Synthesis, structure-activity relationship and biological evaluation of indole derivatives as anti-Candida albicans agents.

In this work, we synthesized a series of indole derivatives to cope with the current increasing fungal infections caused by drug-resistant Candida albicans. All compounds were evaluated for antifungal activities against Candida albicans in vitro, and the structure-activity relationships (SARs) were analyzed. The results indicated that indole derivatives used either alone or in combination with fluconazole showed good activities against fluconazole-resistant Candida albicans. Further mechanisms studies demonstrated that compound 1 could inhibit yeast-to-hypha transition and biofilm formation of Candida albicans, increase the activity of the efflux pump, the damage of mitochondrial function, and the decrease of intracellular ATP content. In vivo studies, further proved the anti-Candida albicans activity of compound 1 by histological observation. Therefore, compound 1 could be considered as a novel antifungal agent.

Routine evaluation of HBV-specific T cell reactivity in chronic hepatitis B using a broad-spectrum T-cell epitope peptide library and ELISpot assay.

BACKGROUND: The clinical routine test of HBV-specific T cell reactivity is still limited due to the high polymorphisms of human leukocyte antigens (HLA) in patient cohort and the lack of universal detection kit, thus the clinical implication remains disputed. METHODS: A broad-spectrum peptide library, which consists of 103 functionally validated CD8+ T-cell epitopes spanning overall HBsAg, HBeAg, HBx and HBpol proteins and fits to the HLA polymorphisms of Chinese and Northeast Asian populations, was grouped into eight peptide pools and was used to establish an ELISpot assay for enumerating the reactive HBV-specific T cells in PBMCs. Totally 294 HBV-infected patients including 203 ones with chronic hepatitis B (CHB), 13 ones in acute resolved stage (R), 52 ones with liver cirrhosis (LC) and 26 ones with hepatocellular carcinoma (HCC) were detected, and 33 CHB patients were longitudinally monitored for 3 times with an interval of 3-5 months. RESULTS: The numbers of reactive HBV-specific T cells were significantly correlated with ALT level, HBsAg level, and disease stage (R, CHB, LC and HCC), and R patients displayed the strongest HBV-specific T cell reactivity while CHB patients showed the weakest one. For 203 CHB patients, the numbers of reactive HBV-specific T cells presented a significantly declined trend when the serum viral DNA load, HBsAg, HBeAg or ALT level gradually increased, but only a very low negative correlation coefficient was defined (r = - 0.21, - 0.21, - 0.27, - 0.079, respectively). Different Nucleotide Analogs (NUCs) did not bring difference on HBV-specific T cell reactivity in the same duration of treatment. NUCs/pegIFN-α combination led to much more reactive HBV-specific T cells than NUCs monotherapy. The dynamic numbers of reactive HBV-specific T cells were obviously increasing in most CHB patients undergoing routine treatment, and the longitudinal trend possess a high predictive power for the hepatitis progression 6 or 12 months later. CONCLUSION: The presented method could be developed into an efficient reference method for the clinical evaluation of cellular immunity. The CHB patients presenting low reactivity of HBV-specific T cells have a worse prognosis for hepatitis progression and should be treated using pegIFN-α to improve host T-cell immunity.

Identification and exploration of aging-related subtypes and distinctive role of SERPINE1 in heart failure based on single-cell and bulk RNA sequencing data.

