Parasitic Leiomyoma at Laparoscopic Trocar Site: A Report of 2 Cases.

BACKGROUND Parasitic leiomyoma refers to leiomyomas outside the uterus, with a prevalence of 0.07%. Patients are initially asymptomatic and may later develop abdominal pain and abdominal distension. Parasitic leiomyomas at a trocar site are extremely rare and lack detailed reporting. Here, we report 2 cases of parasitic leiomyoma at trocar sites. CASE REPORT Case 1. The patient was a 47-year-old woman with parasitic leiomyomas at a left trocar site 4 years after laparoscopic total hysterectomy. After being diagnosed with 3 masses on the surface of the sigmoid colon and 2 in the pelvic cavity, the patient underwent laparoscopic removal of a pelvic lesion and 3 lesions on the surface of the colon, combined with excision of abdominal wall masses. The pathology result indicated that the masses at the left trocar site were multiple leiomyomas, the intestinal mass was multiple leiomyomas with abundant cells, and the pelvic mass was fibrous capsule parietal tissue. This patient received 3 months of gonadotropin-releasing hormone agonist (GnRH-a) treatment, and was followed up for 9 months without recurrence. Case 2. The patient was a 50-year-old woman with parasitic leiomyoma at the right trocar site 15 years after laparoscopic removal of the right ovarian cyst. At admission, she underwent transabdominal total hysterectomy, bilateral fallopian tube resection, and abdominal wall lesion resection. The pathology report showed multiple leiomyomas of the uterus, and the cell-rich parasitic leiomyoma at right trocar site with unclear boundary. She received 3 months of GnRH-a treatment, and was followed up for 6 months without recurrence. CONCLUSIONS For patients with a history of laparoscopy, gynecologists should be alert to the occurrence of parasitic leiomyoma.

A New Orthonairovirus Associated with Human Febrile Illness.

BACKGROUND: In June 2019, a patient presented with persistent fever and multiple organ dysfunction after a tick bite at a wetland park in Inner Mongolia. Next-generation sequencing in this patient revealed an infection with a previously unknown orthonairovirus, which we designated Wetland virus (WELV). METHODS: We conducted active hospital-based surveillance to determine the prevalence of WELV infection among febrile patients with a history of tick bites. Epidemiologic investigation was performed. The virus was isolated, and its infectivity and pathogenicity were investigated in animal models. RESULTS: WELV is a member of the orthonairovirus genus in the Nairoviridae family and is most closely related to the tickborne Hazara orthonairovirus genogroup. Acute WELV infection was identified in 17 patients from Inner Mongolia, Heilongjiang, Jilin, and Liaoning, China, by means of reverse-transcriptase-polymerase-chain-reaction assay. These patients presented with nonspecific symptoms, including fever, dizziness, headache, malaise, myalgia, arthritis, and back pain and less frequently with petechiae and localized lymphadenopathy. One patient had neurologic symptoms. Common laboratory findings were leukopenia, thrombocytopenia, and elevated d-dimer and lactate dehydrogenase levels. Serologic assessment of convalescent-stage samples obtained from 8 patients showed WELV-specific antibody titers that were 4 times as high as those in acute-phase samples. WELV RNA was detected in five tick species and in sheep, horses, pigs, and Transbaikal zokors (Myospalax psilurus) sampled in northeastern China. The virus that was isolated from the index patient and ticks showed cytopathic effects in human umbilical-vein endothelial cells. Intraperitoneal injection of the virus resulted in lethal infections in BALB/c, C57BL/6, and Kunming mice. The Haemaphysalis concinna tick is a possible vector that can transovarially transmit WELV. CONCLUSIONS: A newly discovered orthonairovirus was identified and shown to be associated with human febrile illnesses in northeastern China. (Funded by the National Natural Science Foundation of China and the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences.).

The function of OforOrco in the allogrooming behavior of Odontotermes formosanus (Shiraki) induced by Serratia marcescens Bizio (SM1).

