Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H₂O₂ and divided into four groups: control (NC), 50 μmol/L H₂O₂, 110 μmol/L H₂O₂ and 150 μmol/L H₂O₂. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H₂O₂, and siSIRT3+H₂O₂. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O₂•⁻) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H₂O₂ decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O₂•⁻ did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O₂•⁻ level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H₂O₂ group exhibited decreased cellular ATP levels, elevated mitochondrial O₂•⁻ levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H₂O₂ group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.

ObjectiveTo dynamically observe and evaluate the damage to the pulmonary air-blood barrier in mice during the inflammatory cancer transformation process of ulcerative colitis （UC） based on the "lung-intestine correlation" theory. MethodsSixty-five C57BL/6 mice were divided into a normal group （n=25） and a model group （n=40） using a random number table. Azoxymethane/dextran sodium sulfate （DSS） method was used to establish a mouse model of UC inflammation cancer transformation in the modeling group. According to the tissue collection time points at 5， 8， 11， 13， and 15 weeks， the normal group mice were randomly divided into the normal 5w， 8w， 11w， 13w， and 15w groups. The model group mice， 10 mice of which died after the first cycle of DSS administration， were randomly divided into model 5w， 8w， 11w， 13w， and 15w groups. During the experiment， the general condition of the mice was observed daily， and their body weight was measured weekly. At the corresponding tissue collection time points， the colon length of each group was measured. Histopathology of mouse lung and colon tissues was examined using HE staining. Immunofluorescence was used to detect changes in the positive expression of tight junction protein （ZO-1）， vascular endothelial cadherin （VE-cadherin）， and cytoskeletal protein （F-actin） in lung and colon tissues. RT-PCR was used to detect the mRNA expression of apoptosis regulatory proteins B-cell lymphoma-2 （Bcl-2）， BCL2-associated X protein （Bax）， and Cysteine aspartic acid protease-3 （Caspase-3） in lung tissues. Western Blot was employed to measure protein levels of ZO-1， VE-cadherin， and F-actin in lung tissues. ResultsCompared to the normal group at the same time point， the mice in the model group at each time point generally had poorer conditions， with weight loss and shortened colon length （P＜0.05 or P＜0.01）. In the model 5w group， there was significant inflammatory cell infiltration in the colon tissue； in the model 8w group， there was mild atypical hyperplasia； in the model 11w group， the crypt structure was disordered， and moderate to severe atypical hyperplasia occurred； in the model 13w and 15w groups， tumors appeared. Pulmonary interstitial lesions， inflammation， vasculitis， and fibrosis were observed at all stages of UC inflammation cancer transformation. The protein levels of ZO-1， VE-cadherin， and F-actin， as well as Bcl-2 mRNA expression in lung tissue decreased during the acute inflammatory recovery period， atypical hyperplasia period， and canceration period， while the expressions of Bax and Caspase-3 mRNA increased； the expressions of ZO-1， VE-cadherin， and F-actin proteins in colon tissue decreased during the acute inflammatory recovery period， atypical hyperplasia period， and canceration period （P＜0.01 or P＜0.05）. Compared to the model 5w group， the ZO-1 and F-actin protein levels and Bcl-2 mRNA expression in lung tissue in the other model groups increased in the atypical hyperplasia period and canceration period， while the expressions of Bax and Caspase-3 mRNA decreased； the expression of ZO-1 protein in colon tissue increased in the canceration period， and the expression of VE-cadherin protein decreased in the atypical hyperplasia period （P＜0.01 or P＜0.05）. ConclusionIn the process of "inflammatory response-atypical hyperplasia-carcinogenesis" in UC inflammatory cancer transformation mice， there were damage to air-blood barrier.