Aging is a major risk factor for heart failure (HF) and is the leading cause of death worldwide. Currently, the nature of the relationship between aging and HF is not entirely clear. Herein, this study aimed to explore new diagnostic biomarkers, molecular typing and therapeutic strategies for HF by investigating the biological significance of aging-related genes in HF. A total of 157 differentially expressed genes (DEGs) were screened totally between HF and normal samples, and functional enrichment analysis of DEGs revealed the strong association of HF progression with aging, immune processes and metabolism. Six HF-specific aging-related genes were further identified, and a diagnostic model was developed and validated for good diagnostic efficacy. In addition, we collected blood samples from 10 normal controls and 10 HF patients for RT-qPCR analysis to verify the bioinformation. We also identified two aging-associated subtypes with distinctly different immune infiltration and metabolic microenvironment. Further single-cell sequencing analysis conducted in the study identified SERPINE1 as a key gene in HF. The distinctive role of SERPINE1 fibroblasts was revealed, including three main findings: (I) fibroblasts had a higher proportion and expression of SERPINE1 levels in HF; (II) the ligand-receptor pair MDK-LRP1 made the most contributions in high interactions with other cell types in SERPINE1 fibroblasts; and (III) SERPINE1 fibroblasts were associated with the interaction of extracellular matrix and receptor and may be regulated by the transcription factor EGR1. In conclusion, this study highlights the importance of aging-related genes in diagnosing HF and regulating immune infiltration. We also identified different HF subtypes and a potentially crucial gene, which may provide a better understanding of the molecular-level mechanisms of aging-related HF and aid in developing effective therapeutic strategies.

Chalcones from plants cause toxicity by inhibiting human and rat 11ß-hydroxysteroid dehydrogenase 2: 3D-quantitative structure-activity relationship (3D-QSAR) and in silico docking analysis.

Chalcones from licorice and its related plants have many pharmacological effects. However, the effects of chalcones on the activity of human and rat 11ß-hydroxysteroid dehydrogenase 2 (11ß-HSD2), and associated side effects remain unclear. The inhibition of 11 chalcones on human and rat 11ß-HSD2 were evaluated in microsomes and a 3D-quantitative structure-activity relationship (3D-QSAR) was analyzed. Screening revealed that bavachalcone, echinatin, isobavachalcone, isobavachromene, isoliquiritigenin, licochalcone A, and licochalcone B significantly inhibited human 11ß-HSD2 with IC50 values ranging from 15.62 (licochalcone A) to 38.33 (echinatin) µM. Screening showed that the above chemicals and 4-hydroxychalcone significantly inhibited rat 11ß-HSD2 with IC50 values ranging from 6.82 (isobavachalcone) to 72.26 (4-hydroxychalcone) µM. These chalcones acted as noncompetitive/mixed inhibitors for both enzymes. Comparative analysis revealed that inhibition of 11ß-HSD2 depended on the species. Most chemicals bind to the NAD+ binding site or both the NAD+ and substrate binding sites. Bivariate correlation analysis showed that lipophilicity and molecular weight determine inhibitory strength. Through our 3D-QSAR models, we identified that the hydrophobic region, hydrophobic aliphatic groups, and hydrogen bond acceptors are pivotal factors in inhibiting 11ß-HSD2. In conclusion, many chalcones inhibit human and rat 11ß-HSD2, possibly causing side effects and there is structure-dependent and species-dependent inhibition on 11ß-HSD2.

Estimating residual undifferentiated cells in human chemically induced pluripotent stem cell derived islets using lncRNA as biomarkers.

Human pluripotent stem cells (hPSCs) can generate insulin-producing beta cells for diabetes treatment, but residual undifferentiated cells may cause tumors. We developed a highly sensitive assay to detect these cells in islet cells derived from human chemically induced pluripotent stem cells (hCiPSCs), which are transgene-free and safer. We used RNA-seq data to find protein-coding and non-coding RNAs that were only expressed in hCiPSCs, not in islet cells. We confirmed these biomarkers by RT-qPCR and ddPCR. We chose long non-coding RNA (lncRNA) markers, which performed better than protein-coding RNA markers. We found that LNCPRESS2, LINC00678 and LOC105370482 could detect 1, 1 and 3 hCiPSCs in 106 islet cells by ddPCR, respectively. We tested our method on several hCiPSC lines, which could quantify 0.0001% undifferentiated cell in 106 islet cells by targeting hCiPSCs-specific lncRNA transcripts, ensuring the safety and quality of hCiPSC-derived islet cells for clinical use.