Termites are consistently confronted with a complex microbial environment. In addition to the role of their innate immune system in resisting pathogen infection, social immune behavior also plays a significant role in helping termites withstand the stress caused by pathogenic microorganisms. The allogrooming behavior among different individuals is commonly observed in termites, and it plays a crucial role in the social immune interaction network. In the case of Odontotermes formosanus (Shiraki), Orco is specifically involved in detecting pheromones and volatile chemicals released by termites to communicate with each other. Nonetheless, the function of Orco in the social immunity remains unreported in O. formosanus. Consequently, in this study, we recorded the allogrooming behavior of O. formosanus workers under SM1 stress. The results indicated a significant increase in allogrooming behavior due to SM1 infection. The allogrooming behavior of workers under SM1 stress was significantly increased after the addition of soldiers. Compared with pronotum group treated by SM1, SM1 treatment of workers' heads significantly reduced the allogrooming behavior among workers. In addition, we found that SM1 could greatly increase the expression of OforOrco. Furthermore, interfering with OforOrco could markedly reduce the allogrooming behavior among workers under SM1 stress, and increase the mortality of worker under SM1 stress. This study demonstrated the significant role of OforOrco in the social immunity of O. formosanus, which offers a theoretical foundation for the advancement of research on termite RNA biopesticides, and the integration of RNA interference (RNAi) with pathogens. This study is valuable for elucidating the social immune behavior and interaction network of termites.

Enhancing the sensitivity of immunomagnetic assay for 3-phenoxybenzonic acid: a novel approach utilizing magnetosome-nanobody complexes and gold nanoparticles probes.

The 3-phenoxybenzoic acid (3-PBA) residues in environment are posing a significant challenge to our daily lives. To establish a more sensitive and rapid detection method, anti-3-PBA nanobodies (Nbs) were immobilized onto magnetosomes (bacterial magnetic nanoparticles, termed as BMPs), forming a robust BMP-Nb complex. The 3-PBA derivative was labeled with horseradish peroxidase (HRP) and further associated with gold nanoparticles (AuNPs) to create a highly sensitive probe (3-PBA-HRP-AuNP). An innovative immunoassay that combined BMP-Nb complex with 3-PBA-HRP-AuNP was developed for determinaton of 3-PBA. This method enabled the determination of 3-PBA with a half-maximum signal inhibition concentration (IC50) of 1.03 ng/mL, which was more sensitive than that of using 3-PBA-HRP as tracer with an IC50 of 2.18 ng/mL. The reliability of the assay was evidenced by the quantitative recovery of 3-PBA from water and soil samples ranging from 76.85 to 95.64%. The 3-PBA residues determined by this assay in actual water samples were between < LOD and 2.54 ng/mL and were between < LOD and 11.25 ng/g (dw) in real soils, respectively, which agreed well with those of liquid chromatography mass spectrometry (LC-MS). Collectively, the BMP-Nb and 3-PBA-HRP-AuNP-based immunoassay provides a powerful tool for the precise detection of 3-PBA residues in environment matrices, reinforcing our capacity to monitor and mitigate potential ecological and health impacts associated with this prevalent pollutant.

A stacking ensemble model for predicting the occurrence of carotid atherosclerosis.

Background: Carotid atherosclerosis (CAS) is a significant risk factor for cardio-cerebrovascular events. The objective of this study is to employ stacking ensemble machine learning techniques to enhance the prediction of CAS occurrence, incorporating a wide range of predictors, including endocrine-related markers. Methods: Based on data from a routine health check-up cohort, five individual prediction models for CAS were established based on logistic regression (LR), random forest (RF), support vector machine (SVM), extreme gradient boosting (XGBoost) and gradient boosting decision tree (GBDT) methods. Then, a stacking ensemble algorithm was used to integrate the base models to improve the prediction ability and address overfitting problems. Finally, the SHAP value method was applied for an in-depth analysis of variable importance at both the overall and individual levels, with a focus on elucidating the impact of endocrine-related variables. Results: A total of 441 of the 1669 subjects in the cohort were finally diagnosed with CAS. Seventeen variables were selected as predictors. The ensemble model outperformed the individual models, with AUCs of 0.893 in the testing set and 0.861 in the validation set. The ensemble model has the optimal accuracy, precision, recall and F1 score in the validation set, with considerable performance in the testing set. Carotid stenosis and age emerged as the most significant predictors, alongside notable contributions from endocrine-related factors. Conclusion: The ensemble model shows enhanced accuracy and generalizability in predicting CAS risk, underscoring its utility in identifying individuals at high risk. This approach integrates a comprehensive analysis of predictors, including endocrine markers, affirming the critical role of endocrine dysfunctions in CAS development. It represents a promising tool in identifying high-risk individuals for the prevention of CAS and cardio-cerebrovascular diseases.

