Targeting NKAα1 to treat Parkinson's disease through inhibition of mitophagy-dependent ferroptosis.

Dysfunction of the Na+/K+-ATPase (NKA) has been documented in various neurodegenerative diseases, yet the specific role of NKAα1 in Parkinson's disease (PD) remains incompletely understood. In this investigation, we utilized NKAα1 haploinsufficiency (NKAα1+/-) mice to probe the influence of NKAα1 on dopaminergic (DA) neurodegeneration induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Our findings reveal that NKAα1+/- mice displayed a heightened loss of DA neurons and more pronounced motor dysfunction compared to the control group when exposed to MPTP. Intriguingly, this phenomenon coincided with the activation of ferroptosis and impaired mitophagy both in vivo and in vitro. To scrutinize the role and underlying mechanism of NKAα1 in PD, we employed DR-Ab, an antibody targeting the DR-region of the NKA α subunit. Our study demonstrates that the administration of DR-Ab effectively reinstated the membrane abundance of NKAα1, thereby mitigating MPTP-induced DA neuron loss and subsequent improvement in behavioral deficit. Mechanistically, DR-Ab heightened the formation of the surface NKAα1/SLC7A11 complex, inhibiting SLC7A11-dependent ferroptosis. Moreover, DR-Ab disrupted the cytosolic interaction between NKAα1 and Parkin, facilitating the translocation of Parkin to mitochondria and enhancing the process of mitophagy. In conclusion, this study establishes NKAα1 as a key regulator of ferroptosis and mitophagy, identifying its DR-region as a promising therapeutic target for PD.

Hmgcs2 is the hub gene in diabetic cardiomyopathy and is negatively regulated by Hmgcs2, promoting high glucose-induced cardiomyocyte injury.

BACKGROUND: Diabetic cardiomyopathy (DCM) represents a major cause of heart failure and a large medical burden worldwide. This study screened the potentially regulatory targets of DCM and analyzed their roles in high glucose (HG)-induced cardiomyocyte injury. METHODS: Through GEO database, we obtained rat DCM expression chips and screened differentially expressed genes. Rat cardiomyocytes (H9C2) were induced with HG. 3-hydroxy-3-methylglutarylcoenzyme A synthase 2 (Hmgcs2) and microRNA (miR)-363-5p expression patterns in cells were measured by real-time quantitative polymerase chain reaction or Western blot assay, with the dual-luciferase assay to analyze their binding relationship. Then, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay, lactate dehydrogenase assay, terminal deoxynucleotidyl transferase dUTP nick end labeling assay, enzyme-linked immunosorbent assay, and various assay kits were applied to evaluate cell viability, cytotoxicity, apoptosis, inflammation responses, and oxidative burden. RESULTS: Hmgcs2 was the vital hub gene in DCM. Hmgcs2 was upregulated in HG-induced cardiomyocytes. Hmgcs2 downregulation increased cell viability, decreased TUNEL-positive cell number, reduced HG-induced inflammation and oxidative stress. miR-363-5p is the upstream miRNA of Hmgcs2. miR-363-5p overexpression attenuated HG-induced cell injury. CONCLUSIONS: Hmgcs2 had the most critical regulatory role in DCM. We for the first time reported that miR-363-5p inhibited Hmgcs2 expression, thereby alleviating HG-induced cardiomyocyte injury.

Meta-analysis of the characteristic expression of circulating microRNA in type 2 diabetes mellitus with acute ischemic cerebrovascular disease.