Screening of Efficient Adjuvants for the RBD-Based Subunit Vaccine of SARS-CoV-2.

The variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are more transmissible, with a reduced sensitivity to vaccines targeting the original virus strain. Therefore, developing an effective vaccine against both the original SARS-CoV-2 strain and its variants is an urgent need. It is known that the receptor-binding domain (RBD) in the S protein of SARS-CoV-2 is an important vaccine target, but subunit vaccines usually have lower immunogenicity and efficacy. Thus, selecting appropriate adjuvants to enhance the immunogenicity of protein-based subunit vaccine antigens is necessary. Here, an RBD-Fc subunit vaccine of SARS-CoV-2 has been generated, followed by vaccination in B6 mice, and four adjuvant regimens were investigated, including aluminum salts (Alum) + 3-O-desacyl-4'-monophosphoryl lipid A (MPL), AddaVax, QS21 + MPL, and Imiquimod. The adjuvant potency was evaluated by comparing the elicited polyclonal antibodies titers with measuring binding to RBD and S protein in ELISA and Western blot analysis, and also the cross-neutralizing antibodies titers using a pseudovirus infection assay of hACE2-expressing 293T cells, with pseudoviruses expressing the S protein of the SARS-CoV-2 original strain and Delta strain. The presence of QS21 + MPL adjuvant induced stronger polyclonal antibody response and neutralization potency blocking the original strain and Delta strain, as compared with the non-adjuvant RBD-Fc group and other adjuvant groups. Meanwhile, Imiquimod even had a negative effect in inducing specific antibodies and cross-neutralizing antibody production as an adjuvant.

Environmental Correlates of Physical Activity and Screen-Time in Youth with Autism Spectrum Disorder: A Seven-Country Observational Study.

This cross-sectional observational study sought to examine the environmental correlates of physical activity and screen-time among youth with autism spectrum disorder (ASD). Parents of youth with ASD (n = 1,165) from seven countries/regions provided responses to an online survey form measuring environmental correlates (i.e., physical activity neighborhood environment, social network, social trust and cohesion, bedroom media, social home environment) and outcomes (i.e., physical activity, screen-time). Multiple linear regression analyses were conducted to determine environmental predictors of the outcomes. Physical activity neighborhood environment (B = 0.15, p = 0.047), social network (B = 0.16, p = 0.02), and social home environment (B = 1.07, p < 0.001) were significantly associated with physical activity, whereas social trust and cohesion and bedroom media were not. Further, social trust and cohesion (B = -0.14, p = 0.001), bedroom media (B = 0.10, p = 0.001), and social home environment (B = -0.16, p < 0.001) were significantly associated with screen-time while neighborhood environment and social network were not. The identified environmental attributes of physical activity and screen-time behaviors should be targeted for health promotion among youth with ASD.

Exosomal Vaccine Loading T Cell Epitope Peptides of SARS-CoV-2 Induces Robust CD8+ T Cell Response in HLA-A Transgenic Mice.