Decoding the RNA viromes in shrew lungs along the eastern coast of China.

Shrews being insectivores, serve as natural reservoirs for a wide array of zoonotic viruses, including the recently discovered Langya henipavirus (LayV) in China in 2018. It is crucial to understand the shrew-associated virome, viral diversity, and new viruses. In the current study, we conducted high-throughput sequencing on lung samples obtained from 398 shrews captured along the eastern coast of China, and characterized the high-depth virome of 6 common shrew species (Anourosorex squamipes, Crocidura lasiura, Crocidura shantungensis, Crocidura tanakae, Sorex caecutiens, and Suncus murinus). Our analysis revealed numerous shrew-associated viruses comprising 54 known viruses and 72 new viruses that significantly enhance our understanding of mammalian viruses. Notably, 34 identified viruses possess spillover-risk potential and six were human pathogenic viruses: LayV, influenza A virus (H5N6), rotavirus A, rabies virus, avian paramyxovirus 1, and rat hepatitis E virus. Moreover, ten previously unreported viruses in China were discovered, six among them have spillover-risk potential. Additionally, all 54 known viruses and 12 new viruses had the ability to cross species boundaries. Our data underscore the diversity of shrew-associated viruses and provide a foundation for further studies into tracing and predicting emerging infectious diseases originated from shrews.

The effect of joint mobilization of Maitland on chronic ankle instability: A randomized trial.

BACKGROUND: The aim of study was to observe the therapeutic effect of joint mobilization of Maitland on subjects with chronic ankle instability (CAI). METHODS: 76 subjects with CAI were recruited for this randomized, single-blinded trial and randomized divided into experimental group (EG) and control group (CG). The CG was received conventional rehabilitation, and the EG added 8-weeks treatment of Maitland technology based on the CG. The visual analogue scale, ankle range of motion, Y-balance test, and Foot and Ankle Ability Measure scores (the daily living part of Foot and Ankle Ability Measure scores and the sport part of Foot and Ankle Ability Measure scores) were measured before and 8 weeks after the intervention respectively. RESULTS: There was no significant difference on outcomes between the 2 groups before treatment (P > .05). After 8 weeks of intervention, the visual analogue scale, ankle range of motion (dorsiflexion, plantar flexion, and varus), the value of Y-balance test (forward extension distance, inner extension distance, and posterior extension distance), the daily living part of Foot and Ankle Ability Measure scores, and the sport part of Foot and Ankle Ability Measure scores of the 2 groups were significantly improved (P < .01), and the improvement of the EG showed remarkable than CG (P < .01). CONCLUSION: Maitland therapy is effective in the treatment of CAI. Conventional rehabilitation assisted by Maitland therapy were beneficial to improve pain and functional state in patients with CAI than only routine rehabilitation.

Dynamic prediction of lung cancer suicide risk based on meteorological factors and clinical characteristics：A landmarking analysis approach.

Suicide is a severe public health issue globally. Accurately identifying high-risk lung cancer patients for suicidal behavior and taking timely intervention measures has become a focus of current research. This study intended to construct dynamic prediction models for identifying suicide risk among lung cancer patients. Patients were sourced from the Surveillance, Epidemiology, and End Results database, while meteorological data was acquired from the Centers for Disease Control and Prevention. This cohort comprised 455, 708 eligible lung cancer patients from January 1979 to December 2011. A Cox proportional hazard regression model based on landmarking approach was employed to explore the impact of meteorological factors and clinical characteristics on suicide among lung cancer patients, and to build dynamic prediction models for the suicide risk of these patients. Additionally, subgroup analyses were conducted by age and sex. The model's performance was evaluated using the C-index, Brier score, area under curve (AUC) and calibration plot. During the study period, there were 666 deaths by suicide among lung cancer patients. Multivariable Cox results from the dynamic prediction model indicated that age, marital status, race, sex, primary site, stage, monthly average daily sunlight, and monthly average temperature were significant predictors of suicide. The dynamic prediction model demonstrated well consistency and discrimination capabilities. Subgroup analyses revealed that the association of monthly average daily sunlight and monthly average temperature with suicide remained significant among female and younger lung cancer patients. The dynamic prediction model can effectively incorporate covariates with time-varying to predict lung cancer patients' suicide death. The results of this study have significant implications for assessing lung cancer individuals' suicide risk.

