Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive cognitive decline, functional impairment, and neuropsychiatric symptoms. It represents the most prevalent form of dementia among the elderly population. Accumulating evidence indicates that oxidative stress plays a pivotal role in the pathogenesis of AD. Notably, elevated levels of oxidative stress have been observed in the brains of AD patients, where excessive reactive oxygen species (ROS) can cause extensive damage to lipids, proteins, and DNA, ultimately compromising neuronal structure and function. Amyloid β‑protein (Aβ) has been shown to induce mitochondrial dysfunction and calcium overload, thereby promoting the generation of ROS. This, in turn, exacerbates Aβ aggregation and enhances tau phosphorylation, leading to the formation of two pathological features of AD: extracellular Aβ plaque deposition and intracellular neurofibrillary tangles (NFTs). These events ultimately culminate in neuronal death, forming a vicious cycle. The interplay between oxidative stress and these pathological processes constitutes a core link in the pathogenesis of AD. The signaling pathways mediating oxidative stress in AD include Nrf2, RCAN1, PP2A, CREB, Notch1, NF‑κB, ApoE, and ferroptosis. Nrf2 signaling pathway serves as a key regulator of cellular redox homeostasis, exerts important antioxidant capacity and protective effects in AD. RCAN1 signaling pathway, as a calcineurin inhibitor, and modulates AD progression through multiple mechanisms. PP2A signaling pathway is involved in regulating tau phosphorylation and neuroinflammation processes. CREB signaling pathway contributes to neuroplasticity and memory formation; activation of CREB improves cognitive function and reduce oxidative stress. Notch1 signaling pathway regulates neuronal development and memory, participates in modulation of Aβ production, and interacts with Nrf2 toco-regulate antioxidant activity. NF‑κB signaling pathway governs immune and inflammatory responses; sustained activation of this pathway forms “inflammatory memory”, thereby exacerbating AD pathology. ApoE signaling pathway is associated with lipid metabolism; among its isoforms, ApoE-ε4 significantly increases the risk of AD, leading to elevated oxidative stress, abnormal lipid metabolism, and neuroinflammation. The ferroptosis signaling pathway is driven by iron-dependent lipid peroxidation, and the subsequent release of lipid peroxidation products and ROS exacerbate oxidative stress and neuronal damage. These interconnected pathways form a complex regulatory network that regulates the progression of AD through oxidative stress and related pathological cascades. In terms of therapeutic strategies targeting oxidative stress, among the drugs currently used in clinical practice for AD treatment, memantine and donepezil demonstrate significant therapeutic efficacy and can improve the level of oxidative stress in AD patients. Some compounds with antioxidant effects (such asα-lipoic acid and melatonin) have shown certain potential in AD treatment research and can be used as dietary supplements to ameliorate AD symptoms. In addition, non-drug interventions such as calorie restriction and exercise have been proven to exerted neuroprotective effects and have a positive effect on the treatment of AD. By comprehensively utilizing the therapeutic characteristics of different signaling pathways, it is expected that more comprehensive multi-target combination therapy regimens and combined nanomolecular delivery systems will be developed in the future to bypass the blood-brain barrier, providing more effective therapeutic strategies for AD.

Aim To evaluate the toxicity and mecha-nism of Epimedium sagittatum(Sieb.et Zucc.)Maxim aqueous extract(ESMAE)on HepaRG cells based on serum toxicology.Methods MTT assay was used to detect the activity of HepaRG cells after treatment with the serum containing ESMAE from SD rats.Western blot was used to detect the effects of the serum contai-ning drug on the expression of endoplasmic reticulum stress-related proteins(PERK,eIF-2α,ATF-4,GRP78,CHOP)and pyroptosis related proteins(NL-RP1,caspase-1,cleaved caspase-1,GSDMD,GSDMD-N).MTT assay was used to detect the activity of Hep-aRG cells after treatment with the liver homogenate containing ESMAE from SD rats.Results Twenty percent serum containing drug significantly decreased the viability of HepaRG cells,with the cells exhibiting swelling,rupture,and vesicle-like pyroptosis.The ex-pression levels of endoplasmic reticulum stress-related proteins PERK,eIF-2α,ATF-4,GRP78,and CHOP significantly increased.The expression levels of pro-teins involved in the NLRP1-mediated classical pyrop-tosis pathway were significantly increased.Finally the liver homogenate containing drug decreased the cell ac-tivity,and cells exhibited swelling,rupture,and vesicle-like pyroptosis.Conclusions After administration of ESMAE,the serum containing drug and the liver ho-mogenate containing drug of rats show toxicity to Hep-aRG cells,and can induce endoplasmic reticulum stress and activate the NLRP1/caspase-1/GSDMD pyroptosis pathway in HepaRG cells.

