1.Effect and mechanism of miR-486-5P on hypoxia/reoxygenation induced ferroptosis in H9c2 cardiomyocytes
Guangmei LI ; Wenting XU ; Jiaye ZHAO ; Zeyu ZHOU ; Siming WANG ; Qiyu SUN
International Journal of Laboratory Medicine 2025;46(18):2212-2218
Objective To investigate the effect of miR-486-5P on ferroptosis in H9c2 cardiomyocytes after hypoxia/reoxygenation(H/R),and to analyze its mechanism.Methods Using H9c2 cardiomyocytes as the research object,a H/R injury model was established using cobalt chloride(CoCl2)and fresh culture medium.The cells were divided into control group,H/R group,H/R+miR-486-5P mimic NC group,H/R+miR-486-5P mimic group,H/R+miR-486-5P inhibitor NC group and H/R+miR-486-5P inhibitor group.The relative expression level of miR-486-5P was detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).The cell viability was detected by CCK-8 method.The activities or levels of lactate dehydrogen-ase(LDH),glutathione(GSH),Fe2+and malondialdehyde(MDA)were detected by colorimetric method.The levels of reactive oxygen species(ROS)and mitochondrial membrane potential(MMP)were detected by DCFH-DA fluorescent probe and JC-1 assay,respectively.Western blot was used to detect the levels of AkT/mTOR signaling pathway proteins and ferroptosis related protein solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4)and acyl-coa synthetase long chain family member 4(ACSL4).Results Compared with the control group,the level of miR-486-5P and cell viability in the H/R group de-creased significantly(P<0.05),while LDH activity,MDA,Fe2+level,ROS level and ACSL4 protein level in-creased significantly(P<0.05).The GSH,MMP,SLC7A11 and GPX4 levels and p-Akt/Akt and p-mTOR/mTOR ratios were significantly decreased(P<0.05).After H/R treatment,compared with the H/R+miR-486-5P mimic NC group,the cell viability of the H/R+miR-486-5P mimic group was significantly increased(P<0.05).The LDH activity,MDA,Fe2+level,ROS level and ACSL4 protein level were significantly de-creased(P<0.05),while GSH,MMP,SLC7A11 and GPX4 levels and p-Akt/Akt and p-mTOR/mTOR ratios were significantly increased(P<0.05).Compared with the H/R+miR-486-5P inhibitor NC group,the trend of the above indicators in the H/R+miR-486-5P inhibitor group was opposite.Conclusion miR-486-5P allevi-ates hypoxia/reoxygenation-induced ferroptosis in H9c2 cells by regulating Akt/mTOR signaling pathway,and thus alleviates hypoxia/reoxygenation induced cardiomyocyte injury.
2.Effects of RRM2 on malignant biological behavior and aerobic glycolysis of gastric cancer cells by regulating CDK1
Rongjian TAN ; Wenting OU ; Jiawei ZHAI ; Zhenhao QUAN ; Lijun SUN ; Caijin ZHOU
Journal of International Oncology 2025;52(1):23-30
Objective:To investigate the effect of ribonucleotide reductase regulatory subunit M2 (RRM2) on the malignant biological behavior and aerobic glycolysis of gastric cancer cells by regulating cyclin-dependent kinase (CDK) 1.Methods:Human gastric cancer MKN-45 cells were divided into si-NC group (transfected with blank fragment) , CoCl 2+si-NC group (hypoxia control transfected with blank fragment) , CoCl 2+si-RRM2 group (hypoxia with RRM2 silencing) , CoCl 2+si-RRM2+pcDNA3.1 NC group (hypoxia with RRM2 silencing and blank vector) and CoCl 2+si-RRM2+pcDNA3.1 CDK1 group (hypoxia with RRM2 silencing and CDK1 overexpression) . The mRNA relative expression levels of RRM2 and CDK1 were analyzed by real time fluorescent quantitative reverse transcription PCR. Co-immunoprecipitation (CoIP) was used to analyze the interaction between RRM2 and CDK1 protein. MTT assay was used to analyze the proliferation activity of cells. The cell migration distance was detected by cell scratch assay. Cell apoptosis was detected by flow cytometry. Adenosine triphosphate (ATP) and glucose kit were used to detect ATP production and glucose consumption. The protein expressions of ENO1, RRM2, HK2, PKM2, GLUT1 and p-CDK1/CDK1 were detected by Western blotting. Results:Real time fluorescent quantitative reverse transcription PCR results showed that the relative expression levels of CDK1 mRNA in si-NC group, CoCl 2+si-NC group and CoCl 2+si-RRM2 group were 1.01±0.15, 1.30±0.06 and 0.51±0.18, and the relative expression levels of RRM2 mRNA were 1.03±0.32, 1.59±0.28 and 0.44±0.17, respectively, and there were statistically significant differences ( F=25.52, P=0.001; F=14.47, P=0.005) . The mRNA expressions of RRM2 and CDK1 in CoCl 2+si-NC group were higher than those in si-NC group. Compared with the si-NC group and the CoCl 2+si-NC group, the mRNA expressions of RRM2 and CDK1 were lower in the CoCl 2+si-RRM2 group (all P<0.05) . CoIP results showed that there was interaction between RRM2 and CDK1. MTT assay, cell scratch assay and flow cytometry showed that the cell proliferation activity of si-NC group, CoCl 2+si-NC group, CoCl 2+si-RRM2 group, CoCl 2+si-RRM2+pcDNA3.1 NC group and CoCl 2+si-RRM2+pcDNA3.1 CDK1 group were 1.04±0.01, 1.18±0.04, 0.84±0.03, 0.81±0.03 and 0.93±0.05, respectively. The cell migration distances were (301.83±2.75) , (369.67±0.76) , (176.50±6.38) , (175.83±3.69) , (254.17±1.61) μm, respectively. The