Meta-analysis of the dosage of balance training on ankle function and dynamic balance ability in patients with chronic ankle instability.

OBJECTIVE: To explore and compare the dosage of balance training on ankle function and dynamic balance ability in patients with chronic ankle instability (CAI). METHODS: The PubMed, Embase, Web of Science, Medline, and Cochrane databases were searched up to December 2023. Quality assessment was carried out using the risk-of-bias guidelines of the Cochrane Collaboration, and the standardized mean differences (SMD) or mean differences (MD) for each outcome were compute. RESULTS: Among 20 eligible studies, including 682 participants were analyzed in this meta-analysis. The results of the meta-analysis demonstrated that balance training was effective in enhancing ankle function with self-functional scores (SMD = 1.02; 95% CI, 0.61 to 1.43; p < 0.00001; I2 = 72%) and variables associated with the ability of dynamic balance such as SEBT-A (MD = 5.88; 95% CI, 3.37 to 8.40; p < 0.00001; I2 = 84%), SEBT-PM (MD = 5.47; 95% CI, 3.40 to 7.54; p < 0.00001; I2 = 61%), and SEBT-PL (MD = 6.04; 95% CI, 3.30 to 8.79; p < 0.0001; I2 = 79%) of CAI patients. Meta-regression indicated that the intervention time might be the principal cause of heterogeneity (p = 0.046) in self-functional scores. In subgroup analyses of self-functional score across intervention types, among the intervention time, more than 20 min and less than 30 min had the most favorable effect (MD = 1.21, 95% CI: 0.96 to 1.46, p < 0.00001, I2 = 55%); among the intervention period, 4 weeks (MD = 0.84, 95% CI: 0.50 to 1.19, p < 0.00001, I2 = 78%) and 6 weeks (MD = 1.21, 95% CI: 0.91 to 1.51, p < 0.00001, I2 = 71%) had significant effects; among the intervention frequency, 3 times (MD = 1.14, 95% CI: 0.89 to 1.38), p < 0.00001, I2 = 57%) had significant effects. Secondly, in subgroup analyses of SEBT across intervention types, a 4-week and 6-week intervention with balance training 3 times a week for 20-30 min is the optimal combination of interventions to improve SEBT (dynamic balance) in patients with chronic ankle instability. CONCLUSION: Balance training proves beneficial for ankle function in patients with CAI. Intervention time constitutes a major factor influencing self-function in patients with CAI. It is recommended that the optimal dosage of balance training for CAI involves intervention three times a week, lasting for 20 to 30 min over a period of 4 to 6 weeks for superior rehabilitation.

Lipid droplets-specific fluorescent probe for wash-free imaging and in vivo diagnosis of non-alcoholic fatty liver disease.

Lipid droplets (LDs) dysfunction is closely associated with a multitude of diseases, including nonalcoholic fatty liver disease (NAFLD). Therefore, it is imperative to develop fluorescent probes that specifically target LDs for the early detection and diagnosis of NAFLD. In this study, a series of lipophilic fluorophores CZ1-CZ4 that feature a D-π-A configuration were designed and synthesized based on the carbazole and tricocyanofuran derivatives. The photophysical data revealed that all four probes exhibited large Stokes shifts (~ 120 nm) in high-polarity solvents (e.g., DMSO) and demonstrated enhanced fluorescence in solvents ranging from low-polarity (e.g., 1,4-Dioxane) to high-polarity. Notably, by utilizing probe CZ1, we could specifically visualize LDs and captured high-quality images, even eliminating the need for a time-consuming wash procedure. Moreover, CZ1 enabled monitoring of LDs dynamic changes in-real time within live cells, and importantly, it could be used to effectively distinguish normal and NAFLD tissues at both the organ and in vivo level. This exceptional property of probe CZ1 provides a practical tool for the diagnosis and intervention of NAFLD.

First Genomic Evidence of California Hare Coltivirus from Natural Populations of Ixodes persulcatus Ticks in Northeast China.

