Evanescent hormesis effect induced by environmentally relevant PFOS to marine Chlorella sp.

Perfluorooctanesulfonic acid (PFOS) is widely detected in the aquatic environment. More attentions were paid to its acute biotoxicity at high-dose concentrations, whereas the actual long-term effect (hormesis or inhibition of growth) of PFOS with environmental concentrations on marine phytoplankton remains unclear. In this study, marine Chlorella sp. was exposed to PFOS at low concentrations (100 ng/L, 10 µg/L, and 1 mg/L) for 26 days. The hormesis effect disappeared at the population level on Day 18, but persisted at the molecular and cellular levels on Day 24, suggesting that the stimulatory hormetic effect induced by low-level PFOS (approximating environmental concentrations) does not persist throughout algal life cycle at population level. The 100 ng/L and 1 mg/L PFOS treatments caused algal cell to swell and shrink, respectively. The low-level PFOS treatments could accelerate cells apoptosis and induce cell necrosis at 100 ng/L. Specifically, the energy metabolism associated with carbohydrate metabolism and amino acid metabolism was significantly up-regulated as well as the reduced chlorophyll content (related to the down-regulation of porphyrin metabolism) to combat the 100 ng/L PFOS rather than be engaged in divide and growth. Additionally, the decreased biomass in the 100 ng/L treatment was also attributed to certain proteins associated with down-regulations of carotenoid biosynthesis, thiamine metabolism, non-homologous end-joining, and nitrogen metabolism along with the increased oxidative stress. Our findings provide a new insight into the long-term ecological effect of PFOS at environmental concentrations.

Maize-Tripsacum-Teosinte allopolyploid (MTP), a novel dwarf mutant inducer tool in maize.

Dwarf plant architecture facilitates dense planting, and increased planting densities boost the maize yield. However, breeding applications of dwarfing materials for maize are currently limited. There is an urgent need remove the obstacles to applying dwarf resources. Here, we innovated a new method to add a novel maize dwarf germplasm through the distant hybridization of Maize-Tripsacum-Teosinte allopolyploid (MTP) with maize. We identified ten independent dwarf families with unique characteristics. Five germplasms in our library were controlled by their respective dwarf genes. However, no allele was controlled by Br2. Subsequently, d024 in the library was successfully fine mapped, revealing its linkage to indel-4 in ZmCYP90D1. The indel-4 polymorphism regulates the expression of ZmCYP90D1 and is controlled by an upstream transcription factor (ZmBES1/BZR1-5). The indel-4 of ZmCYP90D1 allele, which reduces plant height, originated from Tripsacum, a wild variety of maize. However, d024 exhibits sensitivity to brassinosteroids (BRs), with lower castasterone levels in the internodes than that in the wild type. Furthermore, ZmCYP90D1 interacted with ZmFDXs and ZmNAD(P)H to positively regulate the downstream BR synthesis pathway. Additionally, we showed that introgressing the indel-4 of the Tripsacum allele into modern hybrids ensures yield potential and improves the harvest index under high-density conditions. Overall, as we begin to manufacture highly engineered dwarf materials using the MTP, this approach will solve the problems faced by corn dwarfs.

Unveiling the hub genes associated with ochratoxin A-induced hepatotoxicity in broiler chickens.

