Microfluidics-Derived Microparticles with Prebiotics and Probiotics for Enhanced In Situ Colonization and Immunoregulation of Colitis.

Oral administration of probiotics orchestrates the balance between intestinal microbes and the immune response. However, effective delivery and in situ colonization are limited by the harsh environment of the gastrointestinal tract. Herein, we provide a microfluidics-derived encapsulation strategy to address this problem. A novel synergistic delivery system composed of EcN Nissle 1917 and prebiotics, including alginate sodium and inulin gel, for treating inflammatory bowel disease and colitis-associated colorectal cancer is proposed. We demonstrated that EcN@AN microparticles yielded promising gastrointestinal resistance for on-demand probiotic delivery and colon-retentive capability. EcN@AN microparticles efficiently ameliorated intestinal inflammation and modulated the gut microbiome in experimental colitis. Moreover, the prebiotic composition of EcN@AN enhanced the fermentation of relative short-chain fatty acid metabolites, a kind of postbiotics, to exert anti-inflammatory and tumor-suppressive effects in murine models. This microfluidcis-based approach for the coordinated delivery of probiotics and prebiotics may have broad implications for gastrointestinal bacteriotherapy applications.

Air Pollutants and Mortality Risk in Patients with Aortic Dissection: Evidence from a Clinical Cohort, Single-Cell Sequencing, and Proteomics.

We aimed to evaluate the association between air pollutants and mortality risk in patients with acute aortic dissection (AAD) in a longitudinal cohort and to explore the potential mechanisms of adverse prognosis induced by fine particulate matter (PM2.5). Air pollutants data, including PM2.5, PM10.0, nitrogen dioxide (NO2), carbon monoxide (CO), sulfur dioxide (SO2), and ozone (O3), were collected from official monitoring stations, and multivariable Cox regression models were applied. Single-cell sequencing and proteomics of aortic tissue were conducted to explore the potential mechanisms. In total, 1,267 patients with AAD were included. Exposure to higher concentrations of air pollutants was independently associated with an increased mortality risk. The high-PM2.5 group carried approximately 2 times increased mortality risk. There were linear associations of PM10, NO2, CO, and SO2 exposures with long-term mortality risk. Single-cell sequencing revealed an increase in mast cells in aortic tissue in the high-PM2.5 exposure group. Enrichment analysis of the differentially expressed genes identified the inflammatory response as one of the main pathways, with IL-17 and TNF signaling pathways being among the top pathways. Analysis of proteomics also identified these pathways. This study suggests that exposure to higher PM2.5, PM10, NO2, CO, and SO2 are associated with increased mortality risk in patients with AAD. PM2.5-related activation and degranulation of mast cells may be involved in this process.

Clinical characteristics and prognostic analysis of postoperative recurrence or metastasis of low-risk gastrointestinal stromal tumors.

BACKGROUND: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the digestive tract. This study aimed to investigate the clinical characteristics and prognosis of postoperative recurrence or metastasis in patients with low-risk stromal tumors, in order to take individualized postoperative management and treatment for patients with low-risk GISTs with relatively high recurrence. METHODS: We retrospectively analyzed the clinicopathological and follow-up data of patients with GISTs who underwent surgical resection in Nanjing Drum Tower Hospital from March 2010 to December 2021. A total of 282 patients with low-risk GISTs were included, none of whom were treated with imatinib. Univariate and multivariate Cox analysis and survival curves were used to explore the relationship between clinical features and recurrence or metastasis in patients with low-risk GISTs. RESULTS: Of the 282 patients with low-risk GISTs who met inclusion criteria, 14 (4.96%) had recurrence or metastasis. There was a correlation between tumor size, primary site, resection type, Ki67 index, neutrophil lymphocyte ratio (NLR) and CD34 expression and postoperative recurrence or metastasis of GISTs (P < 0.05). Subsequently, multifactorial analysis showed that tumor primary site, tumor size, and Ki67 index were independent risk factors affecting postoperative recurrent or metastasis in patients with low-risk GISTs (P < 0.05). Ultimately, According to Kaplan-Meier analysis, non-gastric primary tumors, larger tumors, and high Ki67 index were significantly associated with poor progression-free survival ( PFS ). CONCLUSIONS: Tumor location, tumor size and Ki-67 were independent risk factors for postoperative recurrence and metastasis in patients with low-risk GISTs. Based on the 2008 modified NIH recurrence risk grading system, combined with the above three factors, it can be used to evaluate the prognosis of patients with low-risk GISTs and provide personalized postoperative review and follow-up management recommendations.