Background: California hare coltivirus (CHCV) was isolated in California in 1976 from a hare. Despite its long history, it remained unclear whether CHCV was exclusively distributed in California with limited host ranges. Main body: By next-generation sequencing (NGS), we obtained a complete sequence of CHCV from Ixodes persulcatus collected in 2019 in northeast China. An expanded epidemiological investigation was subsequently performed on ticks belonging to four species (Ix. persulcatus, Haemaphysalis concinna, Devmacentor silvarum, Haemaphysalis longicornis) collected in northeastern China by applying CHCV-specific RT-PCR and sequencing. CHCV RNA-positive results were found in 1.56% of the tick samples. Positive ticks were obtained in three of four sampled locations, with the highest rate observed in Inner Mongolia (2.69%), followed by Heilongjiang (1.94%) and Jilin provinces (0.55%). All positive results were derived from Ix. persulcatus ticks (2.33%), while no positive detection was found in the other tick species, even at the same location. Sequence analysis revealed that the current CHCV showed a high genetic identity (>80% amino acid identity) with the previously reported CHCV in all segments except segment seven (64.59% amino acid identity). Phylogenetic analysis based on RNA-dependent RNA polymerase (RdRp) amino acid sequences demonstrated that both the current and previously reported CHCV strains were grouped phylogenetically into the genus Coltivirus. Both CHCV strains formed a distinct clade, clustering with three human pathogenic coltiviruses (Colorado tick fever virus, Salmon River virus, and Eyach virus), and were distant from the other coltiviruses. Conclusions: We report the identification and characterization of CHCV for the first time in Ix. persulcatus ticks, expanding the currently known geographic scope, host, and genetic heterogeneity in CHCV.

USP7 Promotes TGF-ß1 Signaling by De-Ubiquitinating Smad2/Smad3 in Pulmonary Fibrosis.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a long-term, progressive, and irreversible pulmonary interstitial disease. The activation of Smad family member 2 (Smad2) and Smad3 transcription factors by transforming growth factor ß-1 (TGF-ß1) is a critical event in the pathogenesis of IPF. However, there is still a lack of understanding regarding the molecular mechanisms governing Smad2 and Smad3 proteins. Ubiquitin-specific protease 7 (USP7) is a deubiquitinase that plays a vital role in regulating protein stability within cells. However, its regulation of the TGF-ß signaling pathway and its significance in IPF remain undiscovered. This study aims to clarify the function of USP7 in the TGF-ß signaling pathway, while simultaneously exploring the specific molecular mechanisms involved. Additionally, this study seeks to evaluate the therapeutic potential of targeted USP7 inhibitors in IPF, thereby providing novel insights for the diagnosis and management of IPF. METHODS: We first detected the expression of USP7 in lung tissues of mice with Bleomycin (BLM)-induced pulmonary fibrosis and in Beas-2B cells treated with or without TGF-ß1 through Western blot analysis. Subsequently, we explored the influence of USP7 on fibrotic processes and the TGF-ß1 signaling pathway, utilizing in vitro and in vivo studies. Finally, we assessed the effectiveness of USP7-specific inhibitors in an IPF murine model. RESULTS: In the present study, USP7 was found to de-ubiquitinate Smad2 and Smad3, consequently increasing their stability and promoting the TGF-ß1-induced production of profibrotic proteins including α-smooth muscle actin (α-SMA) and fibronectin 1 (FN-1). Inhibition or knockdown of USP7 resulted in decreased levels of Smad2 and Smad3 proteins, leading to reduced expression of FN-1, Collagen Type I Alpha 1 Chain (Col1A1), and α-SMA induced by TGF-ß1 in human pulmonary epithelial cells. These findings demonstrate that overexpression of USP7 reduces Smad2/3 ubiquitination, whereas inhibition or knockdown of USP7 enhances their ubiquitination. USP7 is abundantly expressed in IPF lungs. The expressions of USP7, Smad2, and Smad3 were upregulated in bleomycin-induced lung injury. The USP7 inhibitor P22077 reduced the expression of FN-1 and type I collagen as well as Smad2/3 and collagen deposition in lung tissue in a model of pulmonary fibrosis induced by bleomycin. CONCLUSIONS: This study demonstrates that USP7 promotes TGF-ß1 signaling by stabilizing Smad2 and Smad3. The contribution of USP7 to the progression of IPF indicates it may be a viable treatment target.

Alterations in the gut microbiota community are associated with childhood obesity and precocious puberty.

