Photoresponsive Hydrogel Dressing Containing Nanoparticles with Excellent Synergetic Photodynamic, Photothermal, and Chemodynamic Therapies for Effective Infected Wound Healing.

Bacterial resistance to antibiotics can negatively affect the treatment of infected skin wounds. The combination of synergistic antibacterial therapies with photodynamic, photothermal, and chemodynamic therapies has been recognized as one of the most promising approaches. In this study, we have developed MSN@Ce6@MnO2-CS/Ag (MCMA) nanoparticles to serve as powerful antibacterial agents when exposed to both 660 nm visible light and 808 nm near-infrared (NIR) light. Through dual-light irradiation, MCMA can induce hyperthermia and generate reactive oxygen species (ROS), leading to a remarkable enhancement in photothermal antibacterial effects and accelerating wound healing. It has a peroxidase-like catalytic activity and promotes the generation of hydroxyl radicals (·OH) by catalyzing the decomposition of H2O2. In vitro antibacterial experiments demonstrated the excellent antibacterial activity of MCMA. The antibacterial efficacy of MCMA at a concentration of 250 µg ml-1 was found to be 99.6 and 100% toward Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli, respectively, under irradiation with an 808 and 660 nm laser. The results of the animal experiments demonstrated that MCMA can effectively accelerate wound healing through wound ulceration inhabitation. These findings substantiate the assertion that synthetic MCMA represents an efficacious strategy for bacterial inhibition and wound healing.

Microbial diversity in earthen site of exhibition Hall of pit no. 1 at the terracotta warriors Museum in Emperor Qinshihuang's mausoleum site museum and its correlation with environmental factors.

Introduction: Earthen sites are essential cultural relic resources, and site museums are a fundamental component of China's cultural heritage protection. The mausoleum of the Qin Shi Huang Emperor is one of the largest, most peculiar, and richest imperial tombs in the world. The exhibition hall of the burial pit No. 1 of the Terra Cotta Warriors is the earliest exhibition hall built and opened to the public. However, after years of excavation and open exhibitions, the earthen site of the Emperor Qinshihuang's Mausoleum Site Museum has deteriorated to varying degrees due to changes in the modern environment. There is an urgent need to control microbial diseases and protect the earthen site. Methods: We analyzed the physical and chemical properties and bioindicators of the collected soil samples. We also established a metagenomic library and conducted a correlation analysis between microbial community composition and environmental factors. Cultivable fungi obtained from air and soil samples were identified, and allicin volatile gas fungistasis test was conducted. Result: Research has found that four different areas of the exhibition hall have different types of microbial diseases owing to their different environments. The main pathogenic fungi in earthen site may lead to potential microbial diseases that affect important cultural relics such as the Terra Cotta Warriors. Penicillium, Aspergillus and Talaromyces showed relatively specific growth in relation to environmental factors and showed a better raw growth advantage.Allicin gas had a inhibitory effect on 12 types of fungi, therefore allicin gas had a potent inhibitory effect on the growth of the most culturable fungal hyphae. Discussion: This study provides basic data for the study of microbial diversity in the exhibition hall of Pit No. 1 at the Terracotta Warriors Museum in Emperor Qinshihuang's Mausoleum Site Museum. It provides a reference for future protection work, which is of great significance.

Comparison of maternal and neonatal outcomes between vaginal birth after cesarean section and normal vaginal birth.