Exploring the Effect of Milk Fat on Fermented Milk Flavor Based on Gas Chromatography-Ion Mobility Spectrometry (GC-IMS) and Multivariate Statistical Analysis.

Milk fat is a premium nutritional health product, yet there is a lack of high-fat dairy products for daily consumption in the current market. This study investigated the influence of different milk fat contents on the physicochemical and textural properties of fermented milk. The research revealed that an increase in milkfat content significantly improved the water-holding capacity, syneresis, color, hardness, springiness, gumminess, and chewiness of fermented milk, while showing minimal changes in pH and total titratable acidity. Response surface analysis indicated that fermented milk with 25% milk fat, 2.5% inoculum, a fermentation time of 16 h, and a fermentation temperature of 30 °C exhibited the highest overall acceptability. Using GC-IMS technology, 36 volatile compounds were identified, with an increase in milk fat content leading to elevated levels of ketone compounds, and 14 compounds were defined as key aroma compounds (ROAV > 1). Electronic nose distinguished samples with different milk fat contents. The results demonstrate that an increase in milk fat content enhances the physicochemical and flavor attributes of fermented milk. This work provides theoretical references for the production and development of high-fat fermented milk.

[Research on mode adjustment control strategy of upper limb rehabilitation robot based on fuzzy recognition of interaction force].

In the process of robot-assisted training for upper limb rehabilitation, a passive training strategy is usually used for stroke patients with flaccid paralysis. In order to stimulate the patient's active rehabilitation willingness, the rehabilitation therapist will use the robot-assisted training strategy for patients who gradually have the ability to generate active force. This study proposed a motor function assessment technology for human upper-limb based on fuzzy recognition on interaction force and human-robot interaction control strategy based on assistance-as-needed. A passive training mode based on the calculated torque controller and an assisted training mode combined with the potential energy field were designed, and then the interactive force information collected by the three-dimensional force sensor during the training process was imported into the fuzzy inference system, the degree of active participation σ was proposed, and the corresponding assisted strategy algorithms were designed to realize the adaptive adjustment of the two modes. The significant correlation between the degree of active participation σ and the surface electromyography signals (sEMG) was found through the experiments, and the method had a shorter response time compared to a control strategy that only adjusted the mode through the magnitude of interaction force, making the robot safer during the training process.

2022 CMICS Expert Consensus on the Management of Isolated Tricuspid Regurgitation after Left-Sided Valve Surgery.

Tricuspid regurgitation (TR) may occur late after left-sided valve surgery (LSVS). Isolated tricuspid regurgitation after left-sided valve surgery (iTR-LSVS) refers to isolated TR without significant lesions in the mitral and/or aortic position late after mitral and/or aortic replacement or repair. Severe TR has a negative impact on long-term prognosis and requires surgical or transcatheter treatment. However, there is no clear recommendation on when and how intervention should be performed for patients with iTR-LSVS in the current guidelines for the management of valvular heart disease. The historically high operative mortality may be reduced by current minimally invasive techniques and transcatheter therapy. To further understand iTR-LSVS, standardize the treatment, improve the prognosis, and promote the collaboration, the Chinese Minimally Invasive Cardiovascular Surgery Committee (CMICS) wrote this expert consensus on the management of iTR-LSVS from the aspects of etiology, preoperative evaluation, indications for intervention, surgical treatment, transcatheter therapy, and postoperative management.