OBJECTIVE: To explore the distribution and differences in the intestinal microbiota in girls with obesity-related precocious puberty and the relationship between intestinal microbiota and obesity-related precocious puberty. METHODS: 16 S rRNA gene amplicons from fecal samples from girls with precocious puberty and obesity-complicated precocious puberty and healthy children were sequenced to define microbial taxa. RESULTS: The α- and ß-diversity indices of the microbiome significantly differed among the three groups. At the phylum level, the proportions of Firmicutes, Actinobacteriota, Bacteroidota, Bacteria, Campylobacterota, and Acidobacteriota were different. At the genus level, there were differences in Bifidobacterium, Bacteroides, Anaerostipes, Fusicatenibacter, Klebsiella, Lachnospiraceae, ErysipelotrichaceaeUCG-003, Prevotella9, Ruminococcus gnavus group, and Lachnoclostridium. Additionally, Bifidobacterium, Anaerostipes, Bacteroides, Candidatus Microthrix, Eubacterium hallii group, Klebsiella, and Erysipelotrichaceae UCG-003 were identified as bacterial biomarkers by LEfSe. Furthermore, Sellimonas, Intestinibacter, Anaerostipes, Ruminococcus gnavus group, and Oscillibacter were identified as the differential biomarkers by random forest. A receiver operating characteristic (ROC) curve was used to evaluate the biomarkers with high predictive value for obesity-related precocious puberty. Spearman correlation analysis confirmed that Anaerostipes levels were negatively correlated with body weight, body mass index (BMI), bone age, luteinizing hormone, follicle-stimulating hormone, and estradiol. CONCLUSIONS: There was a significant correlation between obesity-associated precocious puberty and gut microbiota, especially the functional characteristics of the microbiome and its interactions, which can provide a theoretical basis for the clinical intervention of obesity and precocious puberty through the microbiome.

LCN2 regulates the gut microbiota and metabolic profile in mice infected with Mycobacterium bovis.

Infection with Mycobacterium bovis precipitates a spectrum of pathologies in bovines, notably necrotic pneumonia, mastitis, and arthritis, impinging upon the health and nutritional assimilation of these animals. A pivotal factor, lipocalin 2 (Lcn2), is responsive to microbial invasion, inflammatory processes, and tissue damage, the extent of which Lcn2 modulates the gut environment, however, remains unclear in response to M. bovis-induced alterations. To explore the role of Lcn2 in shaping the gut milieu of mice during a 5-week period post-M. bovis infection, Lcn2 knockout Lcn2-/- mice were scrutinized for changes in the gut microbiota and metabolomic profiles. Results showed that Lcn2-/- mice infected with M. bovis exhibited notable shifts in the operational taxonomic units (OTUs) of gut microbiota, alongside significant disparities in α and ß diversity. Concomitantly, a marked increase was observed during the 5-week period in the abundance of Akkermansia, Oscillospira, and Bacteroides, coupled with a substantial decrease in Ruminococcus within the microbiome of Lcn2 knockout mice. Notably, Akkermansia muciniphila was significantly enriched in the gut flora of Lcn2-/- mice. Furthermore, the absence of Lcn2 significantly altered the gut metabolomic landscape, evidenced by elevated levels of metabolites such as taurodeoxycholic acid, 10-undecenoic acid, azelaic acid, and dodecanedioic acid in Lcn2-/- mice. Our findings demonstrated that the lack of Lcn2 in the context of M. bovis infection profoundly affected the regulation of gut microbiota and metabolomic components, culminating in a transformed gut environment. Our results revealed that Lcn2 may regulate gut microbiota and metabolome components, changing the intestinal environment, thereby affecting the infection status of M. bovis. IMPORTANCE: Our study addresses the critical knowledge gap regarding the specific influence of lipocalin 2 (LCN2) in the context of Mycobacterium bovis infection, particularly focusing on its role in the gut environment. Utilizing LCN2 knockout (Lcn2-/-) mice, we meticulously assessed changes in the gut microbiota and metabolic components following M. bovis infection. Our findings reveal alterations in the gut microbial community, emphasizing the potentially crucial role of LCN2 in maintaining stability. Furthermore, we observed significant shifts in specific microbial communities, including the enrichment of Akkermansia muciniphila, known for its positive impact on intestinal health and immune regulation. The implications of our study extend beyond understanding the dynamics of the gut microbiome, offering insights into the potential therapeutic strategies for gut-related health conditions and microbial dysbiosis.

Mapping the distribution of sandflies and sandfly-associated pathogens in China.

