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.

[Objective] To explore the predictive value of HALP score for prognosis in patients with multiple myeloma (MM). [Methods] A retrospective analysis was conducted on laboratory indicators and related clinical data of newly diagnosed multiple myeloma (NDMM) patients, treated at the Affiliated Hospital of North Sichuan Medical College from January 2016 to October 2023, prior to their first treatment. The HALP score was calculated, and the optimal cutoff value for HALP was determined using X-tile software. Survival analysis was performed using Kaplan-Meier curves for high HALP and low HALP groups. Univariate and multivariate analyses were conducted using the Cox regression model, and a forest plot was generated using Graphpad Prism to illustrate factors that may impact patient prognosis. The predictive ability of HALP score combined with β2-microglobulin and ECOG score for prognosis in MM patients was evaluated using receiver operating characteristic curve (ROC) analysis. [Results] A total of 203 MM patients were included, with the optimal cutoff value for HALP score being 29.15 (P<0.05). Among them, 101 patients were in the low HALP score group, and 102 patients were in the high HALP score group. The results of univariate and multivariate analysis using the Cox regression model showed that a HALP score <29.15 was an independent risk factor for progression-free survival (PFS) and overall survival (OS) (P<0.05). ROC curve analysis indicated that the combination of HALP score with β2-microglobulin and ECOG score had a higher predictive value for prognosis in MM patients compared to using HALP score alone. [Conclusion] The HALP score is closely related to the prognosis of patients with NDMM. A low HALP score indicates a poorer prognosis, while the combination of HALP score with β2-microglobulin and ECOG score provides a higher predictive value when assessed together.

Objective To construct an in vitro"metabolic memory"cell model of HT-22 mouse hippocampal neurons induced by high glucose,and to investigate the effect of"metabolic memory"on apoptosis and histone acetylation in HT-22 cells.Methods HT-22 cells were cultured in high glucose medium(glucose concentration was 55 mmol/L)and conventional glucose medium(glucose concentration was 25 mmol/L),and cells were divided into the control group(NG 4,6 and 8 groups,25 mmol/L glucose was cultured for 4,6 and 8 days,respectively),the high glucose group(HG 4,6 and 8 groups,respectively)and the metabolic memory group(HG2NG2,HG2NG4,HG2NG6,HG4NG2 and HG4NG4 groups,high glucose culture for 2 days to 25 mmol/L glucose culture for 2,4 or 6 days,high glucose culture for 4 days to 25 mmol/L glucose culture for 2 or 4 days).Cell viability was detected by CCK-8 method.The release of lactate dehydrogenase(LDH)in cell culture supernatant was detected,and the optimal time to establish a"metabolic memory"model was selected.Subsequently,cells were divided into the NG4 group,the NG8 group,the HG4 group,the HG4NG4 group and the HG8 group,and the cell morphology of each group was observed by optical microscope.The apoptosis rate was detected by flow cytometry.The activities of deacetylase(HDAC)and histone acetyltransferase(HAT)were detected by enzyme-linked immunosorbent assay(ELISA).Western blot assay was used to detect expression levels of histone deacetylase 4(HDAC4),B lymphocyte tumor 2(Bcl-2),Bcl-2 related X protein(Bax)and Caspase-3 protein.Results The HG4NG4 group was the ideal cell model with high glucose metabolic memory.Cells of the NG4 group and the NG8 group were interwoven into a dense network,growing well,with spindle shaped cells and distinct synaptic structures.However,in the HG4 group and the HG8 group,the cell body became round,synaptic structure disappeared and growth was inhibited.In the HG4NG4 group,the number of cells increased but their morphology was damaged.Results of flow cytometry showed that compared with the NG8 group,the apoptosis rates were significantly increased in the HG8 group and the HG4NG4 group(P＜0.05).ELISA results showed that compared with the NG8 group,the expression levels of HDAC4,Bax,and Caspase-3 proteins increased in the HG8 group and the HG4NG4 group,while the expression level of Bcl-2 protein significantly decreased(P＜0.05).Compared with the HG8 group,there were no significant differences in protein expression levels of HAT and HDAC in the HG4NG4 group.Western blot reslts showed that compared with the NG8 group,the levels of HDAC4,Bax and Caspase-3 protein increased in the HG8 group and the HG4NG4 group(P＜0.05).Compared with the HG8 group,there were no significant differences in protein expression levels in the HG4NG4 group.Conclusion HT-22 mouse hippocampal neurons cultured with 55mmol/L high glucose for 4 days,and then cultured with 25 mmol/L glucose for 4 days are the ideal"metabolic memory"cell model.The mechanism may be related to the increased activity of HDAC,HAT and HDAC4 expression in the hyperglycemic model.