Objective To investigate the effect of leucine carboxyl methyltransferase 1(LCMT1)knockout on fructose-induced lipid deposition in primary mouse hepatocytes.Methods Primary hepatocytes were isolated from wild-type(WT)and hepatocyte-specific LCMT1 knockout(KO)mice via a two-step hepatic portal vein perfusion method.The cells were divided into four groups:WT-control group,WT-fructose group,KO-control group,and KO-fructose group.Cell viability was determined through Alamar-Blue assays.Hepatocyte injury was evaluated based on alanine aminotransferase and aspartate aminotransferase levels.Lipid deposition was visualized via Oil Red O staining and lipid droplet green fluorescence staining,and the cellular triglyceride content was quantified via a GPO-POD assay.The mRNA expression of lipid metabolism-related genes was detected via quantitative real-time PCR,and the protein expression of LCMT1 and PP2Ac was detected via Western blot.Results Fructose treatment did not alter cell viability significantly in any group,and no significant cell damage was observed(P＞0.05).The WT-fructose group exhibited greater accumulation of lipid droplets in hepatocytes than that in the WT-control group(P＜0.001),with significantly elevated triglyceride contents(P＜0.05).The mRNA levels of the de novo lipid synthesis genes ChREBP,SREBP-1c,and ACC1 were increased significantly(P＜0.05,P＜0.001,P＜0.001),whereas FAS expression did not differ significantly between groups(P＞0.05).The mRNA levels of the lipid uptake genes FABP1 and FATP2 also increased significantly(both P＜0.05).In contrast,the KO-fructose group presented a reduced number of lipid droplets(P＜0.01,P＜0.001),decreased triglyceride content(P＜0.05),and decreased mRNA levels of ChREBP,SREBP-1c,ACC1,FABP1,and FATP2(P＜0.01,P＜0.001,P＜0.001,P＜0.001,P＜0.05);CPT1 mRNA levels were markedly increased(P＜0.01).Total PP2Ac expression was significantly higher(P＜0.05)and PP2Ac demethylation was significantly lower(P＜0.01)in the WT-fructose group than in the WT-control group.In the KO-control group,total PP2Ac expression remained unchanged(P＞0.05),whereas PP2Ac demethylation was markedly elevated(P＜0.001).Compared with levels in the WT-fructose group,the KO-fructose group presented markedly lower total PP2Ac expression and significantly higher PP2Ac demethylation levels(P＜0.05,P＜0.01,respectively).Conclusions LCMT1 knockout alleviates fructose-induced lipid deposition in primary hepatocytes by inhibiting lipid uptake,increasing fatty acid oxidation,and downregulating de novo lipid synthesis.These effects are medicated by the LCMT1 knockout-mediated upregulation of PP2Ac demethylation,thereby modulating PP2A activity.

