Cordyceps: Alleviating ischemic cardiovascular and cerebrovascular injury - A comprehensive review.

ETHNOPHARMACOLOGICAL RELEVANCE: Cordyceps has a long medicinal history as a nourishing herb in traditional Chinese medicine (TCM). Ischemic cardio-cerebrovascular diseases (CCVDs), including cerebral ischemic/reperfusion injury (CI/RI) and myocardial ischemic/reperfusion injury (MI/RI), are major contributors to mortality and disability in humans. Numerous studies have indicated that Cordyceps or its artificial substitutes have significant bioactivity on ischemic CCVDs, however, there is a lack of relevant reviews. AIM OF THE STUDY: This review was conducted to investigate the chemical elements, pharmacological effects, clinical application and drug safety of Cordycepson ischemic CCVDs. MATERIALS AND METHODS: A comprehensive search was conducted on the Web of Science, PubMed, Chinese National Knowledge Infrastructure (CNKI), and Wanfang databases using the keywords "Cordyceps", "Cerebral ischemic/reperfusion injury", and "Myocardial ischemic/reperfusion injury" or their synonyms. The retrieved literature was then categorized and summarized. RESULTS: The study findings indicated that Cordyceps and its bioactive components, including adenosine, cordycepin, mannitol, polysaccharide, and protein, have the potential to protect against CI/RI and MI/RI by improving blood perfusion, mitigating damage from reactive oxygen species, suppressing inflammation, preventing cellular apoptosis, and promoting tissue regeneration. Individually, Cordyceps could reduce neuronal excitatory toxicity and blood-brain barrier damage caused by cerebral ischemia. It can also significantly improve cardiac energy metabolism disorders and inhibit calcium overload caused by myocardial ischemia. Additionally, Cordyceps exerts a significant preventive or curative influence on the factors responsible for heart/brain ischemia, including hypertension, thrombosis, atherosclerosis, and arrhythmia. CONCLUSION: This study demonstrates Cordyceps' prospective efficacy and safety in the prevention or treatment of CI/RI and MI/RI, providing novel insights for managing ischemic CCVDs.

Plexin domain-containing 1 may be a biomarker of poor prognosis in hepatocellular carcinoma patients, may mediate immune evasion.

BACKGROUND: For the first time, we investigated the oncological role of plexin domain-containing 1 (PLXDC1), also known as tumor endothelial marker 7 (TEM7), in hepatocellular carcinoma (HCC). AIM: To investigate the oncological profile of PLXDC1 in HCC. METHODS: Based on The Cancer Genome Atlas database, we analyzed the expression of PLXDC1 in HCC. Using immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blotting, we validated our results. The prognostic value of PLXDC1 in HCC was analyzed by assessing its correlation with clinicopathological features, such as patient survival, methylation level, tumor immune microenvironment features, and immune cell surface checkpoint expression. Finally, to assess the immune evasion potential of PLXDC1 in HCC, we used the tumor immune dysfunction and exclusion (TIDE) website and immunohistochemical staining assays. RESULTS: Based on immunohistochemistry, qRT-PCR, and Western blot assays, overexpression of PLXDC1 in HCC was associated with poor prognosis. Univariate and multivariate Cox analyses indicated that PLXDC1 might be an independent prognostic factor. In HCC patients with high methylation levels, the prognosis was worse than in patients with low methylation levels. Pathway enrichment analysis of HCC tissues indicated that genes upregulated in the high-PLXDC1 subgroup were enriched in mesenchymal and immune activation signaling, and TIDE assessment showed that the risk of immune evasion was significantly higher in the high-PLXDC1 subgroup compared to the low-PLXDC1 subgroup. The high-risk group had a significantly lower immune evasion rate as well as a poor prognosis, and PLXDC1-related risk scores were also associated with a poor prognosis. CONCLUSION: As a result of this study analyzing PLXDC1 from multiple biological perspectives, it was revealed that it is a biomarker of poor prognosis for HCC patients, and that it plays a role in determining immune evasion status.

Selective B(3)-H Activation Affording Multinuclear Ir(III) Complexes with (o-Carboranyl)dithioester Ligands.

