SARS-CoV-2 envelope protein-derived extracellular vesicles act as potential media for viral spillover.

Extracellular vesicles (EVs) are shown to be a novel viral transmission model capable of increasing a virus's tropism. According to our earlier research, cells infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or transfected with envelope protein plasmids generate a novel type of EVs that are micrometer-sized and able to encase virus particles. Here, we showed the capacity of these EVs to invade various animals both in vitro and in vivo independent of the angiotensin-converting enzyme 2 receptor. First, via macropinocytosis, intact EVs produced from Vero E6 (monkey) cells were able to enter cells from a variety of animals, including cats, dogs, bats, hamsters, and minks, and vice versa. Second, when given to zebrafish with cutaneous wounds, the EVs showed favorable stability in aqueous environments and entered the fish. Moreover, infection of wild-type (WT) mice with heterogeneous EVs carrying SARS-CoV-2 particles led to a strong cytokine response and a notable amount of lung damage. Conversely, free viral particles did not infect WT mice. These results highlight the variety of processes behind viral transmission and cross-species evolution by indicating that EVs may be possible vehicles for SARS-CoV-2 spillover and raising risk concerns over EVs' potential for viral gene transfer.

Impact of a postpartum care rehabilitation program to prevent postpartum depression at a secondary municipal hospital in Qingdao China: a cross-sectional study.

BACKGROUND: The emerging postpartum rehabilitation (PPR) program in Chinese hospitals characterized by applying ongoing medical care through traditional cultural practices shows a protective effect in early puerperium in China. This study explores the benefit of PPR program practices to postpartum depression (PPD) and the influencing factors for PPD among Chinese women during the first postnatal six weeks. METHODS: The cross-sectional study included 403 participants and was conducted in a Secondary Municipal Hospital in Qingdao, China, from 01 to 2018 to 31 December 2021. Information on this PPR program was collected during the six-weeks postpartum consultation, including the Edinburgh postnatal depression scale (EPDS) scores, the measurement results for diastasis recti abdominis, and the international physical activity questionnaire (long form) (IPAQ-L) scores. Logistic regression models were used to examine the effect of the PPR program on PPD among the local population. The secondary aim of this study was to investigate possible influencing factors for PPD, such as coronavirus disease 2019 (COVID-19), physical exercises, etc. RESULTS: PPR program has shown a positive effect in preventing PPD (p < 0.001) and diastasis recti prevalence (p < 0.001) during the six-weeks postnatal control in Qingdao, China. Better post-pregnancy weight reduction (p = 0.04) and higher metabolic equivalent of task (MET) value (p < 0.001) were noticed in the non-PPR group. Furthermore, lower PPD risk was associated with factors such as longer relationship duration years (2-5 years) (p = 0.04) and exercising one to three times a week (p = 0.01). A higher PPD risk was related to factors such as urinary incontinence during the postpartum period (p = 0.04) and subjective insomnia (p < 0.001). No significant effect was shown between COVID-19 and the EPDS score in this study (p = 0.50). CONCLUSION: Our results suggested that the PPR program provided protection against PPD and diastasis recti during the first six weeks after delivery. Urinary incontinence and subjective insomnia were the main risk factors for PPD, while longer relationship duration years and exercising one to three times a week gave protective effects to PPD. This study emphasized that a comprehensive ongoing medical care program, such as the PPR program, effectively improves women's mental and physical health in the early postpartum in China.

Astragalus polysaccharides affects multidrug resistance gene 1 and P-glycoprotein 170 in adriamycin nephropathy rats via regulating microRNA-16/NF-κB axis. / 黄芪多糖调控微RNA-16/NF-κBè½´å¯¹é˜¿éœ‰ç´ è‚¾ç— å¤§é¼ å¤šè¯è€è¯åŸºå› 1和P-糖蛋白170的影响.

