Background/Aims: Dysregulated cholesterol metabolism is a hallmark of hepatocellular carcinoma (HCC) that drives tumor initiation and progression. However, clinical targeting of cholesterol metabolism has yielded limited benefits due to stringent feedback in tumor cells. Identifying a central mediator capable of restoring cholesterol homeostasis within the cell’s intrinsically fine-tuned regulatory framework is urgently needed. Methods: We integrated a proteomic dataset from patients with cholesterol-dysregulated HCC into a global cholesterol metabolic regulatory network to identify potential therapeutic targets for disrupted cholesterol homeostasis. The prognostic significance of the candidate targets was further validated in an independent cohort through immunohistochemistry. Functional and mechanistic studies were conducted in vitro using HCC cell lines and in vivo using mouse models. The pharmacological efficacy of the candidate agent was evaluated in both subcutaneous and orthotopic HCC mouse models. Results: ER lipid raft-associated 1 (ERLIN1), a pivotal regulator of cholesterol metabolism reprogramming, was identified as an independent favorable prognostic indicator in HCC. ERLIN1 constrains HCC progression both in vitro and in vivo by stabilizing the INSIG1–SCAP–SREBP2 axis and maintaining the metabolic balance of intracellular cholesterol. Under hypoxia, impaired factor-inhibiting hypoxia-1-dependent hydroxylation of ASB11 at asparagine residues 90 and 92 enhances ASB11-mediated ERLIN1 degradation. Pharmacological targeting of this axis using zoledronic acid (ZoA) attenuated HCC progression by weakening the ASB11–ERLIN1 interaction and restoring cholesterol homeostasis. Conclusions ERLIN1 represents a druggable metabolic vulnerability in cholesterol-dysregulated HCC. Targeting the ASB11–ERLIN1 axis with the clinically approved ZoA reestablishes cholesterol homeostasis and offers a promising therapeutic strategy to overcome the current limitations of cholesterol-targeted HCC therapies.

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.

To investigate the prevalence of influenza D virus(IDV)in cattle and swine populations in Jilin Province,China,277 nasopharyngeal swabs were collected from livestock exhibiting influ-enza-like symptoms for IDV detection.Virus isolation was performed using swine testicular(ST)cells for PCR-positive samples,followed by comprehensive analyses including whole-genome se-quencing,phylogenetic analysis,electron microscopic observation of viral morphology,and glycosy-lation site prediction.Two IDV strains were successfully isolated from bovine samples,designated as D/bovine/China/JL22/2024(JL22)and D/bovine/China/JL34/2024(JL34).These strains were demonstrated to have specific hemagglutination activity against turkey red blood cells,while no he-magglutination to chicken,rabbit,or guinea pig erythrocytes.Virus-inoculated ST cells exhibited distinct cytopathic effects(CPE)within 48 h,with a hemagglutination titer of 4 log2 in the culture supernatant.Phylogenetic analysis of the hemagglutinin-esterase-fusion(HEF)gene indicated that these strains were most closely related to the Japanese isolate D/Yamagata2019,belonging to the YAMA2019 lineage.Genomic sequence analysis showed the absence of genetic reassortment in these isolates.In this study,two IDV strains were successfully isolated and characterized,which provides preliminary insights into their genomic sequences and biological properties.The findings confirm the presence of IDV in bovine populations in Jilin Province and provide the fundamental data for future epidemiological surveillance and control strategies of IDV.