Objective: To comprehensively evaluate the characteristics of the circulating microRNA expression profile in type 2 diabetic patients with acute ischemic cerebrovascular disease by systematic evaluation and meta-analysis. Methods: The literatures up to March 2022 related to circulating microRNA and acute ischemic cerebrovascular disease in type 2 diabetes mellitus were searched and screened from multiple databases. The NOS quality assessment scale was used to evaluate methodological quality. Heterogeneity tests and statistical analyses of all data were performed by Stata 16.0. The differences in microRNA levels between groups were illustrated by the standardized mean difference (SMD) and 95% confidence interval (95% CI). Results: A total of 49 studies on 12 circulating miRNAs were included in this study, including 486 cases of type 2 diabetes complicated with acute ischemic cerebrovascular disease and 855 controls. Compared with the control group (T2DM group), miR-200a, miR-144, and miR-503 were upregulated and positively correlated with acute ischemic cerebrovascular disease in type 2 diabetes mellitus patients. Their comprehensive SMD and 95% CI were 2.71 (1.64~3.77), 5.77 (4.28~7.26) and 0.73 (0.27~1.19), respectively. MiR-126 was downregulated and negatively correlated with acute ischemic cerebrovascular disease in type 2 diabetes mellitus patients, its comprehensive SMD and 95% CI were -3.64 (-5.56~-1.72). Conclusion: In type 2 diabetes mellitus patients with acute ischemic cerebrovascular disease, the expression of serum miR-200a, miR-503, plasma and platelet miR-144 was upregulated and the expression of serum miR-126 was downregulated. It may have diagnostic value in the early identification of type 2 diabetes mellitus with acute ischemic cerebrovascular disease.

Feeding Aquilaria sinensis Leaves Modulates Lipid Metabolism and Improves the Meat Quality of Goats.

Aquilaria (A.) sinensis is a medicinal plant widely grown in tropical South China. Given the abundant pruning waste of its leaves, the use of A. sinensis leaves is valuable. In this study, goats were fed a diet containing 20% A. sinensis leaves. Compared with the basal diet, feeding A. sinensis leaves to goats did not affect growth performance but considerably reduced the feeding cost. Strikingly, feeding A. sinensis leaves resulted in a significant decrease in the blood cholesterol levels (2.11 vs. 1.49 mmol/L, p = 0.01) along with a significant increase in the high-density lipoprotein levels (1.42 vs. 1.82 mmol/L, p = 0.01). There was also a tendency to lower the content of low-density lipoprotein levels in goats (0.78 vs. 0.45 mmol/L, p = 0.09). Furthermore, metabolomics analysis demonstrated that the reduction in cholesterol levels occurred in both the serum (0.387-fold change) and muscle (0.382-fold change) of goats during A. sinensis leaf feeding. The metabolic responses to feeding A. sinensis leaves suggest that the activation of lipolysis metabolism might happen in goats. These observed changes would be conducive to improving animal health and meat quality, ultimately benefiting human health.

Bioinformatics analysis identifies immune-related gene signatures and subtypes in diabetic nephropathy.

Background: Systemic inflammation and immune response are involved in the pathogenesis of diabetic nephropathy (DN). However, the specific immune-associated signature during DN development is unclear. Our study aimed to reveal the roles of immune-related genes during DN progression. Methods: The GSE30529 and GSE30528 datasets were acquired from the Gene Expression Omnibus (GEO) database. Then, the intersection between differentially expressed genes (DEGs) and immune score-related genes (ISRGs) was screened. Subsequently, functional enrichment analyses were performed. The different immune phenotype-related subgroups were finally divided using unsupervised clustering. The core genes were identified by WGCNA and the protein-protein interaction (PPI) network. xCell algorithm was applied to assess the proportion of immune cell infiltration. Results: 92 immune score-related DEGs (ISRDEGs) were identified, and these genes were enriched in inflammation- and immune-associated pathways. Furthermore, two distinct immune-associated subgroups (C1 and C2) were identified, and the C1 subgroup exhibited activated immune pathways and a higher percentage of immune cells compared to the C2 subgroup. Two core genes (LCK and HCK) were identified and all up-regulated in DN, and the expressions were verified using GSE30122, GSE142025, and GSE104954 datasets. GSEA indicated the core genes were mainly enriched in immune-related pathways. Correlation analysis indicated LCK and HCK expressions were positively correlated with aDC, CD4+ Tem, CD8+T cells, CD8+ Tem, and mast cells. Conclusions: We identified two immune-related genes and two immune-associated subgroups, which might help to design more precise tailored immunotherapy for DN patients.

Magnetic recyclable heterogeneous catalyst Fe3O4/g-C3N4 for tetracycline hydrochloride degradation via photo-Fenton process under visible light.