BACKGROUND: Understanding and mapping the distribution of sandflies and sandfly-associated pathogens (SAPs) is crucial for guiding the surveillance and control effort. However, their distribution and the related risk burden in China remain poorly understood. METHODS: We mapped the distribution of sandflies and SAPs using literature data from 1940 to 2022. We also mapped the human visceral leishmaniasis (VL) cases using surveillance data from 2014 to 2018. The ecological drivers of 12 main sandfly species and VL were identified by applying machine learning, and their distribution and risk were predicted in three time periods (2021-2040, 2041-2060, and 2061-2080) under three scenarios of climate and socioeconomic changes. RESULTS: In the mainland of China, a total of 47 sandfly species have been reported, with the main 12 species classified into three clusters according to their ecological niches. Additionally, 6 SAPs have been identified, which include two protozoa, two bacteria, and two viruses. The incidence risk of different VL subtypes was closely associated with the distribution risk of specific vectors. The model predictions also revealed a substantial underestimation of the current sandfly distribution and VL risk. The predicted areas affected by the 12 major species of sandflies and the high-risk areas for VL were found to be 37.9-1121.0% and 136.6% larger, respectively, than the observed range in the areas. The future global changes were projected to decrease the risk of mountain-type zoonotic VL (MT-ZVL), but anthroponotic VL (AVL) and desert-type zoonotic VL (DT-ZVL) could remain stable or slightly increase. CONCLUSIONS: Current field observations underestimate the spatial distributions of main sandfly species and VL in China. More active surveillance and field investigations are needed where high risks are predicted, especially in areas where the future risk of VL is projected to remain high or increase.

Novel strategy for Poxviridae prevention: Thermostable combined subunit vaccine patch with intense immune response.

Poxviruses gained international attention due to the sharp rise in monkeypox cases in recent years, highlighting the urgent need for the development of a secure and reliable vaccine. This study involved the development of an innovative combined subunit vaccine (CSV) targeting poxviruses, with lumpy skin disease virus (LSDV) serving as the model virus. To this end, the potential sites for poxvirus vaccines were fully evaluated to develop and purify four recombinant proteins. These proteins were then successfully delivered to the dermis in a mouse model by utilizing dissolvable microneedle patches (DMPs). This approach simplified the vaccination procedure and significantly mitigated the associated risk. CSV-loaded DMPs contained four recombinant proteins and a novel adjuvant, CpG, which allowed DMPs to elicit the same intensity of humoral and cellular immunity as subcutaneous injection. Following immunization with SC and DMP, the mice exhibited notable levels of neutralizing antibodies, albeit at a low concentration. It is noteworthy that the CSV loaded into DMPs remained stable for at least 4 months at room temperature, effectively addressing the storage and transportation challenges. Based on the study findings, CSV-loaded DMPs are expected to be utilized worldwide as an innovative technique for poxvirus inoculation, especially in underdeveloped regions. This novel strategy is crucial for the development of future poxvirus vaccines.

Cellular pharmacokinetic of methotrexate and its modulation by folylpolyglutamate synthetase and γ-glutamyl hydrolase in tumor cells.

BACKGROUND AND PURPOSE: Clinical studies showed that prolonged infusion of methotrexate (MTX) leads to more severe adverse reactions than short infusion of MTX at the same dose. We hypothesized that it is the saturation of folate polyglutamate synthetase (FPGS) at high MTX concentration that limits the intracellular synthesis rate of methotrexate polyglutamate (MTX-PG). Due to a similar accumulation rate, a longer infusion duration may increase the concentration of MTX-PG and, result in more serious adverse reactions. In this study, we validated this hypothesis. EXPERIMENTAL APPROACH: A549, BEL-7402 and MHCC97H cell lines were treated with MTX at gradient concentrations. Liquid chromatograph-mass spectrometer (UPLC-MS/MS) was used to quantify the intracellular concentration of MTX-PG and the abundance of FPGS and γ-glutamyl hydrolase (GGH). High quality data were used to fit the cell pharmacokinetic model. KEY RESULTS: Both cell growth inhibition rate and intracellular MTX-PG concentration showed a nonlinear relationship with MTX concentration. The parameter Vmax in the model, which represents the synthesis rate of MTX-PG, showed a strong correlation with the abundance of intracellular FPGS. CONCLUSION AND IMPLICATIONS: According to the model fitting results, it was confirmed that the abundance of FPGS is a decisive factor limiting the synthesis rate of MTX-PG. The proposed hypothesis was verified in this study. In addition, based on the intracellular metabolism, a reasonable explanation was provided for the correlation between the severity of adverse reactions of MTX and infusion time. This study provides a new strategy for the individualized treatment and prediction of efficacy/side effects of MTX.

