1.Progress in ablation therapy of pulmonary nodules
Xu SHEN ; Cheng SHEN ; Congjia XIAO ; Haonan LIN ; Yunke ZHU ; Feng LIN ; Hu LIAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(03):401-405
In recent years, with the improvement of people's awareness of physical examination and the more accurate detection equipment, the detection rate of pulmonary nodules is getting higher and higher. Surgical resection is the first choice for the treatment of malignant pulmonary nodules, but multiple pulmonary nodules, nodules in complex areas and those with surgical contraindications are not suitable for surgery. As an effective, less invasive and low-cost treatment, ablation has developed rapidly in the treatment of multiple pulmonary nodules. This article introduces the progress of several common ablation techniques (radiofrequency ablation, microwave ablation, cryoablation) in the treatment of multiple pulmonary nodules, the indications and contraindications of ablation techniques, the efficacy evaluation and complications after ablation therapy, and the prospects of ablation techniques in the treatment of multiple pulmonary nodules.
2.The Role of AMPK in Diabetic Cardiomyopathy and Related Intervention Strategies
Fang-Lian LIAO ; Xiao-Feng CHEN ; Han-Yi XIANG ; Zhi XIA ; Hua-Yu SHANG
Progress in Biochemistry and Biophysics 2025;52(10):2550-2567
Diabetic cardiomyopathy is a distinct form of cardiomyopathy that can lead to heart failure, arrhythmias, cardiogenic shock, and sudden death. It has become a major cause of mortality in diabetic patients. The pathogenesis of diabetic cardiomyopathy is complex, involving increased oxidative stress, activation of inflammatory responses, disturbances in glucose and lipid metabolism, accumulation of advanced glycation end products (AGEs), abnormal autophagy and apoptosis, insulin resistance, and impaired intracellular Ca2+ homeostasis. Recent studies have shown that adenosine monophosphate-activated protein kinase (AMPK) plays a crucial protective role by lowering blood glucose levels, promoting lipolysis, inhibiting lipid synthesis, and exerting antioxidant, anti-inflammatory, anti-apoptotic, and anti-ferroptotic effects. It also enhances autophagy, thereby alleviating myocardial injury under hyperglycemic conditions. Consequently, AMPK is considered a key protective factor in diabetic cardiomyopathy. As part of diabetes prevention and treatment strategies, both pharmacological and exercise interventions have been shown to mitigate diabetic cardiomyopathy by modulating the AMPK signaling pathway. However, the precise regulatory mechanisms, optimal intervention strategies, and clinical translation require further investigation. This review summarizes the role of AMPK in the prevention and treatment of diabetic cardiomyopathy through drug and/or exercise interventions, aiming to provide a reference for the development and application of AMPK-targeted therapies. First, several classical AMPK activators (e.g., AICAR, A-769662, O-304, and metformin) have been shown to enhance autophagy and glucose uptake while inhibiting oxidative stress and inflammatory responses by increasing the phosphorylation of AMPK and its downstream target, mammalian target of rapamycin (mTOR), and/or by upregulating the gene expression of glucose transporters GLUT1 and GLUT4. Second, many antidiabetic agents (e.g., teneligliptin, liraglutide, exenatide, semaglutide, canagliflozin, dapagliflozin, and empagliflozin) can promote autophagy, reverse excessive apoptosis and autophagy, and alleviate oxidative stress and inflammation by enhancing AMPK phosphorylation and its downstream targets, such as mTOR, or by increasing the expression of silent information regulator 1 (SIRT1) and peroxisome proliferator-activated receptor‑α (PPAR‑α). Third, certain anti-anginal (e.g., trimetazidine, nicorandil), anti-asthmatic (e.g., farrerol), antibacterial (e.g., sodium houttuyfonate), and antibiotic (e.g., minocycline) agents have been shown to promote autophagy/mitophagy, mitochondrial biogenesis, and inhibit oxidative stress and lipid accumulation via AMPK phosphorylation and its downstream targets such as protein kinase B (PKB/AKT) and/or PPAR‑α. Fourth, natural compounds (e.g., dihydromyricetin, quercetin, resveratrol, berberine, platycodin D, asiaticoside, cinnamaldehyde, and icariin) can upregulate AMPK phosphorylation and downstream targets such as AKT, mTOR, and/or the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), thereby exerting anti-inflammatory, anti-apoptotic, anti-pyroptotic, antioxidant, and pro-autophagic effects. Fifth, moderate exercise (e.g., continuous or intermittent aerobic exercise, aerobic combined with resistance training, or high-intensity interval training) can activate AMPK and its downstream targets (e.g., acetyl-CoA carboxylase (ACC), GLUT4, PPARγ coactivator-1α (PGC-1α), PPAR-α, and forkhead box protein O3 (FOXO3)) to promote fatty acid oxidation and glucose uptake, and to inhibit oxidative stress and excessive mitochondrial fission. Finally, the combination of liraglutide and aerobic interval training has been shown to activate the AMPK/FOXO1 pathway, thereby reducing excessive myocardial fatty acid uptake and oxidation. This combination therapy offers superior improvement in cardiac dysfunction, myocardial hypertrophy, and fibrosis in diabetic conditions compared to liraglutide or exercise alone.
