1.Survey of post-discharge exercise behavior and analysis of factors influencing exercise intensity in patients undergoing lung surgery
Hongyu ZENG ; Xiang WANG ; Tian ZHANG ; Yaqin WANG ; Xing WEI ; Zhen DAI ; Liping ZHANG ; Xiaoqin LIU ; Qiang LI ; Qiuling SHI ; Wei DAI ; Jia LIAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(05):734-742
Objective To investigate the post-discharge exercise behavior and factors influencing moderate to vigorous intensity physical activity (MVPA) in patients undergoing lung surgery. Methods A total of 2874 patients from the large prospective, observational perioperative lung symptom study cohort (CN-PRO-Lung 3) in the Department of Thoracic Surgery at Sichuan Cancer Hospital between April 7, 2021, and January 31, 2024, were selected as the survey subjects. A survey was conducted using the Investigation of Exercise Behavior after Lung Surgery questionnaire and the International Physical Activity Questionnaire-Short Form (IPAQ-SF) among patients who underwent lung surgery. Binary logistic regression was used to analyze the factors influencing patients’ engagement in MVPA. Results A total of 702 patients were surveyed, including 252 males and 450 females, with an average age of (52.4±10.2) years. Patients with lung cancer accounted for 85.9%. Only 36.0% of the patients had regular exercise habits, while 42.3% did not engage in any physical activity. The three main barriers for postoperative exercise were physical discomfort (pain, coughing, shortness of breath, etc, 54.7%), lack of professional guidance (41.7%), and concerns about the surgical wound (28.9%). The proportions of patients engaging in vigorous, moderate, and low-intensity physical activity were 5.7%, 28.2%, and 66.1%, respectively. Multivariate analysis showed that patients with a personal annual income ≥50000 yuan (OR=1.52, 95%CI 1.01-2.29, P=0.044), high school education or above (OR=1.92, 95%CI 1.33-2.76, P<0.001), and lobectomy (OR=1.44, 95%CI 1.02-2.03, P=0.037) engaged in more MVPA. Conclusion Patients undergoing lung surgery have inadequate physical activity after discharge, particularly lacking in MVPA. Patients with higher income, higher educational levels, and lobectomy are more frequently engaged in MVPA. Measures such as symptom control, providing exercise guidance, and enhancing education on wound care may potentially improve the inadequate physical activity in lung surgery patients after discharge.
2.Expert consensus on precise intervention with repetitive transcranial magnetic stimulation for sleep disorders in the elderly
Yuan SHAO ; Jian WANG ; Wei LIANG ; Yingli ZHANG ; Gangqiang HOU ; Xia LI ; Yi XING ; Lu WANG ; Shi TANG ; Yongjun WANG
Sichuan Mental Health 2026;39(2):97-105
In recent years, repetitive transcranial magnetic stimulation (rTMS) has garnered significant attention as a therapeutic approach for sleep disorders in the elderly. However, the prevailing rTMS protocols are predominantly developed based on normative neurophysiological data derived from young adults and fail to incorporate individualized parameters tailored to the brain characteristics of the elderly. To address this gap, the consensus development group synthesized the latest evidence from 2010 to 2025 and established a standardized rTMS protocol specifically for elderly patients with sleep disorders. Adhering to the Appraisal of Guidelines for Research and Evaluation II (AGREE II) framework, systematically screened randomized controlled trials (RCTs) and systematic reviews regarding rTMS in the treatment of sleep disorders across various conditions. Meanwhile, the Grading of Recommendations Assessment, Development and Evaluation (GRADE) system was employed to rigorously grade the quality of evidence and the strength of recommendations. This consensus guideline delineates precise rTMS protocols for the management of sleep disorders in the elderly, highlights the adjustment of stimulation intensity according to scalp-cortex distance recommends either MRI‑guided neuronavigation or the Beam F3/F4 heuristic approach for accurate target localization, thereby providing precise rTMS intervention protocol for sleep disorders in the elderly, aiming to enhance clinical efficacy while ensuring treatment safety. [Funded by National Key Research and Development Program (number, 2023YFC3603200); General Program of Shenzhen Science and Technology Innovation Commission (number, JCYJ20240813112859008, JCYJ20240813112900002); Youth Program of Shenzhen Kangning Hospital (number, KN2023A004); www.guidelines-registry.cn number, PREPARE-2026CN530]
3.Visiting behavior and health management strategy of patients with cardiovascular disease under the family doctor contract service model
Juan LI ; Bingqi XING ; Wei WANG ; Yunfeng TIAN
Journal of Public Health and Preventive Medicine 2026;37(3):176-179