Objective:To investigate the mechanisms by which calpain small subunit 1(CAPN4)regulates cisplatin resistance and cancer stem cell(CSC)stemness in lung adenocarcinoma,and to provide experimental evidence for reversing drug resistance through targeting CSC stemness.Methods:Tissue samples were collected from 10 lung adenocarcinoma patients who underwent surgical resection at Fujian Cancer Hospital from January 2023 to January 2024.Immunohistochemistry(IHC)stining was used to detect the differential expression of CAPN4 in five cisplatin-resistant and five cisplatin-sensitive lung adenocarcinoma tissues,followed with a histological scoring(H-score).CAPN4 gene expression-related survival analysis in lung cancer patients was conducted using The Cancer Genome Atlas(TCGA)database and Gene Expression Profiling Interactive Analysis(GEPIA)platform.Additionally,tissue samples from two cisplatin-resistant and two cisplatin-sensitive lung adenocarcinoma cases were collected to establish lung adenocarcinoma organoid(PDO)models.H-E and IHC staining were used to assess the morphological consistency between PDOs and the primary tumors.CAPN4 gene expression was silenced using lentivirus-mediated shRNA transduction.The expression levels of stem cell markers ALDH1A1,CD133,Nanog,and SOX9 were detected at both the gene and protein levels using quantitative polymerase chain reaction(qPCR)and Western blotting(WB),respectively.The sensitivity of CAPN4-knockdown PDOs to cisplatin was evaluated using the adenosine triphosphate(ATP)assay,and the apoptosis was assessed using the caspase-3 assay.Results:IHC results showed that CAPN4 protein expression was significantly upregulated in cisplatin-resistant lung adenocarcinoma tissues(P<0.05).TCGA cohort analysis revealed that high CAPN4 expression was significantly associated with poor prognosis(reduced OS)in lung adenocarcinoma patients(HR=1.4,P<0.05).PDOs derived from cisplatin-resistant patients exhibited significant upregulation in CAPN4 protein and stemness markers at both gene and protein levels(all P<0.05).Cisplatin sensitivity assays demonstrated that PDOs derived from cisplatin-resistant patients had significantly higher IC50 values than those from cisplatin-sensitive patients(P<0.05).After CAPN4 knockdown,the expression of stem cell makers in PDOs derived from cisplatin-resistant patients were significantly reduced,and their sensitivity to cisplatin was enhanced(P<0.05).Conclusion:Knockdown of CAPN4 reduces stem cell marker expression and enhances cisplatin sensitivity in lung adenocarcinoma PDOs,providing a potential therapeutic target for reversing cisplatin resistance in lung cancer.

Objective:To understand the mental problem in elderly male prisoners and its related factors.Methods:A total of 289 elderly male prisoners from five prisons in Sichuan Province were selected.They were assessed with the self-compiled general situation questionnaire,12-item General Health Questionnaire(GHQ-12,with a total score ≥ 3 indicating a tendency towards mental problem),short-form UCLA Loneliness Scale(ULS-8)and Insomnia Severity Index-Chinese Version(ISI-C).Results:The detection rate of mental problem was 41.52％.Multiple factor logistic regression analysis indicated that having a remaining sentence of≥5 years,educa-tional levels of primary school and below,high school or technical secondary school and above,involvement in drug-related crimes,property crimes,violent crimes,good and poor relationships with spouses,average relationships with fellow inmates,self-assessed blood pressure levels that were low and high,and having insomnia were risk fac-tors for mental problem(OR=9.55,11.75,13.88,82.24,33.96,363.52,5.77,113.27,2.33,1.87,3.80,42.89).Conclusion:The mental problem among elderly male prisoners are related to their sentence,education lev-el,crime type,relationship with spouse and cellmates,self-rated blood pressure and sleep status.