A method was developed to link two or three o-carborane moieties to form a series of carboranyl dithioester bridging ligands via in situ substitution of haloalkanes by tetraphenylphosphonium carboranyldithiocarboxylates. Based on these ligands, direct B-H activation without the assistance of Ag(I) and alkali was successfully achieved with half-sandwich Ir(III) substrate [Cp*IrCl2]2 to yield corresponding bimetallic or trimetallic complexes. Single crystal structure analyses of the B-H activated complexes and corresponding SnCl2-inserted derivatives confirm the selective B(3)-H activation in these complexes.

A preoperative nomogram predicting risk of lymph node metastasis for early-stage cervical cancer.

OBJECTIVE: This study aimed to develop a preoperative nomogram based on clinical and pathological characteristics to provide a more individualized and accurate estimation of lymph node metastasis (LNM) in patients with early-stage cervical cancer. METHODS: A total of 7,349 early-stage cervical cancer patients with pathologically confirmed between 1988 and 2015 were obtained from the Surveillance, Epidemiology, and End Results (SEER) database. All the patients were divided into training (n = 5,500) and validation (n = 1,849) cohorts randomly. A cohort of 455 patients from multicenter was used for the external validation. We established a multivariate logistic regression model based on preoperative clinicopathological data, from which a nomogram was developed and validated. A predicted probability of LNM < 5% was defined as low risk. RESULTS: From multivariate logistic regression analysis, age at diagnosis, histologic subtype, tumor grade, tumor size and FIGO stage were identified as preoperative independent risk factors of LNM. The nomogram incorporating these factors demonstrated good discrimination and calibration (concordance index = 0.723; 95% confidence interval (CI), 0.707-0.738). In the validation cohort, the discrimination accuracy was 0.745 (95% CI, 0.720-0.770) and 0.747 (95% CI, 0.690-0.804), respectively. The nomogram was well calibrated with a high concordance probability. We also established an R-enabled Internet browser for LNM risk assessment, which tool may be convenient for physicians. CONCLUSIONS: We developed an effective preoperative nomogram based on clinical and pathological characteristics to predict LNM for early-stage cervical cancer. This model could improve clinical trial design and help physicians to decide whether to perform lymphadenectomy or not.

Tertiary lymphoid structures in gynecological cancers: prognostic role, methods for evaluating, antitumor immunity, and induction for therapy.

Tertiary lymphoid structures (TLSs), referred to as tertiary lymphoid organs and lymphoid tissue neogenesis, are aggregates of immune cells that occur in nonlymphoid tissues. In recent years, it has been found that TLSs within the tumor microenvironment have been associated with local adaptive immune immunity against cancer and favorable prognosis in several human solid tumors, including gynecological cancers. The issue of the prognosis of gynecological cancers, including endometrial, cervical, and ovarian cancer, is an enormous challenge that many clinical doctors and researchers are now facing. Concerning the predictive prognostic role of TLSs, effective evaluation, and quantification of TLSs in human tissues may be used to assist gynecologists in assessing the clinical outcome of gynecological cancer patients. This review summarizes the current knowledge of TLSs in gynecological cancers, mainly focusing on the potential mechanism of TLS neogenesis, methods for evaluating TLSs, their prognostic value, and their role in antitumor immune immunity. This review also discusses the new therapeutic methods currently being explored in gynecological cancers to induce the formation of TLSs.

A Branched Rutile/Anatase Phase Structure Electrode with Enhanced Electron-Hole Separation for High-Performance Photoelectrochemical DNA Biosensor.

A photoelectrochemical (PEC) detection platform was built based on the branched rutile/anatase titanium dioxide (RA-TiO2) electrode. Theoretical calculations proved that the type-II band alignment of rutile and anatase could facilitate charge separation in the electrode. The self-generated electric field at the interface of two phases can enhance the electron transfer efficiency of the electrode. Carboxylated CdTe quantum dots (QDs) were applied as signal amplification factors. Without the target DNA presence, the CdTe QDs were riveted to the surface of the electrode by the hairpin probe DNA. The sensitization of CdTe QDs increased the photocurrent of the electrode significantly. When the target DNA was present, the structural changes of the hairpin probe DNA resulted in the failure of the sensitized structure. Benefiting from excellent electrode structure design and CdTe QDs sensitization strategy, the PEC assays could achieve highly sensitive and specific detection of target DNA in the range of 1 fM to 1 nM, with a detection limit of 0.23 fM. The electrode construction method proposed in this article can open a new avenue for the preparation of more efficient PEC sensing devices.