OBJECTIVE: This study aimed to compare the outcomes of vaginal birth after cesarean (VBAC) with those of normal vaginal birth (NVB) in a tertiary hospital in China. METHODS: This retrospective cohort study analyzed 1,024 women who birthed vaginally between January 2019 and December 2020. The VBAC group (n = 512) included women with one previous cesarean, while the NVB group (n = 512) had no previous caesareans. All women used epidural analgesia. We assessed maternal and neonatal complications using descriptive statistics, chi-square tests, and logistic regression. Statistical analysis was performed using SPSS version 25.0. RESULTS: The VBAC group had an 87.5% success rate for vaginal birth under epidural analgesia, whereas the NVB group had a 100% success rate. A primary focus of the study was uterine rupture. Vaginal birth after cesarean was associated with a higher incidence of uterine rupture (0.8% vs 0%, p = 0.031), postpartum hemorrhage (6.6% vs 3.5%, p = 0.021) and the need for blood transfusions (2.7% vs 0.8%, p = 0.012) compared with NVB. There were no substantial differences in maternal infections, wound infections or perineal lacerations between the groups. Although neonatal outcomes were generally similar, the VBAC group experienced higher rates of 5-minute Apgar scores <7 (2.3% vs 0.6%, p = 0.009) and admissions to neonatal intensive care units (3.1% vs 1.2%, p = 0.016 Even after adjusting for confounders, VBAC remained an independent risk factor for several complications. CONCLUSION: Although VBAC is feasible and mostly safe, it is associated with a higher risk of specific complications compared with NVB. Careful selection of candidates and close monitoring are essential for optimizing outcomes in VBAC cases.

Association of circulating cytokine levels and tissue-infiltrating myeloid cells with achalasia: results from Mendelian randomization and validation through clinical characteristics and single-cell RNA sequencing.

BACKGROUND: Achalasia is a rare motility disorder of the esophagus often accompanied by immune dysregulation, yet specific underlying mechanisms remain poorly understood. METHODS: We utilized Mendelian randomization (MR) to explore the causal effects of cytokine levels on achalasia, with cis-expression/protein quantitative trait loci (cis-eQTLs/pQTLs) for 47 cytokines selected from a genome-wide association study (GWAS) meta-analysis and GWAS data for achalasia obtained from FinnGen. For cytokines significantly linked to achalasia, we analyzed their plasma concentrations and expression differences in the lower esophageal sphincter (LES) using enzyme-linked immunosorbent assay and single-cell RNA sequencing (scRNA-seq) profiling, respectively. We further employed bioinformatics approaches to investigate underlying mechanisms. RESULTS: We revealed positive associations of circulating Eotaxin, macrophage inflammatory protein-1b (MIP1b), soluble E-selectin (SeSelectin) and TNF-related apoptosis-inducing ligand (TRAIL) with achalasia. When combining MR findings with scRNA-seq data, we observed upregulation of TRAIL (OR = 2.70, 95% CI, 1.20-6.07), encoded by TNFSF10, in monocytes and downregulation of interleukin-1 receptor antagonist (IL-1ra) (OR = 0.70, 95% CI 0.59-0.84), encoded by IL1RN, in FOS_macrophages in achalasia. TNFSF10high monocytes in achalasia displayed activated type I interferon signaling, and IL1RNlow FOS_macrophages exhibited increased intercellular communications with various lymphocytes, together shaping the proinflammatory microenvironment of achalasia. CONCLUSIONS: We identified circulating Eotaxin, MIP1b, SeSelectin and TRAIL as potential drug targets for achalasia. TNFSF10high monocytes and IL1RNlow macrophages may play a role in the pathogenesis of achalasia.

Mutational Landscape of Gastric Adenocarcinoma of the Fundic Gland Type Revealed by Whole Genome Sequencing.

BACKGROUND: Gastric adenocarcinoma of the fundic gland type (GA-FG) is a newly described variant of gastric adenocarcinoma with lack of knowledges regarding its genetic features. METHODS: We performed whole-genome sequencing (WGS) in formalin-fixed paraffin-embedded (FFPE) tumor tissues and matched adjacent noncancerous specimens from 21 patients with GA-FG, and integrated published datasets from 1105 patients with traditional gastric adenocarcinoma with the purpose of dissecting genetic determinants both common to conventional gastric adenocarcinoma and unique to GA-FG disease. RESULTS: We characterized the genomic architecture of GA-FG disease, revealing the predominant proportion of C > T substitution among the six types of SNVs. GNAS was the most significantly mutated driver gene (14.29%). 42.8% of samples harbored "Kataegis." Distinct genomic alterations between GA-FG and conventional gastric cancer were identified. Specifically, low mutational burden and relatively moderate mutational frequencies of significantly mutated driver genes, coupled with the absence of non-silent alterations of formerly well-known drivers such as TP53, PIK3CA and KRAS were identified in GA-FG patients. Oncogenic signaling pathway analysis revealed mutational processes associated with focal adhesions and proteoglycans in cancer, highlighting both common and specific procedures during the development of GA-FG and conventional gastric cancer. CONCLUSION: Our study is the first to comprehensively depict the genomic landscape highlighting the multidimensional perturbations in GA-FG patients. These discoveries offered mechanistic insights for novel diagnostic and therapeutic strategies for patients with such disease.