Purpose: Current vaccines for the SARS-CoV-2 virus mainly induce neutralizing antibodies but overlook the T cell responses. This study aims to generate an exosomal vaccine carrying T cell epitope peptides of SARS-CoV-2 for the induction of CD8+ T cell response. Methods: Thirty-one peptides presented by HLA-A0201 molecule were conjugated to the DMPE-PEG-NHS molecules, and mixed with DSPE-PEG to form the peptide-PEG-lipid micelles, then fused with exosomes to generate the exosomal vaccine, followed by purification using size-exclusion chromatography and validation by Western blotting, liquid nuclear magnetic resonance (NMR) test and transmission electron microscopy. Furthermore, the exosomal vaccine was mixed with Poly (I:C) adjuvant and subcutaneously administered for three times into the hybrid mice of HLA-A0201/DR1 transgenic mice with wild-type mice. Then, the epitope-specific T cell responses were detected by ex vivo ELISPOT assay and intracellular cytokine staining. Results: The exosomal vaccine was purified from the Peak 2 fraction of FPLC and injected into the hybrid mice for three times. The IFN-γ spot forming units and the frequencies of IFN-γ+/CD8+ T cells were 10-82-fold and 13-65-fold, respectively, higher in the exosomal vaccine group compared to the Poly (I:C) control group, without visible organ toxicity. In comparison with the peptides cocktail vaccine generated in our recent work, the exosomal vaccine induced significantly stronger T cell response. Conclusion: Exosomal vaccine loading T cell epitope peptides of SARS-CoV-2 virus was initially generated without pre-modification for both peptides and exosomes, and elicited robust CD8+ T cell response in HLA-A transgenic mice.

A socio-ecological approach to inclusive physical education in China: A systematic review.

Background: Since the implementation of inclusive education in China, students with special education needs (SEN) have increasingly been integrating into mainstream schools, like physical education classes. However, inclusive physical education (IPE) in China has developed slowly, and gaps can be found in the knowledge of the factors that inhibit or promote the participation in IPE of students with SEN. Objectives: The purpose of this systematic review was to provide a comprehensive summary of the factors related to inclusion in IPE of students with SEN, by applying a socio-ecological model (SEM). Five databases were searched: ERIC, SPORTDiscus with Full Text, Education Full Text (H.W.Wilson), PsychINFO and CNKI in March 2022, to find studies that identify factors regarding IPE in China. Two researchers independently screened studies and summarized relevant data. Results: Fourteen studies were included in the detailed review. By applying the SEM, multi-level factors were identified, ranging from intrapersonal to societal levels that positively or negatively influenced IPE participation in students with SEN. This review indicates that multi-level factors affect the IPE participation of students with SEN in China. Conclusion: The findings will help assist educators and policymakers to develop effective IPE for Chinese students with SEN.

Observation of Anesthetic Effect of Dexmedetomidine Combined With Intraspinal Anesthesia in Hip Arthroplasty and its Effect on Postoperative Delirium and Stress Response.

Objective: To observe the anesthetic effect of dexmedetomidine combined with spinal anesthesia in hip arthroplasty, and to analyze the effects of dexmedetomidine on postoperative stress response, incidence of delirium, immune function and inflammatory indicators. Methods: A total of 42 patients who underwent hip replacement in our hospital from March 2020 to June 2021 were selected as the research subjects and randomly divided into the control group and the observation group, 21 cases in each group. The control group was given intraspinal anesthesia, and the observation group was given dexmedetomidine on this basis. The onset time and maintenance time of sensory and motor nerve block were recorded. Stress response indexes [cortisol (Cor), blood glucose (Glu), adrenaline (E), noadrenaline (NE)], T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+), inflammatory indexes [tumor necrosis factor -α (TNF-α) and interleukin-6 (IL-6)] were detected before and after operation, and the incidence of postoperative delirium in both groups was recorded. Results: The onset time of sensory nerve block and motor block in the observation group were lower than those in the control group, and the retention time of sensory nerve block and motor nerve block were higher than those in the control group (P < 0.05). After surgery, the levels of Cor, Glu, E and NE in the observation group were lower than those in the control group (P < 0.05). After surgery, the incidence of postoperative delirium in the observation group (4.79%) was lower than that in the control group (28.57%) (P < 0.05). After surgery, the levels of CD3+, CD4+, CD8+, and CD4+/CD8+ in the observation group were higher than those in the control group (P < 0.05). After surgery, the levels of TNF-α and IL-6 in the observation group were lower than those in the control group (P < 0.05). Conclusion: The combined use of dexmedetomidine and intraspinal anesthesia has good anesthesia effect in hip joint replacement, which can greatly reduce the stress response of patients, reduce the incidence of postoperative delirium, and effectively restore the immune function of patients, reduce the level of inflammatory response, and has high clinical application value.