CD32 Expression by CD4+ T and CD8+ T Lymphocytes Is Increased in Patients with Chronic Hepatitis B Virus Infection.

FcγR is expressed by many immune cells and plays an important role in the immune response to hepatitis B virus (HBV) infection. CD32 belongs to the FcγR family. This study aimed to observe changes in CD32 expression by CD4+ T and CD8+ T lymphocytes in chronic HBV infection patients and evaluate the clinical utility of CD4+ T and CD8+ T CD32 expression to assess the severity of liver injury in chronic HBV-infected patients. A total of 68 chronic HBV patients and 40 healthy individuals were recruited, and the median fluorescence intensity (MFI) of CD32 expression on CD4+ T, CD8+ T lymphocytes was measured using flow cytometry and the CD4+ T, CD8+ T CD32 index was calculated. The reactivity of the healthy individual lymphocytes to mixed patients' plasma containing HBV was observed. Finally, the correlation between CD4+ T, CD8+ T lymphocytes CD32 MFI and liver function indicator levels was analyzed. The CD4+ T, CD8+ T CD32 MFI and index were significantly elevated in HBV patient groups than in normal control group (p < 0.001, for all). Furthermore, the CD32 MFI of healthy persons' CD4+ T and CD8+ T lymphocytes were remarkably increased when stimulated with mixed patients' plasma containing high HBV copies (p < 0.001; P < 0.001). More importantly, in HBV patients, there was a significant positive correlation between CD4+ T, CD8+ T CD32 MFI and the level of serum aspartate aminotransferase (p < 0.05, p < 0.05). In conclusion, the increased expression of CD32 on CD4+ T and CD8+ T lymphocytes might be potential promising biomarkers for the severity of liver function impairment in chronic HBV patients.

In situ growth of TiO2/Ti3C2 MXene Schottky heterojunction as a highly sensitive photoelectrochemical biosensor for DNA detection.

In this work, a heterojunction composed of a TiO2 nanosheet and layered Ti3C2 was synthesized by directly growing TiO2 in Ti3C2 MXene. Compared with pure TiO2, TiO2/Ti3C2 composites had increased surface area, and a light absorption range that extended from ultraviolet to visible light, which greatly extended the life of photogenerated carriers. A photoelectrochemical biosensor for DNA detection was constructed based on the TiO2/Ti3C2 heterogeneous structure, which was comprehensively studied based on photocurrent responses. In the absence of the target, the CdSe QDs were close to the surface of the electrode, resulting in enhanced sensitization and increased photocurrent. In the presence of the target, the photocurrent decreases due to the formation of rigid double strands with the probe DNA, which caused the CdSe QDs to be far away from the electrode surface. The sensor had stability and sensitivity for DNA detection in the range of 10 nM-10 fM, and the lower detection limit was 6 fM. Its outstanding characteristics also provided ideas for detecting various other target DNA for early diagnosis of various diseases.

Carbon Dot-Modified Branched TiO2 Photoelectrochemical Glucose Sensors with Visible Light Response.

The development of a photoelectrochemical (PEC) sensor for the sensitive and rapid detection of glucose is highly desirable. In PEC enzyme sensors, inhibition of the charge recombination of electrode materials is an efficient technique, and detection in visible light can prevent enzyme inactivation due to ultraviolet irradiation. In this study, a visible light-driven PEC enzyme biosensor was proposed, using CDs/branched TiO2 (B-TiO2) as the photoactive material and glucose oxidase (GOx) as the identification element. The CDs/B-TiO2 composites were produced via a facile hydrothermal method. Carbon dots (CDs) can not only act as photosensitizers but also inhibit photogenerated electron and hole recombination of B-TiO2. Under visible light, electrons in the carbon dots flowed to B-TiO2 and further to the counter electrode through the external circuit. In the presence of glucose and dissolved oxygen, H2O2 generated through the catalysis of GOx could consume electrons in B-TiO2, causing a decrease in photocurrent intensity. Ascorbic acid was added to ensure the stability of the CDs during the test. Based on the variation of the photocurrent response, the CDs/B-TiO2/GOx biosensor presented a good sensing performance of glucose in visible light, its detection range was from 0 to 9.00 mM, and the detection limit was 0.0430 mM.

