Human mesenchymal stem cell therapy in severe COVID-19 patients: 2-year follow-up results of a randomized, double-blind, placebo-controlled trial.

BACKGROUND: Long-term effects of human mesenchymal stem cell (MSC) treatment on COVID-19 patients have not been fully characterized. The aim of this study was to evaluate the safety and efficacy of a MSC treatment administered to severe COVID-19 patients enrolled in our previous randomized, double-blind, placebo-controlled clinical trial (NCT04288102). METHODS: A total of 100 patients experiencing severe COVID-19 received either MSC treatment (n = 65, 4 × 107 cells per infusion) or a placebo (n = 35) combined with standard of care on days 0, 3, and 6. Patients were subsequently evaluated 18 and 24 months after treatment to evaluate the long-term safety and efficacy of the MSC treatment. Outcomes measured included: 6-min walking distance (6-MWD), lung imaging, quality of life according to the Short Form 36 questionnaire (SF-36), COVID-19-related symptoms, titers of SARS-CoV-2 neutralizing antibodies, tumor markers, and MSC-related adverse events (AEs). FINDINGS: Two years after treatment, a marginally smaller proportion of patients had a 6-MWD below the lower limit of the normal range in the MSC group than in the placebo group (OR = 0.19, 95% CI: 0.04-0.80, Fisher's exact test, p = 0.015). At month 18, the general health score from the SF-36 was higher in the MSC group than in the placebo group (50.00 vs. 35.00, 95% CI: 0.00-20.00, Wilcoxon rank sum test, p = 0.018). Total severity score of lung imaging and the titer of neutralizing antibodies were similar between the two groups at months 18 and 24. There was no difference in AEs or tumor markers at the 2-year follow-up between the two groups. INTERPRETATION: Long-term safety was observed for the COVID-19 patients who received MSC treatment. However, efficacy of MSC treatment was not significantly sustained through the end of the 2-year follow-up period. FUNDING: The National Key Research and Development Program of China (2022YFA1105604, 2020YFC0860900, 2022YFC2304401), the specific research fund of The Innovation Platform for Academicians of Hainan Province (YSPTZX202216) and the Fund of National Clinical Center for Infectious Diseases, PLA General Hospital (NCRC-ID202105,413FZT6).

Carryover Contamination-Controlled Amplicon Sequencing Workflow for Accurate Qualitative and Quantitative Detection of Pathogens: a Case Study on SARS-CoV-2.

Carryover contamination during amplicon sequencing workflow (AMP-Seq) put the accuracy of the high-throughput detection for pathogens at risk. The purpose of this study is to develop a carryover contaminations-controlled AMP-Seq (ccAMP-Seq) workflow to enable accurate qualitative and quantitative detection for pathogens. By using the AMP-Seq workflow to detect SARS-CoV-2, Aerosols, reagents and pipettes were identified as potential sources of contaminations and ccAMP-Seq was then developed. ccAMP-Seq used filter tips and physically isolation of experimental steps to avoid cross contamination, synthetic DNA spike-ins to compete with contaminations and quantify SARS-CoV-2, dUTP/uracil DNA glycosylase system to digest the carryover contaminations, and a new data analysis procedure to remove the sequencing reads from contaminations. Compared to AMP-Seq, the contamination level of ccAMP-Seq was at least 22-folds lower and the detection limit was also about an order of magnitude lower-as low as one copy/reaction. By testing the dilution series of SARS-CoV-2 nucleic acid standard, ccAMP-Seq showed 100% sensitivity and specificity. The high sensitivity of ccAMP-Seq was further confirmed by the detection of SARS-CoV-2 from 62 clinical samples. The consistency between qPCR and ccAMP-Seq was 100% for all the 53 qPCR-positive clinical samples. Seven qPCR-negative clinical samples were found to be positive by ccAMP-Seq, which was confirmed by extra qPCR tests on subsequent samples from the same patients. This study presents a carryover contamination-controlled, accurate qualitative and quantitative amplicon sequencing workflow that addresses the critical problem of pathogen detection for infectious diseases. IMPORTANCE Accuracy, a key indicator of pathogen detection technology, is compromised by carryover contamination in the amplicon sequencing workflow. Taking the detection of SARS-CoV-2 as case, this study presents a new carryover contamination-controlled amplicon sequencing workflow. The new workflow significantly reduces the degree of contamination in the workflow, thereby significantly improving the accuracy and sensitivity of the SARS-CoV-2 detection and empowering the ability of quantitative detection. More importantly, the use of the new workflow is simple and economical. Therefore, the results of this study can be easily applied to other microorganism, which has great significance for improving the detection level of microorganism.