Ochratoxin A (OTA) widely exists in raw food and feed materials and can induce liver damage and toxicity. However, the mechanisms of OTA-induced hepatotoxicity were largely unknown. Thus, our study aimed to uncover the vital genes relevant to OTA-induced hepatotoxicity in broiler chickens. Gene expression data of chicken embryo primary hepatocytes (CEPHs) in OTA-treated and control groups were obtained from the GEO database. Totally 1407 differentially expressed genes (DEGs) were selected, of which 850 and 557 genes were up- and downregulated in OTA-treated CEPHs. Gene ontology (GO) enrichment revealed that the DEGs were in connection with various biological processes, such as signal transduction, extracellular matrix organization, axon guidance, cell division, cholesterol homeostasis, proteolysis, microtubule cytoskeleton organization, and chromosome segregation. Pathway enrichment showed that the DEGs were related to metabolic pathways, ferroptosis, calcium, FoxO, Wnt, cell cycle, apoptosis, calcium, and cell adhesion molecules signaling pathways. Furthermore, the hub genes, including CDK1, DLGAP5, KIF2C, VCL, ITGB3, and ZYX, were identified as hub genes potentially contributing to OTA-induced hepatotoxicity. Taken together, this study provides valuable insights into the mechanisms underlying OTA-induced hepatotoxicity in broiler chickens.

Genetic Variants in METTL16 Affect the Risk of Non-Syndromic Orofacial Clefts.

OBJECTIVE: N6-methyladenosine (m6A) is the most prevalent modification of RNA in eukaryotes which is associated with many cellular processes and diseases. Here, our objective is to explore whether genetic variants in m6A modification genes are associated with the risk of non-syndrome orofacial clefts (NSOCs). METHODS: The transmission disequilibrium test (TDT) was performed to calculate the association between single nucleotide polymorphisms (SNPs) in m6A modification genes and NSOCs risk in 944 case-parent trios. The function of SNP was predicted by HaploReg, RegulomeDB and histone enrichment data. The expression quantitative trait locus (eQTL) analysis was examined using Genotype-Tissue Expression (GTEx) and eQTLGen. The role of gene in the development of NSOCs was assessed with correlation and enrichment analysis based on gene expression data in mice craniofacial tissue and zebrafish embryo. RESULTS: We identified that rs8078195 (A > C) in METTL16 was suggestively associated with the increased risk of NSOCs (OR = 1.32, p = 1.80E - 03). The region surrounding rs8078195 was subjected to deoxyribonuclease hypersensitivity and enriched with multiple histone modifications. In addition, it had a significant eQTL effect with METTL16 in skin tissue and human peripheral blood, which played an important role in NSOCs development. Bioinformatic analysis indicated that METTL16 contributed to the development of NSOCs probably by regulating cell cycle process. CONCLUSIONS: Rs8078195 in METTL16 was associated with the occurrence of NSOCs.

An accumulated mutation gained in mosquito cells enhances Zika virus virulence and fitness in mice.

Zika virus (ZIKV) remains a significant public health threat worldwide. A number of adaptive mutations have accumulated within the genome of ZIKV during global transmission, some of which have been linked to specific phenotypes. ZIKV maintains an alternating cycle of replication between mosquitoes and vertebrate hosts, but the role of mosquito-specific adaptive mutations in ZIKV has not been well investigated. In this study, we demonstrated that serial passaging of ZIKV in mosquito Aag2 cells led to the emergence of critical amino acid substitutions, including A94V in the prM protein and V153D and H401Y in the E protein. Further characterization via reverse genetics revealed that the H401Y substitution in the E protein did not augment viral replication in mosquitoes but significantly enhanced neurovirulence and lethality compared with those of the wild-type (WT) virus in mice. More importantly, the H401Y mutant maintained its virulence phenotype in mice after propagation in mosquitoes in mosquito-mouse cycle model. In particular, recombinant ZIKV harboring the H401Y substitution showed enhanced competitive fitness over WT ZIKV in various mammalian cells and mouse brains, but not in mosquito cells. Notably, the H401Y substitution in the ZIKV E protein has been detected in recent isolates derived from both mosquitoes and humans in Asia and the Americas. In summary, our findings not only identify a novel virulence determinant of ZIKV but also highlight the complexity of the relationship between the evolution of vector-borne viruses and their clinical outcome in nature. IMPORTANCE: Zika virus (ZIKV) is an important arbovirus with a global impact. Experimental evolution by serial passaging of ZIKV in susceptible cells has led to the identification of a panel of critical amino acid substitutions with specific functions. Herein, we identified a mosquito cell-derived substitution, H401Y, in the ZIKV E protein via experimental evolution. The H401Y substitution significantly enhanced viral virulence and fitness in mammal cells and mice. Notably, the H401Y substitution has been detected in recent mosquito and human isolates from regions spanning Asia to the Americas. Our work elucidates unrecognized virulence determinant in the ZIKV genome that warrants urgent attention. Moreover, the findings underscore the critical need for extensive molecular surveillance and rigorous clinical observation to establish the potential impact in natural circulation. These endeavors are crucial for unraveling the potential of mutation to act as a catalyst for future epidemics, thereby preempting the public health challenges it may pose.

