Mir22hg facilitates ferritinophagy-mediated ferroptosis in sepsis by recruiting the m6A reader YTHDC1 and enhancing Angptl4 mRNA stability.

BACKGROUND: Ferritinophagy-mediated ferroptosis plays a crucial role in fighting pathogen aggression. The long non-coding RNA Mir22hg is involved in the regulation of ferroptosis and aberrantly overexpression in lipopolysaccharide (LPS)-induced sepsis mice, but whether it regulates sepsis through ferritinophagy-mediated ferroptosis is unclear. METHODS: Mir22hg was screened by bioinformatics analysis. Ferroptosis was assessed by assaying malondialdehyde (MDA), reactive oxygen species (ROS), and Fe2+ levels, glutathione (GSH) activity, as well as ferroptosis-related proteins GPX4 and SLC3A2 by using matched kits and performing western blot. Ferritinophagy was assessed by Lyso tracker staining and FerroOrange staining, immunofluorescence analysis of Ferritin and LC-3, and western blot analysis of LC-3II/I, p62, FTH1, and NCOA4. The bind of YTH domain containing 1 (YTHDC1) to Mir22hg or angiopoietin-like-4 (Angptl4) was verified by RNA pull-down and/or immunoprecipitation (RIP) assays. RESULTS: Mir22hg silencing lightened ferroptosis and ferritinophagy in LPS-induced MLE-12 cells and sepsis mouse models, as presented by the downregulated MDA, ROS, Fe2+, NCOA4, and SLC3A2 levels, upregulated GPX4, GSH, and FTH1 levels, along with a decrease in autophagy. Mir22hg could bind to the m6A reader YTHDC1 without affecting its expression. Mechanistically, Mir22hg enhanced Angptl4 mRNA stability through recruiting the m6A reader YTHDC1. Furthermore, Angptl4 overexpression partly overturned Mir22hg inhibition-mediated effects on ferroptosis and ferritinophagy in LPS-induced MLE-12 cells. CONCLUSION: Mir22hg contributed to in ferritinophagy-mediated ferroptosis in sepsis via recruiting the m6A reader YTHDC1 and strengthening Angptl4 mRNA stability, highlighting that Mir22hg may be a potential target for sepsis treatment based on ferroptosis.

TRPM7 mediates endoplasmic reticulum stress and ferroptosis in sepsis-induced myocardial injury.

Transient receptor potential melastatin 7 (TRPM7), a non-selective cation channel, was significantly upregulated in the blood of patients with sepsis. This study focuses on the preliminary exploration of the probable regulatory mechanism of TRPM7 in sepsis-induced myocardial injury (SIMI). HL-1 cardiac muscle cell line was treated with lipopolysaccharide (LPS) to mimic SIMI in vitro, and TRPM7 level was assessed. The impacts of TRPM7 knockdown on cellular inflammation response, oxidative stress, apoptosis, endoplasmic reticulum (ER) stress, and ferroptosis were identified. In order to explore the mechanism, ER stress agonist tunicamycin (TM) or ferroptosis inducer erastin was applied to treat HL-1 cells. The influences of TM and erastin on the aforementioned aspects were evaluated. TRPM7 was elevated in response to LPS stimulation, and its knockdown reduced the secretion of inflammatory factors and oxidative stress degree. Moreover, TRPM7 knockdown significantly suppressed cell apoptosis, ER stress, and ferroptosis. TM and erastin reversed the functions of TRPM7 knockdown, indicating ER stress and ferroptosis mediated in the regulation of TRPM7. This research proposes the possibility of TRPM7 as a marker or target for SIMI, and provides theoretical support for follow-up research.

Comparative global B cell receptor repertoire difference induced by SARS-CoV-2 infection or vaccination via single-cell V(D)J sequencing.