The relationship between intolerance of uncertainty, coping style, resilience, and anxiety during the COVID-19 relapse in freshmen: A moderated mediation model.

Background: The repeated outbreaks of COVID-19 and the rapid increase in uncertainty have had many negative effects on the public's mental health, especially on emotional aspects such as anxiety and depression. However, in previous studies, there are few studies exploring the positive factors between uncertainty and anxiety. The innovation of this study is the first to explore the mechanism of coping style and resilience as people's psychological protective factors between uncertainty and anxiety caused by the COVID-19 pandemic. Methods: This study explored the relationship between intolerance of uncertainty and anxiety of freshmen with coping style as mediating variable and resilience as moderating variable. A total of 1049 freshmen participated in the study and completed the Intolerance of Uncertainty Scale (IUS-12), Self-rating Anxiety Scale (SAS), Simplified Coping Style Questionnaire (SCSQ), and Connor-Davidson Resilience Scale (CD-RISC). Results: SAS score of the surveyed students (39.56 ± 10.195) was significantly higher than that of the Normal Chinese score (29.78 ± 10.07, p < 0.001). Intolerance of uncertainty was significantly positively correlated with anxiety (ß = 0.493, p < 0.001). Positive coping style has a significant negative impact on anxiety (ß = -0.610, p < 0.001), negative coping style has a significant positive impact on anxiety (ß = 0.951, p < 0.001). Resilience moderates the second half of the influence of negative coping style on anxiety (ß = 0.011, t = 3.701, p < 0.01). Conclusion: The findings suggest that high levels of intolerance of uncertainty had negative effects mental burden during the COVID-19 pandemic. The knowledge of the mediating role of coping style and the moderating role of resilience may be used by health care workers when consulting freshmen with physical health complaints and psychosomatic disorders.

Rumination and "hot" executive function of middle school students during the COVID-19 pandemic: A moderated mediation model of depression and mindfulness.

Background: The outbreak of COVID-19 had a widely negative effect on adolescents' academics, stress, and mental health. At a critical period of cortical development, adolescents' cognition levels are highly developed, while the ability of emotion control is not developed at the same pace. Faced with negative emotions such as stress and social loneliness caused by COVID-19, adolescents' "hot" executive function encounters severer emotional regulation challenges than ever before. Objective: The present study established a moderated mediation model to investigate the impact of rumination on "hot" execution function among Chinese middle school students during the COVID-19 pandemic, and the specific role of depression and mindfulness in the association. Materials and methods: This cross-sectional study was conducted on 650 students recruited from a province in central China. The participants completed questionnaires and experiment between July 2021 and August 2021. Rumination Responses Scales, Self-rating Depression Scale, and Mindful Attention Awareness Scale were used to measure the level of rumination, depression, and mindfulness. The reaction time and accuracy of the emotional conflict experiment were recorded to reflect the "hot" executive function. Results: The results of the moderated mediation model indicated that rumination of middle school students significantly and positively predicted depression in adolescents (ß = 0.26, p < 0.001). Meanwhile, the indirect effect of depression on the relationship between rumination and "hot" executive function was significant; depression partially mediated this relationship (word-face congruent condition: ß = -0.09, p < 0.01; word-face incongruent condition: ß = -0.07, p < 0.05). Furthermore, mindfulness buffered the association between rumination and depression, according to moderated mediation analysis (ß = -0.11, p < 0.001). For adolescents with low levels of mindfulness, the relationship was substantially stronger. Conclusion: In the context of the COVID-19 pandemic, middle school students' rumination would lead to depression, which can negatively impact their "hot" executive function. Besides, mindfulness could resist the adverse effect of rumination on depression. The educators should pay more attention to students' mental health, provide targeted strategies that boost mindfulness to promote their cognitive flexibility, and thus protect the normal development of their executive function during crisis events.

Human mesenchymal stem cells treatment for severe COVID-19: 1-year follow-up results of a randomized, double-blind, placebo-controlled trial.