Biomimetic mineralization of collagen from fish scale to construct a functionally gradient lamellar bone-like structure for guided bone regeneration.

Wide used guided bone regeneration (GBR) membrane materials, such as collagen, Teflon, and other synthesized polymers, present a great challenge in term of integrating the mechanical property and degradation rate when addressing critical bone defects. Therefore, inspired by the distinctive architecture of fish scales, this study utilized epigallocatechin gallate to modify decellularized fish scales following biomimetic mineralization to fabricate a GBR membrane that mimics the structure of lamellar bone. The structure, physical and chemical properties, and biological functions of the novel GBR membrane were evaluated. Results indicate that the decellularized fish scale with 5 remineralization cycles (5R-E-DCFS) exhibited a composite and structure similar to natural bone and had a special functionally gradient mineral contents character, demonstrating excellent mechanical properties, hydrophilicity, and degradation properties. In vitro, the 5R-E-DCFS membrane exhibited excellent cytocompatibility promoting Sprague-Dawley (SD) rat bone marrow mesenchymal stem cell proliferation and differentiation up-regulating the expression of osteogenic-related genes and proteins. Furthermore, in vivo, the 5R-E-DCFS membrane promoted the critical skull bone defects of SD rats repairment and regeneration. Therefore, this innovative biomimetic membrane holds substantial clinical potential as an ideal GBR membrane with mechanical properties for space-making and suitable degradation rate for bone regeneration to manage bone defects.

N-(4-methoxyphenyl) quinoline-8-sulfonamide reduces the inflammatory response of fibroblast-like synoviocytes by targeting receptor (calcitonin) activity modifying protein 1 in rheumatoid arthritis.

BACKGROUND: Rheumatoid arthritis (RA) is characterized by chronic inflammation of the synovium of joints. Fibroblast-like synoviocytes (FLS) play an important role in RA pathogenesis. We aimed to investigate the effect of N-(4-methoxyphenyl) quinoline-8-sulfonamide (QS-3g) on the inflammatory response of FLS and explore the potential underlying mechanisms. METHODS: We screened and found that QS-3g exhibits the best anti-inflammatory response against FLS using the CCK8 assay. To investigate the therapeutic effects of QS-3g on K/BxN STA mice, we used H&E staining, immunofluorescence, immunohistochemistry, micro-CT, and other techniques. Additional investigations, including RNA-seq, molecular docking, and CETSA, revealed that QS-3g binds to RAMP1. RESULTS: Among a series of 8-quinoline sulfonyl amide derivatives, QS-3g reduced the inflammatory response in TNF-α stimulated FLS, such as the release of interleukin (IL)-1ß and IL-6. H&E staining and micro-CT showed that QS-3g inhibited synovial hypertrophy, inflammatory cell infiltration, and bone destruction. RNA-seq and CETSA analyses revealed the targeted inhibition of RAMP1 by QS-3g. Inhibition of RAMP1 expression could reduce IL-6 and IL-1ß levels. Compared with RAMP1-si, combined administration of QS-3g and RAMP1-si reduced the TNF-α-induced inflammation in TNF-α stimulated FLS without statistically significant differences. Finally, the results of in vitro experiments showed that QS-3g could restore the balance of Gαi/Gαs by inhibiting Gαi and activating Gαs and up-regulate the expression of cAMP protein, thus inhibiting the RAMP1-mediated inflammatory response in FLS. CONCLUSION: QS-3g could inhibit RAMP1 activity and mediate the Gαs/Gαi-cAMP pathway to reduce FLS inflammatory response. Therefore, QS-3g may serve as a novel anti-inflammatory compound for treating RA.