A case report of adolescent myofibrillar myopathy due to a de novo R406W pathogenic variant in desmin with symptoms of "hypertrophic cardiomyopathy".

Objective: Myofibrillar myopathies (MFM) are a group of sporadic and inherited progressive skeletal muscle disorders that can lead to physical disability and premature death. To date, pathogenic variants in different genes are associated with MFM. MFM induced by variants in the Desmin (DES) gene is the most common subtype of MFM. Case presentation: A 15-year-old boy with MFM was described, whose symptoms first presented as cardiac symptoms. Enlarged right and left atria, thickened ventricular septal (IVS) and mild mitral (MR) and tricuspid regurgitation (TR) in the echocardiography were found. Atrial fibrillation, intermittent atrioventricular (AV) block, ST-T changes in the dynamic electrocardiogram (ECG) were shown. Mild myopathic changes in the electromyographic exam were detected. Ultrastructural analysis found slight Z-line changes and a few small myolysis lesions, but no abnormal inclusion bodies. Genetic testing detected a heterozygous missense variant (c.1216C > T) of DES, and 2 rare variants: TNNI3K (c.1102C > G) and PRDM16 (c.3074G > A). The patient's parents didn't show skeletal and cardiac muscle disorders. DNA sequencing analysis showed no variant of DES was carried by them. Thus, we detected a case of MFM caused by de novo DES variant c.1216C > T/p.Arg406Trp with predominantly myocardial alterations.

A case of hypertrophic cardiomyopathy with previous aortic valve replacement.

We describe a 45-year-old patient who was diagnosed with hypertrophic obstructive cardiomyopathy (HOCM) after the aortic valve replacement surgery. Enlarged left atria, thickened ventricular septum, left ventricular outflow tract stenosis, moderate mitral regurgitation and mild tricuspid regurgitation in the echocardiography were found. We offered the patient the new minimally invasive treatment modality: percutaneous intra-myocardial septal radiofrequency ablation (PIMSRA). We demonstrate the safety and efficacy with pictures. One month after surgery, the patient recovered well with improved symptoms of chest tightness, and no LVOT obstruction or arrhythmia.

Inter-arm blood pressure difference in post-stroke patients with hemiparesis.

The aim of this study was to investigate that inter-arm blood pressure (BP) difference (IAD) and reference arm in 420 post-stroke patients with hemiparesis. Synchronous bilateral-arm BP was measured with two automatic BP devices, and the systolic BP difference of ≥10 mm Hg was recorded as increased sIAD. The arm with higher systolic BP (SBP) was assigned as the reference arm. Our results showed that the prevalence of sIAD was 18.1% in the total group. The paretic arms had similar mean SBP levels (133.6±18.4 vs. 133.8±18.4 mm Hg, NS) and DBP (77.8±11.5 vs. 77.2±10.9 mm Hg, NS) as compared with the unaffected arms. The detection rate of hypertension or uncontrolled hypertension on the SBP values of the reference arm was higher than that on the unaffected arm (41.8% vs. 36.3%). It is concluded that in the post-stroke patients with hemiparesis in the rehabilitation period, the prevalence of sIAD ≥10 mmHg was relatively higher, and using the unaffected arm, rather than the unaffected arm, for BP measurement could induce correctly detection of hypertension.

Early and Mid-Term Outcomes of Using the Chimney Technique in Redo Mitral Valve Replacement in Patients with a Small Mitral Annulus.