Meeting the 24-h movement guidelines and health-related outcomes among youth with autism spectrum disorder: a seven-country observational study.

BACKGROUND: Meeting daily guidelines for physical activity, screen time, and sleep duration is associated with a host of health indicators for youth. In this cross-sectional observational study, we investigated the associations between adherence to the movement guidelines and health-related outcomes among youth with autism spectrum disorder (ASD). METHODS: Parents of youth with ASD (10-17 years) from seven countries and regions were invited to provide online proxy-reports for child's movement behaviors (i.e., physical activity, sleep and screen time), and health-related outcomes (i.e., body mass index [BMI], general health, and quality of life). A series of multiple linear regression analyses were used to examine the associations between meeting movement guidelines and health-related outcomes, adjusted for covariates. RESULTS: The final sample consisted of 1165 youth with ASD. Compared with youth meeting all three guidelines, a higher BMI z-score was observed in those who met no guidelines (B = 0.62, P = 0.04), "sedentary time only" (B = 0.60, P = 0.047), and "physical activity plus sleep only" (B = 0.85, P = 0.04). Compared with meeting all three guidelines, meeting no guidelines was associated with poorer general health (B = - 0.46, P = 0.02). Further, compared with youth meeting all three guidelines, a lower quality of life score was observed in those who met no guidelines (B = - 0.47, P = 0.02) and "physical activity only" (B = - 0.62, P = 0.03). Lastly, there were dose-response associations between the number of guidelines met and all three health-related outcomes (all Ptrend < 0.05). CONCLUSIONS: In conclusion, meeting more 24-h movement guidelines was generally associated with more favorable health-related outcomes in youth with ASD. The low level of adherence to all three guidelines (2.0%) suggests the urgent need to promote the adoption of all the guidelines in this group.

Synthesis and in vitro synergistic antifungal activity of analogues of Panax stipulcanatus saponin against fluconazole-resistant Candida albicans.

Oleanolic acid 3-O-ß-d-glucopyranosyl-(1 â†’ 3)-ß-d-glucopyranoside (1) and oleanolic acid 3-O-ß-d-glucopyranosyl-(1 â†’ 3)-[α-l-arabinofuranosyl-(1 â†’ 4)]-ß-d-glucopyranoside (2), novel Panax stipulcanatus saponin analogues, were synthesized for the first time starting from commercially available oleanolic acid, d-glucose, and L-(+)-arabinose. Glycosyl N-phenyltrifluoroacetimidates as donors and two-step consecutive glycosylation reactions are crucial in the synthesis. In vitro antifungal activity results indicated that analogue 2 combined with fluconazole showed synergistic antifungal activity against fluconazole-resistant Candida albicans, with MIC50 values 31.80 µg/mL and FICI values 0.32. We also found that intermediate compounds 16 and 17 revealed synergistic antifungal activity against susceptible Candida albicans when combined with fluconazole, with MIC50 values 1.43 µg/mL and 1.59 µg/mL, FICI values 0.29 and 0.32, respectively.

Screening and Identification of HBV Epitopes Restricted by Multiple Prevalent HLA-A Allotypes.