Dynamic changes of the paired heavy and light chain B cell receptor (BCR) repertoire provide an essential insight into understanding the humoral immune response post-SARS-CoV-2 infection and vaccination. However, differences between the endogenous paired BCR repertoire kinetics in SARS-CoV-2 infection and previously recovered/naïve subjects treated with the inactivated vaccine remain largely unknown. We performed single-cell V(D)J sequencing of B cells from six healthy donors with three shots of inactivated SARS-CoV-2 vaccine (BBIBP-CorV), five people who received the BBIBP-CorV vaccine after having recovered from COVID-19, five unvaccinated COVID-19 recovered patients and then integrated with public data of B cells from four SARS-CoV-2-infected subjects. We discovered that BCR variable (V) genes were more prominently used in the SARS-CoV-2 exposed groups (both in the group with active infection and in the group that had recovered) than in the vaccinated groups. The VH gene that expanded the most after SARS-CoV-2 infection was IGHV3-33, while IGHV3-23 in the vaccinated groups. SARS-CoV-2-infected group enhanced more BCR clonal expansion and somatic hypermutation than the vaccinated healthy group. A small proportion of public clonotypes were shared between the SARS-CoV-2 infected, vaccinated healthy, and recovered groups. Moreover, several public antibodies had been identified against SARS-CoV-2 spike protein. We comprehensively characterize the paired heavy and light chain BCR repertoire from SARS-CoV-2 infection to vaccination, providing further guidance for the development of the next-generation precision vaccine.

Depression, anxiety and post-traumatic growth among COVID-19 survivors six-month after discharge.

Background: Pre-hospitalisation, hospitalisation and post-hospitalisation factors may significantly affect depression, anxiety and post-traumatic growth (PTG) among COVID-19 survivors. Objective: Our study investigated depression, anxiety and PTG and their correlates among COVID-19 survivors. Method: A cross-sectional telephone survey recruited 199 COVID-19 patients (Mean age = 42.7; 53.3% females) at six-month follow-up after hospital discharge in five Chinese cities (i.e. Wuhan, Shenzhen, Zhuhai, Dongguan and Nanning). Their demographic information, clinical records and experiences during (e.g. severity of covid-19 symptoms, treatment and exposure to other patients' suffering) and after hospitalisation (e.g. perceived impact of covid-19, somatic symptoms after hospitalisation), and psychosocial factors (e.g. perceived discrimination, self-stigma, affiliate stigma, resilience and social support) were investigated. Depressive and anxiety symptoms were measured by the Patient Health Questionnaire (PHQ-9) and the Generalised anxiety disorder (GAD-7) scale, respectively. PTG was examined by the Post-traumatic Growth Inventory (PTGI) instrument. Results: The proportion of depressive symptoms <5, ≥5 and <10, ≥10 were 76.9%, 12.0% and 11.1%, respectively. The proportion of anxiety symptoms <5, ≥5 and <10, ≥10 were 77.4%, 15.1% and 7.5%, respectively. Multivariate logistic regression showed that receiving mental health care services during hospitalisation, somatic symptoms after discharge, perceived affiliate stigma and perceived impact of being infected with COVID-19 were significantly and positively associated with probable depression. Significant correlates of probable anxiety also included permanent residents of the city, somatic symptoms after discharge, perceived impact of being infected with COVID-19 and self-stigma. Social support, self-stigma and receiving mental health care services during hospitalisation were positively associated with PTG.Conclusions: The results suggest that post-hospitalisation and psychosocial factors had relatively stronger associations with depression, anxiety and PTG than pre-hospitalisation and hospitalisation factors. Promoting social support and social inclusion may be useful strategies to improve the mental health of COVID-19 survivors. HIGHLIGHTS: • Post-hospitalisation and psychosocial factors had relatively stronger associations with depression, anxiety and PTG than pre-hospitalisation and hospitalisation factors, promoting social support and social inclusion may be useful strategies to improve mental health of COVID-19 survivors.