BACKGROUND: The long-term consequences of human umbilical cord-derived mesenchymal stem cell (UC-MSC) treatment for COVID-19 patients are yet to be reported. This study assessed the 1-year outcomes in patients with severe COVID-19, who were recruited in our previous UC-MSC clinical trial. METHODS: In this prospective, longitudinal, cohort study, 100 patients enrolled in our phase 2 trial were prospectively followed up at 3-month intervals for 1 year to evaluate the long-term safety and effectiveness of UC-MSC treatment. The primary endpoint was an altered proportion of whole-lung lesion volumes measured by high-resolution CT. Other imaging outcomes, 6 min walking distance (6-MWD), lung function, plasma biomarkers, and adverse events were also recorded and analyzed. This trial was registered with ClinicalTrials.gov (NCT04288102). FINDINGS: MSC administration improved in whole-lung lesion volume compared with the placebo with a difference of -10.8% (95% CI: -20.7%, -1.5%, p = 0.030) on day 10. MSC also reduced the proportion of solid component lesion volume compared with the placebo at each follow-up point. More interestingly, 17.9% (10/56) of patients in the MSC group had normal CT images at month 12, but none in the placebo group (p = 0.013). The incidence of symptoms was lower in the MSC group than in the placebo group at each follow-up time. Neutralizing antibodies were all positive, with a similar median inhibition rate (61.6% vs. 67.6%) in both groups at month 12. No difference in adverse events at the 1-year follow-up and tumor markers at month 12 were observed between the two groups. INTERPRETATION: UC-MSC administration achieves a long-term benefit in the recovery of lung lesions and symptoms in COVID-19 patients. FUNDING: The National Key R&D Program of China, the Innovation Groups of the National Natural Science Foundation of China, and the National Science and Technology Major Project.

Psychological experience of patients with confirmed COVID-19 at the initial stage of pandemic in Wuhan, China: a qualitative study.

BACKGROUND: Coronavirus disease 2019 (COVID-19) quickly developed into a global pandemic and affected patients' mental health. However, little is known about psychological experience of patients with COVID-19. The aim was to elucidate the psychological experience of patients with confirmed COVID-19 in Wuhan, at the initial stage of the pandemic. METHODS: This study was conducted using a phenomenological approach in a qualitative study. Thirteen patients with confirmed COVID-19 from a COVID-19-designated hospital in Wuhan, were recruited between March 15th and April 20th, 2020 via purposive sampling. Semi-structured in-depth interviews were conducted face-to-face. The interview data were analyzed using inductive thematic analysis. RESULTS: The psychological experience of patients was summarized into three themes: mental distress related to COVID-19, expectations of life scenarios after discharge, and making sense of the experience. These themes were classified into 10 sub-themes. Patients experienced confusion, uncertainty, worry, guilt and concern. Both positive and negative expectations of life scenarios after discharge were reported, manifested as expectations about making up for lost time with family, anxiety about social discrimination and feelings of helplessness about poor financial security. Moreover, patients perceived strength of abundant social support and awareness of social responsibility from their unique experience to cope with their condition. CONCLUSIONS: This study demonstrated that patients with confirmed COVID-19 in Wuhan underwent complex psychological experience, both positive and negative at the initial stage of the pandemic. These findings will contribute to the delivery of effective mental health care to safeguard patients' wellbeing.

Mesenchymal stem cell therapy for severe COVID-19.

The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has placed a global public burden on health authorities. Although the virological characteristics and pathogenesis of COVID-19 has been largely clarified, there is currently no specific therapeutic measure. In severe cases, acute SARS-CoV-2 infection leads to immune disorders and damage to both the adaptive and innate immune responses. Having roles in immune regulation and regeneration, mesenchymal stem cells (MSCs) serving as a therapeutic option may regulate the over-activated inflammatory response and promote recovery of lung damage. Since the outbreak of the COVID-19 pandemic, a series of MSC-therapy clinical trials has been conducted. The findings indicate that MSC treatment not only significantly reduces lung damage, but also improves patient recovery with safety and good immune tolerance. Herein, we summarize the recent progress in MSC therapy for COVID-19 and highlight the challenges in the field.

Abnormal Coagulation Function of Patients With COVID-19 Is Significantly Related to Hypocalcemia and Severe Inflammation.