Although host T cell immune responses to hepatitis B virus (HBV) have been demonstrated to have important influences on the outcome of HBV infection, the development of T cell epitope-based vaccine and T cell therapy and the clinical evaluation of specific T cell function are currently hampered markedly by the lack of validated HBV T cell epitopes covering broad patients. This study aimed to screen T cell epitopes spanning overall HBsAg, HBeAg, HBx and HBpol proteins and presenting by thirteen prevalent human leukocyte antigen (HLA)-A allotypes which gather a total gene frequency of around 95% in China and Northeast Asia populations. 187 epitopes were in silico predicted. Of which, 62 epitopes were then functionally validated as real-world HBV T cell epitopes by ex vivo IFN-γ ELISPOT assay and in vitro co-cultures using peripheral blood mononuclear cells (PBMCs) from HBV infected patients. Furthermore, the HLA-A cross-restrictions of each epitope were identified by peptide competitive binding assay using transfected HMy2.CIR cell lines, and by HLA-A/peptide docking as well as molecular dynamic simulation. Finally, a peptide library containing 105 validated epitopes which cross-binding by 13 prevalent HLA-A allotypes were used in ELISPOT assay to enumerate HBV-specific T cells for 116 patients with HBV infection. The spot forming units (SFUs) was significantly correlated with serum HBsAg level as confirmed by multivariate linear regression analysis. This study functionally validated 62 T cell epitopes from HBV main proteins and elucidated their HLA-A restrictions and provided an alternative ELISPOT assay using validated epitope peptides rather than conventional overlapping peptides for the clinical evaluation of HBV-specific T cell responses.

A Systematic Review of T Cell Epitopes Defined from the Proteome of Hepatitis B Virus.

Hepatitis B virus (HBV) infection remains a worldwide health problem and no eradicative therapy is currently available. Host T cell immune responses have crucial influences on the outcome of HBV infection, however the development of therapeutic vaccines, T cell therapies and the clinical evaluation of HBV-specific T cell responses are hampered markedly by the lack of validated T cell epitopes. This review presented a map of T cell epitopes functionally validated from HBV antigens during the past 33 years; the human leukocyte antigen (HLA) supertypes to present these epitopes, and the methods to screen and identify T cell epitopes. To the best of our knowledge, a total of 205 CD8+ T cell epitopes and 79 CD4+ T cell epitopes have been defined from HBV antigens by cellular functional experiments thus far, but most are restricted to several common HLA supertypes, such as HLA-A0201, A2402, B0702, DR04, and DR12 molecules. Therefore, the currently defined T cell epitope repertoire cannot cover the major populations with HLA diversity in an indicated geographic region. More researches are needed to dissect a more comprehensive map of T cell epitopes, which covers overall HBV proteome and global patients.

Meeting the 24-Hour Movement Guidelines and Outcomes in Adolescents with ADHD: A Cross-Sectional Observational Study.

According to the 24-Hour Movement Guidelines, meeting daily recommendations for physical activity, sleep, and screen time is important for obtaining optimal health benefits. This cross-sectional observational study aimed to examine (a) the prevalence of meeting the movement guidelines; and (b) the associations between meeting the guidelines and selected outcomes in adolescents with attention-deficit/hyperactivity disorder (ADHD). Data from the 2018-2019 National Survey for Children's Health dataset was used. Participants were adolescents (10-17 years) with ADHD and without other chronic conditions. Outcomes were flourishing, school engagement, and body weight status. Exposures of interest were adherence to the movement guidelines. The frequency of the participants' adherence to the guidelines was estimated, and regression analyses were conducted to examine the associations between adherence to the guidelines and outcomes, adjusting for potential confounders. Complete observations were available for 634 adolescents with ADHD. Overall, 46.8% of the participants met at least one movement guideline, but only 6.5% met all three. The number of guidelines met had a significant and positive association with flourishing and school engagement (ß = 0.21/0.17, ptrend < 0.001). Compared with meeting all three guidelines, significant associations with lower flourishing levels were found in participants who met none, sleep only, and sedentary time only (ß = -0.38--0.13, p < 0.05). Similar findings were identified in the school engagement outcome. Adherence to the guidelines was, however, not significantly associated with the odds of being overweight or obese. Collectively, the findings suggest the movement guidelines may be appropriate for extending to adolescents with ADHD and there is a need to increase adherence to the guidelines in this group.

Identification of HLA-A2 restricted epitopes of glypican-3 and induction of CTL responses in HLA-A2 transgenic mice.