Objective To explore the protective effects of genetically modified porcine erythrocyte suspension as a subnormothermic normoxic mechanical perfusate on hypoxic-ischemic brain injury in cynomolgus monkeys caused by traumatic hemorrhage.Methods Cynomolgus monkeys were randomly divided into positive and negative control groups(a total of 3 monkeys,with 3 left cerebral hemispheres as the positive control group and 3 right cerebral hemispheres as the negative control group)and the subnormothermic perfusion group(n=3).The positive control group was directly sampled 1 hour after circulatory arrest,while the negative control group was placed at subnormothermic conditions for 6 hours after circulatory arrest.The subnormothermic perfusion group underwent 6 hours of subnormothermic normoxic mechanical perfusion of the bilateral common carotid arteries of the cynomolgus monkey hypoxic-ischemic brain injury model using genetically modified porcine erythrocyte suspension 1 hour after circulatory arrest.Before perfusion,cross-matching experiments were conducted between the six genetically modified pig and the cynomolgus monkeys.After the start of perfusion,the levels of routine blood indicators in the perfusate were detected at 0,1,2,3,4,5 and 6 hours.Blood oxygen saturation was recorded,and the levels of Na+,K+,Ca2+,glucose and blood pH in the perfusate were measured,as well as the levels of IgG and IgM in the perfusate.After 6 hours of perfusion,the water content of the brain tissue was measured.Nissl staining was performed on the frontal cortex and hippocampal regions,and immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP),ionized calcium-binding adapter molecule 1(Iba1)and neuronal nuclear antigen(NEUN).Results The cross-matching results between the six genetically modified pig and the cynomolgus monkeys were negative.The number of red blood cells in the perfusate decreased significantly at 3 hours of perfusion,and the hemoglobin level showed a downward trend at 1,3,5 and 6 hours.The number of white blood cells and platelets decreased at all time points.The blood oxygen saturation in the subnormothermic perfusion group remained stable at 95%-98%,and the levels of blood oxygen saturation,Na+,Ca2+,glucose and pH were stable,while the K+level first increased and then decreased.There was no significant difference in the levels of IgG and IgM before and after perfusion.The water content of brain tissue at the end of perfusion in the subnormothermic perfusion group was significantly higher than that in the positive control group(P<0.001).Nissl staining results showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex of the subnormothermic perfusion group maintained better morphological integrity,with no significant increase in enlarged and deformed cells.In the hippocampal CA1 region,there was a slight increase in enlarged and deformed cells,and a few cells with undamaged structures showed reduced cell size.In the hippocampal dentate gyrus,fewer granule neurons had compromised structural integrity,with increased cell edema.NEUN immunofluorescence staining showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex and hippocampal CA1 region of the subnormothermic perfusion group had better morphological states,with clear axons.The granule cells in the hippocampal dentate gyrus were well preserved,but the nuclei were less well protected.GFAP immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had sparser protrusions that were more tightly associated with neurons.Iba1 immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had thicker and fewer protrusions.Conclusions Compared with the positive control group,subnormothermic normoxic mechanical perfusion with genetically modified porcine erythrocyte perfusate increases brain tissue edema in cynomolgus monkeys,but better preserves the morphological integrity of neurons and glial cells.The protective effects may be related to the continuous oxygen and energy supply,maintenance of ion homeostasis and perfusate pH,reduced rejection,and low metabolic state of the whole brain.

Background and Aims:Single-incision laparoscopic cholecystectomy(LC)has become increasingly popular due to its minimal invasiveness and cosmetic advantages,but challenges remain in biliary identification and limited operative space.Indocyanine green(ICG)fluorescence navigation enables real-time visualization of the biliary tract and may enhance surgical safety and efficiency.This study aimed to evaluate the clinical application value of ICG fluorescence navigation in single-incision LC.Methods:A retrospective analysis was conducted on 59 patients with benign gallbladder diseases who underwent elective single-incision LC at Yixing Hospital affiliated to Jiangsu University from January 2023 to December 2024.Patients were divided into a fluorescence group(n=27)and a white-light group(n=32)according to whether ICG fluorescence navigation was applied.The two groups were compared in terms of Calot's triangle dissection time,operative time,intraoperative blood loss,surgeon satisfaction,hospital stay,and postoperative complications.Results:No significant differences were observed in baseline clinical characteristics between the two groups(all P＞0.05).In the fluorescence group,the cystic duct,common hepatic duct,and common bile duct were all successfully visualized.Compared with the white-light group,the fluorescence group had significantly shorter Calot's triangle dissection time[(25.56±3.49)min vs.(38.81±5.59)min],shorter operative time[(44.67±3.06)min vs.(61.31±4.96)min],and less intraoperative blood loss[(13.44±1.70)mL vs.(14.50±2.11)mL](all P＜0.05),with significantly higher surgeon satisfaction(P＜0.05).No intraoperative bile duct injury,conversion to three-port surgery,or postoperative complications occurred in either group.Conclusion:ICG fluorescence navigation can significantly improve biliary identification efficiency in single-incision LC,shorten operative time,reduce blood loss,and enhance surgeon satisfaction,demonstrating promising clinical application prospects.