Objective To explore the visiting behavior and health management strategy of patients with cardiovascular disease (CVD) and the industry management strategies under the family doctor contract service model. Methods A total of 351 patients with CVD from January 2022 to January 2025 were included in the case investigation by random sampling. Taking the number of visits in community health service institutions/total number of visits ≥90% as high compliance, the influencing factors of the visiting behavior of CVD patients were analyzed. Results Finally, 344 valid questionnaires were collected, with an effective recovery rate of 98.01%. The study subjects were categorized into high compliance group (271 cases) and low compliance group (73 cases). The scores of policy cognition understanding, institutional trust and medical service understanding in the high compliance group were higher compared with those in the low compliance group (P<0.05). There was no significant difference in the understanding of medical expenses between the two groups (P>0.05). Multivariate logistic regression model analysis revealed that CVD duration [OR (95% CI): 1.72 (1.21-2.44)], policy cognition understanding [OR (95% CI): 2.16 (1.20-3.89)] and institutional trust [OR (95% CI): 1.88 (1.08-3.25)] were independently associated with medical treatment behavior in CVD patients (P<0.05). Conclusion Enhancing the policy cognition understanding and institutional trust of CVD patients on family doctor contract service can promote patients to seek medical treatment and promote the high-quality development of family doctor contract service.
4.Jianpi Xiao'ai Prescription Inhibits Colorectal Cancer Progression by Inducing Mitochondrial Dysfunction via Modulation of iNOS-ARG1 Axis
Xing LUO ; Bo PAN ; Jianfeng FU ; Jia HUANG ; Wei PENG ; Fang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):99-111
ObjectiveTo investigate the mechanism by which Jianpi Xiao'ai prescription (JPXAP) inhibits colorectal cancer progression by regulating the inducible nitric oxide synthase-arginase 1 (iNOS-ARG1) metabolic axis and inducing mitochondrial reactive oxygen species (mito-ROS)-mediated mitochondrial structural and functional impairment. MethodsAn arginine metabolism disorder model of human colorectal cancer HCT116 cells was established by combined treatment with recombinant human interferon-γ (IFN-γ, 10 μg·L-1) and N(ω)-hydroxy-L-arginine (Nor-NOHA, 200 μmol·L-1) for 24 h, followed by intervention with 5%, 10%, or 20% JPXAP-containing serum. Cell proliferation was assessed using cell counting kit-8 (CCK-8), 5-ethynyl-2′-deoxyuridine (EdU) staining, and colony formation assays. Cell invasion and migration were evaluated using Transwell chamber and wound healing assays. Mitochondrial membrane potential (MMP) and ROS levels were assessed by JC-1 and MitoSOX staining, respectively. Mitochondrial ultrastructure was observed by transmission electron microscopy (TEM). The expression of iNOS, ARG1, and mitochondrial dynamics-related proteins, including mitofusin 2 (MFN2) and dynamin-related protein 1 (DRP1), was analyzed by Western blot and immunofluorescence. The levels of L-arginine, citrulline, and urea were determined by colorimetric methods and enzyme-linked immunosorbent assay (ELISA). ResultsCompared with the blank group, the model group exhibited significantly upregulated iNOS expression, downregulated ARG1 expression, a decreased ARG1/iNOS ratio, reduced L-arginine and urea levels, and increased citrulline levels (P<0.05). Meanwhile, mito-ROS accumulation was significantly increased, the JC-1 red/green fluorescence ratio was decreased, and mitochondria showed swelling and cristae disruption, indicating that metabolic disorder induced mitochondrial injury. Compared with the model group, all JPXAP-treated groups further decreased the ARG1/iNOS ratio, enhanced nitric oxide (NO) and reactive nitrogen species accumulation, further reduced L-arginine and urea levels, and increased citrulline levels (P<0.01). EdU-positive rate, colony formation rate, wound healing rate, and Transwell invasion number all decreased significantly with increasing serum concentration (P<0.01). Mito-ROS levels were further elevated, and the JC-1 red/green ratio further decreased. TEM revealed aggravated mitochondrial swelling and vacuolization. MFN2 expression was downregulated and DRP1 expression was upregulated (P<0.01),in a dose-dependent manner. ConclusionJPXAP further activates NO-mediated oxidative/nitrosative stress under arginine metabolism imbalance, inducing mito-ROS accumulation, MMP collapse, and mitochondrial dynamics imbalance, thereby inhibiting colorectal cancer cell proliferation and migration. These findings reveal an antitumor mechanism of JPXAP based on coordinated targeting of the "metabolism-mitochondria" axis.