Hepatocellular carcinoma (HCC) is a malignant tumor with high mortality, but lacks effective treatments. Carcinoembryonic antigen glypican-3 (GPC3) is a tumor-associated antigen overexpressed in HCC but rarely expressed in healthy individuals and thus is one of the most promising therapeutic targets. T cell epitope-based vaccines may bring light to HCC patients, especially to the patients at a late stage. However, few epitopes from GPC3 were identified to date, which limited the application of GPC3-derived epitopes in immunotherapy and T cell function detection. In this study, a total of 25 HLA-A0201 restricted GPC3 epitopes were in silico predicted and selected as candidate epitopes. Then, HLA-A0201+/GPC3+ HCC patients' PBMCs were collected and co-stimulated with the candidate epitope peptides in ex vivo IFN-γ Elispot assay, by which five epitopes were identified as real-world epitopes. Their capacity to elicit specific CD8+ T cells activation and proliferation was further confirmed by in vitro co-cultures of patients' PBMCs with peptide, in vitro co-cultures of healthy donors' PBLs with DCs and peptide, T2 cell binding assay as well as HLA-A2 molecule stability assay. Moreover, the in vivo immunogenicity of the five validated epitopes was confirmed by peptides cocktail/poly(I:C) vaccination in HLA-A0201/DR1 transgenic mice. Robust epitope-specific CD8+ T cell responses and cytotoxicity targeting HepG2 cells were observed as detected by IFN-γ Elispot, intracellular IFN-γ staining and cytolysis assay. This study provided novel GPC3 CTL epitopes for the development of T cell epitope vaccines and evaluation of GPC3 specific T cell responses.

Receptor-Binding Domain Proteins of SARS-CoV-2 Variants Elicited Robust Antibody Responses Cross-Reacting with Wild-Type and Mutant Viruses in Mice.

Multiple variants of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) have spread around the world, but the neutralizing effects of antibodies induced by the existing vaccines have declined, which highlights the importance of developing vaccines against mutant virus strains. In this study, nine receptor-binding domain (RBD) proteins of the SARS-CoV-2 variants (B.1.1.7, B.1.351 and P.1 lineages) were constructed and fused with the Fc fragment of human IgG (RBD-Fc). These RBD-Fc proteins contained single or multiple amino acid substitutions at prevalent mutation points of spike protein, which enabled them to bind strongly to the polyclonal antibodies specific for wild-type RBD and to the recombinant human ACE2 protein. In the BALB/c, mice were immunized with the wild-type RBD-Fc protein first and boosted twice with the indicated mutant RBD-Fc proteins later. All mutant RBD-Fc proteins elicited high-level IgG antibodies and cross-neutralizing antibodies. The RBD-Fc proteins with multiple substitutions tended to induce higher antibody titers and neutralizing-antibody titers than the single-mutant RBD-Fc proteins. Meanwhile, both wild-type RBD-Fc protein and mutant RBD-Fc proteins induced significantly decreased neutralization capacity to the pseudovirus of B.1.351 and P.1 lineages than to the wild-type one. These data will facilitate the design and development of RBD-based subunit vaccines against SARS-COV-2 and its variants.

Changes of recovery quality and early cognitive function after treatment of sevoflurane combined with propofol in the elderly undergoing cholecystectomy.