Binding characteristics of Pb and Zn to low-temperature feces-based biochar-derived DOM revealed by EEM-PARAFAC combined with general and moving-window two-dimensional correlation analyses.

Pyrolysis carbonization of human feces has shown potential for converting feces biomass into a soil amendment. However, little is known about the interactions of DOM derived from feces-based biochar produced at low-temperature with heavy metals (HMs). In this study, the binding properties of Pb(II) and Zn(II) with DOM derived from feces-based biochar produced at low pyrolysis temperatures were investigated using EEM-PARAFAC combined with general, and moving-window two-dimensional correlation analyses (2D-COS). The results revealed that DOM from biochar produced at 280 °C exhibited a higher Pb(II) and Zn(II) affinity and more binding sites than DOM produced at 380 °C. The fulvic-like and humic-like components exhibited obvious fluorescence quenching after the heavy metal addition, and the complexes formed with Pb(II) and Zn(II) were more stable. C-H groups exhibited the fastest response to Pb(II) and Zn(II) binding in the FB280 DOM, while the COO- groups of carboxylic acids in the FB380 DOM exhibited the fastest response to Pb(II) and Zn(II). Moreover, the mutation concentration range of components and functional groups in DOM, as analyzed by MW2D-COS, was greater for Zn(II) than for Pb(II). These results provide a more detailed molecular-level understanding of the interaction mechanisms between heavy metals and feces-based biochar-derived DOM and the effect of HM concentration on DOM binding. Further, these results will help to provide a reasonable reference for feces management and feces-based biochar in controlling soil HMs.

The association of dietary resistance starch intake with all-cause and cause-specific mortality.

Background: Several studies have estimated daily intake of resistant starch (RS), but no studies have investigated the relationship of RS intake with mortality. Objective: We aimed to examine associations between RS intake and all-cause and cause-specific mortality. Methods: Data from US National Health and Nutrition Examination Survey (NHANES) from 1999 to 2018 with 24-h dietary recall data was used in current study. The main exposure in this study was RS intake, and the main outcome was the mortality status of participants until December 31, 2019. The multivariable Cox proportional hazards regression models were developed to evaluate the hazard ratios (HRs) and 95% confidence interval (95% CI) of cardiovascular disease (CVD), cancer, and all-cause mortality associated with RS intake. Results: A total of 42,586 US adults [mean (SD) age, 46.91 (16.88) years; 22,328 (52.43%) female] were included in the present analysis. During the 454,252 person-years of follow-up, 7,043 all-cause deaths occurred, including 1,809 deaths from CVD and 1,574 deaths from cancer. The multivariable-adjusted HRs for CVD, cancer, and all-cause mortality per quintile increase in RS intake were 1 (95%CI, 0.97-1.04), 0.96 (95%CI, 0.93-1), and 0.96 (95%CI, 0.95-0.98), respectively. The associations remained similar in the subgroup and sensitivity analyses. Conclusion: Higher RS intake is significantly associated with lower cancer and all-cause mortality, but not significantly with CVD mortality. Future studies focusing on other populations with different food sources of RS and RS subtypes are needed to access the dose-response relationship and to improve global dietary recommendations.

Boosting the Photocatalytic Ability of TiO2 Nanosheet Arrays for MicroRNA-155 Photoelectrochemical Biosensing by Titanium Carbide MXene Quantum Dots.