Objective:To explore the effect of getting out of bed activity 6 hours after silicone breast augmentation surgery on the comfort of patients.Methods:From May 2019 to August 2020, 84 female patients aged 20-37 (28.34±6.27) years who underwent silicone breast augmentation surgery at the Department of Plastic Surgery and Cosmetology at the Second People′s Hospital of Guangdong Province were randomly divided into a control group and an observation group, with 42 patients in each group, using a random number table method. The control group got out of bed 24 hours after surgery, while the observation group got out of bed 6 hours after surgery. The chest drainage and fixation patency, comfort, silicone prosthesis displacement, and incision bleeding rate between the two groups of patients were compared 24 hours after surgery.Results:The extraction time of the thoracic drainage tube in the observation group was (82.64±11.78 hours), which was shorter than that in the control group (90.67±12.44 hours), and the difference was statistically significant ( t=5.369, P<0.01). The comfort scores of the observation group patients at 6 hours after surgery (95.07±4.14) and 24 hours after surgery (97.52±4.07) were higher than those of the control group at 6 hours (91.14±4.03) and 24 hours (94.19±3.93), and the difference was statistically significant ( t=4.413, 3.832, both P<0.01); 7 days after surgery, there were 2 cases of silicone prosthesis displacement in the observation group, with an incidence rate of 4.76% (2/42); there were 2 cases of silicone prosthesis displacement in the control group, with an incidence rate of 4.76% (2/42); there were 6 cases of incision bleeding, with an incidence rate of 14.29% (6/42); there were 5 cases of incision bleeding in the control group, with an incidence rate of 11.90% (5/42). There was no statistically significant difference in the incidence of silicone implant displacement and incision bleeding between the two groups (χ 2=0.21, 0.10, P>0.05). Conclusions:Getting out of bed and moving around 6 hours after silicone breast augmentation surgery can help improve the patient′s physical comfort, ensure smooth fixation of the chest drainage tube, and do not increase the occurrence of incision bleeding in patients.

Clinicians need to focus on various points in the diagnosis and treatment of insomnia.This article prescribed the treatment protocol based on the unique features,such as insomnia in the elderly,women experiencing specific physiologi-cal periods,children insomnia,insomnia in sleep-breathing disorder patients,insomnia in patients with chronic liver and kidney dysfunction.It pro-vides some reference for clinicians while they make decision on diagnosis,differentiation and treat-ment methods.

Cognitive frailty is a reversible neurodegeneration, and its early identification and prevention are crucial. This paper summarizes the current situation, risk factors, and risk prediction models of cognitive frailty in elderly patients with chronic diseases, aiming to provide a reference for clinical medical and nursing staff to identify the risk of cognitive frailty in elderly patients with chronic diseases as early as possible and formulate intervention measures.

Objective To explore and validate the mechanism of Prepared Radix Polygoni Multiflori in the treatment of NAFLD based on network pharmacology and animal non-alcoholic fatty liver disease(NAFLD)model experiments.Methods Consult the literature to compare the differences between Radix Polygoni Multiflori and Prepared Radix Polygoni Multiflori(PRPM).Herb database and SwissADME database were used to screen the active ingredients of Prepared Radix Polygoni Multiflori,SwissTargetPrediction database was used to predict its targets,OMIM,DISGENET and GEENCARDS databases were used to screen the NAFLD-related targets,conduct GO functional enrichment analysis and KEGG pathway enrichment analysis.The active ingredient-target-KEGG signaling pathway-NAFLD network was mapped later.The mice with NAFLD were treated with Prepared Radix Polygoni Multiflori by gavage for 8 weeks;serum triglyceride level and alanine aminotransferase(ALT)activity were measured;the liver lesions were observed by HE staining;the potential mechanism of action of Radix Polygoni Multiflori in the treatment of NAFLD was verified by Western blot.Results The differences between Radix Polygoni Multiflori and PRPM were consulted.Six pharmacological components and 32 potential action targets of Radix Polygoni Multiflori for the treatment of NAFLD were screened by network pharmacology,GO and KEGG pathways were enriched to lipid and atherosclerosis-related pathways,AMPK signaling pathway,etc.;HE staining verified that Prepared Radix Polygoni Multiflori has the function of improving NAFLD and is associated with the alteration of FASN,ACC,SCD protein of AMPK signaling pathway.Conclusion Radix Polygoni Multiflori has the potential to improve NAFLD by regulating FASN,ACC and SCD.