Bisphenol AP inhibits mouse oocyte maturation in vitro by disrupting cytoskeleton architecture and cell cycle processes.

Bisphenol A (BPA) is among the extensively researched environmental endocrine-disrupting chemicals (EDCs), and its utilization is restricted owing to the detrimental impacts it has on human health. Bisphenol AP (BPAP) is one of the alternatives to BPA, but the influence of BPAP on human health has not been elucidated. The objective of the current research was to determine the influence of BPAP exposure on the in vitro maturation of mouse oocytes and to explore its potential reproductive toxicity. BPAP exposure was found to inhibit polar body extrusion during mouse oocyte maturation, resulting in an arrest at the metaphase I stage of meiosis. Exposure to BPAP led to sustained activation of BubR1, preventing the degradation of both Securin and Cyclin B1. Mechanistically, BPAP exposure disrupts spindle assembly and chromosome alignment. Levels of acetylated α-tubulin were significantly elevated in BPAP-treated oocytes, reflecting decreased spindle stability. Exposure to BPAP also induced DNA damage and impaired DNA damage repair. In addition, BPAP exposure altered histone modification levels. In summary, this investigation suggests that exposure to BPAP can influence cytoskeletal assembly, interfere with cell cycle progression, induce DNA damage, alter histone modifications, and ultimately impede oocyte meiotic maturation. This investigation enhances understanding of the impact of bisphenol analogs on female gametes, underscoring that BPAP cannot be considered a reliable replacement for BPA.

Palladium-Catalyzed Cyclization and Direct Arylation of 1-[2-(2,2-Dibromoethenyl)phenyl]-1H-pyrrole with Allenes.

A novel and efficient palladium-catalyzed highly regioselective reaction of 1-[2-(2,2-dibromoethenyl)phenyl]-1H-pyrrole with allenes was realized to synthesize pyrrolo[1,2-a]quinolones. The tandem process involves intermolecular cyclization and intramolecular direct arylation, leading to the formation three new C-C bonds and two new rings. Notably, this transformation exhibits broad substrate scope and high functional group tolerance.

Early pregnancy SARS-COV-2 infection and fetal cardiac and hemodynamic changes.

AIM: Our objective was to investigate the impact on fetal cardiac function and fetal hemodynamics after recovery from severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection in early pregnancy. METHODS: A prospective study involving 60 women in pregnancy who had recovered from a previous SARS-CoV-2 infection and 20 control wemen was performed. Between 11 and 14 weeks of pregnancy, women recovering from infection and controls underwent fetal ultrasound evaluation. Ultrasound parameters assessing cardiac function (TAPSE, MAPSE, E/A ratio) and hemodynamics (DV/S, DV-D, DV-A, DV-TAMV, DV-PI, DV-PLI, DV-PVIV) were measured. RESULTS: Based on ultrasound measurements, the median gestation age of the groups recovering from SARS-CoV-2 infection (RSI) was 12 (0.5) weeks, while the control group's was 12 (0.7) weeks (p = 0.76). The RSI group and the control group didn't indicate statistically significant differences in ultrasound measurements of cardiac function and hemodynamics (p > 0.05). CONCLUSIONS: According to our findings, the infection of SARS-CoV-2 in early pregnancy has no substantial influence on fetal cardiac function and fetal hemodynamics in pregnant women. However, the effect on mid-pregnancy to late-pregnancy is not yet known. Future studies will help elucidate the overall impact on fetal cardiac function of SARS-CoV-2 infection.