The aim of the experiment was to study the effects of Echinacea purpurea extract on the digestive ability,immune function and intestinal barrier of the Bamei ternary pigs.Forty-eight healthy pigs of 50 days old were randomly divided into four groups,each with three replicates and four pigs in each replicate.The control group(CON)was fed a basal diet,and the experimental group was divided into 5 g/kg(T1),10 g/kg(T2)and 15 g/kg(T3)of Echinacea extract in the basal diet.The pre-test period was 5 days and the main test period was 35 days with free feeding and drinking during the test period.The results showed that:There was no significant effect on the apparent digestibility of nutrients and jejunum morphology in the test group compared to the CON group(P＞0.05).Amylase and trypsin activities were significantly higher in the experimental group and lipase activities were significantly higher in T2 and T3 groups compared to CON group(P＜0.05).GSH,CAT and T-AOC activities were significantly higher in the experimental group compared to CON group(P＜0.05).In comparison with CON group,the levels of IL-1 and IL-6 were significantly lower in the experimental group(P＜0.05),and the levels of TNF-α were significantly lower in the T2 and T3 groups(P＜0.05);and the levels of IL-10 were signifi-cantly higher in the T3 group(P＜0.05).The mRNA and protein expression levels of TLR4,MyD88,TRAF6,and NF-κB were significantly lower in the experimental group(P＜0.05).The mRNA and protein expression levels of CLDN-1,OCLN and ZO-1 were significantly increased in the test group(P＜0.05).The results between the test groups showed that the protein expression levels of CLDN-1 and OCLN were significantly increased in the T3 group compared with the T1 group(P＜0.05).In conclusion,the addition of Echinacea extract to the ration can,to a certain ex-tent,improve the digestive ability,balance the immune ability,and improve the intestinal barrier function of the Bamei ternary pigs,thus promoting the intestinal health of pigs.

Background and Aims:Single-incision laparoscopic cholecystectomy(LC)has become increasingly popular due to its minimal invasiveness and cosmetic advantages,but challenges remain in biliary identification and limited operative space.Indocyanine green(ICG)fluorescence navigation enables real-time visualization of the biliary tract and may enhance surgical safety and efficiency.This study aimed to evaluate the clinical application value of ICG fluorescence navigation in single-incision LC.Methods:A retrospective analysis was conducted on 59 patients with benign gallbladder diseases who underwent elective single-incision LC at Yixing Hospital affiliated to Jiangsu University from January 2023 to December 2024.Patients were divided into a fluorescence group(n=27)and a white-light group(n=32)according to whether ICG fluorescence navigation was applied.The two groups were compared in terms of Calot's triangle dissection time,operative time,intraoperative blood loss,surgeon satisfaction,hospital stay,and postoperative complications.Results:No significant differences were observed in baseline clinical characteristics between the two groups(all P＞0.05).In the fluorescence group,the cystic duct,common hepatic duct,and common bile duct were all successfully visualized.Compared with the white-light group,the fluorescence group had significantly shorter Calot's triangle dissection time[(25.56±3.49)min vs.(38.81±5.59)min],shorter operative time[(44.67±3.06)min vs.(61.31±4.96)min],and less intraoperative blood loss[(13.44±1.70)mL vs.(14.50±2.11)mL](all P＜0.05),with significantly higher surgeon satisfaction(P＜0.05).No intraoperative bile duct injury,conversion to three-port surgery,or postoperative complications occurred in either group.Conclusion:ICG fluorescence navigation can significantly improve biliary identification efficiency in single-incision LC,shorten operative time,reduce blood loss,and enhance surgeon satisfaction,demonstrating promising clinical application prospects.

In elderly patients with malignant tumor,sarcopenia frequently co-occurs,which is largely attributed to the cachexia state observed in cancer patients.Although both sarcopenia and cachexia involve muscle loss,there are significant distinctions between the two conditions.Current research indicates that sarcopenia is a multifactorial condition,influenced by factors such as aging,endocrine dysfunction,motor neuron loss,cachexia,inflammation,and malnutrition.With the increasing depth of research into the correlation between sarcopenia and the nervous and immune systems,this article reviews the research progress on sarcopenia in patients with malignant tumors,with a focus on neuroimmunological aspects.It systematically analyzes the mechanisms underlying its onset and progression,as well as the influencing factors and potential intervention strategies for sarcopenia in this patient population.

