two genotypes including C. viatorum (n = 11); C. occultus (n = 2); C. muris (n = 1); and C. erinacei (n = 1); rat genotypes III (n = 13) and IV (n = 47). Three novel subtypes of C. viatorum were identified in 6 of the 11 infected Edward's long-tailed rats: XVcA2G1a (n = 4), XVcA2G1b (n = 1) and XVdA3 (n = 1). The identification of human pathogenic C. viatorum and zoonotic C. occultus, C. muris and C. erinacei, suggested that wild rats infected with Cryptosporidium pose a threat to human health. Taken together, these findings highlight the need to control the rat population in Hainan, China. The need to improve the public awareness of the risk of disease transmission from wild rats to humans is also highlighted.