Antecedentes: Los factores pre-hospitalización, durante la hospitalización y post-hospitalización pueden afectar significativamente la depresión, la ansiedad y el crecimiento postraumático (CPT) en los sobrevivientes de COVID-19.Objetivo: Nuestro estudio investigó la depresión, la ansiedad y el CPT y sus correlatos en sobrevivientes de COVID-19.Método: Una encuesta telefónica transversal reclutó a 199 pacientes con COVID-19 (edad promedio = 42,7; 53,3% mujeres) a los seis meses de seguimiento después del alta hospitalaria en cinco ciudades chinas (Wuhan, Shenzhen, Zhuhai, Dongguan y Nanning). Su información demográfica, registros clínicos y experiencias durante la hospitalización (e.g. gravedad de los síntomas de COVID-19, tratamiento, exposición al sufrimiento de otros pacientes) y después de la hospitalización (e.g. impacto percibido de COVID-19, síntomas somáticos después de la hospitalización) y factores psicosociales (e.g. discriminación percibida, autoestigma, estigma de afiliación, resiliencia, apoyo social) fueron investigados. Los síntomas depresivos y de ansiedad se midieron mediante el Cuestionario de Salud del Paciente (PHQ-9 en su sigla en inglés) y la escala de trastorno de ansiedad generalizada (GAD-7 en su sigla en inglés) respectivamente, el CPT se examinó mediante el instrumento Inventario de Crecimiento Postraumático (PTGI en su sigla en inglés).Resultados: La proporción de síntomas depresivos <5, ≥5 y <10, y ≥10 fue 76,9%, 12,0% y 11,1% respectivamente. La proporción de síntomas de ansiedad <5, ≥5 y <10, y ≥10 fue del 77,4%, 15,1% y 7,5% respectivamente. La regresión logística multivariante mostró que recibir servicios de atención de salud mental durante la hospitalización, los síntomas somáticos después del alta, el estigma de afiliación percibido y el impacto percibido de estar infectado con COVID-19 se asociaron significativa y positivamente con una probable depresión. Los correlatos significativos de ansiedad probable también incluyeron ser residente permanente de la ciudad, síntomas somáticos después del alta, impacto percibido de estar infectado con COVID-19 y autoestigma. El apoyo social, el autoestigma y recibir servicios de salud mental durante la hospitalización se asociaron positivamente con el CPT.Conclusiones: Los resultados sugieren que los factores psicosociales y posteriores a la hospitalización tuvieron asociaciones relativamente más fuertes con la depresión, la ansiedad y el CPT que los factores previos a la hospitalización y hospitalización. Promover el apoyo social y la inclusión social pueden ser estrategias útiles para mejorar la salud mental de los sobrevivientes de COVID-19.

Hospitalization, interpersonal and personal factors of social anxiety among COVID-19 survivors at the six-month follow-up after hospital treatment: the minority stress model.

Background: As a highly infectious disease with human-to-human transmission characteristics, COVID-19 has caused panic in the general public. Those who have recovered from COVID-19 may experience discrimination and internalized stigma. They may be more likely to worry about social interaction and develop social anxiety. Objectives: This study investigated the associations among hospitalization factors, social/interpersonal factors, personal factors, and social anxiety to reveal the mechanism of social anxiety in COVID-19 survivors. Methods: A cross-sectional, multicenter telephone survey was conducted from July to September 2020 in five Chinese cities (i.e. Wuhan, Nanning, Shenzhen, Zhuhai, and Dongguan); adult COVID-19 survivors were recruited 6 months after they were discharged from the hospital. Linear regressions and path analysis based on the minority stress model were conducted to test the relationships among hospitalization, social/interpersonal factors, personal factors, and social anxiety. Results: The response rate was 74.5% (N = 199, 55.3% females). Linear regression analyses showed that various hospitalization, social/interpersonal, and personal factors were statistically significantly associated with social anxiety. Path analysis showed that the proposed model fit the data well (χ2(df) = 3.196(3), p = .362, CFI = .999, NNFI = .996, RMSEA = .018). Internalized stigma fully mediated the association between perceived discrimination/social support and social anxiety, while it partially mediated the association between perceived affiliate stigma and social anxiety. Conclusions: The results suggest that social/interpersonal and personal factors have a stronger association with social anxiety than hospitalization factors and highlight the importance of internalized stigma in understanding the mechanisms of these relationships. Clinical psychologists can refer to these modifiable psychosocial factors to develop efficient interventions for mental health promotion.