The prevention and control of emerging and reemerging infectious diseases are crucial for national biosecurity, and surveillance and reporting of pneumonia of unknown etiology are main ways for the early detection of these diseases and mitigation of their severity. This paper summaries the surveillance methods for pneumonia of unknown etiology and emerging and reemerging infectious diseases globally, indicating that such surveillance is mainly conducted based on hospital. Western countries primarily combine active and passive surveillance while utilizing artificial intelligence technology to rapidly identify cases. China mainly use passive surveillance based on the surveillance system for pneumonia of unknown etiology, with the function of early warning in the identification of emerging infectious diseases, such as avian influenza. However, with the improvement in the surveillance system operation, the overlap with other surveillance disease systems, such as influenza, has occurred, and the improvements in case definition and operational protocol are needed. It is recommended to improve the specificity of the case definition, strenthen training in hospital staff, inclduing clinical workers and office workers, and formulate incentive mechanisms. It is necessary to emphasize the responsibility of clinicians as the main force for the detection and reporting of pneumonia of unknown etiology and emerging infectious diseases, improve the appilication of artifical intelligent technique and conduct multi-source surveillance, such as third-party testing.

Objective To investigate the effect of leucine carboxyl methyltransferase 1(LCMT1)knockout on fructose-induced lipid deposition in primary mouse hepatocytes.Methods Primary hepatocytes were isolated from wild-type(WT)and hepatocyte-specific LCMT1 knockout(KO)mice via a two-step hepatic portal vein perfusion method.The cells were divided into four groups:WT-control group,WT-fructose group,KO-control group,and KO-fructose group.Cell viability was determined through Alamar-Blue assays.Hepatocyte injury was evaluated based on alanine aminotransferase and aspartate aminotransferase levels.Lipid deposition was visualized via Oil Red O staining and lipid droplet green fluorescence staining,and the cellular triglyceride content was quantified via a GPO-POD assay.The mRNA expression of lipid metabolism-related genes was detected via quantitative real-time PCR,and the protein expression of LCMT1 and PP2Ac was detected via Western blot.Results Fructose treatment did not alter cell viability significantly in any group,and no significant cell damage was observed(P＞0.05).The WT-fructose group exhibited greater accumulation of lipid droplets in hepatocytes than that in the WT-control group(P＜0.001),with significantly elevated triglyceride contents(P＜0.05).The mRNA levels of the de novo lipid synthesis genes ChREBP,SREBP-1c,and ACC1 were increased significantly(P＜0.05,P＜0.001,P＜0.001),whereas FAS expression did not differ significantly between groups(P＞0.05).The mRNA levels of the lipid uptake genes FABP1 and FATP2 also increased significantly(both P＜0.05).In contrast,the KO-fructose group presented a reduced number of lipid droplets(P＜0.01,P＜0.001),decreased triglyceride content(P＜0.05),and decreased mRNA levels of ChREBP,SREBP-1c,ACC1,FABP1,and FATP2(P＜0.01,P＜0.001,P＜0.001,P＜0.001,P＜0.05);CPT1 mRNA levels were markedly increased(P＜0.01).Total PP2Ac expression was significantly higher(P＜0.05)and PP2Ac demethylation was significantly lower(P＜0.01)in the WT-fructose group than in the WT-control group.In the KO-control group,total PP2Ac expression remained unchanged(P＞0.05),whereas PP2Ac demethylation was markedly elevated(P＜0.001).Compared with levels in the WT-fructose group,the KO-fructose group presented markedly lower total PP2Ac expression and significantly higher PP2Ac demethylation levels(P＜0.05,P＜0.01,respectively).Conclusions LCMT1 knockout alleviates fructose-induced lipid deposition in primary hepatocytes by inhibiting lipid uptake,increasing fatty acid oxidation,and downregulating de novo lipid synthesis.These effects are medicated by the LCMT1 knockout-mediated upregulation of PP2Ac demethylation,thereby modulating PP2A activity.