5.A study on the latent profile analysis and influencing factors of public acceptance of palliative care in Hainan Province
Ling ZHANG ; Xiaoting ZHAO ; Wenling LIU ; Shiyuan WANG ; Wei LIU ; Hongjiao CHEN ; Xing GAO
Chinese Medical Ethics 2026;39(5):669-677
ObjectiveTo explore the potential categories and characteristics of the public hospice care demand in Hainan Province, and analyze different potential types of influencing factors, so as to provide reference for relevant departments to improve the public awareness and demand of hospice care. MethodsUsing convenience sampling method, select 6484 cities of the public as the survey object, using the general data questionnaire, the hospice care demand questionnaire of the potential profile analysis, and analyze the influencing factors of the public hospice care demand category. ResultsThe characteristics of the hospice care demand in Hainan Province were divided into three potential categories: low demand group (14.19%), medium demand group (49.99%) and high demand group (35.82%). Multivariate analysis showed that gender, age, education level, cultural belief, and life-death education experience were the main influencing factors of public hospice care demand (p<0.05). Males, those aged 41-60 years, and those with high school education or below had relatively lower hospice care demand, while those with life-death education experience had relatively higher demand. ConclusionRelevant departments should focus on hospice care knowledge popularization and demand enhancement for males, middle-aged groups, and people with low education levels, while strengthening universal life-death education through stratified and classified publicity strategies and educational interventions to improve different populations’ awareness and acceptance of hospice care.
6.Regulatory Effect of Danhe Granules on Oxidative Stress in Rats with Mixed Hyperlipidemia
Jingke MENG ; Susu LIU ; Pan GAO ; Mingjiao JIA ; Bochao JIA ; Qingzheng XING ; Yulong CHEN ; Wei WANG ; Xinlou CHAI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):112-122
ObjectiveTo investigate the therapeutic mechanism of Danhe granules in treating mixed hyperlipidemia based on network pharmacology, as well as animal and cell experiments. MethodsThe active compounds and targets of Danhe granules were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the Encyclopedia of Traditional Chinese Medicine (ETCM). Related targets for mixed hyperlipidemia were obtained from the GeneCards database. The intersecting targets were subjected to Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. A high-fat model was established in human hepatocellular carcinoma cells (HepG2) induced by palmitic acid (PA), followed by intervention with Danhe granules to assess intracellular lipid accumulation and oxidative stress levels. A mixed hyperlipidemia rat model was also established and divided into low-, medium-, and high-dose Danhe granules groups (1.134, 2.268, and 4.536 g·kg-1, respectively), as well as a positive control group treated with pravastatin sodium (4.020 mg·kg-1). After eight weeks of intervention, serum lipid levels, inflammatory factors, oxidative stress indices, and the expression of key hepatic lipid metabolism-related proteins were determined. ResultsNetwork pharmacology identified 93 intersecting targets between Danhe granules and mixed hyperlipidemia, with peroxisome proliferator-activated receptor gamma (PPARG), peroxisome proliferator-activated receptor alpha (PPARA), tumor necrosis factor (TNF), interleukin-6 (IL-6), and IL-1B among the key nodes. The PPAR signaling pathway, AGE/RAGE signaling pathway, lipid metabolism, atherosclerosis and non-alcoholic fatty liver disease (NAFLD) were among the most significantly enriched pathways. Cellular experiments demonstrated that Danhe granules significantly reduced reactive oxygen species (ROS) and malondialdehyde (MDA) levels while increasing catalase (CAT) activity (P<0.05), thereby alleviating intracellular lipid accumulation and triglyceride (TG) content in HepG2. In animal experiments, Danhe granules markedly decreased serum total cholesterol (TC), TG, and low-density lipoprotein cholesterol (LDL-C) levels (P<0.05), reduced hepatic MDA levels, and elevated superoxide dismutase (SOD) and CAT levels. Histological analysis showed alleviation of hepatic steatosis, upregulation of hepatic PPARA and lipoprotein lipase (LPL) expressions, and downregulation of sterol regulatory element-binding protein 1 (SREBP1) expression (P<0.05, P<0.01). ConclusionDanhe granules improve lipid metabolism disorders in mixed hyperlipidemia by reducing MDA levels, enhancing SOD and CAT activities, scavenging excessive ROS, inhibiting oxidative stress, and mitigating liver injury. The underlying mechanism may involve the upregulation of PPARA and LPL and the suppression of SREBP1 expression.