To investigate the prevalence of influenza D virus(IDV)in cattle and swine populations in Jilin Province,China,277 nasopharyngeal swabs were collected from livestock exhibiting influ-enza-like symptoms for IDV detection.Virus isolation was performed using swine testicular(ST)cells for PCR-positive samples,followed by comprehensive analyses including whole-genome se-quencing,phylogenetic analysis,electron microscopic observation of viral morphology,and glycosy-lation site prediction.Two IDV strains were successfully isolated from bovine samples,designated as D/bovine/China/JL22/2024(JL22)and D/bovine/China/JL34/2024(JL34).These strains were demonstrated to have specific hemagglutination activity against turkey red blood cells,while no he-magglutination to chicken,rabbit,or guinea pig erythrocytes.Virus-inoculated ST cells exhibited distinct cytopathic effects(CPE)within 48 h,with a hemagglutination titer of 4 log2 in the culture supernatant.Phylogenetic analysis of the hemagglutinin-esterase-fusion(HEF)gene indicated that these strains were most closely related to the Japanese isolate D/Yamagata2019,belonging to the YAMA2019 lineage.Genomic sequence analysis showed the absence of genetic reassortment in these isolates.In this study,two IDV strains were successfully isolated and characterized,which provides preliminary insights into their genomic sequences and biological properties.The findings confirm the presence of IDV in bovine populations in Jilin Province and provide the fundamental data for future epidemiological surveillance and control strategies of IDV.

Porcine arthritis,one of the common chronic diseases in large-scale pig farms,can signifi-cantly reduce the production performance of meat pigs.In this study,a strain of Haemophilus pa-rasuis(HPS)was isolated from the joint fluid of a lame pig.The HPS was analyzed in terms of se-rotypes,virulence genes,and resistance genes.Additionally,it was treated with sensitive antibiotics to provide a theoretical basis for the comprehensive prevention and treatment of arthritis in meat pigs in future production settings.A strain of HPS type 14 was isolated from the joint fluid of dis-eased pigs.The HPS isolate demonstrated sensitivity to β-lactams and tetracyclines,while florfeni-col and polymyxin effectively inhibited its growth at low concentrations.However,the bacteria ex-hibited resistance to sulfonamides and ciprofloxacin.The treatment of affected pigs with clinical ar-thritis using doxycycline and enrofloxacin injections proved effective.Compared to the infected group,in which the sick pigs experienced difficulty flexing their carpal and tarsal joints and exhibi-ted significant lameness,the pigs in the treatment group showed marked improvement.Their joints were only slightly swollen,and the clinical symptoms of arthropathy were alleviated.

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.

Objective:To evaluate the relationship between the mechanism of buprenorphine in attenuating neuroinflammation in microglia and the MAM domain-containing glycosylphosphatidylinositol anchor gene 1 ( MDGA1). Methods:The human microglial cell line HMC-3 was cultured in vitro and divided into 4 groups ( n=6 each) using a table of random numbers: control group (Con group), lipopolysaccharide(LPS)group, buprenorphine + LPS group (Bup+ LPS group) and buprenorphine + LPS + MDGA1 knockdown group (Bup+ LPS+ shMDGA1 group). LPS group was incubated with LPS at a final concentration of 1 μg/ml for 4 h. Bup+ LPS group was incubated with buprenorphine at a final concentration of 100 ng/ml for 1 h, followed by incubation with LPS at a final concentration of 1 μg/ml for 4 h. Bup+ LPS+ shMDGA1 group was transfected with MDGA1-specific shorthairpin RNA for knockdown, and the remaining treatment was similar to those previously described in Bup+ LPS group. The expression of MDGA1 in microglia was detected using real-time quantitative polymerase chain reaction, and the concentrations of interleukin (IL)-6, IL-1β, tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS) in the supernatant were measured using enzyme-linked immunosorbent assay. Results:Compared with Con group, the concentrations of IL-6, IL-1β, TNF-α and iNOS in the supernatant were significantly increased, and the expression of MDGA1 in microglia was down-regulated in LPS group ( P<0.05). Compared with LPS group, the concentrations of IL-6, IL-1β, TNF-α and iNOS in the supernatant were significantly decreased, and the expression of MDGA1 in microglia was up-regulated in Bup+ LPS group ( P<0.05). Compared with Bup+ LPS group, the concentrations of IL-6, IL-1β, TNF-α and iNOS in the supernatant were significantly increased, and the expression of MDGA1 in microglia was down-regulated in Bup+ LPS+ sh MDGA1 group ( P<0.05). Conclusions:The mechanism by which buprenorphine alleviates neuroinflammation in microglia may be related to the up-regulation of the expression of MDGA1.