3.Multi-omics analysis for ferroptosis-related genes as prognostic factors in cutaneous melanoma
Meng WU ; Ke LI ; Yangying LIAO ; Lan LI ; Xiao XIAO ; Yongjian CHEN ; Junweichen GUO ; Feng HU ; Jing QU ; Zheng WANG ; Hao FENG
Journal of Central South University(Medical Sciences) 2024;49(2):159-174
Objective:Melanoma is highly malignant and heterogeneous.It is essential to develop a specific prognostic model for improving the patients'survival and treatment strategies.Recent studies have shown that ferroptosis results from the overproduction of lipid peroxidation and is an iron-dependent form of programmed cell death.Despite this,ferroptosis-related genes(FRGs)and their clinical significances remain unknown in malignant melanoma.This study aims to assess the role of FRGs in melanoma,with the goal of developing a novel prognostic model that provides new insights into personalized treatment and improvement of therapeutic outcomes for melanoma. Methods:We systematically characterized the genetic alterations and mRNA expression of 73 FRGs in The Cancer Genome Atlas(TCGA)-skin cutaneous melanoma(SKCM)dataset in this study.The results were validated with real-time RT-PCR and Western blotting.Subsequently,a multi-gene feature model was constructed using the TCGA-SKCM cohort.Melanoma patients were classified into a high-risk group and a low-risk group based on the feature model.As a final step,correlations between ferroptosis-related signatures and immune features,immunotherapy efficacy,or drug response were analyzed. Results:By analyzing melanoma samples from TCGA-SKCM dataset,FRGs exhibited a high frequency of genetic mutations and copy number variations(CNVs),significantly impacting gene expression.Additionally,compared with normal skin tissue,30 genes with significantly differential expression were identified in melanoma tissues.A prognostic model related to FRGs,constructed using the LASSO Cox regression method,identified 13 FRGs associated with overall survival prognosis in patients and was validated with external datasets.Finally,functional enrichment and immune response analysis further indicated significant differences in immune cell infiltration,mutation burden,and hypoxia status between the high-risk group and the low-risk group,and the model was effective in predicting responses to immunotherapy and drug sensitivity. Conclusion:This study develops a strong ferroptosis-related prognostic signature model which could put forward new insights into target therapy and immunotherapy for patients with melanoma.