Identification and field verification of the aggregation pheromone components produced by male Holotrichia parallela Motschulsky (Coleoptera: Scarabaeidae).

BACKGROUND: The chafer beetle, Holotrichia parallela, causes damage to numerous economically significant crops worldwide. Adult beetles exhibit aggregation behavior likely mediated by a male-produced pheromone. Advancements in biological research technology have facilitated the identification of insect aggregation pheromones and promoted their applications as bait for trapping and monitoring pests. Currently, only a few active components of aggregation pheromones from Holotrichia species have been identified. However, the specific components of aggregation pheromones produced by H. parallela remain unknown. RESULT: In this study, we initially observed from Y-tube olfactometer assays that both male and female H. parallela were significantly attracted to volatiles emitted by males, but not to those from females. We then collected hindgut crude extracts of male adults and carried out gas chromatography-mass spectrometry analysis to identify potential aggregation pheromone components. Pentadecyl acetate, cis-13-docosenol, and behenic acid were identified as male-specific compounds in comparison to female extracts, serving as components of the aggregation pheromone in H. parallela. We further evaluated their attractiveness to H. parallea in both laboratory and field experiments. In laboratory settings, pentadecyl acetate, cis-13-docosenol, and behenic acid evoked significant responses to both males and females at specific concentrations, as evidenced by both electroantennography tests and behavioral bioassays. Under field conditions, traps baited with these three compounds captured significantly more H. parallela adults compared to control traps. CONCLUSION: In this study, we found that pentadecyl acetate, cis-13-docosenol, and behenic acid are specifically present in male H. parallela, serving as aggregation pheromones. Both laboratory and field-trapping experiments suggest their potential as monitoring and controlling tools against H. parallela adults. © 2024 Society of Chemical Industry.

Association of urinary albumin excretion with all-cause and cardiovascular mortality among patients with rheumatoid arthritis: a national prospective study.

Background: Rheumatoid arthritis (RA) patients suffering from chronic renal insufficiency tend to exhibit subtle manifestations at the beginning. Urine albumin to creatinine ratio (ACR) is a sensitive indicator for early assessment of renal function. However, it is unclear whether it serves as an independent risk factor influencing the prognosis of RA patients. Methods: National Health and Nutrition Examination Survey (NHANES) data from 2009-2018 were included. Kaplan-Meier (K-M) curves were plotted to compare the cumulative survival probability of RA patients with different urinary albumin excretion. The association of ACR with mortality among RA patients was investigated with Cox regression model, restricted cubic spline (RCS) and stratified analyses. The prognostic efficacy of ACR and estimated glomerular filtration rate (eGFR) was evaluated by receiver operating characteristic (ROC) curves. Results: The Cox regression model adjusted with covariates showed a 53% (HR 1.53, 95% CI 1.06-2.21) increase in all-cause mortality and a statistically non-significant increase in cardiovascular disease (CVD) mortality in RA patients with microalbuminuria (30mg/g ≤ACR<300mg/g). ACR≥300mg/g was associated with an increase in all-cause mortality (HR 2.62, 95% CI 1.55-4.45) and CVD mortality (HR 5.67, 95% CI 1.96-16.39). RCS demonstrated a nonlinear correlation between ACR and all-cause mortality in RA patients with microalbuminuria. Subgroup analysis showed that CVD mortality was higher in RA patients with microalbuminuria characterized by the following features: female, other ethnicity, eGFR≥60 ml/min/1.73 m2, hypertension or hyperlipidemia. Compared with eGFR, ACR provided better prognostic efficacy than eGFR with higher values of the area under the curve (AUC) for all-cause mortality (AUC=0.683, 95% CI 0.613-0.754) and CVD mortality (AUC=0.681, 95% CI 0.541-0.820). Conclusion: ACR is an independent risk factor affecting the prognosis of RA patients. The all-cause mortality was increased in RA patients with albuminuria. There was an upward trend in the CVD mortality of those with macroalbuminuria when ACR increased.