The outcomes of redo mitral valve replacement (Re-MVR) in a small mitral annulus with the use of the chimney technique are not well documented. The purpose of this study is to present our early experience with this group of patients, illustrating the periop-erative complications and mortality outcomes. From 2019 to 2020, 77 consecutive patients underwent Re-MVR with the use of the chimney technique because of a small mitral annulus. To evaluate heart structural integrity and clinical outcomes, postoperative clinical data and echocardiograms were examined. The mean age was 56.7 ± 15.98 years. All patients underwent mitral valve surgery, of which 62 were mitral valve replacements, 7 mitral valve repairs, and 8 double valve replacements. The preoperative mitral valve mean gradient was 18.07 ± 9.40 mmHg, and the postoperative mitral prosthesis size was 28.51 ± 1.22 mm. The median increment of mitral size enlargement was 4 (0, 6) valve sizes. The mean mitral gradient coming out of the operating room was 10.34 ± 2.12 mmHg, and at the follow-up echocardiogram performed at 3 years after the procedure, it was 10.36 ± 1.70 mmHg. One-year survival was 93.3%, while the 4-year survival rate was 89.3%, with no reoperation. The use of the chimney technique in small mitral valve re-mitral valve replacement results in larger valve sizes. Moreover, the mean gradients over the mitral valve are acceptable both intraoperatively and over time.

Signs of front-line healthcare professionals' information anxiety during the COVID-19 pandemic: Grounded theory study in a Wuhan hospital.

AIM: Being front-line healthcare professionals is associated with possible severe information anxiety during the COVID-19 pandemic. Investigating signs of information anxiety is the first and key step of its targeted medical intervention. This study aims to explore the signs of front-line healthcare professionals' information anxiety during the COVID-19 pandemic. DESIGN: This study is qualitative research. Grounded theory was used to classify information anxiety signs of front-line healthcare professionals. METHODS: Twenty-four front-line healthcare professionals from a general hospital with over 5000 beds in Wuhan were recruited to participate in semi-structured interviews. According to the frequency and frequency variation of signs appearing in interviews, the trends of signs during the virus encounter, lockdown, flattening and second wave were compared. Based on the interviews, those signs that were conceptually related to each other were extracted to construct a conceptual model. RESULTS: Psychological signs (emotion, worry, doubt, caution, hope), physical signs (insomnia, inattention, memory loss, appetite decreased) and behavioural signs (panic buying of goods, be at a loss, pay attention to relevant information, change habits) could be generalized from 13 subcategories of information anxiety signs. Psychological signs were the most in every period of the pandemic. Furthermore, psychological signs decreased significantly during lockdown, while behavioural and physical signs increased. Finally, severe psychological and behavioural signs were associated with physical signs.

Molecular basis of TMPRSS2 recognition by Paeniclostridium sordellii hemorrhagic toxin.

Hemorrhagic toxin (TcsH) is a major virulence factor produced by Paeniclostridium sordellii, which is a non-negligible threat to women undergoing childbirth or abortions. Recently, Transmembrane Serine Protease 2 (TMPRSS2) was identified as a host receptor of TcsH. Here, we show the cryo-EM structures of the TcsH-TMPRSS2 complex and uncover that TcsH binds to the serine protease domain (SPD) of TMPRSS2 through the CROP unit-VI. This receptor binding mode is unique among LCTs. Five top surface loops of TMPRSS2SPD, which also determine the protease substrate specificity, constitute the structural determinants recognized by TcsH. The binding of TcsH inhibits the proteolytic activity of TMPRSS2, whereas its implication in disease manifestations remains unclear. We further show that mutations selectively disrupting TMPRSS2-binding reduce TcsH toxicity in the intestinal epithelium of the female mice. These findings together shed light on the distinct molecular basis of TcsH-TMPRSS2 interactions, which expands our knowledge of host recognition mechanisms employed by LCTs and provides novel targets for developing therapeutics against P. sordellii infections.

Pore structure characterization and its influence on aqueous phase trapping damage in tight gas sandstone reservoirs.