This study aimed to detect, analyze, and correlate the clinical characteristics, blood coagulation functions, blood calcium levels, and inflammatory factors in patients with mild and severe COVID-19 infections. The enrolled COVID-19 infected patients were from Wuhan Jin Yin-tan Hospital (17 cases, Wuhan, China), Suzhou Infectious Disease Hospital (87 cases, Suzhou, China), and Xuzhou Infectious Disease Hospital (14 cases, Xuzhou, China). After admission, basic information was collected; X-ray and chest CT images were obtained; and data from routine blood tests, liver and kidney function, myocardial enzymes, electrolytes, blood coagulation function, (erythrocyte sedimentation rate) ESR, C-reactive protein (CRP), IL-6, procalcitonin (PCT), calcitonin, and other laboratory tests were obtained. The patients were grouped according to the clinical classification method based on the pneumonia diagnosis and treatment plan for new coronavirus infection (trial version 7) in China. The measurements from mild (56 cases) and severe cases (51 cases) were compared and analyzed. Most COVID-19 patients presented with fever. Chest X-ray and CT images showed multiple patchy and ground glass opacities in the lungs of COVID 19 infected patients, especially in patients with severe cases. Compared with patients with mild infection, patients with severe infection were older (p = 0.023) and had a significant increase in AST and BUN. The levels of CK, LDH, CK-MB, proBNP, and Myo in patients with severe COVID-19 infection were also increased significantly compared to those in patients with mild cases. Patients with severe COVID-19 infections presented coagulation dysfunction and increased D-dimer and fibrin degradation product (FDP) levels. Severe COVID-19 patients had low serum calcium ion (Ca2+) concentrations and high calcitonin and PCT levels and exhibited serious systemic inflammation. Ca2+ in COVID-19 patients was significantly negatively correlated with PCT, calcitonin, D-dimer, PFDP, ESR, CRP and IL-6. D-dimer in COVID-19 patients was a significantly positively correlated with CRP and IL-6. In conclusion, patients with severe COVID-19 infection presented significant metabolic dysfunction and abnormal blood coagulation, a sharp increase in inflammatory factors and calcitonin and procalcitonin levels, and a significant decrease in Ca2+. Decreased Ca2+ and coagulation dysfunction in COVID-19 patients were significantly correlated with each other and with inflammatory factors.

How Does Digital Competence Preserve University Students' Psychological Well-Being During the Pandemic? An Investigation From Self-Determined Theory.

This study conceptualized digital competence in line with self-determined theory (SDT) and investigated how it alongside help-seeking and learning agency collectively preserved university students' psychological well-being by assisting them to manage cognitive load and academic burnout, as well as increasing their engagement in online learning during the coronavirus disease 2019 (COVID-19) pandemic. Moreover, students' socioeconomic status and demographic variables were examined. Partial least square modeling and cluster analysis were performed on the survey data collected from 695 students. The findings show that mental load and mental effort were positively related to academic burnout, which was significantly negatively associated with student engagement in online learning. Digital competence did not directly affect academic burnout, but indirectly via its counteracting effect on cognitive load. However, help-seeking and agency were not found to be significantly negatively related to cognitive load. Among the three SDT constructs, digital competence demonstrated the greatest positive influence on student engagement. In addition, female students from humanities and social sciences disciplines and lower-income families seemed to demonstrate the weakest digital competence, lowest learning agency, and least help-seeking behaviors. Consequently, they were more vulnerable to high cognitive load and academic burnout, leading to the lowest learning engagement. This study contributes to the ongoing arguments related to the psychological impact of the COVID-19 pandemic and informs the development of efficient interventions that preserve university students' psychological well-being in online learning.

Epidemiological and clinical characteristics of coronavirus disease 2019 in non-epidemic areas: report of 57 cases

Objective: To investigate the epidemiological and clinical characteristics of coronavirus disease 2019 (COVID-19) in non-endemic areas.

Effective treatment of severe COVID-19 patients with tocilizumab.

After analyzing the immune characteristics of patients with severe coronavirus disease 2019 (COVID-19), we have identified that pathogenic T cells and inflammatory monocytes with large amount of interleukin 6 secreting may incite the inflammatory storm, which may potentially be curbed through monoclonal antibody that targets the IL-6 pathways. Here, we aimed to assess the efficacy of tocilizumab in severe patients with COVID-19 and seek a therapeutic strategy. The patients diagnosed as severe or critical COVID-19 in The First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Hospital) and Anhui Fuyang Second People's Hospital were given tocilizumab in addition to routine therapy between 5 and 14 February 2020. The changes of clinical manifestations, computerized tomography (CT) scan image, and laboratory examinations were retrospectively analyzed. Fever returned to normal on the first day, and other symptoms improved remarkably within a few days. Within 5 d after tocilizumab, 15 of the 20 patients (75.0%) had lowered their oxygen intake, and 1 patient needed no oxygen therapy. CT scans manifested that the lung lesion opacity absorbed in 19 patients (90.5%). The percentage of lymphocytes in peripheral blood, which decreased in 85.0% of patients (17/20) before treatment (mean, 15.52 ± 8.89%), returned to normal in 52.6% of patients (10/19) on the fifth day after treatment. Abnormally elevated C-reactive protein decreased significantly in 84.2% of patients (16/19). No obvious adverse reactions were observed. All patients have been discharged on average 15.1 d after giving tocilizumab. Preliminary data show that tocilizumab, which improved the clinical outcome immediately in severe and critical COVID-19 patients, is an effective treatment to reduce mortality.