AIM: To investigate and clarify the intervention mechanism of trigonelline(TRG)in preventing ferroptosis in ARPE-19 cells based on the Nrf2/HO-1/GPX4 pathway.METHODS: The ARPE-19 cells were cultured and subsequently treated with varying concentrations of trigonelline to ascertain the most effective concentration for modulating the cells. Then the cells were categorized into distinct groups, including normal control(NC)group, high glucose(HG)group, Fer-1 group, TRG group based on the determined concentration. Samples from each group were then gathered to assess relevant indicators. The intracellular levels of glutathione(GSH), malondialdehyde(MDA), and Ferrion were quantified in accordance with the protocols provided by the GSH, MDA, and Ferrion detection kits. Flow cytometry was employed to measure the ROS levels within each group. Additionally, Western blot analysis was conducted to examine the expression of nuclear factor erythroid 2-related factor 2(Nrf2), heme oxygenase-1(HO-1), glutathione peroxidase(GPX4), and acyl-CoA synthetase long-chain family member 4(ACSL4)across the different groups.RESULTS: The preconditioning intervention with 40 μg/mL TRG effectively mitigated the decline in cell activity induced by high glucose levels. The levels of reactive oxygen species(ROS)and MDA in the HG group were markedly elevated compared to the NC group; and the TRG group exhibited significantly reduced levels of ROS and MDA compared to those of the HG group, with the antioxidant stress index GSH showing opposite trends to those of ROS and MDA across all the groups. Whereas the Fer-1 and TRG groups showed decreased expression levels of ACSL4 protein and iron ions, and the expression levels of Nrf2, HO-1 and GPX4 in the Fer-1 and TRG groups were increased.CONCLUSION: TRG protects ARPE-19 cells from the detrimental effects of high glucose by targeting the Nrf2/HO-1/GPX4 signaling pathway to counter ferroptosis.

Objective:To evaluate the relationship between the mechanism of buprenorphine in attenuating neuroinflammation in microglia and the MAM domain-containing glycosylphosphatidylinositol anchor gene 1 ( MDGA1). Methods:The human microglial cell line HMC-3 was cultured in vitro and divided into 4 groups ( n=6 each) using a table of random numbers: control group (Con group), lipopolysaccharide(LPS)group, buprenorphine + LPS group (Bup+ LPS group) and buprenorphine + LPS + MDGA1 knockdown group (Bup+ LPS+ shMDGA1 group). LPS group was incubated with LPS at a final concentration of 1 μg/ml for 4 h. Bup+ LPS group was incubated with buprenorphine at a final concentration of 100 ng/ml for 1 h, followed by incubation with LPS at a final concentration of 1 μg/ml for 4 h. Bup+ LPS+ shMDGA1 group was transfected with MDGA1-specific shorthairpin RNA for knockdown, and the remaining treatment was similar to those previously described in Bup+ LPS group. The expression of MDGA1 in microglia was detected using real-time quantitative polymerase chain reaction, and the concentrations of interleukin (IL)-6, IL-1β, tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS) in the supernatant were measured using enzyme-linked immunosorbent assay. Results:Compared with Con group, the concentrations of IL-6, IL-1β, TNF-α and iNOS in the supernatant were significantly increased, and the expression of MDGA1 in microglia was down-regulated in LPS group ( P<0.05). Compared with LPS group, the concentrations of IL-6, IL-1β, TNF-α and iNOS in the supernatant were significantly decreased, and the expression of MDGA1 in microglia was up-regulated in Bup+ LPS group ( P<0.05). Compared with Bup+ LPS group, the concentrations of IL-6, IL-1β, TNF-α and iNOS in the supernatant were significantly increased, and the expression of MDGA1 in microglia was down-regulated in Bup+ LPS+ sh MDGA1 group ( P<0.05). Conclusions:The mechanism by which buprenorphine alleviates neuroinflammation in microglia may be related to the up-regulation of the expression of MDGA1.

[This corrects the article DOI: 10.1016/j.apsb.2021.10.012.].