The electrodes of two-dimensional (2D) titanium dioxide (TiO2) nanosheet arrays were successfully fabricated for microRNA-155 detection. The (001) highly active crystal face was exposed to catalyze signaling molecules ascorbic acid (AA). Zero-dimensional (0D) titanium carbide quantum dots (Ti3C2Tx QDs) were modified to the electrode as co-catalysts and reduced the recombination rate of the charge carriers. Spectroscopic methods were used to determine the band structure of TiO2 and Ti3C2Tx QDs, showing that a type Ⅱ heterojunction was built between TiO2 and Ti3C2Tx QDs. Benefiting the advantages of materials, the sensing platform achieved excellent detection performance with a wide liner range, from 0.1 pM to 10 nM, and a low limit of detection of 25 fM (S/N = 3).

Exosomes derived from bone marrow mesenchymal stem cells inhibit neuroinflammation after traumatic brain injury.

Exosomes derived from bone marrow mesenchymal stem cells can inhibit neuroinflammation through regulating microglial phenotypes and promoting nerve injury repair. However, the underlying molecular mechanism remains unclear. In this study, we investigated the mechanism by which exosomes derived from bone marrow mesenchymal stem cells inhibit neuroinflammation. Our in vitro co-culture experiments showed that bone marrow mesenchymal stem cells and their exosomes promoted the polarization of activated BV2 microglia to their anti-inflammatory phenotype, inhibited the expression of proinflammatory cytokines, and increased the expression of anti-inflammatory cytokines. Our in vivo experiments showed that tail vein injection of exosomes reduced cell apoptosis in cortical tissue of mouse models of traumatic brain injury, inhibited neuroinflammation, and promoted the transformation of microglia to the anti-inflammatory phenotype. We screened some microRNAs related to neuroinflammation using microRNA sequencing and found that microRNA-181b seemed to be actively involved in the process. Finally, we regulated the expression of miR181b in the brain tissue of mouse models of traumatic brain injury using lentiviral transfection. We found that miR181b overexpression effectively reduced apoptosis and neuroinflamatory response after traumatic brain injury and promoted the transformation of microglia to the anti-inflammatory phenotype. The interleukin 10/STAT3 pathway was activated during this process. These findings suggest that the inhibitory effects of exosomes derived from bone marrow mesenchymal stem cells on neuroinflamation after traumatic brain injury may be realized by the action of miR181b on the interleukin 10/STAT3 pathway.

Ferrostatin­1 alleviates oxalate­induced renal tubular epithelial cell injury, fibrosis and calcium oxalate stone formation by inhibiting ferroptosis.

The present study aimed to evaluate the role and mechanism of ferrostatin­1 (Fer­1) in oxalate (Ox)­induced renal tubular epithelial cell injury, fibrosis, and calcium oxalate (CaOx) stone formation. A CaOx model in mice kidneys was established via intraperitoneal injection of 80 mg/kg glyoxylic acid for 14 days. The mice were randomly divided into three groups (n=6), namely, the control (Con), the CaOx group, and the CaOx + Fer­1 group. Cultured human renal tubular epithelial cells (HK­2 cells) were randomly divided into three groups (n=3), namely, the control (Con), the Ox group, and the Ox + Fer­1 group. The levels of heme oxygenase 1 (HO­1), superoxide dismutase 2 (SOD2), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) were assessed by immunofluorescence and western blot analysis. Renal tubular injury and apoptosis were evaluated by H&E and TUNEL staining. Kidney interstitial fibrosis was evaluated by Masson and Sirius red staining, and the levels of E­cadherin, vimentin and α­SMA were detected by immunofluorescence or western blot analysis. Mitochondrial structure was observed using a transmission electron microscope. The levels of reactive oxygen species (ROS) were determined by flow cytometry and CaOx stone formation was evaluated by von Kossa staining. The results revealed that in comparison with the Con group, mitochondrial injury under glyoxylic acid treatment was observed by TEM. The expression of GPX4 and SLC7A11 in the CaOx and Ox groups was downregulated (P<0.05), whereas the expression of HO­1 and SOD2 was upregulated (P<0.05). Renal tissue damage, apoptosis of renal tubular epithelial cells, and interstitial fibrosis were increased in the CaOx and Ox groups (P<0.05). In comparison with the CaOx or Ox group, the expression of GPX4 and SLC7A11 in the CaOx + Fer­1 or Ox + Fer­1 group was upregulated (P<0.05), whereas that of HO­1 and SOD2 was downregulated (P<0.05). Renal tissue damage, apoptosis of renal tubular epithelial cells and interstitial fibrosis were decreased following Fer­1 treatment (P<0.05). The ROS level was also decreased following Fer­1 treatment. Moreover, CaOx stone formation was decreased in the CaOx + Fer­1 group (P<0.05). In conclusion, Fer­1 alleviated Ox­induced renal tubular epithelial cell injury, fibrosis, and CaOx stone formation by inhibiting ferroptosis.