Aqueous phase trapping (APT), which is one of the most prominent damages, seriously restricts the natural gas production in tight gas sandstone with low permeability. Pore size and microscopic pore structures are the most important factors to determine the water blocking damage. In this paper, 9 core samples from tight gas sandstone with various physical properties were employed, and the pore size distribution (PSD) of the core samples were investigated by high pressure mercury intrusion tests (HPMI). Results showed that the porosity of core samples ranges from 5.68% to 13.7%, and the permeability ranges from 0.00456 to 7.86 mD, which is a typical tight reservoir with strong heterogeneity. According to the HPMI capillary curve, the cores can be divided into two types: Type I and Type II, and the pore sizes of type I are larger than that of type II. Fractal distributions were obtained using HPMI data to further determine the pore structure characteristics of tight reservoirs. The pore structures of tight sandstones display the multifractal fractal feature: D1 corresponding to macro-pores, and D2 corresponding to fractal dimension of micro-pores. Furthermore, APT damage was determined by the permeability recovery ratios (Kr) after gas flooding tests. The correlation of Kr and PSD and fractal dimensions were jointly analyzed in tight gas sandstone. Although positive correlations between pore size parameters and the permeability recovery ratios were observed with relatively weak correlations, for those core samples with very close permeability, pore size parameters (both permeability and PSD) is inadequate in clarifying this damage. The fractal dimension can well describe the complexity and heterogeneity of flow channels in pores, which can become the determining factor to distinguish the flow capacity of tight sandstone. The D2 for samples of type I and type II exhibited a good negative relation with Kr with a correlation coefficient of 0.9878 and 0.7723, respectively. The significance of this finding is that for tight gas sandstone, fractal dimensions, especially the small pore fractal dimension (D2), can be used to predict the possible APT damage very well.

Fight or flee, a vital choice for Clostridioides difficile.

Clostridioides difficile is a leading cause of healthcare-associated infections, causing billions of economic losses every year. Its symptoms range from mild diarrhea to life-threatening damage to the colon. Transmission and recurrence of C. difficile infection (CDI) are mediated by the metabolically dormant spores, while the virulence of C. difficile is mainly due to the two large clostridial toxins, TcdA and TcdB. Producing toxins or forming spores are two different strategies for C. difficile to cope with harsh environmental conditions. It is of great significance to understand the molecular mechanisms for C. difficile to skew to either of the cellular processes. Here, we summarize the current understanding of the regulation and connections between toxin production and sporulation in C. difficile and further discuss the potential solutions for yet-to-be-answered questions.

Multi-scale 3D-CRU for EEG emotion recognition.

In this paper, we propose a novel multi-scale 3D-CRU model, with the goal of extracting more discriminative emotion feature from EEG signals. By concurrently exploiting the relative electrode locations and different frequency subbands of EEG signals, a three-dimensional feature representation is reconstructed wherein the Delta (δ) frequency pattern is included. We employ a multi-scale approach, termed 3D-CRU, to concurrently extract frequency and spatial features at varying levels of granularity within each time segment. In the proposed 3D-CRU, we introduce a multi-scale 3D Convolutional Neural Network (3D-CNN) to effectively capture discriminative information embedded within the 3D feature representation. To model the temporal dynamics across consecutive time segments, we incorporate a Gated Recurrent Unit (GRU) module to extract temporal representations from the time series of combined frequency-spatial features. Ultimately, the 3D-CRU model yields a global feature representation, encompassing comprehensive information across time, frequency, and spatial domains. Numerous experimental assessments conducted on publicly available DEAP and SEED databases provide empirical evidence supporting the enhanced performance of our proposed model in the domain of emotion recognition. These findings underscore the efficacy of the features extracted by the proposed multi-scale 3D-GRU model, particularly with the incorporation of the Delta (δ) frequency pattern. Specifically, on the DEAP dataset, the accuracy of Valence and Arousal are 93.12% and 94.31%, respectively, while on the SEED dataset, the accuracy is 92.25%.

Influence of Supercritical CO2 Fracturing Fluid on the Permeability of Shale Reservoir and Mechanism Analysis.