4.Effects of different dose prediction models of sufentanil during hip arthroplasty with bow tie fascia iliac block
Hong CHEN ; Ming-Feng LIAO ; Shi-Yong LI ; Ai-Lin LUO ; Xiao-Hui CHI
Journal of Regional Anatomy and Operative Surgery 2024;33(11):949-954
Objective To compare the effects of different dose prediction models of sufentanil in elderly hip arthroplasty with ultrasound-guided bow tie fascia iliac block.Methods A total of 90 elderly patients who underwent ultrasound-guided bow tie fascia iliac during hip arthroplasty in our hospital from June 2018 to June 2022 were selected.The dose of sufentanil was determined by a linear regression(ALR),multiple linear regression(MLR)or deep belief networks(DBN).Patients were randomly divided into the ALR group,the MLR group and the DBN group,with 30 cases in each group.The analgesic effect,hemodynamic index,inflammatory factor level and surgical index were compared among all groups.Kaplan-Meier method was used to analyze the incidence of hip infection 2 weeks after surgery in each group.Results There was no significant difference in the gender,age,body weight,American Society of Anesthesiologists(ASA)classification or complications among all groups(P>0.05).The Harris score 24 hours after surgery and duration of sensory block in the DBN group were higher/longer than those in the ALR group,numerical rating scale(NRS)score 24 hours after surgery,mean arterial pressure(MAP)and heart rate(HR)at the beginning of surgery and 30 minutes after surgery,levels of interleukin-17(IL-17)and tumor necrosis factor-α(TNF-α)24 hours after surgery,onset time of sensory block,and number of patient controlled analgesia(PCA)compression were lower/earlier/less than those in the ALR group,the modified observer's assessment of alertness/sedation scale(OAA/S)score 24 hours after surgery was significantly lower than those in the ALR group and the MLR group,with statistically significant differences(P<0.05).Harris score 24 hours after surgery and duration of sensory block in the MLR group were higher/longer than those in the ALR group,modified OAA/S score 12 hours after surgery,MAP 30 minutes after surgery,levels of IL-17 and TNF-α 24 hours after surgery,onset time of sensory block,and the number of PCA compressions were lower/longer/less than those in the ALR group,with statistically significant differences(P<0.05).Kaplan-Meier analysis showed that there were significant differences in the incidences of hip infection 2 weeks after surgery among all groups(P<0.05),and there were significant differences in the incidences of hip infection 2 weeks after surgery between the ALR group and the DBN group(P<0.05).Conclusion The effectiveness of calculating the dose of sufentanil using the MLR and DBN in elderly hip arthroplasty with bow tie fascia iliac block is better than that of ALR,and the advantage of DBN is more obvious.
5.Changing resistance profiles of Staphylococcus isolates in hospitals across China:results from the CHINET Antimicrobial Resistance Surveillance Program,2015-2021
Yuling XIAO ; Mei KANG ; Yi XIE ; Yang YANG ; Fupin HU ; Demei ZHU ; Yingchun XU ; Xiaojiang ZHANG ; Ping JI ; Fengbo ZHANG ; Chuanqing WANG ; Pan FU ; Yuanhong XU ; Ying HUANG ; Ziyong SUN ; Zhongju CHEN ; Yuxing NI ; Jingyong SUN ; Yunzhuo CHU ; Sufei TIAN ; Zhidong HU ; Jin LI ; Yunsong YU ; Jie LIN ; Bin SHAN ; Yan DU ; Sufang GUO ; Lianhua WEI ; Fengmei ZOU ; Hong ZHANG ; Chun WANG ; Yunjian HU ; Xiaoman AI ; Chao ZHUO ; Danhong SU ; Dawen GUO ; Jinying ZHAO ; Hua YU ; Xiangning HUANG ; Wen'en LIU ; Yanming LI ; Yan JIN ; Chunhong SHAO ; Xuesong XU ; Chao YAN ; Shanmei WANG ; Yafei CHU ; Lixia ZHANG ; Juan MA ; Shuping ZHOU ; Yan ZHOU ; Lei ZHU ; Jinhua MENG ; Fang DONG ; Zhiyong LÜ ; Fangfang HU ; Han SHEN ; Wanqing ZHOU ; Wei JIA ; Gang LI ; Jinsong WU ; Yuemei LU ; Jihong LI ; Jinju DUAN ; Jianbang KANG ; Xiaobo MA ; Yanping ZHENG ; Ruyi GUO ; Yan ZHU ; Yunsheng CHEN ; Qing MENG ; Shifu WANG ; Xuefei HU ; Jilu SHEN ; Wenhui HUANG ; Ruizhong WANG ; Hua FANG ; Bixia YU ; Yong ZHAO ; Ping GONG ; Kaizhen WEN ; Yirong ZHANG ; Jiangshan LIU ; Longfeng LIAO ; Hongqin GU ; Lin JIANG ; Wen HE ; Shunhong XUE ; Jiao FENG ; Chunlei YUE
Chinese Journal of Infection and Chemotherapy 2024;24(5):570-580