Antibiotic pollution of water resources is a global problem, and the development of new treatments for destroying antibiotics in water is a priority research. We successfully manufactured recyclable magnetic Fe3O4/g-C3N4 through the electrostatic self-assembly method. Selecting tetracycline (TC) as the target pollutant, using Fe3O4/g-C3N4 and H2O2 developed a heterogeneous optical Fenton system to remove TC under visible light. Fe3O4/g-C3N4 was systematically characterized by SEM, TEM, XRD, FTIR, XPS, DRS, and electrochemical methods. The removal efficiency of 7% Fe3O4/g-C3N4 at pH = 3, H2O2 = 5 mM, and catalyst dosage of 1.0 g/L can reach 99.8%. After magnetic separation, the Fe3O4/g-C3N4 photocatalyst can be recycled five times with minimal efficiency loss. The excellent degradation performance of the prepared catalyst may be attributed to the proper coupling interface between Fe3O4 and g-C3N4 which promotes the separation and transfer of photogenerated electrons. Photogenerated electrons can also accelerate the conversion of Fe3+ to Fe2+, thereby producing more ˙OH. The new Fe3O4/g-C3N4 can be used as a raw material for advanced oxidation of water contaminated by refractory antibiotics.

Decreased expression of serum semaphorin 3B is associated with poor prognosis of patients with hepatocellular carcinoma.

Semaphorin 3B (SEMA-3B), which belongs to the semaphorin family, has an important role in cell apoptosis and inhibition of angiogenesis. A previous study by our group revealed that SEMA-3B was downregulated in tumor tissues of patients with hepatocellular carcinoma (HCC) and exerts anti-motility and anti-invasion effects on tumor cells. However, the serum levels of SEMA-3B and their clinical significance have remained elusive; therefore, the aim of the present study was to monitor its expression in HCC and investigate its clinical significance. ELISA was used to determine the serum levels of SEMA-3B in 132 patients with HCC and 57 healthy individuals. The association between SEMA-3B and clinicopathological parameters was investigated. Serum SEMA-3B was indicated to be significantly decreased in patients with HCC as compared with that in the controls (P<0.05) and it was negatively associated with tumor size (P=0.039), encapsulation (P=0.002) and TNM stage (P=0.034). The prognosis of patients with low expression of SEMA-3B was poor. In conclusion, the results of the present study revealed that serum SEMA-3B is decreased in HCC and is negatively associated with prognosis; therefore, it may be used as a prognostic marker in HCC.

Association between IL-1ß and recurrence after the first epileptic seizure in ischemic stroke patients.

To analyze the association of IL-1ß with recurrence after the first epileptic seizure in ischemic stroke patients and evaluate its predictive value. 238 patients with the first epileptic seizure after ischemic stroke were included in this study. IL-1ß expression levels were detected through quantitative Real-Time PCR. Kaplan-Meier method was used to perform univariate analysis with log-rank test. The variables with P < 0.1 were then included in multivariate analysis. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value. Among all 238 patients, 107 patients (44.96%) had seizure recurrence and 131 patients (55.04%) had no recurrence. Kaplan-Meier analysis showed that high expression of IL-1ß, low age (< 65 years), male, cortical involvement, large lesion size, late onset, severe neurological impairment and partial seizure type were associated with seizure recurrence. Multivariate analysis showed that IL-1ß expression level (hazard ratio 2.057, 95% confidence interval 1.296-3.318) was independently associated with seizure recurrence. The area under ROC curve (AUC) was 0.803 (SE 0.030, 95% confidence interval 0.744-0.862) when IL-1ß expression levels were applied in predicting seizure recurrence. IL-1ß might be a useful biomarker for early discovery of recurrence after the first epileptic seizure in ischemic stroke patients.

Sevoflurane attenuates brain damage through inhibiting autophagy and apoptosis in cerebral ischemia­reperfusion rats.