Ultrasound strain elastography to improve diagnostic performance of breast lesions by reclassifying BI-RADS 3 and 4a lesions: a multicenter diagnostic study.

OBJECTIVE: To investigate the added value of strain elastography (SE) by recategorizing ultrasound (US) Breast Imaging Reporting and Data System (BI-RADS) 3 and 4a lesions. METHODS: A total of 4371 patients underwent US and SE with BI-RADS 2-5 categories solid breast lesions were included from thirty-two hospitals. We evaluated the elastographic images according to elasticity scores (ES) and strain ratios (SR). Three combined methods (BI-RADS+ES, BI-RADS+SR, BI-RADS+ES+SR) and two reclassified methods were used (method one: upgrading BI-RADS 3 and downgrading BI-RADS 4a, method two: downgrading BI-RADS 4a alone). The diagnostic performance and the potential reduction of unnecessary biopsies were evaluated. RESULTS: Combining BI-RADS with SE had a higher area under the curve (AUC) than BI-RADS alone (0.822-0.898 vs. 0.794, P<0.01). For reclassified method one, the sensitivity, specificity, and accuracy were 99.36%, 66.70%, 78.36% for BI-RADS+ES and 98.01%, 66.45%, 77.72% for BI-RADS+SR, and 99.42%, 66.70%, 78.38% for BI-RADS+ES+SR, respectively. For reclassified method two, the sensitivity, specificity, and accuracy were 99.17%, 70.72%, 80.87% for BI-RADS+ES and 97.76%, 81.75%, 87.46% for BI-RADS+SR, and 99.23%, 69.83%, 80.32% for BI-RADS+ES+SR, respectively. Downgrading BI-RADS 4a alone had higher AUC, specificity, and accuracy (P<0.01) and similar sensitivity (P>0.05) to upgrading BI-RADS 3 and downgrading BI-RADS 4a. Combining SE with BI-RADS could help reduce unnecessary biopsies by 17.64%-55.20%. CONCLUSIONS: Combining BI-RADS with SE improved the diagnostic performance in distinguishing benign from malignant lesions and could decrease false-positive breast biopsy rates. Downgrading BI-RADS 4a lesions alone might be sufficient for achieving good diagnostic performance. ADVANCES IN KNOWLEDGE: Downgrading BI-RADS category 4a lesions alone had higher AUC, specificity, and accuracy, and similar sensitivity to upgrading or downgrading BI-RADS category 3 and 4a lesions.

Mitigating greenhouse gas emission and enhancing fermentation by phosphorus slag addition during sewage sludge composting.

During the composting of sewage sludge (SS), a quantity of greenhouse gases has been produced. This study aimed to clarify the microbial mechanisms associated with the addition of industrial solid waste phosphorus slag (PS) to SS composting, specifically focusing on its impact on greenhouse gas emissions and the humification. The findings indicated that the introduction of PS increased the temperature and extended the high-temperature phase. Moreover, the incorporation of 10% and 15% PS resulted in a decrease of N2O emissions by 68.9% and 88.6%, respectively. Microbial diversity analysis indicated that PS improved waste porosity, ensuring the aerobic habitat. Therefore, the environmental factors of the system were altered, leading to the enrichment of various functional bacterial species, such as Firmicutes and Chloroflexi, and a reduction of pathogenic bacterium Dokdonella. Consequently, incorporating PS into SS composting represents an effective waste treatment strategy, exhibiting economic feasibility and promising application potential.

A robust myoelectric pattern recognition framework based on individual motor unit activities against electrode array shifts.