Objective To investigate the changing distribution and antibiotic resistance profiles of clinical isolates of Staphylococcus in hospitals across China from 2015 to 2021.Methods Antimicrobial susceptibility testing was conducted for the clinical isolates of Staphylococcus according to the unified protocol of CHINET(China Antimicrobial Surveillance Network)using disk diffusion method and commercial automated systems.The CHINET antimicrobial resistance surveillance data from 2015 to 2021 were interpreted according to the 2021 CLSI breakpoints and analyzed using WHONET 5.6.Results During the period from 2015 to 2021,a total of 204,771 nonduplicate strains of Staphylococcus were isolated,including 136,731(66.8%)strains of Staphylococcus aureus and 68,040(33.2%)strains of coagulase-negative Staphylococcus(CNS).The proportions of S.aureus isolates and CNS isolates did not show significant change.S.aureus strains were mainly isolated from respiratory specimens(38.9±5.1)%,wound,pus and secretions(33.6±4.2)%,and blood(11.9±1.5)%.The CNS strains were predominantly isolated from blood(73.6±4.2)%,cerebrospinal fluid(12.1±2.5)%,and pleural effusion and ascites(8.4±2.1)%.S.aureus strains were mainly isolated from the patients in ICU(17.0±7.3)%,outpatient and emergency(11.6±1.7)%,and department of surgery(11.2±0.9)%,whereas CNS strains were primarily isolated from the patients in ICU(32.2±9.7)%,outpatient and emergency(12.8±4.7)%,and department of internal medicine(11.2±1.9)%.The prevalence of methicillin-resistant strains was 32.9%in S.aureus(MRSA)and 74.1%in CNS(MRCNS).Over the 7-year period,the prevalence of MRSA decreased from 42.1%to 29.2%,and the prevalence of MRCNS decreased from 82.1%to 68.2%.MRSA showed higher resistance rates to all the antimicrobial agents tested except trimethoprim-sulfamethoxazole than methicillin-susceptible S.aureus(MSSA).Over the 7-year period,MRSA strains showed decreasing resistance rates to gentamicin,rifampicin,and levofloxacin,MRCNS showed decreasing resistance rates to gentamicin,erythromycin,rifampicin,and trimethoprim-sulfamethoxazole,but increasing resistance rate to levofloxacin.No vancomycin-resistant strains were detected.The prevalence of linezolid-resistant MRCNS increased from 0.2%to 2.3%over the 7-year period.Conclusions Staphylococcus remains the major pathogen among gram-positive bacteria.MRSA and MRCNS were still the principal antibiotic-resistant gram-positive bacteria.No S.aureus isolates were found resistant to vancomycin or linezolid,but linezolid-resistant strains have been detected in MRCNS isolates,which is an issue of concern.
6.Mechanism of Xuebijing Injection in treatment of sepsis-associated ARDS based on network pharmacology and in vitro experiment.
Wei-Chao DING ; Juan CHEN ; Hao-Yu LIAO ; Jing FENG ; Jing WANG ; Yu-Hao ZHANG ; Xiao-Hang JI ; Qian CHEN ; Xin-Yao WU ; Zhao-Rui SUN ; Shi-Nan NIE
China Journal of Chinese Materia Medica 2023;48(12):3345-3359
The aim of this study was to investigate the effect and molecular mechanism of Xuebijing Injection in the treatment of sepsis-associated acute respiratory distress syndrome(ARDS) based on network pharmacology and in vitro experiment. The active components of Xuebijing Injection were screened and the targets were predicted by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). The targets of sepsis-associated ARDS were searched against GeneCards, DisGeNet, OMIM, and TTD. Weishengxin platform was used to map the targets of the main active components in Xuebijing Injection and the targets of sepsis-associated ARDS, and Venn diagram was established to identify the common targets. Cytoscape 3.9.1 was used to build the "drug-active components-common targets-disease" network. The common targets were imported into STRING for the building of the protein-protein interaction(PPI) network, which was then imported into Cytoscape 3.9.1 for visualization. DAVID 6.8 was used for Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment of the common targets, and then Weishe-ngxin platform was used for visualization of the enrichment results. The top 20 KEGG signaling pathways were selected and imported into Cytoscape 3.9.1 to establish the KEGG network. Finally, molecular docking and in vitro cell experiment were performed to verify the prediction results. A total of 115 active components and 217 targets of Xuebijing Injection and 360 targets of sepsis-associated ARDS were obtained, among which 63 common targets were shared by Xuebijing Injection and the disease. The core targets included interleukin-1 beta(IL-1β), IL-6, albumin(ALB), serine/threonine-protein kinase(AKT1), and vascular endothelial growth factor A(VEGFA). A total of 453 GO terms were annotated, including 361 terms of biological processes(BP), 33 terms of cellular components(CC), and 59 terms of molecular functions(MF). The terms mainly involved cellular response to lipopolysaccharide, negative regulation of apoptotic process, lipopolysaccharide-mediated signaling pathway, positive regulation of transcription from RNA polyme-rase Ⅱ promoter, response to hypoxia, and inflammatory response. The KEGG enrichment revealed 85 