Analysis of risk factors for prolonged stay in the intensive care unit after cardiac surgery in children with pneumonia.

AIM: Preoperative pneumonia in children with CHD may lead to longer stays in the ICU after surgery. However, research on the associated risk factors is limited. This study aims to evaluate the pre-, intra-, and postoperative risk factors contributing to extended ICU stays in these children. METHODS: This retrospective cohort study collected data from 496 children with CHD complicated by preoperative pneumonia who underwent cardiac surgery following medical treatment at a single centre from 2017 to 2022. We compared the clinical outcomes of patients with varying ICU stays and utilised multivariate logistic regression analysis and multiple linear regression analyses to evaluate the risk factors for prolonged ICU stays. RESULTS: The median ICU stay for the 496 children was 7 days. Bacterial infection, severe pneumonia, and Risk Adjustment for Congenital Heart Surgery-1 were independent risk factors for prolonged ICU stays following cardiac surgery (P < 0.05). CONCLUSION: CHD complicated by pneumonia presents a significant treatment challenge. Better identification of the risk factors associated with long-term postoperative ICU stays in these children, along with timely diagnosis and treatment of respiratory infections in high-risk populations, can effectively reduce ICU stays and improve resource utilisation.

A core microbiome signature as an indicator of health.

The gut microbiota is crucial for human health, functioning as a complex adaptive system akin to a vital organ. To identify core health-relevant gut microbes, we followed the systems biology tenet that stable relationships signify core components. By analyzing metagenomic datasets from a high-fiber dietary intervention in type 2 diabetes and 26 case-control studies across 15 diseases, we identified a set of stably correlated genome pairs within co-abundance networks perturbed by dietary interventions and diseases. These genomes formed a "two competing guilds" (TCGs) model, with one guild specialized in fiber fermentation and butyrate production and the other characterized by virulence and antibiotic resistance. Our random forest models successfully distinguished cases from controls across multiple diseases and predicted immunotherapy outcomes through the use of these genomes. Our guild-based approach, which is genome specific, database independent, and interaction focused, identifies a core microbiome signature that serves as a holistic health indicator and a potential common target for health enhancement.

Interface-Strengthened Ru-Based Electrocatalyst for High-Efficiency Proton Exchange Membrane Water Electrolysis at Industrial-Level Current Density.

Developing an OER electrocatalyst that balances high performance with low cost is crucial for widely adopting PEM water electrolyzers. Ru-based catalysts are gaining attention for their cost-effectiveness and high activity, positioning them as promising alternatives to Ir-based catalysts. However, Ru-based catalysts can be prone to oxidation at high potentials, compromising their durability. In this study, we utilize a simple synthesis method to synthesize a SnO2, Nb2O5, and RuO2 composite catalyst (SnO2/Nb2O5@RuO2) with multiple interfaces and abundant oxygen vacancies. The large surface area and numerous active sites of the SnO2/Nb2O5@RuO2 catalyst lead to outstanding acidic oxygen evolution reaction (OER) performance, achieving current densities of 10, 50, and 200 mA cm-2 at ultralow overpotentials of 287, 359, and 534 mV, respectively, significantly surpassing commercial IrO2. Moreover, incorporating Nb2O5 into the SnO2/Nb2O5@RuO2 alters the electronic structure at the interfaces and generates a high density of oxygen vacancies, markedly enhancing durability. Consequently, the membrane electrode composed of SnO2/Nb2O5@RuO2 and commercial Pt/C demonstrated stable operation in the PEM cell for 25 days at an industrial current density of 1 A cm-2. This research presents a convenient approach for developing a highly efficient and durable Ru-based electrocatalyst, underscoring its potential for proton exchange membrane water electrolysis.