BACKGROUND AND OBJECTIVE: Electrode shift is always one of the critical factors to compromise the performance of myoelectric pattern recognition (MPR) based on surface electromyogram (SEMG). However, current studies focused on the global features of SEMG signals to mitigate this issue but it is just an oversimplified description of the human movements without incorporating microscopic neural drive information. The objective of this work is to develop a novel method for calibrating the electrode array shifts toward achieving robust MPR, leveraging individual motor unit (MU) activities obtained through advanced SEMG decomposition. METHODS: All of the MUs from decomposition of SEMG data recorded at the original electrode array position were first initialized to train a neural network for pattern recognition. A part of decomposed MUs could be tracked and paired with MUs obtained at the original position based on spatial distribution of their MUAP waveforms, so as to determine the shift vector (describing both the orientation and distance of the shift) implicated consistently by these multiple MU pairs. Given the known shift vector, the features of the after-shift decomposed MUs were corrected accordingly and then fed into the network to finalize the MPR task. The performance of the proposed method was evaluated with data recorded by a 16 × 8 electrode array placed over the finger extensor muscles of 8 subjects performing 10 finger movement patterns. RESULTS: The proposed method achieved a shift detection accuracy of 100 % and a pattern recognition accuracy approximating to 100 %, significantly outperforming the conventional methods with lower shift detection accuracies and lower pattern recognition accuracies (p < 0.05). CONCLUSIONS: Our method demonstrated the feasibility of using decomposed MUAP waveforms' spatial distributions to calibrate electrode shift. This study provides a new tool to enhance the robustness of myoelectric control systems via microscopic neural drive information at an individual MU level.

A Visual Distance-Based Capillary Immunoassay Using Biomimetic Polymer Nanoparticles for Highly Sensitive and Specific C-Reactive Protein Quantification.

The low-cost daily monitoring of C-reactive protein (CRP) levels is crucial for screening acute inflammation or infections as well as managing chronic inflammatory diseases. In this study, we synthesized novel 2-Methacryloyloxy ethyl phosphorylcholine (MPC)-based biomimetic nanoparticles with a large surface area to develop a visual CRP-quantification assay using affordable glass capillaries. The PMPC nanoparticles, synthesized via reflux precipitation polymerization, demonstrated multivalent binding capabilities, enabling rapid and specific CRP capture. In the presence of CRP, PMPC nanoparticles formed sandwich structures with magnetic nanoparticles functionalized with CRP antibodies, thereby enhancing detection sensitivity and specificity. These sandwich complexes were magnetically accumulated into visible and quantifiable stacks within the glass capillaries, allowing for the rapid, sensitive, and specific quantification of CRP concentrations with a detection limit of 57.5 pg/mL and a range spanning from 0 to 5000 ng/mL. The proposed visual distance-based capillary biosensor shows great potential in routine clinical diagnosis as well as point-of-care testing (POCT) in resource-limited settings.

Association of novel DNAH11 variants with asthenoteratozoospermia lead to male infertility.

BACKGROUND: Bi-allelic variants in DNAH11 have been identified as causative factors in Primary Ciliary Dyskinesia, leading to abnormal respiratory cilia. Nonetheless, the specific impact of these variants on human sperm flagellar and their involvement in male infertility remain largely unknown. METHODS: A collaborative effort involving two Chinese reproductive centers conducted a study with 975 unrelated infertile men. Whole-exome sequencing was employed for variant screening, and Sanger sequencing confirmed the identified variants. Morphological and ultrastructural analyses of sperm were conducted using Scanning Electron Microscopy and Transmission Electron Microscopy. Western Blot Analysis and Immunofluorescence Analysis were utilized to assess protein levels and localization. ICSI was performed to evaluate its efficacy in achieving favorable pregnancy outcomes for individuals with DNAH11 variants. RESULTS: In this study, we identified seven novel variants in the DNAH11 gene in four asthenoteratozoospermia subjects. These variants led the absence of DNAH11 proteins and ultrastructure defects in sperm flagella, particularly affecting the outer dynein arms (ODAs) and adjacent structures. The levels of ODA protein DNAI2 and axoneme related proteins were down regulated, instead of inner dynein arms (IDA) proteins DNAH1 and DNAH6. Two out of four individuals with DNAH11 variants achieved clinical pregnancies through ICSI. The findings confirm the association between male infertility and bi-allelic deleterious variants in DNAH11, resulting in the aberrant assembly of sperm flagella and contributing to asthenoteratozoospermia. Importantly, ICSI emerges as an effective intervention for overcoming reproductive challenges caused by DNAH11 gene variants.