pathways. After diseases and generalized pathways were eliminated, hypoxia-inducible factor-1(HIF-1), tumor necrosis factor(TNF), nuclear factor-kappa B(NF-κB), Toll-like receptor, and NOD-like receptor signaling pathways were screened out. Molecular docking showed that the main active components of Xuebijing Injection had good binding activity with the core targets. The in vitro experiment confirmed that Xuebijing Injection suppressed the HIF-1, TNF, NF-κB, Toll-like receptor, and NOD-like receptor signaling pathways, inhibited cell apoptosis and reactive oxygen species generation, and down-regulated the expression of TNF-α, IL-1β, and IL-6 in cells. In conclusion, Xuebijing Injection can regulate apoptosis and response to inflammation and oxidative stress by acting on HIF-1, TNF, NF-κB, Toll-like receptor, and NOD-like receptor signaling pathways to treat sepsis-associated ARDS.
Humans
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Network Pharmacology
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Vascular Endothelial Growth Factor A
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NF-kappa B
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Interleukin-6
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Lipopolysaccharides
;
Molecular Docking Simulation
;
Respiratory Distress Syndrome
;
Tumor Necrosis Factor-alpha
;
Sepsis/genetics*
;
NLR Proteins
7.Organizing Pneumonia in A Patient Double-Positive for ANCA and Anti-GBM Antibodies: A Case Report.
Fang-Yuan WANG ; Xiang-Ning YUAN ; Dan-Ni SUN ; Gong XIAO ; Cheng-Huan HU ; Zhong-Hua LIAO ; Jian-Ping NING ; Hui XU ; Jun-Tao FENG ; Hong-Ling YIN ; Xiao-Zhao LI
Chinese Medical Sciences Journal 2023;38(1):66-69
Both anti-glomerular basement membrane (GBM) disease and the anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) are common causes of pulmonary-renal syndrome. Organizing pneumonia (OP), a special pattern of interstitial lung disease, is extremely rare either in AAV or anti-GBM disease. We report an old woman presented with OP on a background of co-presentation with both ANCA and anti-GBM antibodies.
Female
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Humans
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Antibodies, Antineutrophil Cytoplasmic
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Organizing Pneumonia
;
Autoantibodies
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Glomerulonephritis
;
Anti-Glomerular Basement Membrane Disease
;
Pneumonia
;
Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis/complications*
8.A descriptive analysis on hypertension in adult twins in China.
Yu Tong WANG ; Wei Hua CAO ; Jun LYU ; Can Qing YU ; Sheng Feng WANG ; Tao HUANG ; Dian Jian Yi SUN ; Chun Xiao LIAO ; Yuan Jie PANG ; Zeng Chang PANG ; Min YU ; Hua WANG ; Xian Ping WU ; Zhong DONG ; Fan WU ; Guo Hong JIANG ; Xiao Jie WANG ; Yu LIU ; Jian DENG ; Lin LU ; Wen Jing GAO ; Li Ming LI
Chinese Journal of Epidemiology 2023;44(4):536-543
Objective: To describe the distribution characteristics of hypertension among adult twins in the Chinese National Twin Registry (CNTR) and to provide clues for exploring the role of genetic and environmental factors on hypertension. Methods: A total of 69 220 (34 610 pairs) of twins aged 18 and above with hypertension information were selected from CNTR registered from 2010 to 2018. Random effect models were used to describe the population and regional distribution of hypertension in twins. To estimate the heritability, the concordance rates of hypertension were calculated and compared between monozygotic twins (MZ) and dizygotic twins (DZ). Results: The age of all participants was (34.1±12.4) years. The overall self-reported prevalence of hypertension was 3.8%(2 610/69 220). Twin pairs who were older, living in urban areas, married, overweight or obese, current smokers or ex-smokers, and current drinkers or abstainers had a higher self-reported prevalence of hypertension (P<0.05). Analysis within the same-sex twin pairs found that the concordance rate of hypertension was 43.2% in MZ and 27.0% in DZ, and the difference was statistically significant (P<0.001). The heritability of hypertension was 22.1% (95%CI: 16.3%- 28.0%). Stratified by gender, age, and region, the concordance rate of hypertension in MZ was still higher than that in DZ. The heritability of hypertension was higher in female participants. Conclusions: There were differences in the distribution of hypertension among twins with different demographic and regional characteristics. It is indicated that genetic factors play a crucial role in hypertension in different genders, ages, and regions, while the magnitude of genetic effects may vary.