Rapid and sensitive detection of glucocorticoids using engineered magnetosomes functionalized protein A conjugated broad-spectrum monoclonal antibody.

Engineered bacterial magnetic nanoparticles (BMPs) fused with protein A (BMP-PA) can bind antibodies, creating immunomagnetic beads that offer an attractive tool for targets screening. In the study, BMP-PA-IgG was formed by attaching broad-spectrum monoclonal antibodies against glucocorticoids (GCs) to BMP-PA. Immunomagnetic assay was developed for analysis of GCs, using the BMP-PA-IgG and hydrocortisone-horseradish peroxidase. The developed assay exhibited broad specificity for GCs, including hydrocortisone (HCS), betamethasone (BMS), dexamethasone (DMS), prednisolone (PNS), beclomethasone (BCMS), cortisone (CS), 6-α-methylprednisone (6-α-MPNS), and fludrocortisone acetate (HFCS), with half inhibitory concentrations (IC50) ranging from 0.88 to 6.57 ng/mL. The proposed assay showed average recoveries of HCS and DMS ranging from 75.6% to 105.2% in chicken and pork samples, which were correlated well with those obtained by LC-MS/MS. This study indicated that the integration of engineered immunomagnetic beads into immunoassay systems offer possibilities for the sensitive and selective detection of GCs.

Bacteriophage defends murine gut from Escherichia coli invasion via mucosal adherence.

Bacteriophage are sophisticated cellular parasites that can not only parasitize bacteria but are increasingly recognized for their direct interactions with mammalian hosts. Phage adherence to mucus is known to mediate enhanced antimicrobial effects in vitro. However, little is known about the therapeutic efficacy of mucus-adherent phages in vivo. Here, using a combination of in vitro gastrointestinal cell lines, a gut-on-a-chip microfluidic model, and an in vivo murine gut model, we demonstrated that a E. coli phage, øPNJ-6, provided enhanced gastrointestinal persistence and antimicrobial effects. øPNJ-6 bound fucose residues, of the gut secreted glycoprotein MUC2, through domain 1 of its Hoc protein, which led to increased intestinal mucus production that was suggestive of a positive feedback loop mediated by the mucus-adherent phage. These findings extend the Bacteriophage Adherence to Mucus model into phage therapy, demonstrating that øPNJ-6 displays enhanced persistence within the murine gut, leading to targeted depletion of intestinal pathogenic bacteria.

Distinct characteristics of the gut virome in patients with osteoarthritis and gouty arthritis.

BACKGROUND/PURPOSE(S): The gut microbiota and its metabolites play crucial roles in pathogenesis of arthritis, highlighting gut microbiota as a promising avenue for modulating autoimmunity. However, the characterization of the gut virome in arthritis patients, including osteoarthritis (OA) and gouty arthritis (GA), requires further investigation. METHODS: We employed virus-like particle (VLP)-based metagenomic sequencing to analyze gut viral community in 20 OA patients, 26 GA patients, and 31 healthy controls, encompassing a total of 77 fecal samples. RESULTS: Our analysis generated 6819 vOTUs, with a considerable proportion of viral genomes differing from existing catalogs. The gut virome in OA and GA patients differed significantly from healthy controls, showing variations in diversity and viral family abundances. We identified 157 OA-associated and 94 GA-associated vOTUs, achieving high accuracy in patient-control discrimination with random forest models. OA-associated viruses were predicted to infect pro-inflammatory bacteria or bacteria associated with immunoglobulin A production, while GA-associated viruses were linked to Bacteroidaceae or Lachnospiraceae phages. Furthermore, several viral functional orthologs displayed significant differences in frequency between OA-enriched and GA-enriched vOTUs, suggesting potential functional roles of these viruses. Additionally, we trained classification models based on gut viral signatures to effectively discriminate OA or GA patients from healthy controls, yielding AUC values up to 0.97, indicating the clinical utility of the gut virome in diagnosing OA or GA. CONCLUSION: Our study highlights distinctive alterations in viral diversity and taxonomy within gut virome of OA and GA patients, offering insights into arthritis etiology and potential treatment and prevention strategies.