Reduction of the reservoir permeability during supercritical CO2 fracturing caused significant reservoir damage, which directly affects the crude oil recovery rate. This investigation explored a novel thickener for supercritical CO2 fracturing fluid, this CO2 thickener not only effectively improves the viscosity and rheological properties of CO2 fracturing fluid but also contributes to reduce reservoir damage and improve permeability. The research results indicated that the synthesized CO2 thickener (3 wt %) can increase the apparent viscosity of supercritical CO2 fracturing fluid to 7 mPa·s, and a 9% matrix permeability damage rate and a 0.5 mD permeability decrease value are shown in a 3% CO2 fracturing fluid. However, 3 wt % of commercial CO2 thickener only increases the apparent viscosity of supercritical CO2 fracturing fluid to 3 mPa·s, while the reservoir damage rate increases to 13%. Two thickeners exhibit completely different damage capabilities to the reservoir, and the synthesized CO2 thickener shows excellent characteristics of reducing reservoir permeability and is also conducive to protecting shale reservoirs. Furthermore, supercritical CO2 fracturing fluid containing synthetic thickeners has better temperature and shear resistance compared with commercial thickeners. This may be because the synthesized thickener and the micro grid formed by supercritical CO2 reduce the adsorption in shale crevices, but a large amount of commercial thickeners can adsorb on the surface of shale.

miR-129-2-3p inhibits colon cancer cell proliferation by down-regulating the expression of BZW1.

BACKGROUND AND STUDY AIMS: MicroRNA (miRNA) is involved in diverse biological and physiological processes of tumors. Dysregulation of miRNA will induce a series of human diseases. miR-129-2-3p has vital effects in the pathogenesis of various tumors. However, the regulatory function of miR-129-2-3p in colon cancer remains to be clarified. This study investigated the role of miR-129-2-3p targeting BZW1 in proliferation, apoptosis, migration, and invasion of colon cancer. PATIENTS AND METHODS: Here, RT-qPCR was applied to measure the miR-129-2-3p levels in colon cancer tissues. The predicted targets of miR-129-2-3p were identified by bioinformatics and verified using luciferase reporter assay. The effects of miR-129-2-3p on colon cancer were detected by CCK-8, colony formation, transwell chamber test, wound healing, and flow cytometry assays. Finally, the influence of miR-129-2-3p on tumor growth was studied. Nude mice were xenografted with transfected Lovo cells by subcutaneous injection of 5 × 105 cells in 100 µl. HE staining and TUNEL were used to assess metastasis ability. RESULTS: miR-129-2-3p level in colon cancer tissue was significantly reduced. Furthermore, it was verified that BZW1 was a target of miR-129-2-3p, and its expression in colon cancer cells was inhibited by miR-129-2-3p. Additionally, miR-129-2-3p inhibited colon cancer cell proliferation, colony formation, mobility ability and tumor growth, and promoted cell apoptosis by targeting BZW1. miR-129-2-3p overexpression in tumor xenografts in vivo decreased BZW1 expression, and suppressed tumor growth. CONCLUSION: Collectively, these findings indicated that miR-129-2-3p exerts a suppressive role in colon cancer cells by directly targeting BZW1, and may have significant therapeutic implications for patients with colon cancer.

The TTN p. Tyr4418Ter mutation causes cardiomyopathy in human and mice.