The present study aimed to investigate the effects of sevoflurane post­conditioning in a rat brain cerebral ischemia­reperfusion (I/R) model and examine its possible mechanism. Rats were randomly divided into six groups: Sham control group (Sham), I/R group, sevoflurane group (Se), Toll­like receptor­4 (TLR4) inhibitor group (Tak­242), nuclear factor (NF)­κB inhibitor group (QNZ) and Sevoflurane post­conditioning combined with TLR4­NF­κB signaling pathway inhibitor group (Se + Tak­242). Morris water maze test and tetrazolium chloride staining were used to investigate the I/R injury. The nerve cell apoptosis and autophagy in cortical tissue were detected by TUNEL and transmission electron microscopy, respectively. The expression of TLR4 protein in cortical tissue was observed by immunohistochemical staining. The expression of autophagy and apoptotic associated proteins in cortical tissues and the activity of TLR4­NF­κB signaling pathway were assayed by western blot analysis. Sevoflurane post­conditioning improved the learning and memory dysfunction caused by cerebral I/R injury. The cerebral infarction area, nerve cell apoptosis and formation of autophagic vacuoles were reduced after sevoflurane administration. The expression of light chain 3II/I, Beclin­1, Bad and Cleaved­Caspase­3 proteins were inhibited and the expression of Bcl­2 protein was upregulated after sevoflurane administration. Sevoflurane post­conditioning also inhibited the TLR4 protein and NF­κB phosphorylation, and increased inhibitor of kBα phosphorylation. The treatment effect of Tak­242 and QNZ groups were not significantly different compared with the Se group (P>0.05), and the Se + Tak­242 group had the best results. The present study demonstrated that sevoflurane post­conditioning could protect middle cerebral artery occlusion­induced brain injury rats by inhibiting autophagy and apoptosis, and that its mechanism is related to the TLR4­NF­κB signaling pathway.

Construction of a synthetic Araneus ventricosus dragline silk gene multimer and its expression in Escherichia coli.

One of the most representative core gene sequence of Araneus ventricosus dragline silk protein partial cDNA monomer (JN857964.2) was selected and multimerized using a "head-to-tail" strategy by compatible but nonregenerable sites at both ends resulting in a concatemer of 16 contiguous monomers. This concatemer was cloned into pET-28a(+) expression vector and transformed into Escherichia coli. A 52.6 kDa silk protein was successfully expressed and detected by SDS-PAGE and confirmed by Western blotting. A maximum yield of the silk protein was expressed with 7.06 mM IPTG after 5 h incubation. This is the first report on the construction and overexpression of a A. ventricosus dragline silk multimeric gene construct and the results from our study will provide a reference point for further exploration and development of large-scale production of spider silk protein.

Differentially methylated genomic fragments related with sexual dimorphism of rice pests, Sogatella furcifera.

Sogatella furcifera (Hovarth) is a major rice pest with sexual dimorphism. The objective of the current research was to monitor differentially cytosine methylation at CCGG sequences in male and female adults of S. furcifera to determine the association between gene methylation and sexual phenotypes using methylation-sensitive representational difference analysis. After the second subtractive hybridization, four differentially methylated DNA bands were obtained and sequenced. Ten different fragments were found. One fragment from the positive hybridization was 120 bp, and highly similar to the tramtrack genes from Nasonia vitripennis. Another fragment from the reverse hybridization was 414 bp, and homologous to the 28S rRNA gene of S. furcifera with a similarity rate as high as 99%. We also discussed how DNA methylation of tramtrack and 28S rRNA genes produced effects on sexual differentiation and development. These results provide potential evidence that DNA methylation of some genes may be related to sexual phenotype variations in S. furcifera and will facilitate future studies on the epigenetic mechanisms of insect sexual dimorphism.

[Imaging assessment of neonatal necrotizing enterocolitis].