Targeted degradation of VEGF with bispecific aptamer-based LYTACs ameliorates pathological retinal angiogenesis.

Rationale: Neovascular ocular diseases (NODs) represent the leading cause of visual impairment globally. Despite significant advances in anti-angiogenic therapies targeting vascular endothelial growth factor (VEGF), persistent challenges remain prevalent. As a proof-of-concept study, we herein demonstrate the effectiveness of targeted degradation of VEGF with bispecific aptamer-based lysosome-targeting chimeras (referred to as VED-LYTACs). Methods: VED-LYTACs were constructed with three distinct modules: a mannose-6-phosphate receptor (M6PR)-binding motif containing an M6PR aptamer, a VEGF-binding module with an aptamer targeting VEGF, and a linker essential for bridging and stabilizing the two-aptamer structure. The degradation efficiency of VED-LYTACs via the autophagy-lysosome system was examined using an enzyme-linked immunosorbent assay (ELISA) and immunofluorescence staining. Subsequently, the anti-angiogenic effects of VED-LYTACs were evaluated using in vitro wound healing assay, tube formation assay, three-dimensional sprouting assay, and ex vivo aortic ring sprouting assay. Finally, the potential therapeutic effects of VED-LYTACs on pathological retinal neovascularization and vascular leakage were tested by employing mouse models of NODs. Results: The engineered VED-LYTACs promote the interaction between M6PR and VEGF, consequently facilitating the translocation and degradation of VEGF through the lysosome. Our data show that treatment with VED-LYTACs significantly suppresses VEGF-induced angiogenic activities both in vitro and ex vivo. In addition, intravitreal injection of VED-LYTACs remarkably ameliorates abnormal vascular proliferation and leakage in mouse models of NODs. Conclusion: Our findings present a novel strategy for targeting VEGF degradation with an aptamer-based LYTAC system, effectively ameliorating pathological retinal angiogenesis. These results suggest that VED-LYTACs have potential as therapeutic agents for managing NODs.

Electroporation Delivery of Cas9 sgRNA Ribonucleoprotein-Mediated Genome Editing in Sheep IVF Zygotes.

The utilization of electroporation for delivering CRISPR/Cas9 system components has enabled efficient gene editing in mammalian zygotes, facilitating the development of genome-edited animals. In this study, our research focused on targeting the ACTG1 and MSTN genes in sheep, revealing a threshold phenomenon in electroporation with a voltage tolerance in sheep in vitro fertilization (IVF) zygotes. Various poring voltages near 40 V and pulse durations were examined for electroporating sheep zygotes. The study concluded that stronger electric fields required shorter pulse durations to achieve the optimal conditions for high gene mutation rates and reasonable blastocyst development. This investigation also assessed the quality of Cas9/sgRNA ribonucleoprotein complexes (Cas9 RNPs) and their influence on genome editing efficiency in sheep early embryos. It was highlighted that pre-complexation of Cas9 proteins with single-guide RNA (sgRNA) before electroporation was essential for achieving a high mutation rate. The use of suitable electroporation parameters for sheep IVF zygotes led to significantly high mutation rates and heterozygote ratios. By delivering Cas9 RNPs and single-stranded oligodeoxynucleotides (ssODNs) to zygotes through electroporation, targeting the MSTN (Myostatin) gene, a knock-in efficiency of 26% was achieved. The successful generation of MSTN-modified lambs was demonstrated by delivering Cas9 RNPs into IVF zygotes via electroporation.

Vegetation restoration enhancing soil carbon sequestration in karst rocky desertification ecosystems: A meta-analysis.