OBJECTIVES: Nephrotic syndrome is a common disease of the urinary system. The aim of this study is to explore the effect of astragalus polysaccharides (APS) on multidrug resistance gene 1 (MDR1) and P-glycoprotein 170 (P-gp170) in adriamycin nephropathy rats and the underlying mechanisms. METHODS: A total of 72 male Wistar rats were divided into a control group, a model group, an APS low-dose group, an APS high-dose group, an APS+micro RNA (miR)-16 antagomir group and an APS+miR-16 antagomir control group, with 12 rats in each group. Urine protein (UP) was detected by urine analyzer, and serum cholesterol (CHOL), albumin (ALB), blood urea nitrogen (BUN), and creatinine (SCr) were detected by automatic biochemical analyzer; serum interleukin-6 (IL-6), IL-1ß, tumor necrosis factor α (TNF-α) levels were detected by ELISA kit; the morphological changes of kidney tissues were observed by HE staining; the levels of miR-16 and MDR1 mRNA in kidney tissues were detected by real-time RT-PCR; the expression levels of NF-κB p65, p-NF-κB p65, and P-gp170 protein in kidney tissues were detected by Western blotting; and dual luciferase was used to verify the relationship between miR-16 and NF-κB. RESULTS: The renal tissue structure of rats in the control group was normal without inflammatory cell infiltration. The renal glomeruli of rats in the model group were mildly congested, capillary stenosis or occlusion, and inflammatory cell infiltration was obvious. The rats in the low-dose and high-dose APS groups had no obvious glomerular congestion, the proliferation of mesangial cells was significantly reduced, and the inflammatory cells were reduced. Compared with the high-dose APS group and the APS+miR-16 antagomir control group, there were more severe renal tissue structure damages in the APS + miR-16 antagomir group. Compared with the control group, the levels of UP, CHOL, BUN, SCr, IL-6, IL-1ß, TNF-α, and MDR1 mRNA, and the protein levels of p-NF-κB p65 and P-gp170 in the model group were significantly increased (all P<0.05); the levels of ALB and miR-16 were significantly decreased (both P<0.05). Compared with the model group, the levels of UP, CHOL, BUN, SCr, IL-6, IL-1ß, TNF-α, and MDR1 mRNA, and the protein levels of pNF-κB p65 and P-gp170 in the low-dose and high-dose APS groups were significant decreased (all P<0.05); and the levels of ALB and miR-16 were significantly increased (both P<0.05). Compared with APS+miR-16 antagomir control group, the UP, CHOL, BUN, SCr, IL-6, IL-1ß, and TNF-α levels, MDR1 mRNA, and the protein levels of p-NF-κB p65 and P-gp170 were significantly increased (all P<0.05). The levels of ALB and miR-16 were significantly decreased in the APS+miR-16 antagomir group compared with the APS+miR-16 antagomir control group (both P<0.05). CONCLUSIONS: APS can regulate the miR-16/NF-κB signaling pathway, thereby affecting the levels of MDR1 and P-gp170, and reducing the inflammation in the kidney tissues in the adriamycin nephropathy rats.

The histone methyltransferase SETD2 is required for expression of acrosin-binding protein 1 and protamines and essential for spermiogenesis in mice.

Spermatogenesis is precisely controlled by complex gene expression programs and involves epigenetic reprogramming, including histone modification and DNA methylation. SET domain-containing 2 (SETD2) is the predominant histone methyltransferase catalyzing the trimethylation of histone H3 lysine 36 (H3K36me3) and plays key roles in embryonic stem cell differentiation and somatic cell development. However, its role in male germ cell development remains elusive. Here, we demonstrate an essential role of Setd2 for spermiogenesis, the final stage of spermatogenesis. Using RNA-seq, we found that, in postnatal mouse testes, Setd2 mRNA levels dramatically increase in 14-day-old mice. Using a germ cell-specific Setd2 knockout mouse model, we also found that targeted Setd2 knockout in germ cells causes aberrant spermiogenesis with acrosomal malformation before step 8 of the round-spermatid stage, resulting in complete infertility. Furthermore, we noted that the Setd2 deficiency results in complete loss of H3K36me3 and significantly decreases expression of thousands of genes, including those encoding acrosin-binding protein 1 (Acrbp1) and protamines, required for spermatogenesis. Our findings thus reveal a previously unappreciated role of the SETD2-dependent H3K36me3 modification in spermiogenesis and provide clues to the molecular mechanisms in epigenetic disorders underlying male infertility.