7.Regulatory Effect of Danhe Granules on Oxidative Stress in Rats with Mixed Hyperlipidemia
Jingke MENG ; Susu LIU ; Pan GAO ; Mingjiao JIA ; Bochao JIA ; Qingzheng XING ; Yulong CHEN ; Wei WANG ; Xinlou CHAI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):112-122
ObjectiveTo investigate the therapeutic mechanism of Danhe granules in treating mixed hyperlipidemia based on network pharmacology, as well as animal and cell experiments. MethodsThe active compounds and targets of Danhe granules were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the Encyclopedia of Traditional Chinese Medicine (ETCM). Related targets for mixed hyperlipidemia were obtained from the GeneCards database. The intersecting targets were subjected to Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. A high-fat model was established in human hepatocellular carcinoma cells (HepG2) induced by palmitic acid (PA), followed by intervention with Danhe granules to assess intracellular lipid accumulation and oxidative stress levels. A mixed hyperlipidemia rat model was also established and divided into low-, medium-, and high-dose Danhe granules groups (1.134, 2.268, and 4.536 g·kg-1, respectively), as well as a positive control group treated with pravastatin sodium (4.020 mg·kg-1). After eight weeks of intervention, serum lipid levels, inflammatory factors, oxidative stress indices, and the expression of key hepatic lipid metabolism-related proteins were determined. ResultsNetwork pharmacology identified 93 intersecting targets between Danhe granules and mixed hyperlipidemia, with peroxisome proliferator-activated receptor gamma (PPARG), peroxisome proliferator-activated receptor alpha (PPARA), tumor necrosis factor (TNF), interleukin-6 (IL-6), and IL-1B among the key nodes. The PPAR signaling pathway, AGE/RAGE signaling pathway, lipid metabolism, atherosclerosis and non-alcoholic fatty liver disease (NAFLD) were among the most significantly enriched pathways. Cellular experiments demonstrated that Danhe granules significantly reduced reactive oxygen species (ROS) and malondialdehyde (MDA) levels while increasing catalase (CAT) activity (P<0.05), thereby alleviating intracellular lipid accumulation and triglyceride (TG) content in HepG2. In animal experiments, Danhe granules markedly decreased serum total cholesterol (TC), TG, and low-density lipoprotein cholesterol (LDL-C) levels (P<0.05), reduced hepatic MDA levels, and elevated superoxide dismutase (SOD) and CAT levels. Histological analysis showed alleviation of hepatic steatosis, upregulation of hepatic PPARA and lipoprotein lipase (LPL) expressions, and downregulation of sterol regulatory element-binding protein 1 (SREBP1) expression (P<0.05, P<0.01). ConclusionDanhe granules improve lipid metabolism disorders in mixed hyperlipidemia by reducing MDA levels, enhancing SOD and CAT activities, scavenging excessive ROS, inhibiting oxidative stress, and mitigating liver injury. The underlying mechanism may involve the upregulation of PPARA and LPL and the suppression of SREBP1 expression.