Objective:To understand the mental problem in elderly male prisoners and its related factors.Methods:A total of 289 elderly male prisoners from five prisons in Sichuan Province were selected.They were assessed with the self-compiled general situation questionnaire,12-item General Health Questionnaire(GHQ-12,with a total score ≥ 3 indicating a tendency towards mental problem),short-form UCLA Loneliness Scale(ULS-8)and Insomnia Severity Index-Chinese Version(ISI-C).Results:The detection rate of mental problem was 41.52％.Multiple factor logistic regression analysis indicated that having a remaining sentence of≥5 years,educa-tional levels of primary school and below,high school or technical secondary school and above,involvement in drug-related crimes,property crimes,violent crimes,good and poor relationships with spouses,average relationships with fellow inmates,self-assessed blood pressure levels that were low and high,and having insomnia were risk fac-tors for mental problem(OR=9.55,11.75,13.88,82.24,33.96,363.52,5.77,113.27,2.33,1.87,3.80,42.89).Conclusion:The mental problem among elderly male prisoners are related to their sentence,education lev-el,crime type,relationship with spouse and cellmates,self-rated blood pressure and sleep status.

Alzheimer's disease(AD)is the most common neurodegenerative disorder worldwide,characterized primarily by cognitive impairment and memory dysfunction.Its pathological mechanisms involve the toxic accumulation of amyloid β-protein(Aβ),hyperphosphorylation of Tau protein leading to neurofibrillary tangles,mitochondrial dysfunction,synaptic impairment,cholinergic system dysfunction,neuroinflammation,and oxidative stress.Current clinical treatments for AD include acetylcholinesterase inhibitors and N-methyl-D-aspartate antagonists,which can improve cognitive function but fail to slow disease progression and often have side effects.Research indicates that traditional Chinese medicine(TCM)may regulate the phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)signaling pathway,promoting neuronal survival,inhibiting neuroinflammation,reducing oxidative stress,preventing apoptosis,and decreasing Aβ deposition,thus improving the symptoms of AD.This review summarizes the mechanisms by which individual TCM components,extracts,and formulas may regulate the PI3K/AKT pathway in the treatment of AD,with the aim of providing a theoretical foundation for the application of TCM in AD therapy.

BACKGROUND:As the end bearing joint of the human body,the ankle joint bears the top-down pressure of the body,which leads to the ankle joint is easy to be damaged in the movement,can induce functional ankle instability,which negatively affects daily life.The study of lower extremity biomechanics in patients with functional ankle instability during counter movement jump is of great significance for scientific training,prevention of ankle injury,and clinical rehabilitation after injury. OBJECTIVE:To investigate the kinetics and kinematics of lower limbs in the longitudinal jumping of functional ankle instability population. METHODS:From March to September 2023,15 male patients with functional ankle instability and 15 healthy people,aged 22-28 years old,were recruited in Soochow University.All subjects completed counter movement jump experiment.Vicon infrared high-speed motion capture system and Kistler three-dimensional force measuring table were used to simultaneously collect the lower limb kinematics and kinetics indexes of the two groups of subjects at the take-off stage of counter movement jump,the instant off the ground,the initial landing moment and the peak moment of vertical ground reaction force. RESULTS AND CONCLUSION:(1)At the instant off the ground,the affected side of the functional ankle instability group showed smaller knee internal rotation moment(P=0.020)and smaller ankle internal rotation moment(P=0.009)compared with the affected side of the healthy control group.(2)At the moment of landing,the affected side of the functional ankle instability group showed a smaller hip flexion angle than the affected side of the healthy control group(P=0.039).Compared with the healthy control group,functional ankle instability group showed smaller hip abduction angle(P=0.022),smaller knee varus angle(P=0.010),larger knee external rotation angle(P=0.021),smaller ankle varus angle(P=0.004),and smaller external ankle rotation angle(P=0.008).(3)At the peak of vertical ground reaction force,functional ankle instability group showed a smaller ankle varus angle than healthy control group(P=0.044).(4)The results showed that the lower limb biomechanical characteristics of the patients with functional ankle instability were abnormal compared with the healthy people during counter movement jump,which mainly showed the changes of the kinematics and kinetics indexes of the lower limb joints in the sagittal plane and the frontal plane at the moment of lift-off and landing.These changes reflect that people with functional ankle instability adopt rigid take-off and landing patterns when performing counter movement jump,tend to transfer the load of the affected ankle joint to other joints of the lower limb,and show compensatory phenomenon of the healthy lower limb.Therefore,detection and correction of abnormal biomechanical features should be a part of rehabilitation training for those with functional ankle instability.