OBJECTIVE: To improve the understanding of recognizing and diagnosis of neonatal necrotizing enterocolitis (NEC), imaging assessment of neonates with NEC was analyzed retrospectively. METHOD: Data of 211 cases of NEC were retrospectively collected from the Department of Neonatology, Children's Hospital of Chongqing Medical University between Jan.1(st) 2006-Dec.31(st) 2011. RESULT: Analysis of abdominal X-ray of 211 cases showed that there were 40 cases (19.0%) who had no changes on each X-ray, 47 cases (22.3%) had improvement and 23 cases (10.9%) became worse. In the group of no changes, positive rate with good prognosis was 97.5% and with poor prognosis, it was 2.5%. In the group of improvement, positive rate with good prognosis was 97.9%, and the contrary was 2.1%. Positive rate with good prognosis was 56.5%, and the contrary was 43.5% in worse group. Chi-square analysis of the three groups showed χ(2) = 31.742, P < 0.01. Comparison of detection rate of pneumoperitoneum on abdominal X-ray (16.0%, 12/75) and Doppler US (1.3%, 1/75), χ(2) = 10.191, P < 0.05, portal pneumatosis on abdominal X-ray(1.3%, 1/75) versus Doppler US (12.0%,9/75), χ(2) = 6.857, P < 0.05. Surgical timing mostly corresponded to pneumoperitoneum (OR = 19.543) and intestinal obstruction (OR = 19.527) of abdominal X-ray. The logistic regression equation is y = -2.915-1.588x1+2.972x4+2.973x7 + 1.711x9 (χ(2) = 101.705, P < 0.01). CONCLUSION: Abdominal X-ray is the most important method of diagnosis of NEC, the group of deterioration of abdominal X-ray has obvious bad prognosis differ from no change group and better group. Comparison with abdominal X-ray and Doppler US, the former in pneumoperitoneum positive rate was higher than the latter, at the same time, portal pneumatosis on Doppler US is more sensitive to abdominal X-ray, the value of two imaging assessments both supplement each other. Surgical timing mostly corresponds to pneumoperitoneum and intestinal obstruction.

The effect of gamma sterilization on the insecticidal toxicity of engineered and conventional Bacillus thuringiensis strains.

This study evaluates the effect of gamma radiation on the spore activity, toxicity, and crystal structures of two engineered Bacillus thuringiensis (Bt) strains, TnX and TnY, and the reference Bt strain HD-1. We attempted to identify dosages of cobalt-60 gamma radiation that would inactivate Bt spores but not affect its toxicity. In the radiation dosage range of 10-15 kilogray, no viable spore formation and no significant reduction of the efficiency of Bt against lepidopteran larvae were observed. However, further sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results show that the components of the protoxin are affected by gamma radiation and that some bands are absent after treatment compared with the controls; the change in the protoxin band pattern depends on the type of Bt strain. Furthermore, the spore crystal structure of three Bt strains was studied with scanning electron microscopy and transmission electron microscopy. The results show that there are no changes in the size or shape of the treated Bt spores and crystals compared with the controls. The use of gamma radiation is effective to inactivate the spores of engineered Bt strains while preserving stable Bt toxicity against the target insect larvae.

Interspecific interaction between Spodoptera exigua multiple nucleopolyhedrovirus and Microplitis bicoloratus (Hymenoptera: Braconidae: Microgastrina) in Spodoptera exigua (Lepidoptera: Noctuidae) larvae.

Baculoviruses and parasitoids are important biological control factors of insects in integrated pest management. Microplitis bicoloratus Chen (Hymenoptera: Braconidae: Microgastrina) is a solitary endoparasitoid of beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) larvae. M. bicoloratus parasitized first to third instars of S. exigua and most effectively parasitized second instars. The survival rate of parasitoids emerging from S. exigua multiple nucleopolyhedrovirus (SeMNPV)-infected hosts decreased with increasing SeMNPV doses at second to fourth instars. In addition, the mortality rate of the host was > 80% when coinfected with SeMNPV and M. bicoloratus, regardless of virus doses or the timing of virus treatment. Occlusion body production was significantly reduced in parasitized hosts. A comparison of host weights showed that hosts coinfected with SeMNPV and M bicoloratus were significantly lighter than those infected with SeMNPV alone, suggesting that the decrease in virus yield resulted from a reduction in larval growth. The viral genome copy number in parasitized host was significantly lower than that in nonparasitized host at 48 and 72 h postinfection. These results suggest that SeMNPV and M. bicoloratus are compatible as S. exigua control agents.