8.Strategic Optimization of CHO Cell Expression Platforms for Biopharmaceutical Manufacturing
Rui-Ming ZHANG ; Meng-Lin LI ; Hong-Wei ZHU ; Xing-Xiao ZHANG
Progress in Biochemistry and Biophysics 2026;53(2):327-341
Chinese hamster ovary (CHO) cells are the most established and versatile mammalian expression system for the large-scale production of recombinant therapeutic proteins, owing to their genetic stability, adaptability to serum-free suspension culture, and ability to perform human-like post-translational modifications. More than 70% of biologics approved by the U.S. Food and Drug Administration rely on CHO-based production platforms, underscoring their central role in modern biopharmaceutical manufacturing. Despite these advantages, CHO systems continue to face three persistent bottlenecks that limit their potential for high-yield, reproducible, and cost-efficient production: excessive metabolic burden during high-density culture, heterogeneity of glycosylation patterns, and progressive loss of long-term expression stability. This review provides an integrated analysis of recent advances addressing these challenges and proposes a forward-looking framework for constructing intelligent and sustainable CHO cell factories. In terms of metabolic regulation, excessive lactate and ammonia accumulation disrupts energy balance and reduces recombinant protein synthesis efficiency. Optimization of culture parameters such as temperature, pH, dissolved oxygen, osmolarity, and glucose feeding can effectively alleviate metabolic stress, while supplementation with modulators including sodium butyrate, baicalein, and S-adenosylmethionine promotes specific productivity (qP) by modulating apoptosis and chromatin structure. Furthermore, genetic engineering strategies—such as overexpression of MPC1/2, HSP27, and SIRT6 or knockout of Bax, Apaf1, and IGF-1R—have demonstrated significant improvements in cell viability and product yield. The combination of multi-omics metabolic modeling with artificial intelligence (AI)-based prediction offers new opportunities for building self-regulating CHO systems capable of dynamic adaptation to environmental stress. Regarding glycosylation uniformity, which determines therapeutic efficacy and immunogenicity, gene editing-based glycoengineering (e.g., FUT8 knockdown or ST6Gal1 overexpression) has enabled the humanization of CHO glycan profiles, minimizing non-human sugar residues and enhancing drug stability. Process-level strategies such as galactose or manganese co-feeding and fine control of temperature or osmolarity further allow rational regulation of glycosyltransferase activity. Additionally, in vitro chemoenzymatic remodeling provides a complementary route to construct human-type glycans with defined structures, though industrial applications remain constrained by cost and scalability. The integration of model-driven process design and AI feedback control is expected to enable real-time prediction and correction of glycosylation deviations, ensuring batch-to-batch consistency in continuous biomanufacturing. Long-term expression stability, another critical challenge, is often impaired by promoter silencing, chromatin condensation, and random genomic integration. Molecular optimization—such as the use of improved promoters (CMV, EF-1α, or CHO endogenous promoters), Kozak and signal peptide refinement, and incorporation of chromatin-opening elements (UCOE, MAR, STAR)—helps maintain durable transcriptional activity, while site-specific integration systems including Cre/loxP, Flp/FRT, φC31, and CRISPR/Cas9 can enable single-copy, position-independent gene insertion at genomic safe-harbor loci, ensuring stable, predictable expression. Collectively, this review highlights a paradigm shift in CHO system optimization driven by the convergence of genome editing, synthetic biology, and artificial intelligence. The transition from empirical optimization to rational, data-driven design will facilitate the development of programmable CHO platforms capable of autonomous regulation of metabolic flux, glycosylation fidelity, and transcriptional activity. Such intelligent cell factories are expected to accelerate the transformation from laboratory-scale research to industrial-scale, high-consistency, and economically sustainable biopharmaceutical manufacturing, thereby supporting the next generation of efficient and customizable biologics manufacturing.
9.Strategic Optimization of CHO Cell Expression Platforms for Biopharmaceutical Manufacturing
Rui-Ming ZHANG ; Meng-Lin LI ; Hong-Wei ZHU ; Xing-Xiao ZHANG
Progress in Biochemistry and Biophysics 2026;53(2):327-341
Chinese hamster ovary (CHO) cells are the most established and versatile mammalian expression system for the large-scale production of recombinant therapeutic proteins, owing to their genetic stability, adaptability to serum-free suspension culture, and ability to perform human-like post-translational modifications. More than 70% of biologics approved by the U.S. Food and Drug Administration rely on CHO-based production platforms, underscoring their central role in modern biopharmaceutical manufacturing. Despite these advantages, CHO systems continue to face three persistent bottlenecks that limit their potential for high-yield, reproducible, and cost-efficient production: excessive metabolic burden during high-density culture, heterogeneity of glycosylation patterns, and progressive loss of long-term expression stability. This review provides an integrated analysis of recent advances addressing these challenges and proposes a forward-looking framework for constructing intelligent and sustainable CHO cell factories. In terms of metabolic regulation, excessive lactate and ammonia accumulation disrupts energy balance and reduces recombinant protein synthesis efficiency. Optimization of culture parameters such as temperature, pH, dissolved oxygen, osmolarity, and glucose feeding can effectively alleviate metabolic stress, while supplementation