Antecedentes: Como una enfermedad altamente infecciosa con características de transmisión de persona a persona, el COVID-19 ha causado pánico en el público en general. Aquellos que se han recuperado del COVID-19 pueden experimentar discriminación y estigma internalizado. Es más probable que se preocupen por la interacción social y desarrollen ansiedad social.Objetivos: Este estudio investigó las asociaciones entre factores de hospitalización, factores sociales /interpersonales, factores personales y ansiedad social para revelar el mecanismo de ansiedad social en sobrevivientes de COVID-19.Métodos: Se realizó una encuesta telefónica transversal multicentro de julio a septiembre de 2020 en cinco ciudades chinas (es decir, Wuhan, Nanning, Shenzhen, Zhuhai y Dongguan). Se reclutaron sobrevivientes adultos de COVID-19 seis meses después de ser dados de alta del hospital. Se realizaron regresiones lineales y análisis de ruta basados en el modelo de estrés de minoría para probar las relaciones entre la hospitalización, los factores sociales/interpersonales, los factores personales y la ansiedad social.Resultados: La tasa de respuesta fue del 74,5% (N = 199, 55,3% mujeres). Los análisis de regresión lineal mostraron que varios factores de hospitalización, sociales/interpersonales y personales se asociaron de manera estadísticamente significativa con la ansiedad social. El análisis de ruta mostró que el modelo propuesto se ajustaba bien a los datos (χ2 (df) = 3.196 (3), p = .362, CFI = .999, NNFI = .996, RMSEA = .018). El estigma internalizado medió completamente la asociación entre discriminación/apoyo social percibido y ansiedad social, mientras que medió parcialmente la asociación entre el estigma percibido de afiliados y ansiedad social.Conclusiones: Los resultados sugieren que los factores sociales/interpersonales y personales tienen una asociación más fuerte con la ansiedad social que los factores de hospitalización y resaltan la importancia del estigma internalizado en la comprensión de los mecanismos de estas relaciones. Los psicólogos clínicos pueden referirse a estos factores psicosociales modificables para desarrollar intervenciones eficientes para la promoción de la salud mental.

Associations between COVID-19 related stigma and sleep quality among COVID-19 survivors six months after hospital discharge.

BACKGROUND: Many COVID-19 survivors reported stigmatization after recovery. This study investigated the association between stigma (discrimination experiences, self-stigma and perceived affiliate stigma) and sleep quality among COVID-19 survivors six months after hospital discharge. METHODS: Participants were recovered adult COVID-19 survivors discharged between February 1 and April 30, 2020. Medical staff of five participating hospitals approached all discharged COVID-19 period during this period. A total of 199 participants completed the telephone interview during July to September, 2020. Structural equation modeling was performed to test the hypothesize that resilience and social support would mediate the associations between stigma and sleep quality. RESULTS: The results showed that 10.1% of the participants reported terrible/poor sleep quality, 26.1% reported worse sleep quality in the past week when comparing their current status versus the time before COVID-19. After adjusting for significant background characteristics, participants who had higher number of discrimination experience, perceived stronger self-stigma and stronger perceived affiliate stigma reported poorer sleep quality. Resilience and social support were positively and significantly associated with sleep quality. The indirect effect of self-stigma on sleep quality through social support and resilience was significant and negative. Perceived affiliate stigma also had a significant and negative indirect effect on sleep quality through social support and resilience. CONCLUSIONS: Various types of stigma after recovery were associated with poor sleep quality among COVID-19 survivors, while social support and resilience were protective factors. Resilience and social support mediated the associations between self-stigma/perceived affiliate stigma and sleep quality.

Pre-hospital, in-hospital and post-hospital factors associated with sleep quality among COVID-19 survivors 6 months after hospital discharge: cross-sectional survey in five cities in China.

Background: Understanding factors associated with post-discharge sleep quality among COVID-19 survivors is important for intervention development. Aims: This study investigated sleep quality and its correlates among COVID-19 patients 6 months after their most recent hospital discharge. Method: Healthcare providers at hospitals located in five different Chinese cities contacted adult COVID-19 patients discharged between 1 February and 30 March 2020. A total of 199 eligible patients provided verbal informed consent and completed the interview. Using score on the single-item Sleep Quality Scale as the dependent variable, multiple linear regression models were fitted. Results: Among all participants, 10.1% reported terrible or poor sleep quality, and 26.6% reported fair sleep quality, 26.1% reported worse sleep quality when comparing their current status with the time before COVID-19, and 33.7% were bothered by a sleeping disorder in the past 2 weeks. After adjusting for significant background characteristics, factors associated with sleep quality included witnessing the suffering (adjusted B = -1.15, 95% CI = -1.70, -0.33) or death (adjusted B = -1.55, 95% CI = -2.62, -0.49) of other COVID-19 patients during hospital stay, depressive symptoms (adjusted B = -0.26, 95% CI = -0.31, -0.20), anxiety symptoms (adjusted B = -0.25, 95% CI = -0.33, -0.17), post-traumatic stress disorders (adjusted B = -0.16, 95% CI = -0.22, -0.10) and social support (adjusted B = 0.07, 95% CI = 0.04, 0.10). Conclusions: COVID-19 survivors reported poor sleep quality. Interventions and support services to improve sleep quality should be provided to COVID-19 survivors during their hospital stay and after hospital discharge.