Induction of Broadly Cross-Reactive Antibody Responses to SARS-CoV-2 Variants by S1 Nanoparticle Vaccines.

Despite the rapid deployment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines, the emergence of SARS-CoV-2 variants and reports of their immune evasion characteristics have led to an urgent need for novel vaccines that confer potent cross-protective immunity. In this study, we constructed three different SARS-CoV-2 spike S1-conjugated nanoparticle vaccine candidates that exhibited high structural homogeneity and stability. Notably, these vaccines elicited up to 50-times-higher neutralizing antibody titers than the S1 monomer in mice. Crucially, it was found that the S1-conjugated nanoparticle vaccine could elicit comparable levels of neutralizing antibodies against wild-type or emerging variant SARS-CoV-2, with cross-reactivity to SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV), the effect of which could be further enhanced using our designed nanoparticles. Our results indicate that the S1-conjugated nanoparticles are promising vaccine candidates with the potential to elicit potent and cross-reactive immunity against not only wild-type SARS-CoV-2, but also its variants of concern, variants of interest, and even other pathogenic betacoronaviruses. IMPORTANCE The emergence of SARS-CoV-2 variants led to an urgent demand for a broadly effective vaccine against the threat of variant infection. The spike protein S1-based nanoparticle designed in our study could elicit a comprehensive humoral response toward different SARS-CoV-2 variants of concern and variants of interest and will be helpful to combat COVID-19 globally.

Quadrivalent mosaic HexaPro-bearing nanoparticle vaccine protects against infection of SARS-CoV-2 variants.

Emerging SARS-CoV-2 variants of concern (VOCs) harboring multiple mutations in the spike protein raise concerns on effectiveness of current vaccines that rely on the ancestral spike protein. Here, we design a quadrivalent mosaic nanoparticle vaccine displaying spike proteins from the SARS-CoV-2 prototype and 3 different VOCs. The mosaic nanoparticle elicits equivalent or superior neutralizing antibodies against variant strains in mice and non-human primates with only small reduction in neutralization titers against the ancestral strain. Notably, it provides protection against infection with prototype and B.1.351 strains in mice. These results provide a proof of principle for the development of multivalent vaccines against pandemic and potential pre-emergent SARS-CoV-2 variants.

microRNA-486-5p is implicated in the cisplatin-induced apoptosis and acute inflammation response of renal tubular epithelial cells by targeting HAT1.

The proinflammatory property of cisplatin is potentially destructive and contributes to the pathogenesis of acute kidney injury (AKI). The role and upstream regulatory mechanism of histone acetyltransferase 1 (HAT1) in acute kidney inflammation are still unknown. We performed RNA sequencing to filter differentially expressed microRNAs (miRNAs) in the kidney tissue of mice with AKI induced by cisplatin and ischemia-reperfusion. Here, we found that miR-486-5p was upregulated and that the expression of HAT1 was reduced in AKI mouse models and injured human renal proximal tubular epithelial cell (HK-2) model induced by cisplatin. miR-486-5p is implicated in cisplatin-induced kidney damage in vivo. Bioinformatics analysis predicted a potential binding site between miR-486-5p and HAT1. The Luciferase reporter assay and Western blot confirmed that miR-486-5p directly targeted the 3'-untranslated region of HAT1 mRNA and inhibited its expression in the cytoplasm of HK-2 cells. In the in vitro study, inhibiting miR-486-5p reduced apoptosis, and the expression of proinflammatory mediators was induced by cisplatin in HK-2 cells. Simultaneously, the downregulation of miR-486-5p inhibited the activation of the toll-like receptor 4 (TLR4) and nuclear factor-kappa B (NF-κB). We further found that HAT1 could inhibit apoptosis and the activation of cisplatin on the TLR4/NF-κB pathway and that the upregulation of miR-486-5p reversed this effect. Therefore, the upregulation of miR-486-5p targeting HAT1 promoted the cisplatin-induced apoptosis and acute inflammation response of renal tubular epithelial cells by activating the TLR4/NF-κB pathway, providing a new basis to highlight the potential intervention of regulating the miR-486-5p/HAT1 axis.