OBJECTIVE To establish the fingerprint and multi-component content determination method of Crataegus pinnatifida leaves from different producing areas， and to evaluate the quality of C. pinnatifida leaves and screen the differential markers. METHODS Seventy-eight batches of C. pinnatifida leaves were collected from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province. High-performance liquid chromatography （HPLC） and Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints （2012 edition） were used to draw the fingerprints and conduct similarity evaluation. Grey correlation analysis， cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed by using SPSS 19.0， MetaboAnalyst 5.0 and SIMCA 14.1 software. The differential markers affecting the quality of C. pinnatifida leaves were screened with variable importance in the projection （VIP） value greater than 1 and the error line not exceeding the origin as the criterion. Using vitexin rhamnoside as an internal reference， the contents of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin in 78 batches of C. pinnatifida leaves were determined by the same HPLC combined with quantitative analysis of multi- components by single-marker （QAMS）， and the results were compared with external standard method. RESULTS Eight common peaks were calibrated in the fingerprints for 78 batches of C. pinnatifida leaves from 4 producing areas. Five known components were identified， including chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， vitexin rhamnoside （peak 4）， hypericin （peak 7） and isoquercetin （peak 8）； their similarities ranged from 0.871 to 0.998. Average relative correlations of samples from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province were 0.538， 0.528， 0.462 and 0.435， respectively. CA and PCA showed that the samples from Chengde of Hebei Province and Huludao of Liaoning Province were roughly classified into one category， while the samples from Linyi of Shandong Province and Yuncheng of Shanxi Province were roughly classified into one category； VIP values of peak 1， 2， 3 and 5 were all greater than 1. By QAMS， the relative correction factors of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin were 0.401， 0.993， 1.670 and 1.615 （RSD＜2%）. Compared with external standard method， except for isoquercetin in the two batches of samples （S39 and S41）， there was no significant difference in the content of each component in other batches of samples （the relative deviations≤ 5%）. CONCLUSIONS The established fingerprint and QAMS method are simple to operate and can be used to evaluate the quality of C. pinnatifida leaves. The sample from Chengde of Hebei Province is relatively good in quality. Chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， and the corresponding components of peaks 2 and 5 may be differential markers affecting the quality of C. pinnatifida leaves.

Objective:To investigate the reasons for efficacy heterogeneity analysis of 3 patients with pantothenate kinase associated neurodegeneration (PKAN) from one family after deep brain stimulation (DBS).Methods:Three PKAN patients with the same PANK2 gene mutation from one family, admitted to and accepted bilateral subthalamic nucleus (STN)-DBS in our hospital from May to August 2021, were selected. The clinical manifestations, including general condition, medical history, symptoms, and signs, of these 3 patients were collected; Burkefahn-Marsden Dystonia Movement Rating Scale (BFMDRS) scores and surface electromyography results were analyzed and compared before DBS and 2 weeks and 6 months after DBS. Results:The clinical phenotype of these 3 patients had obvious heterogeneity: patient 1 had the latest onset age and shortest duration, and the main manifestation included abnormal body activity and forced postures, without obvious body deformation; preoperative surface electromyography suggested that the involuntary muscle contractions intensity was the smallest, and involuntary movements characterized by alternating patterns of contractile discharge activity were the predominant. The patient 2 and patient 3 had early onset age and long course of disease with gradually aggravated disease, and manifested as lower limb claudication and involuntary limb torsion; patient 3 also had marked limb deformities; preoperative surface electromyography showed high intensity of involuntary muscle contraction and torsional spastic dystonia characterized by co-contractile firing activity. The BFMDRS scores and surface electromyography results of the 3 patients after DBS were significantly improved as compared with those before surgery, with obvious heterogeneity; the improvement rates of BFMDRS scores of patient 1, 2 and 3 were 88.1%, 60.5%, and 43.2%, and the improvement rates of surface electromyography were 82.36%, 63.79% and 72.25%, respectively,at 6 months after surgery as compared with those before surgery.Conclusion:PKAN exhibits complicated clinical heterogeneity, which is one of the reasons for efficacy heterogeneity for PKAN after DBS.

Objective:To investigate the reasons for efficacy heterogeneity analysis of 3 patients with pantothenate kinase associated neurodegeneration (PKAN) from one family after deep brain stimulation (DBS).Methods:Three PKAN patients with the same PANK2 gene mutation from one family, admitted to and accepted bilateral subthalamic nucleus (STN)-DBS in our hospital from May to August 2021, were selected. The clinical manifestations, including general condition, medical history, symptoms, and signs, of these 3 patients were collected; Burkefahn-Marsden Dystonia Movement Rating Scale (BFMDRS) scores and surface electromyography results were analyzed and compared before DBS and 2 weeks and 6 months after DBS. Results:The clinical phenotype of these 3 patients had obvious heterogeneity: patient 1 had the latest onset age and shortest duration, and the main manifestation included abnormal body activity and forced postures, without obvious body deformation; preoperative surface electromyography suggested that the involuntary muscle contractions intensity was the smallest, and involuntary movements characterized by alternating patterns of contractile discharge activity were the predominant. The patient 2 and patient 3 had early onset age and long course of disease with gradually aggravated disease, and manifested as lower limb claudication and involuntary limb torsion; patient 3 also had marked limb deformities; preoperative surface electromyography showed high intensity of involuntary muscle contraction and torsional spastic dystonia characterized by co-contractile firing activity. The BFMDRS scores and surface electromyography results of the 3 patients after DBS were significantly improved as compared with those before surgery, with obvious heterogeneity; the improvement rates of BFMDRS scores of patient 1, 2 and 3 were 88.1%, 60.5%, and 43.2%, and the improvement rates of surface electromyography were 82.36%, 63.79% and 72.25%, respectively,at 6 months after surgery as compared with those before surgery.Conclusion:PKAN exhibits complicated clinical heterogeneity, which is one of the reasons for efficacy heterogeneity for PKAN after DBS.