Research on Influencing Factors and Classification of Patients With Mild and Severe COVID-19 Symptoms.

Objective: To analyze the epidemiological history, clinical symptoms, laboratory testing parameters of patients with mild and severe COVID-19 infection, and provide a reference for timely judgment of changes in the patients' conditions and the formulation of epidemic prevention and control strategies. Methods: A retrospective study was conducted in this research, a total of 90 patients with COVID-19 infection who received treatment from January 21 to March 31, 2020 in the Ninth People's Hospital of Dongguan City were selected as study subject. We analyzed the clinical characteristics of laboratory-confirmed patients with COVID-19, used the oversampling method (SMOTE) to solve the imbalance of categories, and established Lasso-logistic regression and random forest models. Results: Among the 90 confirmed COVID-19 cases, 79 were mild and 11 were severe. The average age of the patients was 36.1 years old, including 49 males and 41 females. The average age of severe patients is significantly older than that of mild patients (53.2 years old vs 33.7 years old). The average time from illness onset to hospital admission was 4.1 days and the average actual hospital stay was 18.7 days, both of these time actors were longer for severe patients than for mild patients. Forty-eight of the 90 patients (53.3%) had family cluster infections, which was similar among mild and severe patients. Comorbidities of underlying diseases were more common in severe patients, including hypertension, diabetes and other diseases. The most common symptom was cough [45 (50%)], followed by fever [43 (47.8%)], headache [7 (7.8%)], vomiting [3 (3.3%)], diarrhea [3 (3.3%)], and dyspnea [1 (1.1%)]. The laboratory findings of patients also included leukopenia [13(14.4%)] and lymphopenia (17.8%). Severe patients had a low level of creatine kinase (median 40.9) and a high level of D-dimer. The median NLR of severe patients was 2.82, which was higher than that of mild patients. Logistic regression showed that age, phosphocreatine kinase, procalcitonin, the lymphocyte count of the patient on admission, cough, fatigue, and pharynx dryness were independent predictors of COVID-19 severity. The classification of random forest was predicted and the importance of each variable was displayed. The variable importance of random forest indicates that age, D-dimer, NLR (neutrophil to lymphocyte ratio) and other top-ranked variables are risk factors. Conclusion: The clinical symptoms of COVID-19 patients are non-specific and complicated. Age and the time from onset to admission are important factors that determine the severity of the patient's condition. Patients with mild illness should be closely monitored to identify those who may become severe. Variables such as age and creatine phosphate kinase selected by logistic regression can be used as important indicators to assess the disease severity of COVID-19 patients. The importance of variables in the random forest further complements the variable feature information.

Rapid isolation and immune profiling of SARS-CoV-2 specific memory B cell in convalescent COVID-19 patients via LIBRA-seq.

B cell response plays a critical role against SARS-CoV-2 infection. However, little is known about the diversity and frequency of the paired SARS-CoV-2 antigen-specific BCR repertoire after SARS-CoV-2 infection. Here, we performed single-cell RNA sequencing and VDJ sequencing using the memory and plasma B cells isolated from five convalescent COVID-19 patients, and analyzed the spectrum and transcriptional heterogeneity of antibody immune responses. Via linking BCR to antigen specificity through sequencing (LIBRA-seq), we identified a distinct activated memory B cell subgroup (CD11chigh CD95high) had a higher proportion of SARS-CoV-2 antigen-labeled cells compared with memory B cells. Our results revealed the diversity of paired BCR repertoire and the non-stochastic pairing of SARS-CoV-2 antigen-specific immunoglobulin heavy and light chains after SARS-CoV-2 infection. The public antibody clonotypes were shared by distinct convalescent individuals. Moreover, several antibodies isolated by LIBRA-seq showed high binding affinity against SARS-CoV-2 receptor-binding domain (RBD) or nucleoprotein (NP) via ELISA assay. Two RBD-reactive antibodies C14646P3S and C2767P3S isolated by LIBRA-seq exhibited high neutralizing activities against both pseudotyped and authentic SARS-CoV-2 viruses in vitro. Our study provides fundamental insights into B cell response following SARS-CoV-2 infection at the single-cell level.