An ac34 deletion mutant of Autographa californica nucleopolyhedrovirus exhibits delayed late gene expression and a lack of virulence in vivo.

Ac34 and its homologs are highly conserved in all sequenced alphabaculoviruses. In this paper, we show that ac34 transcripts were detected from 6 to 48 h postinfection (p.i.) in Autographa californica nucleopolyhedrovirus (AcMNPV)-infected Sf9 cells. Ac34 localized to both the cytoplasm and the nuclei of infected cells but was not a viral structural protein. To determine the function of ac34 in the viral life cycle, an ac34 knockout AcMNPV (vAc34KO) was constructed. Compared with wild-type and repair viruses, vAc34KO exhibited an approximately 100-fold reduction in infectious virus production. Further investigations showed that the ac34 deletion did not affect the replication of viral DNA, polyhedron formation, or nucleocapsid assembly but delayed the expression of late genes, such as vp39, 38k, and p6.9. Bioassays revealed that vAc34KO was unable to establish a fatal infection in Trichoplusia ni larvae via per os inoculation. Few infectious progeny viruses were detected in the hemolymph of the infected larvae, indicating that the replication of vAc34KO was attenuated. These results suggest that Ac34 is an activator protein that promotes late gene expression and is essential for the pathogenicity of AcMNPV.

Comparison of bacterial diversity in wheat bran and in the gut of larvae and newly emerged adult of Musca domestica (Diptera: Muscidae) by use of ethidium monoazide reveals bacterial colonization.

The objective of the current study is to investigate the bacterial colonization within the gut of the house fly, Musca domestica L. (Diptera: Muscidae), at the larval stage and the bacterial community of the gut of the house fly at the newly emerged adult stage. After using ethidium monoazide to inhibit recovery of nucleic acids from dead bacteria, three polymerase chain reaction (PCR)-amplified 16S rDNA libraries from wheat bran, larvae, and newly emerged adults was constructed, analyzed, and compared. In total, 24, 11, and four phylotypes in the 16S rDNA libraries of wheat bran and the gut of larvae and adults, respectively, were found and assigned to three phylogenetic phyla of the domain Bacteria: Firmicutes, Proteobacteria, and Bacteroidetes. In the wheat bran library, 76% of the total number of sequences were affiliated to the genera Pseudomonas, Halomonas, Providencia, and Ignatzschineria. The three genera Morganella (79.05%), Providencia (8.78%), and Ignatzschineria (9.46%) dominated the library of the larval gut. Compared with the wheat bran library, the relative abundance of Morganella morganii (Winslow) was significantly higher (79.05 versus 0.8%), whereas that of Ignatzschineria larvae and of Providencia spp. was similar. These results demonstrate that M. morganii, Providencia spp., and I. larvae colonized the gut of the house fly larvae. Live bacteria of M. morganii, Providencia spp., and Proteus spp. were found in the gut of newly emerged adults. Therefore, the bacteria M. morganii and Providencia spp. colonized the larval gut could survive in the gut from larval metamorphosis to adult eclosion of the house fly.

Cryopreservation of the late stage embryos of Spodoptera exigua (Lepidoptera: Noctuidae).

Genetic devolution, genetic drift and contamination are all threats to maintain germplasm stability during mass rearing of many insects. Cryopreservation of beet armworm (Spodoptera exigua) embryos was studied to provide information to improve mass rearing. A series of experiments was conducted on late-stage embryos (45-48 h at 27 degree C) of the beet armyworm, which included evaluation of cryoprotectants (CPAs), their toxicity and glass-forming tendency and optimization of experimental procedures. The results showed that ethylene glycol (EG) was the best CPA with comparatively low toxicity compared to the other six CPAs tested (methanol, 1,3-propanediol, glycerol, 2-amino-1-ethanol, 3-amino-1-propanol 3-methoxy-1 and 2-propanediol). The highest hatching rate of 8.8 degree was attained after freezing with a 3-step loading procedure and a 1-step unloading procedure, but the hatched larvae from frozen-thawed embryos did not actively feed and could not develop to a later stage. This was attributed to injuries from freezing in late stage embryos of S. exigua which had formed midguts.