Uncovering hub genes and immunological characteristics for heart failure utilizing RRA, WGCNA and Machine learning.

Background: Heart failure (HF) is a major public health issue with high mortality and morbidity. This study aimed to find potential diagnostic markers for HF by the combination of bioinformatics analysis and machine learning, as well as analyze the role of immune infiltration in the pathological process of HF. Methods: The gene expression profiles of 124 HF patients and 135 nonfailing donors (NFDs) were obtained from six datasets in the NCBI Gene Expression Omnibus (GEO) public database. We applied robust rank aggregation (RRA) and weighted gene co-expression network analysis (WGCNA) method to identify critical genes in HF. To discover novel diagnostic markers in HF, three machine learning methods were employed, including best subset regression, regularization technique, and support vector machine-recursive feature elimination (SVM-RFE). Besides, immune infiltration was investigated in HF by single-sample gene set enrichment analysis (ssGSEA). Results: Combining RRA with WGCNA method, we recognized 39 critical genes associated with HF. Through integrating three machine learning methods, FCN3 and SMOC2 were determined as novel diagnostic markers in HF. Differences in immune infiltration signature were also found between HF patients and NFDs. Moreover, we explored the potential associations between two diagnostic markers and immune response in the pathogenesis of HF. Conclusions: In summary, FCN3 and SMOC2 can be used as diagnostic markers of HF, and immune infiltration plays an important role in the initiation and progression of HF.

Integrative analysis of bioinformatics and machine learning to identify cuprotosis-related biomarkers and immunological characteristics in heart failure.

Backgrounds: Cuprotosis is a newly discovered programmed cell death by modulating tricarboxylic acid cycle. Emerging evidence showed that cuprotosis-related genes (CRGs) are implicated in the occurrence and progression of multiple diseases. However, the mechanism of cuprotosis in heart failure (HF) has not been investigated yet. Methods: The HF microarray datasets GSE16499, GSE26887, GSE42955, GSE57338, GSE76701, and GSE79962 were downloaded from the Gene Expression Omnibus (GEO) database to identify differentially expressed CRGs between HF patients and nonfailing donors (NFDs). Four machine learning models were used to identify key CRGs features for HF diagnosis. The expression profiles of key CRGs were further validated in a merged GEO external validation dataset and human samples through quantitative reverse-transcription polymerase chain reaction (qRT-PCR). In addition, Gene Ontology (GO) function enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and immune infiltration analysis were used to investigate potential biological functions of key CRGs. Results: We discovered nine differentially expressed CRGs in heart tissues from HF patients and NFDs. With the aid of four machine learning algorithms, we identified three indicators of cuprotosis (DLAT, SLC31A1, and DLST) in HF, which showed good diagnostic properties. In addition, their differential expression between HF patients and NFDs was confirmed through qRT-PCR. Moreover, the results of enrichment analyses and immune infiltration exhibited that these diagnostic markers of CRGs were strongly correlated to energy metabolism and immune activity. Conclusions: Our study discovered that cuprotosis was strongly related to the pathogenesis of HF, probably by regulating energy metabolism-associated and immune-associated signaling pathways.

Study characteristical and regional influences on postpartum depression before vs. during the COVID-19 pandemic: A systematic review and meta-analysis.