Post-traumatic Stress Disorder Symptoms in COVID-19 Survivors 6 Months After Hospital Discharge: An Application of the Conservation of Resource Theory.

COVID-19 survivors who had acute respiratory symptoms might experience prolonged post-traumatic stress disorder (PTSD) due to further rehabilitation, somatic symptoms and related distress. The conservation of resource (COR) theory is a well-developed theory to understand how people develop PTSD symptoms in traumatic events. The current study aimed to examine the potential factors of PTSD symptoms and interrelationships among this factors among COVID-19 survivors based on the COR theory. This cross-sectional telephone survey enrolled 199 COVID-19 patients (Mean age = 42.7; 53.3% females) 6 months after their hospital discharge in five Chinese cities (i.e., Wuhan, Shenzhen, Zhuhai, Dongguan, and Nanning). The results showed that 7% of participants were classified as having probable PTSD. The significant potential factors relating to PTSD symptoms included socio-demographic status, hospitalization experiences, post-hospitalization experiences, and psychological status. Besides, the proposed statistical mediation model based on the COR framework showed good model fit, χ2(df) = 17.286 (5), p = 0.004, CFI = 0.962, NNFI = 0.951, RMSEA = 0.077. Perceived resource loss/gain fully mediated the association between exposure to other patients' suffering during hospitalization and PTSD symptoms, and partially mediated the relationships from somatic symptoms/perceived impact of being infected with COVID-19 after discharge to PTSD symptoms. On the other hand, resilience was a full mediator in the relationship from ICU experience to PTSD symptoms and a partial mediator in the relationship from perceived impact to PTSD symptoms. The results provide preliminary support on applying the COR theory to understand the factors of PTSD symptoms among COVID-19 survivors. Interventions to reduce PTSD symptoms in this population can be developed based on the modifiable psychosocial mediators.

Phylogeographic analyses strongly suggest cryptic speciation in the giant spiny frog (Dicroglossidae: Paa spinosa) and interspecies hybridization in Paa.

Species identification is one of the most basic yet crucial issues in biology with potentially far-reaching implications for fields such as conservation, population ecology, and epidemiology. The widely distributed but threatened frog Paa spinosa has been speculated to represent a complex of multiple species. In this study, 254 individuals representing species of the genus Paa were investigated along the entire range of P. spinosa: 196 specimens of P. spinosa, 8 specimens of P. jiulongensis, 5 specimens of P. boulengeri, 20 specimens of P. exilispinosa, and 25 specimens of P. shini. Approximately 1333 bp of mtDNA sequence data (genes 12S rRNA and 16S rRNA) were used. Phylogenetic analyses were conducted using maximum parsimony, maximum likelihood and Bayesian inference. BEAST was used to estimate divergence dates of major clades. Results suggest that P. spinosa can be divided into three distinct major lineages. Each major lineage totally corresponds to geographical regions, revealing the presence of three candidate cryptic species. Isolation and differentiation among lineages are further supported by the great genetic distances between the lineages. The bifurcating phylogenetic pattern also suggests an east-west dispersal trend during historic cryptic speciation. Dating analysis estimates that P. spinosa from Western China split from the remaining P. spinosa populations in the Miocene and that P. spinosa from Eastern China diverged from Central China in the Pliocene. We also found that P. exilispinosa from Mainland China and Hong Kong might have a complex of multiple species. After identifying cryptic lineages, we then determine the discrepancy between the mtDNA and the morphotypes in several individuals. This discrepancy may have been caused by introgressive hybridization between P. spinosa and P. shini.