Objective To investigate the role of the O-6-methylguanine-DNA methyltransferase (MGMT) gene-3′ untranslated region (UTR) microRNA-associated single nucleotide polymorphism (miRSNP) (rs7896488 G>A) in affecting miR-4297-targeted modulation of MGMT in senescence of lung cells with polymorphic genotypes induced by fractionated low dose ionizing radiation (LDIR). Methods i) MiRSNPs were predicted and screened using bioinformatics, and DNA from two types of lung cells, A549 cells and human bronchial epithelioid cells (HBE cells), was extracted for target gene sequencing. After co-transfection of pGL3c-MGMT-3′UTR-rs7896488 G>A reporter gene recombinant plasmid, pRL-TK Vector with micrON mimic NC #22 or micrON hsa-miR-4297 mimic (set up as the mimic NC group and the miR-4297 mimic group) in these two types of lung cells, dual luciferase reporter gene assay was performed. The relative expression of MGMT mRNA was detected by real-time fluorescence quantitative polymerase chain reaction, and the relative expression of MGMT protein was detected by Western blotting. ii) These two types of lung cells were randomly divided into the control group and irradiation group, which received either 0 or 100 mGy X-rays irradiation seven times. After irradiation, the cells were transfected with either micrON mimic NC #22 or micrON hsa-miR-4297 mimic, resulting in mimic NC + control group, miR-4297 mimic + control group, mimic NC + irradiation group, and miR-4297 mimic + irradiation group. Cells were collected for senescence-associated-β-galactosidase (SA-β-Gal) staining, and the relative expression of matrix metalloproteinase-9 (MMP-9) and chemokine (C-X-C motif) ligand-1 (CXCL-1) proteins was detected via Western blotting. Results i) The rs7896488 G>A was the miRSNP located in the conserved binding region targeted by miR-4297 in the MGMT gene 3′UTR. A549 cells were the rs7896488 GG wild-type homozygous genotype, while HBE cells were the rs7896488 GA heterozygous mutant genotype. In the miR-4297 mimic group, A549 and HBE cells carrying the rs7896488 G allele showed significantly lower dual-luciferase activity compared with that in the mimic NC group (both P<0.01). However, there was no significant difference in dual-luciferase activity between the two groups in both A549 and HBE cells carrying the rs7896488 A allele (both P>0.05). The relative expression levels of MGMT mRNA and MGMT protein of A549 cells in the miR-4297 mimic group were lower than those in the mimic NC group (both P<0.05). However, there was no significant difference in MGMT mRNA and MGMT protein of HBE cells between these two groups (both P>0.05). ii) The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of A549 cells in the miR-4297 mimic+irradiation group were higher than those in the mimic NC + control group, the miR-4297 mimic + control group, and the mimic NC + irradiation group (all P<0.05). The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of HBE cells in the miR-4297 mimic + irradiation group were higher than those in the mimic NC + control group and the miR-4297 mimic + control group (all P<0.05), while there was no significant difference compared with those in the mimic NC + irradiation group (all P>0.05). Conclusion MGMT-3′UTR-miRSNP rs7896488 G>A plays a role in LDIR-induced senescence of lung cells with different polymorphic genotypes by affecting miR-4297-targeted regulation of MGMT.

The glymphatic system is an important pathway for fluid drainage and metabolic waste clearance in the central nervous system. Its core mechanism involves the active cerebrospinal fluid-interstitial fluid circulation process mediated by perivascular spaces and aquaporin-4 channels located on astrocytic endfeet. This process is crucial for eliminating neurotoxic substances such as β-amyloid and tau proteins, as well as maintaining homeostasis in the central nervous system. Recent studies have shown that dynamic changes in the glymphatic system are associated with recovery after ischemic stroke. This article elaborates on the role of the glymphatic system in ischemic stroke and evaluates its potential value as a novel therapeutic target, providing new insights for post-stroke treatment strategies.