Background: While the public is under serious pressure from the coronavirus disease 2019 (COVID-19), the final impact and possible contributing factors to postpartum depression symptoms (PPDS) remain unknown. Therefore, a meta-analysis to investigate the association between PPDS and the COVID-19 pandemic was carried out by comparing the data between pre-pandemic and post-pandemic timeframes and exploring the influencing factors. Methods: This systematic review was prospectively registered and recorded in a study protocol (Prospero CRD42022336820, http://www.crd.york.ac.uk/PROSPERO). A comprehensive search of PubMed, Embase, Web of Science, CINALH, Cochrane and Scopus was cmpleted on June 6, 2022. Studies that compared the prevalence of PPD before and during the COVID-19 pandemic period were included. Results: Of 1766 citations identified, 22 studies were included with 15,098 participates before the COVID-19 pandemic and 11,836 participants during the COVID-19 pandemic. Overall, the analysis showed that the epidemic crisis was associated with an increased prevalence of PPDS (OR: 0.81 [0.68, 0.95], P = 0.009, I 2 = 59%). Subgroup analysis was conducted according to the study characteristics and regions. Within the study characteristics classification, results showed an obvious increase in the prevalence of PPDS during the COVID-19 pandemic if PPDS cutoff was defined as Edinburgh postpartum depression score (EPDS) ≥13 points (OR: 0.72 [0.52, 0.98], P = 0.03, I 2 = 67%) and an increased prevalence in follow-ups that happened after 2 weeks (≥ 2 weeks postpartum) (OR: 0.81 [0.68, 0.97], P = 0.02, I 2 = 43%). Selected studies that were high-quality (OR: 0.79 [0.64, 0.97], P = 0.02, I 2 = 56%) demonstrated an increased prevalence of PPDS during the COVID-19 pandemic period. Sorting by regional factors, studies conducted in Asia (OR: 0.81 [0.70, 0.93], P = 0.003, I 2 = 0%) showed an increase of PPDS prevalence rates during the COVID-19 period, while studies conducted in Europe (OR: 0.82 [0.59, 1.13], P = 0.23, I 2 = 71%) and North America (OR: 0.66 [0.42, 1.02], P = 0.06, I 2 = 65%) showed no significant difference. All studies conducted in the developed (OR: 0.79 [0.64, 0.98], P = 0.03, I 2 = 65%) and developing countries (OR: 0.81 [0.69, 0.94], P = 0.007, I 2 = 0%) showed an increase of PPDS during the COVID-19 period. Conclusions: The COVID-19 pandemic is associated with an increased prevalence of PPDS, especially after long-term follow-up and among the group with a high possibility of depression. The negative influence from the pandemic, causing more PPDS was significant in studies from Asia.

Extracellular vesicles mediate antibody-resistant transmission of SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic. Antibody resistance dampens neutralizing antibody therapy and threatens current global Coronavirus (COVID-19) vaccine campaigns. In addition to the emergence of resistant SARS-CoV-2 variants, little is known about how SARS-CoV-2 evades antibodies. Here, we report a novel mechanism of extracellular vesicle (EV)-mediated cell-to-cell transmission of SARS-CoV-2, which facilitates SARS-CoV-2 to escape from neutralizing antibodies. These EVs, initially observed in SARS-CoV-2 envelope protein-expressing cells, are secreted by various SARS-CoV-2-infected cells, including Vero E6, Calu-3, and HPAEpiC cells, undergoing infection-induced pyroptosis. Various SARS-CoV-2-infected cells produce similar EVs characterized by extra-large sizes (1.6-9.5 µm in diameter, average diameter > 4.2 µm) much larger than previously reported virus-generated vesicles. Transmission electron microscopy analysis and plaque assay reveal that these SARS-CoV-2-induced EVs contain large amounts of live virus particles. In particular, the vesicle-cloaked SARS-CoV-2 virus is resistant to neutralizing antibodies and able to reinfect naïve cells independent of the reported receptors and cofactors. Consistently, the constructed 3D images show that intact EVs could be taken up by recipient cells directly, supporting vesicle-mediated cell-to-cell transmission of SARS-CoV-2. Our findings reveal a novel mechanism of receptor-independent SARS-CoV-2 infection via cell-to-cell transmission, provide new insights into antibody resistance of SARS-CoV-2 and suggest potential targets for future antiviral therapeutics.

Pyroptosis-Related Risk Signature Exhibits Distinct Prognostic, Immune, and Therapeutic Landscapes in Hepatocellular Carcinoma.