with modulators including sodium butyrate, baicalein, and S-adenosylmethionine promotes specific productivity (qP) by modulating apoptosis and chromatin structure. Furthermore, genetic engineering strategies—such as overexpression of MPC1/2, HSP27, and SIRT6 or knockout of Bax, Apaf1, and IGF-1R—have demonstrated significant improvements in cell viability and product yield. The combination of multi-omics metabolic modeling with artificial intelligence (AI)-based prediction offers new opportunities for building self-regulating CHO systems capable of dynamic adaptation to environmental stress. Regarding glycosylation uniformity, which determines therapeutic efficacy and immunogenicity, gene editing-based glycoengineering (e.g., FUT8 knockdown or ST6Gal1 overexpression) has enabled the humanization of CHO glycan profiles, minimizing non-human sugar residues and enhancing drug stability. Process-level strategies such as galactose or manganese co-feeding and fine control of temperature or osmolarity further allow rational regulation of glycosyltransferase activity. Additionally, in vitro chemoenzymatic remodeling provides a complementary route to construct human-type glycans with defined structures, though industrial applications remain constrained by cost and scalability. The integration of model-driven process design and AI feedback control is expected to enable real-time prediction and correction of glycosylation deviations, ensuring batch-to-batch consistency in continuous biomanufacturing. Long-term expression stability, another critical challenge, is often impaired by promoter silencing, chromatin condensation, and random genomic integration. Molecular optimization—such as the use of improved promoters (CMV, EF-1α, or CHO endogenous promoters), Kozak and signal peptide refinement, and incorporation of chromatin-opening elements (UCOE, MAR, STAR)—helps maintain durable transcriptional activity, while site-specific integration systems including Cre/loxP, Flp/FRT, φC31, and CRISPR/Cas9 can enable single-copy, position-independent gene insertion at genomic safe-harbor loci, ensuring stable, predictable expression. Collectively, this review highlights a paradigm shift in CHO system optimization driven by the convergence of genome editing, synthetic biology, and artificial intelligence. The transition from empirical optimization to rational, data-driven design will facilitate the development of programmable CHO platforms capable of autonomous regulation of metabolic flux, glycosylation fidelity, and transcriptional activity. Such intelligent cell factories are expected to accelerate the transformation from laboratory-scale research to industrial-scale, high-consistency, and economically sustainable biopharmaceutical manufacturing, thereby supporting the next generation of efficient and customizable biologics manufacturing.
10.Effect and mechanism of Wnt5a knockdown on the efficacy of M1 bone marrow-derived macrophage in treatment of liver cirrhosis
Feifei XING ; Danyang WANG ; Xinrui ZHENG ; Yannan XU ; Shihao ZHANG ; Junyi ZHAN ; Wei LIU ; Gaofeng CHEN ; Jiamei CHEN ; Ping LIU ; Yongping MU
Journal of Clinical Hepatology 2026;42(3):618-628
ObjectiveTo observe the effect of M1 bone marrow-derived macrophages (M1-BMDM) with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis, and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDMWnt5a-KD cells. A rat model of liver cirrhosis induced by CCl4/2-AAF was established, and at the end of week 8, rats were randomly divided into model group, M1-BMDM group, M1-BMDM Wnt5a-knockdown empty vector group (M1-BMDMKD-EV group), and M1-BMDM Wnt5a-knockdown group (M1-BMDMWnt5a-KD group), with 6 rats in each group. On the first day of week 9, the rats in each group were given a single injection of the corresponding cells via the caudal vein, along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology, serum liver function parameters, hepatic stellate cell activation, and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group, all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase (AST) in serum (all P<0.01), and the M1-BMDMWnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group (P<0.05). The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group, all cell treatment groups had a significant reduction in the percentage of CD68-positive area (all P<0.05), and compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area (both P<0.05). Compared with the model group, all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α (all P<0.05) and the protein expression level of CD68 (all P<0.01); compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 (both P<0.05), significant reductions in the protein and mRNA expression levels of CD68 (both P<0.05), and a significant reduction in the protein expression level of tumor necrosis factor-α (P<0.01). Sirius Red collagen staining and alpha-smooth muscle actin (α-SMA) immunohistochemical staining showed that compared with the model group, all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area, with the most significant changes in the M1-BMDMWnt5a-KD group, and compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue (all P<0.05). Compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β (all P<0.05). Compared with the model group, all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue (all P<0.05), and the M1-BMDMWnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDMKD-EV group (both P<0.05). The M1-BMDMWnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDMKD-EV group (P<0.01). Immunofluorescence co-staining showed that compared with the model group, all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 (all P<0.01), and the M1-BMDMWnt5a-KD group had a significantly higher number of such cells than the M1-BMDMKD-EV group (P<0.05). ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl4/2-AAF, which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.


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