Objective:To explore the effects of carcinoma-associated fibroblasts (CAF) in gastric cancer microenvironment on the proliferation of gastric cancer organoids and the sensitivity to chemotherapeutic drugs, as well as the underlying mechanisms.Methods:Tissue specimens of 6 patients with gastric cancer who underwent surgical resection in Fujian Cancer Hospital from June 2022 to January 2024 were collected from the biobank. Organoids derived from gastric cancer and isolated tumor tissue CAF and paracancerous tissue normal fibroblasts (NF) were constructed by the primary tissue culture method. The morphology of organoids and fibroblasts was observed under microscope. Immunohistochemistry (IHC) method was used to detect the expressions of gastric cancer markers in organoids. The expression levels of fibroblast related markers were detected by using immunofluorescence, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting methods. A co-culture model of gastric cancer organoids and fibroblasts was constructed by using the Transwell chamber and then was divided into 3 groups: NF and gastric cancer organoids co-culture group (NF co-culture group), CAF and gastric cancer organoids co-culture group (CAF co-culture group), and gastric cancer organoid separate culture group (separate culture group). Different concentrations of oxaliplatin, paclitaxel, and 5-fluorouracil (5-FU) were applied to the co-culture system of fibroblasts and gastric cancer organoids. The morphology of gastric cancer organoids in each group was observed under microscope, and the relative viability of the organoids was detected by using the CellTiter-Glo 3D luminescence method. CAF and NF of 3 patients who successfully constructed organoids were collected for transcriptome sequencing, and the differentially expressed genes (with |log 2 fold change| > 0 and P < 0.05) between CAF and NF were analyzed. Pathway enrichment analysis of the upregulated differentially expressed genes in CAF was performed by using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Results:The organoids of 4 patients were successfully constructed, among which 3 organoids with relatively consistent results and covering the heterogeneity of the disease were performed for further analysis. After HE staining, microscopic observation showed that the morphological characteristics of the organoids were homogeneous with those of gastric cancer tissues. IHC method detection showed that both organoids and primary tumor tissues both expressed gastric cancer markers CDX2, Villin, and CK20. Inverted microscopic observation revealed that NF was morphologically similar to CAF, but NF had fewer cytoplasmic protrusions. RT-qPCR method, immunofluorescence and Western blotting detections showed that the mRNA and protein expression levels of α-smooth muscle actin (α-SMA), fibroblast activation protein (FAP) and Vimentin in CAF were all higher than those in NF, and the differences were statistically significant (all P < 0.05). Inverted microscope observation showed that after co-cultured for 96 h, the size of organoids in CAF co-culture group [(308±61) μm] was larger than that in NF co-culture group [(155±33) μm] and separate culture group [(91±17) μm], and the differences were statistically significant (all P < 0.05). CellTiter-Glo 3D luminescence assay showed that the relative viability of organoids in the CAF co-culture group was enhanced after 96 h of co-culture compared with NF co-culture group and separate culture group, and the differences were statistically significant (all P < 0.05); there was no statistically significant difference in the relative viability between NF co-culture group and separate culture group ( P > 0.05). With the increase of the concentrations of oxaliplatin, 5-FU and paclitaxel, the relative activity of organoids after drug effect for 96 h in the 3 groups gradually decreased. The relative viability of the organoids in CAF co-culture groups treated with 6.25, 12.5, 25, 50, and 100 μmol/L oxaliplatin, 12.5, 25, 50, and 100 μmol/L 5-FU, and 0.625, 1.25, 2.5, and 5 μmol/L paclitaxel was higher than that of the other 2 groups, and the differences were statistically significant (all P < 0.05). Transcriptome sequencing analysis revealed that compared with NF, there were 893 differentially expressed genes with upregulated mRNA expression and 424 differentially expressed genes with downregulated mRNA expression in CAF. KEGG pathway enrichment analysis showed that the upregulated differentially expressed genes were mainly involved in the calcium signaling pathway. Conclusions:CAF can promote the proliferation ability of gastric cancer organoids and decrease the sensitivity to chemotherapy drugs, which may be related to the calcium signaling pathway.