OBJECTIVE: To explore the application value of sevoflurane + propofol in elderly patients undergoing cholecystectomy. METHODS: A total of 121 elderly patients undergoing cholecystectomy in our hospital from February 2017 to March 2020 were enrolled. Among them, 58 patients were assigned to Group A given, anesthesia with sevoflurane during operation, and 63 patients to Group B who were given anesthesia with sevoflurane + propofol during the operation. The Mini-Mental State Examination (MMSE) was adopted to evaluate the cognitive function of the two groups at 1 hour (T1), 3 hours (T2) and 12 hours (T3) after operation, and enzyme-linked immuno-sorbent assay (ELISA) was used to determine inflammatory factors. The incidence of postoperative adverse reactions was compared between the two groups. RESULTS: The heart rate (HR) of patients at T2 was significantly lower than that at T1 and T3, and their HR at T3 was lower than that at T1 (P<0.05). There were differences in systolic blood pressure (SBP) and diastolic blood pressure (DBP) at different time points in each group (both P<0.001). The mean artistic pressure (MAP) of patients at T2 was significantly lower than that at T1 and T3, and their MAP at T3 was lower than that at T1 (P<0.05). Additionally, oxygen saturation (SpO2) of patients at T2 was also significantly lower than that at T1 and T3, and their SpO2 at T3 was lower than that at T1 (P<0.05). Moreover, Group B showed significantly lower levels of serum inflammatory factors than Group A at T2 and T3 (P<0.05), and also got greatly lower Observer Assessment of Sedation (OAA/S) scores than Group A (P<0.05). CONCLUSION: Sevoflurane + propofol can effectively improve the recovery quality and cognitive function and reduce inflammation after cholecystectomy in the elderly, so it is worthy of clinical promotion.

Screening HLA-A-restricted T cell epitopes of SARS-CoV-2 and the induction of CD8+ T cell responses in HLA-A transgenic mice.

Since severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-specific T cells have been found to play essential roles in host immune protection and pathology in patients with coronavirus disease 2019 (COVID-19), this study focused on the functional validation of T cell epitopes and the development of vaccines that induce specific T cell responses. A total of 120 CD8+ T cell epitopes from the E, M, N, S, and RdRp proteins were functionally validated. Among these, 110, 15, 6, 14, and 12 epitopes were highly homologous with SARS-CoV, OC43, NL63, HKU1, and 229E, respectively; in addition, four epitopes from the S protein displayed one amino acid that was distinct from the current SARS-CoV-2 variants. Then, 31 epitopes restricted by the HLA-A2 molecule were used to generate peptide cocktail vaccines in combination with Poly(I:C), R848 or poly (lactic-co-glycolic acid) nanoparticles, and these vaccines elicited robust and specific CD8+ T cell responses in HLA-A2/DR1 transgenic mice as well as wild-type mice. In contrast to previous research, this study established a modified DC-peptide-PBL cell coculture system using healthy donor PBMCs to validate the in silico predicted epitopes, provided an epitope library restricted by nine of the most prevalent HLA-A allotypes covering broad Asian populations, and identified the HLA-A restrictions of these validated epitopes using competitive peptide binding experiments with HMy2.CIR cell lines expressing the indicated HLA-A allotype, which initially confirmed the in vivo feasibility of 9- or 10-mer peptide cocktail vaccines against SARS-CoV-2. These data will facilitate the design and development of vaccines that induce antiviral CD8+ T cell responses in COVID-19 patients.

Intron-targeted mutagenesis reveals roles for Dscam1 RNA pairing architecture-driven splicing bias in neuronal wiring.

Drosophila melanogaster Down syndrome cell adhesion molecule (Dscam1) can generate 38,016 different isoforms through largely stochastic, yet highly biased, alternative splicing. These isoforms are required for nervous functions. However, the functional significance of splicing bias remains unknown. Here, we provide evidence that Dscam1 splicing bias is required for mushroom body (MB) axonal wiring. We generate mutant flies with normal overall protein levels and an identical number but global changes in exon 4 and 9 isoform bias (DscamΔ4D-/- and DscamΔ9D-/-), respectively. In contrast to DscamΔ4D-/-, DscamΔ9D-/- exhibits remarkable MB defects, suggesting a variable domain-specific requirement for isoform bias. Importantly, changes in isoform bias cause axonal defects but do not influence the self-avoidance of axonal branches. We conclude that, in contrast to the isoform number that provides the molecular basis for neurite self-avoidance, isoform bias may play a role in MB axonal wiring by influencing non-repulsive signaling.