Detection of the Novel HLA-B*15:358 Allele by Sanger Dideoxy Nucleotide Sequencing.

HLA-B*15:358 differs from HLA-B*15:02:01:01 by one nucleotide substitution at position 685 in exon 4.

Circular RNA TAF4B targeting MFN2 accelerates cell growth in bladder cancer through p27 depression and AKT activation.

Introduction: Bladder cancer (BCa) is a common malignancy in the urinary tract. It has high recurrence rates and often requires microscopic examination, which presents significant challenges in clinical treatment. Previous research has shown that circular TAF4B (circTAF4B) is significantly upregulated in BCa and is associated with a poor prognosis. However, the specific targets and molecular mechanisms by which circTAF4B functions in BCa are still not well - understood. Methods: In this study, an RNA pull - down assay and mass spectrometry were utilized to identify MFN2 as a binding protein of circTAF4B. Additionally, siRNA was used to silence MFN2 to observe the amplification of the inhibitory effects of circTAF4B overexpression on cell growth and migration in BCa cells. Moreover, circTAF4B shRNA lentiviral particles were employed to study their impact on BCa progression by examining the regulation of p27 and the blocking of AKT signaling. Results: It was found that MFN2 is a binding protein of circTAF4B. Silencing MFN2 with siRNA enhanced the inhibitory effects of circTAF4B overexpression on cell growth and migration in BCa cells. Also, circTAF4B shRNA lentiviral particles inhibited BCa progression by upregulating p27 and blocking AKT signaling. Discussion: In conclusion, the physical binding of circTAF4B to MFN2 is a crucial process in the tumorigenesis and progression of BCa. Targeting circTAF4B or its complexes may have potential as a therapeutic strategy for BCa diagnosis and treatment.

The abnormal expression of peripheral blood CD4+ T lymphocyte subsets are correlated with primary Sjögren's syndrome complicated with haematological involvement.

OBJECTIVES: Complicated primary Sjögren's syndrome (pSS) with haematological involvement (HI) is not uncommon; however, the aetiology of this condition remains obscure. The clinical characteristics, cytokine levels, and expression of peripheral blood lymphocyte subsets (CD4+ T lymphocyte subsets in particular) of patients with pSS-HI were investigated in this study. METHODS: The pSS-HI group (n = 43), the pSS complicated without HI (pSS-non-HI) group (n = 94), and the healthy controls (HCs) group (n = 40) were enrolled in the Second Hospital of Shanxi Medical University. The clinical data were gathered, and cytokines and peripheral blood lymphocyte subsets were quantified using flow cytometry and the Cytometric Bead Array (CBA), respectively. RESULTS: Patients with pSS-HI were more likely than those without pSS-HI to develop skin involvement, had a higher positive rate of anti-SSA antibody, and had elevated levels of IgA, IgG, and ESR. Compared to the pSS-non-HI group, the number of all lymphocyte subsets was lower in the pSS-HI group. However, the proportion of Th2 cells in the pSS-HI group was higher than those in the pSS-non-HI group. In contrast to the pSS-non-HI group, the pSS-HI group exhibited elevated levels of IL-10 and decreased levels of IL-4. A significant correlation was observed between IL-10 and the number of total T cells, CD4+ T cells, CD8+ T cells, NK cells, Th1 cells, Th2 cells, and Th17 cells. In the context of pSS-HI, protective factors may include the number of Treg cells and CD4+ T cells, whereas risk factors may include IgA and the number of Th2 cells. CONCLUSIONS: An immunological mechanism potentially implicated in the development of pSS-HI may be the elevation of IL-10 and the reduction of peripheral blood CD4+ T cell subsets (particularly Treg cells) and serum IL-4 levels.