Vegetation restoration measures have been increasingly employed to alleviate rocky desertification in karst ecosystems. However, the comprehensive effects of these interventions on soil properties and soil organic carbon (SOC) remain poorly understood. Herein, we gathered 644 paired observations from 68 studies and conducted a meta-analysis to quantify the performance of different vegetation restoration measures including moss (MS), grassland (GL), cash crop (CP), shrub (SH), and secondary forest (SF) through soil properties and SOC. Our results demonstrated significant effects of MS, GL, CP, SH, and SF on soil biotic and abiotic factors, each with distinct response characteristics. Particularly, MS significantly enhanced all soil properties (excluding a slight decrease in soil pH by 10.8%). Moreover, MS, GL, CP, SH, and SF could elevate SOC by 32.1%, 17.6%, 24.9%, 59.2%, and 48.7% respectively. Utilizing random forest and linear regression models, we identified primary drivers for SOC in MS, GL, CP, SH, and SF as soil moisture content, arbuscular mycorrhizal fungi, soil microbial phosphorus, total nitrogen, and ß-1,4-glucosidase, respectively. This meta-analysis underlined the varied effects of vegetation restoration measures on soil properties and advocates for restoration measures that prioritize plant productivity and reduce soil temperature during the karst rocky desertification restoration process. Additionally, this study underscores the pivotal role of vegetation rehabilitation in environmental conservation and carbon sequestration of ecologically vulnerable regions.

Clinical characteristics, diagnosis and short-term outcomes of COVID-19-associated acute myocarditis in China.

AIMS: Acute myocarditis (AM) has been recognized as a rare complication of coronavirus disease 2019 (COVID-19) infection. This study was conducted to present the clinical characteristics, disease courses and short-term prognoses of Omicron variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induced AM in China, which has been unavailable so far. METHODS AND RESULTS: Data from 28 patients diagnosed with definite COVID-19-associated AM from 6 hospitals in China between 1 December 2022 and 30 June 2023 were collected and analysed. The diagnosis of AM was based on increased troponin level plus typical findings of AM on cardiac magnetic resonance (CMR) imaging and/or endomyocardial biopsy. Among 28 patients with definite COVID-19-related AM, median age was 37 years (Q1-Q3: 22-52) and 53.6% were men. Twenty-three patients occurred within 2 weeks of the onset of COVID-19 infection, 10 patients underwent endomyocardial biopsy and CMR was performed in all patients. Seven (25.0%) patients developed fulminant myocarditis that required inotropic agents or temporary mechanical circulatory support. Of the nine patients (32.1%) with left ventricular ejection fraction (LVEF) below 50% on admission, five had fully recovered LVEF and two demonstrated improvement but to levels below normal at discharge. The comparison of CMR parameters between the baseline and first follow-up showed that ECV was decreased at the first follow-up [28.95 (25.38, 32.55)% vs. 33.65 (31.58, 37.55)%, P = 0.028), while other CMR parameters had no significant changes. Eighteen patients (64.3%) were prescribed with corticosteroids, and seven patients (25.0%) underwent temporary mechanical circulatory support. Only two patients died during hospitalization. CONCLUSIONS: The majority of COVID-19-associated AM occurred within 2 weeks of Omicron variant infection. Fulminant myocarditis complicated by hemodynamic instability requiring temporary mechanical circulatory support was not uncommon. However, short-term outcome was generally good and most AM patients fully recovered.

Changes of CD8+CD28- Tregs and Gamma-Delta-T-cells in a Neonate with Intrauterine Cytomegalovirus Infection: A Case Report.