Background: Hepatocellular carcinoma (HCC) is a common abdominal cancer. The existing therapeutic approaches often fail to achieve satisfactory results. Pyroptosis, an inflammatory form of programmed cell death, provides new ideas for anticancer treatment. However, the roles of pyroptosis-related (PR) genes (PRGs) in HCC remain elusive. Methods: Differentially expressed genes (DEGs) (n = 22) were screened out using TCGA and GTEx databases. A novel PR risk signature was constructed through Lasso regression analysis. Its prognostic value was evaluated through a series of survival analyses and was tested in ICGC and GSE14520 cohorts. CIBERSORT, ssGSEA, and ESTIMATE methods were employed to determine the effects of the PR risk score on the tumor immune microenvironment (TIM). The TIDE scoring system, IMvigor210 cohort, GSE109211 dataset, and GSDC database were applied to explore the associations of the PR risk score with therapeutic effects. The biofunctions of WNK1 in hepatocellular cancer (HC) cells were confirmed through qPCR, colony formation, and Transwell assays. Results: Overall, 22 of 45 PRGs (48.9%) were abnormally expressed in HCC samples. Then, a PR risk signature consisting of eight PRGs was constructed. A high PR risk score led to an unfavorable prognosis. The PR risk score was identified as an independent prognostic factor of HCC and could increase the decision-making benefit of the traditional TNM model. In addition, we established a nomogram containing the clinical stage and PR risk score to predict the survival rates of HCC patients. The prognostic value of the PR model was successfully validated in ICGC and GSE14520 cohorts. Moreover, high PR risk conferred the decreased infiltration level of CD8+ T cells and weakened the activities of "cytolytic activity" pathways. As for therapeutic correlation, a high PR risk score seemed to imply a poor efficacy of PD-1/L1 inhibitors and sorafenib. Finally, the overexpression of WNK1 could promote the proliferation, migration, and invasion of HC cells. Conclusions: The PR risk score was closely related to the prognosis, antitumor immune process, therapeutic outcomes, and malignant progression of HCC. WNK1, the core regulator of pyroptosis, possesses pro-oncogenic abilities, showing promise as a novel treatment target.

Inhibitors and facilitators to the utilization of postpartum care in China: an integrative review.

BACKGROUND: Postpartum care is an expanding concept in China, and it is gaining vast attention in Chinese society. However, due to some Chinese traditions and rituals during the postpartum period, the utilization of modern postpartum care should be improved on both individual and community levels from different aspects. This integrative review outlined the inhibitors and facilitators of postpartum care utilization in China. METHODS: Writing an integrative review, a literature search was conducted in Chinese and English databases including Wan Fang, China National Knowledge infrastructure, Medline, Web of Science, and Embase till 31 October 2021 to capture citations covering 'postpartum care', 'utilization' and 'China'. Titles and abstracts were screened independently by three reviewers. Included studies were critically appraised using tools and checklists independently for both qualitative and quantitative studies by two different reviewers who also performed thematic synthesis. RESULTS: Of the 4359 citations screened, 41 studies (450,788 patients) were selected. Categorization of the factors influencing postpartum care utilization revealed five components: sociocultural (25 studies); educational (24 studies); organizational (12 studies); economic (19 studies); and physical (6 studies). Factors influencing postpartum care utilization both on individual and community levels were identified. They included facilitated factors such as higher mother's and partner's education level, higher socioeconomic status, lower parity, better insurance coverage, urban geographical location, Han ethnicity, and better transportation. Inhibitory factors such as under-managed policy regulation, migrants without domicile, and lower quality of care were also reported. CONCLUSION: This review has identified the inhibitors and facilitators of postpartum care utilization in China. Five major aspects including sociocultural, educational, organizational, economic, and physical components have been analysed. Results can be used to improve the utilization of modern postpartum care on both individual and community levels in Chinese society.

SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target.

Cytokine storm and multi-organ failure are the main causes of SARS-CoV-2-related death. However, the origin of excessive damages caused by SARS-CoV-2 remains largely unknown. Here we show that the SARS-CoV-2 envelope (2-E) protein alone is able to cause acute respiratory distress syndrome (ARDS)-like damages in vitro and in vivo. 2-E proteins were found to form a type of pH-sensitive cation channels in bilayer lipid membranes. As observed in SARS-CoV-2-infected cells, heterologous expression of 2-E channels induced rapid cell death in various susceptible cell types and robust secretion of cytokines and chemokines in macrophages. Intravenous administration of purified 2-E protein into mice caused ARDS-like pathological damages in lung and spleen. A dominant negative mutation lowering 2-E channel activity attenuated cell death and SARS-CoV-2 production. Newly identified channel inhibitors exhibited potent anti-SARS-CoV-2 activity and excellent cell protective activity in vitro and these activities were positively correlated with inhibition of 2-E channel. Importantly, prophylactic and therapeutic administration of the channel inhibitor effectively reduced both the viral load and secretion of inflammation cytokines in lungs of SARS-CoV-2-infected transgenic mice expressing human angiotensin-converting enzyme 2 (hACE-2). Our study supports that 2-E is a promising drug target against SARS-CoV-2.