Adult
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Female
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Humans
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Male
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Alcohol Drinking
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Diseases in Twins/genetics*
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Hypertension/genetics*
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Twins, Dizygotic/genetics*
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Twins, Monozygotic/genetics*
9.A descriptive analysis of hyperlipidemia in adult twins in China.
Ke MIAO ; Wei Hua CAO ; Jun LYU ; Can Qing YU ; Sheng Feng WANG ; Tao HUANG ; Dian Jian Yi SUN ; Chun Xiao LIAO ; Yuan Jie PANG ; Zeng Chang PANG ; Min YU ; Hua WANG ; Xian Ping WU ; Zhong DONG ; Fan WU ; Guo Hong JIANG ; Xiao Jie WANG ; Yu LIU ; Jian DENG ; Lin LU ; Wen Jing GAO ; Li Ming LI
Chinese Journal of Epidemiology 2023;44(4):544-551
Objective: To describe the distribution characteristics of hyperlipidemia in adult twins in the Chinese National Twin Registry (CNTR) and explore the effect of genetic and environmental factors on hyperlipidemia. Methods: Twins recruited from the CNTR in 11 project areas across China were included in the study. A total of 69 130 (34 565 pairs) of adult twins with complete information on hyperlipidemia were selected for analysis. The random effect model was used to characterize the population and regional distribution of hyperlipidemia among twins. The concordance rates of hyperlipidemia were calculated in monozygotic twins (MZ) and dizygotic twins (DZ), respectively, to estimate the heritability. Results: The age of all participants was (34.2±12.4) years. This study's prevalence of hyperlipidemia was 1.3% (895/69 130). Twin pairs who were men, older, living in urban areas, married,had junior college degree or above, overweight, obese, insufficient physical activity, current smokers, ex-smokers, current drinkers, and ex-drinkers had a higher prevalence of hyperlipidemia (P<0.05). In within-pair analysis, the concordance rate of hyperlipidemia was 29.1% (118/405) in MZ and 18.1% (57/315) in DZ, and the difference was statistically significant (P<0.05). Stratified by gender, age, and region, the concordance rate of hyperlipidemia in MZ was still higher than that in DZ. Further, in within-same-sex twin pair analyses, the heritability of hyperlipidemia was 13.04% (95%CI: 2.61%-23.47%) in the northern group and 18.59% (95%CI: 4.43%-32.74%) in the female group, respectively. Conclusions: Adult twins were included in this study and were found to have a lower prevalence of hyperlipidemia than in the general population study, with population and regional differences. Genetic factors influence hyperlipidemia, but the genetic effect may vary with gender and area.
Adult
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Female
;
Humans
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Male
;
Middle Aged
;
Young Adult
;
China/epidemiology*
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Diseases in Twins/genetics*
;
Hyperlipidemias/genetics*
;
Metabolic Diseases
;
Twins, Dizygotic
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Twins, Monozygotic/genetics*
10.Synchronization isolation method for multiple types of cells from mouse liver.
Jian GAN ; Cui Feng JI ; Xiao Rong MAO ; Jiang Tao WANG ; Chun Yan LYU ; Yi Fan SHI ; Yao LIAO ; Ya Li HE ; Lian SHU ; Long LI ; Jun Feng LI
Chinese Journal of Hepatology 2023;31(5):532-537
Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.
Male
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Mice
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Animals
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Keratin-18
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Actins
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Desmin
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Liver
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Hepatocytes
;
Hepatic Stellate Cells

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