BACKGROUND: Congenital cytomegalovirus (cCMV) infection can lead to a range of adverse outcomes. The majority of cCMV neonates with clinical symptoms are infected postnatally; however, established cases of intrauterine infection are uncommon, resulting in a paucity of reports on clinical findings and lymphocytes expression in CMV-infected neonates. CASE PRESENTATION: We followed a neonate with cCMV infection from the onset of hospitalization to several months of follow-up. This infant was intrauterine CMV-positive in the amniotic fluid of the mother at 21 weeks' gestation and received intravenous ganciclovir infusion and sequential oral valganciclovir after birth. The typical clinical signs manifested in the nervous system, liver, and peripheral blood and were documented during the hospitalizaion period and up to the follow-up visit. Flow cytometry was employed to examine the expression of T cells, their subsets, and the associated cytokines in peripheral blood samples at various time points. The flow data for the cCMV neonate were compared with those of the controls at each time point. Following treatment, clinical symptoms improved and the infant became CMV negative. However, developmental delays occurred later in life. The proportion of CD8+CD28- Tregs in the peripheral blood of the neonate with congenital CMV infection was higher than that in the controls at the three time points. The expression levels of perforin and granzyme B secreted by γδ T cells (Vδ1 and Vδ2 T cells), increased during the course of hospitalization until follow-up and were higher than those in the controls at the three time points. CONCLUSIONS: Despite the alleviation of clinical symptoms, developmental delay in later life remains inevitable in this intrauterine cCMV neonate. CD8+CD28- Tregs and Vδ1 and Vδ2 T cells secreting perforin and granzyme B may be involved in congenital CMV infection, although this hypothesis requires validation in a larger study. This report may contribute to our understanding of the effect of current treatment and the immune status of intrauterine cCMV-infected neonates.

CALR3 defects disrupt sperm-zona pellucida binding in humans: new insights into male factor fertilization failure and relevant clinical therapeutic approaches.

STUDY QUESTION: Do biallelic deleterious variants of Calreticulin 3 (CALR3) cause fertilization failure (FF), resulting in male infertility in humans? SUMMARY ANSWER: Biallelic mutations in CALR3 were identified in two infertile men from unrelated families and were shown to cause FF associated with failed sperm-zona pellucida (ZP) binding. WHAT IS KNOWN ALREADY: In male mice, the Calr3-knockout has been reported to cause male infertility and FF. However, the mechanism behind this remains unclear in humans. STUDY DESIGN, SIZE, DURATION: Sequencing studies were conducted in a research hospital on samples from Han Chinese families with primary infertility and sperm head deformations to identify the underlying genetic causes. PARTICIPANTS/MATERIALS, SETTING, METHODS: Data from two infertile probands characterized by sperm head deformation were collected through in silico analysis. Sperm cells from the probands were characterized using light and electron microscopy and used to verify the pathogenicity of genetic factors through functional assays. Subzonal insemination (SUZI) and IVF assays were performed to determine the exact pathogenesis of FF. ICSI were administered to overcome CALR3-affected male infertility. MAIN RESULTS AND THE ROLE OF CHANCE: Novel biallelic deleterious mutations in CALR3 were identified in two infertile men from unrelated families. We found one homozygous frameshift CALR3 mutation (M1: c.17_27del, p.V6Gfs*34) and one compound heterozygous CALR3 mutation (M2: c.943A>G, p.N315D; M3: c.544T>C, p.Y182H). These mutations are rare in the general population and cause acrosomal ultrastructural defects in affected sperm. Furthermore, spermatozoa from patients harbouring the CALR3 mutations were unable to bind to the sperm-ZP or they disrupted gamete fusion or prevented oocyte activation. Molecular assays have revealed that CALR3 is crucial for the maturation of the ZP binding protein in humans. Notably, the successful fertilization via SUZI and ICSI attempts for two patients, as well as the normal expression of PLCζ in the mutant sperm, suggests that ICSI is an optimal treatment for CALR3-deficient FF. LIMITATIONS, REASONS FOR CAUTION: The results are based on sperm-related findings from two patients. Further studies are required to gain insight into the developmental stage and function of CALR3 in human testis. WIDER IMPLICATIONS OF THE FINDINGS: Our findings highlight the underlying risk of FF associated with sperm defects and provide a valuable reference for personalized genetic counselling and clinical treatment of these patients. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by the National Key R&D Program of China (2021YFC2700901), Hefei Comprehensive National Science Center Medical-Industrial Integration Medical Equipment Innovation Research Platform Project (4801001202), the National Natural Science Foundation of China (82201803, 82371621, 82271639), Foundation of the Education Department of Anhui Province (gxgwfx2022007), Key Project of Natural Science Research of Anhui Educational Committee (2023AH053287), and the Clinical Medical Research Transformation Project of Anhui Province (202204295107020037). The authors declare no competing interests. TRIAL REGISTRATION NUMBER: N/A.