[Effect of pneumatic tourniquet on perioperative blood loss in total knee arthroplasty].

OBJECTIVE: To evaluate the effect of pneumatic tourniquet on perioperative period of total knee arthroplasty (TKA). METHODS: The perioperative period data of 116 patients over 60 years old with severe knee osteoarthritis treated with TKA between January 2018 and January 2019 were retrospectively analyzed. According to whether pneumatic tourniquet was used during operation, the patients were divided into trial group (49 cases, pneumatic tourniquet was not used during operation) and control group (67 cases, pneumatic tourniquet was used during operation). There was no significant difference in gender, age, body mass index, lesion side, disease duration, and preoperative hemoglobin between the two groups ( P>0.05). The operation time, actual total blood loss, overt blood loss, hidden blood loss, and percentage of hidden blood loss, knee swelling at 3 days after operation, and range of motion of knee at 2 weeks after operation were recorded and compared between the two groups. RESULTS: The operation time of the trial group was significantly longer than that of the control group ( t=14.013, P=0.000). The actual total blood loss, hidden blood loss, and percentage of hidden blood loss in the trial group were significantly lower than those in the control group ( P<0.05); there was no significant difference in the overt blood loss between the two groups ( t=-1.293, P=0.200). The knee swelling degree in the trial group was significantly slighter than that in the control group at 3 days after operation, and the range of motion of knee in the trial group was significantly better than that in the control group at 2 weeks after operation ( P<0.05). CONCLUSION: Pneumatic tourniquet can reduce the operation time of TKA significantly. However, it may increase the hidden blood loss and knee swelling, and negatively impact the recovery of knee function in the early postoperative stage of TKA.

Co-expression of pregnane X receptor and ATP-binding cassette sub-family B member 1 in peripheral blood: A prospective indicator for drug resistance prediction in non-small cell lung cancer.

The aim of the present study was to investigate the protein expression profiling of pregnane X receptor (PXR) and ATP-binding cassette sub-family B member 1 (ABCB1; also known as MDR1 or P-gp), present in the peripheral blood mononuclear cells (PBMCs) and cancerous tissues of cases of non-small cell lung cancer (NSCLC). Furthermore, the study aimed to assess the feasibility of predicting drug resistance through the medium of PBMCs. Of the subjects included in the study, 37 were histopathologically diagnosed with NSCLC and 17 were control patients without cancer. ThinPrep liquid-based smears with cytosine were applied in the examination of the PBMCs and proved quite effective in preserving the morphology and surface antigens of the lymphocytes. Measurements of expression levels in the PBMCs and cancerous tissues were obtained by immunohistochemical means. The results showed that, with the exception of the selective PXR expression in the normal lung tissues, the two types of proteins existed extensively throughout the PBMCs, normal tissues and tumors. Among the cancer patients, prior to chemotherapy, a significant rise in ABCB1 expression could be observed in the PBMCs, together with a similar rise in ABCB1 and PXR expression in the tumor specimens. Marked upregulation of the two proteins was detected in the PBMCs following 1 cycle of first-line chemotherapy. ABCB1 expression, correlated with PXR, persisted mostly in the PBMCs and tissue samples. When bound to and activated by ligands, PXR translocates from the cytoplasm to the nucleus of the cells. PXR subsequently binds to its DNA response elements as a heterodimer with the retinoid X receptor. A PXR translocation of moderate or low differentiation was identified in 3 cases of adenocarcinoma, which were co-expressing the two genes in the PBMCs prior to chemotherapy. During follow-up visits, tumor recurrence was observed within 3 months in 5 cases, which were characterized by PXR translocation. These findings indicate that the combined expression of PXR and ABCB1 in PBMCs may be used as a prospective indicator in diagnosis prior to histopathological diagnosis, and therefore may function as a novel biomarker for the prediction of drug resistance.