OBJECTIVE: To generate a mouse model carrying TTNtv Y4370* simulating the newly discovered human heterozygous nonsense TTNtv c.13254T>G (p.Tyr4418Ter) to supplement and improve the functional evidence of pathogenic mutation TTNtv c.13254T>G on the pathogenic type of dilated cardiomyopathy. METHODS: We generated 4 mice carrying TTNtv p. Y4370* through CRISPR/Cas-mediated genome engineering. Monthly serological detection, bimonthly echocardiography, and histology evaluation were carried out to observe and compare alterations of cardiac structure and function between 4 TTN+/- mice and 4 wild-type (WT) mice. RESULTS: For the two-month-old TTN+/- mice, serum glutamic-oxalacetic transaminase (AST), lactic dehydrogenase (LDH), and creatine kinase (CK) were significantly increased, the diastolic Left Ventricular Systolic Anterior Wall (LVAW), and the LV mass markedly rose, with the left ventricular volume displaying an increasing trend and Ejection Fraction (EF) and Fractional Shortening (FS) showing a decreasing trend. Besides, the histological evaluation showed that cardiac fibrosis level and positive rate of cardiac mast cell of TTN+/- mice were obviously increased compared with WT mice. CONCLUSIONS: TTNtv Y4370* could lead to cardiac structure and function alterations in mice, supplementing the evidence of TTNtv c.13254T>G pathogenicity in human.

Identification of a hemorrhagic determinant in Clostridioides difficile TcdA and Paeniclostridium sordellii TcsH.

Paeniclostridium sordellii hemorrhagic toxin (TcsH) and Clostridioides difficile toxin A (TcdA) are two major members of the large clostridial toxin (LCT) family. These two toxins share ~87% similarity and are known to cause severe hemorrhagic pathology in animals. Yet, the pathogenesis of their hemorrhagic toxicity has been mysterious for decades. Here, we examined the liver injury after systemic exposure to different LCTs and found that only TcsH and TcdA induce overt hepatic hemorrhage. By investigating the chimeric and truncated toxins, we demonstrated that the enzymatic domain of TcsH alone is not sufficient to determine its potent hepatic hemorrhagic toxicity in mice. Likewise, the combined repetitive oligopeptide (CROP) domain of TcsH/TcdA alone also failed to explain their strong hemorrhagic activity in mice. Lastly, we showed that disrupting the first two short repeats of CROPs in TcsH and TcdA impaired hemorrhagic toxicity without causing overt changes in cytotoxicity and lethality. These findings lead to a deeper understanding of toxin-induced hemorrhage and the pathogenesis of LCTs and could be insightful in developing therapeutic avenues against clostridial infections. IMPORTANCE: Paeniclostridium sordellii and Clostridioides difficile infections often cause hemorrhage in the affected tissues and organs, which is mainly attributed to their hemorrhagic toxins, TcsH and TcdA. In this study, we demonstrate that TcsH and TcdA, but not other related toxins. including Clostridioides difficile toxin B and TcsL, induce severe hepatic hemorrhage in mice. We further determine that a small region in TcsH and TcdA is critical for the hemorrhagic toxicity but not cytotoxicity or lethality of these toxins. Based on these results, we propose that the hemorrhagic toxicity of TcsH and TcdA is due to an uncharacterized mechanism, such as the presence of an unknown receptor, and future studies to identify the interactive host factors are warranted.

Revealing the Development Patterns of the Mandibular Glands of Apis mellifera carnica Based on Transcriptomics and Morphology.

The mandibular gland in worker bees synthesizes and secretes the organic acids present in royal jelly, and its development directly affects yield and quality. Therefore, we aimed to analyze the differences in morphology and gene expression in the mandibular glands of Apis mellifera carnica worker bees of different ages (3, 6, 9, 12, and 16 d). We dissected their mandibular glands and performed morphological and transcriptomic analyses to investigate the development of the mandibular gland and the molecular regulatory mechanisms involved in royal jelly secretion. Microscopy revealed that mandibular gland development is likely completed in the early stages. There were no significant differences in the structural morphology or organelles involved in the secretion of royal jelly at different ages. Transcriptomics revealed a total of 1554 differentially expressed genes, which were mainly involved in fat metabolism, lipid transport, and energy metabolism. The extracellular matrix-receptor interaction pathway was significantly enriched and contributed to the royal jelly secretion process. These results elucidate the genetic basis of the role of the mandibular gland in royal jelly secretion in A. mellifera and provide a reference for the genetic improvement of bees with high royal jelly production in the future.