Risk factors for type 2 diabetes mellitus in Chinese rheumatoid arthritis patients from 2018 to 2022: a real-world, single-center, retrospective study.

Introduction: In patients with rheumatoid arthritis (RA), the increased risk of concomitant type 2 diabetes mellitus (T2D) is an important contributor to increased mortality and decreased quality of life; however, the mechanisms and pathogenetic factors remain unknown. Methods: In this study, we aimed to assess the risk factors for T2D in patients with RA. We recruited 206 healthy controls and 488 patients with RA, 160 of whom had comorbid T2D. General clinical information, disease characteristics, and circulating lymphocyte levels detected using modified flow cytometry were collected from all participants. Logistic regression models adjusted for confounders were fitted to estimate the risk factors of T2D in patients with RA. Results: The incidence of RA in patients with T2D was 15.6%. Patients with RA and T2D had a longer disease duration, higher BMI, and a higher incidence of hypertension and a family history of diabetes than those with RA but no T2D. The absolute numbers of T helper 2 cell (Th2) and Regulatory T cells (Treg) decreased in patients with RA and T2D, which led to an increase in the ratios of Th1/Th2 and Th17/Treg cells. Multivariate logistic regression analysis showed that a family history of diabetes, a higher incidence of hypertension, higher neutrophil-lymphocyte ratio (NLR) levels, lower platelet-lymphocyte ratio (PLR) levels, and fewer circulating Th2 and Treg cells were associated with an increased risk of T2D in patients with RA. Discussion: The levels of peripheral lymphocytes, especially Th2 and Treg cells, are closely related to the occurrence of T2D in patients with RA; however, the influence of body mass index (BMI), family history of diabetes, and systemic inflammation should not be ignored.

Orexin A protects against cerebral ischemia-reperfusion injury by enhancing reperfusion in ischemic cortex via HIF-1α-ET-1/eNOS pathway.

The purpose of this study was to investigate the protective effect and underlying mechanism of orexin A on cerebral ischemia-reperfusion injury, specifically through vasodilation mediated by the hypoxia inducible factor-1α (HIF-1α)-Endothelin-1(ET-1)/endothelial nitric oxide synthase (eNOS) pathway. A model of middle cerebral artery occlusion was established in both wild-type SD rats with exogenous orexin A intervention and in orexin A transgenic rats. Neurological deficit scores and cerebral infarction areas were assessed, and ischemic cortical blood flow was monitored. Gene and protein expression levels of HIF-1α, HIF-2α, ET-1, and three types of NOS were detected using real-time RT-qPCR and Western blot analysis, respectively. Additionally, nitric oxide (NO) levels in the cortex were analyzed through biochemical detection methods. Orexin A demonstrated a protective effect by reducing cerebral infarction and improving neurological deficits, which was achieved by increasing cortical blood flow during reperfusion. This protective mechanism was associated with upregulated HIF-1α expression, downregulated ET-1 expression, upregulated eNOS expression, and increased NO production. This study demonstrates the protective effect of orexin A on cerebral ischemia-reperfusion injury, achieved by regulating the release of vasomotor substances to enhance cortical blood flow during reperfusion. These findings suggest that orexin A may represent a potential therapeutic strategy for ischemic stroke.

Identification of the Novel HLA-A*02:407 Allele by Sanger Dideoxy Nucleotide Sequencing.

HLA-A*02:407 differs from HLA-A*02:01:01:01 by one nucleotide substitution in codon 109 in exon 3.

Characterisation of the Novel HLA-A*02:406 Allele by Sequencing-Based Typing in a Chinese Individual.

HLA-A*02:406 differs from HLA-A*02:01:01:01 by one nucleotide substitution in codon 116 in exon 3.

A Novel HLA-B*15 Variant Allele, HLA-B*15:359, Identified by Sequencing-Based Typing in a Chinese Individual.

HLA-B*15:359 differs from HLA-B*15:01:01:01 by one nucleotide substitution in exon 3.