Design of a mutation-integrated trimeric RBD with broad protection against SARS-CoV-2.

The continuous emergence of SARS-CoV-2 variants highlights the need of developing vaccines with broad protection. Here, according to the immune-escape capability and evolutionary convergence, the representative SARS-CoV-2 strains carrying the hotspot mutations were selected. Then, guided by structural and computational analyses, we present a mutation-integrated trimeric form of spike receptor-binding domain (mutI-tri-RBD) as a broadly protective vaccine candidate, which combined heterologous RBDs from different representative strains into a hybrid immunogen and integrated immune-escape hotspots into a single antigen. When compared with a homo-tri-RBD vaccine candidate in the stage of phase II trial, of which all three RBDs are derived from the SARS-CoV-2 prototype strain, mutI-tri-RBD induced significantly higher neutralizing antibody titers against the Delta and Beta variants, and maintained a similar immune response against the prototype strain. Pseudo-virus neutralization assay demonstrated that mutI-tri-RBD also induced broadly strong neutralizing activities against all tested 23 SARS-CoV-2 variants. The in vivo protective capability of mutI-tri-RBD was further validated in hACE2-transgenic mice challenged by the live virus, and the results showed that mutI-tri-RBD provided potent protection not only against the SARS-CoV-2 prototype strain but also against the Delta and Beta variants.

Identification of inflammatory response and alternative splicing in acute kidney injury and experimental verification of the involvement of RNA­binding protein RBFOX1 in this disease.

An increasing number of inflammatory responses and alternative splicing (AS) have been recently reported to be associated with various kidney diseases. The effect of inflammatory response on acute kidney injury (AKI) has not been fully clarified. In the present study, a mouse model of AKI induced by cisplatin and ischemia­reperfusion (IR) was established and genome­wide profiling analysis and identification of differentially expressed genes (DEGs) in kidney tissue was conducted by Gene Ontology (GO) functional analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, protein­protein interaction (PPI) network analysis and RT­qPCR. The results revealed that common DEGs in AKI induced by cisplatin and IR were enriched in the inflammatory response pathway, including hub genes CSF­1, CXCL1, CXCL10, IL­1ß, IL­34, IL­6 and TLR2. AS in AKI was initially reported. Cisplatin­induced AS was enriched in the phosphorylation pathway, involving regulated AS genes CSNK1A1, PAK2, CRK, ADK and IKBKB. IR­induced AS was enriched in apoptosis and proliferation pathways, including DEGs ZDHHC16, BCL2L1 and FGF1 regulated by AS. The ability of RNA­binding proteins (RBPs) to regulate AS was coordinated with the function of context­dependent genetic mechanisms. A total of 49 common differentially expressed RBP genes were screened. RNA binding fox­1 homolog 1 (RBFOX1) was revealed to be the top downregulated gene. The relative levels of RBFOX1 in the nuclei of mouse renal tubular epithelial cells in mRNA and proteins were downregulated by cisplatin and IR. Moreover, the biological functions of RBFOX1 were investigated in human renal proximal tubular epithelial cells (HK­2 cells). Results of in vitro experiments revealed that exogenous RBFOX1 inhibited inflammation and oxidative stress to reduce hypoxia/reoxygenation­induced apoptosis of HK­2 cells. This phenomenon may be related to the inhibition of NF­κB and the activation of the NRF2/HO­1 signaling pathway. In conclusion, the inflammatory cytokines, AS and RBPs in AKI were analyzed in the present study via whole transcriptome sequencing. It was revealed that the RBP gene RBFOX1 was involved in the pathogenesis of AKI. Thus, the present study provided novel insights into the mechanism of AKI pathogenesis.