apoptosis rates were 8.05%±0.21%, 5.75%± 0.20%, 28.28%±0.04%, 30.18%±1.51% and 17.79%±0.22%, respectively, all with statistically significant differences ( F=73.82, P<0.001; F=1 600.01, P<0.001; F=787.15, P<0.001) . Compared with the si-NC group and CoCl 2+si-NC group, the proliferation and migration ability of cells in the CoCl 2+si-RRM2 group, CoCl 2+si-RRM2+pcDNA3.1 NC group and CoCl 2+si-RRM2+pcDNA3.1 CDK1 group were weaker, and the apoptosis rates were higher (all P<0.05) . Compared with the CoCl 2+si-RRM2+pcDNA3.1 NC group, the proliferation and migration ability of cells in the CoCl 2+si-RRM2+pcDNA3.1 CDK1 group were stronger, and the apoptosis rate was lower (all P<0.05) . The results of ATP and glucose detection showed that there were statistically significant differences in the amount of ATP production and glucose consumption among the above five groups ( F=12.53, P<0.001; F=19.21, P<0.001) . Compared with the si-NC group, the glucose consumption of cells was lower in the CoCl 2+si-RRM2+pcDNA3.1 CDK1 group ( P<0.05) . Compared with the CoCl 2+si-NC group, the ATP production and glucose consumption of cells in the CoCl 2+si-RRM2+pcDNA3.1 CDK1 group were lower (both P<0.05) . Compared with the CoCl 2+si-RRM2+pcDNA3.1 NC group, the ATP production and glucose consumption of the CoCl 2+si-RRM2+pcDNA3.1 CDK1 group were higher (both P<0.05) . Western blotting showed that there were statistically significant differences in the protein expressions of ENO1, RRM2, HK2, PKM2, GLUT1, and p-CDK1/CDK1 among the above five groups (all P<0.001) . Compared with the si-NC group and the CoCl 2+si-NC group, the protein expressions of ENO1, RRM2, HK2, PKM2, GLUT1 and p-CDK1/CDK1 in the CoCl 2+si-RRM2+pcDNA3.1 CDK1 group were lower (all P<0.05) . Compared with the CoCl 2+si-RRM2+pcDNA3.1 NC group, the protein expressions of ENO1, RRM2, PKM2, GLUT1 and p-CDK1/CDK1 in the CoCl 2+si-RRM2+pcDNA3.1 CDK1 group were higher (all P<0.05) . Conclusions:Silencing RRM2 can inhibit the malignant biological behavior of gastric cancer cells and the occurrence of aerobic glycolysis by regulating CDK1.
3.Role of microglial polarization in age-related macular degeneration
Yichi* ZHANG ; Xiuxia* YANG ; Pingping LIU ; Mengjie LIU ; Wenting LUO ; Yang LIU ; Chengcheng YANG
International Eye Science 2024;24(12):1863-1872
Microglia, originating from primitive macrophages in the yolk sac, serves as both immune system defenders and regulators of homeostasis. These cells exhibit two primary polarization states: conventionally activated(M1)and alternatively activated(M2). The polarization of microglia plays a crucial role in influencing inflammatory disorders, metabolic imbalances, and neural degeneration. This process is implicated in various aspects of ocular diseases, especially age-related macular degeneration(AMD), including inflammation, oxidative stress and pathological angiogenesis. The distinct functional phenotypes of microglia impact disease progression and prognosis. Thus, regulating the polarization or functional phenotype of microglia at different stages of AMD holds promise for personalized therapeutic approaches. This comprehensive review outlines the involvement of microglia polarization in both physiological and pathological conditions, emphasizing its relevance in AMD. The discussion underscores the potential of polarization as a foundation for personalized treatment strategies for AMD.
4.Effect of gender difference on the model of rheumatoid arthritis induced by bovine type Ⅱ collagen
Wenting SUN ; Yingying WAN ; Jiaxi YANG ; Wenqian WANG ; Haonan WANG ; Wanting YE ; Qiuai KOU
Chinese Journal of Comparative Medicine 2024;34(2):25-34
Objective To establish female and male bovine collagen-induced arthritis(CIA)models and compare the effects of gender differences on joint and extra-articular manifestations of the CIA model.Methods The CIA model was induced by injection of bovine type Ⅱ collagen and Freund's complete adjuvant into female and male SD rats.The general condition,arthritis index,foot swelling,serum proinflammatory factors and plasminogen activator inhibitor levels,spleen index,knee and ankle joint pathologies,right rear paw bone destruction,and pulmonary interstitial lesions were evaluated.Results The arthritis index of female CIA rats was significantly higher than that of male CIA rats on day 21 after initial immunization(P<0.05),but no significant difference was found in the degree of foot swelling between the two groups at any time point(P>0.05).Serum tumor necrosis factor α,interleukin-1β,and the spleen index of female CIA rats were significantly higher than those of male CIA rats(P<0.05,P<0.001).No significant difference was found in plasminogen activator inhibitor levels(P>0.05).The scores of inflammatory cell infiltration and synovial hyperplasia in the knee and ankle pathologies of female CIA rats were significantly higher than those of male CIA rats(P<0.05),and cartilage damage of the knee joint and bone damage of the right rear paw of female CIA rats were significantly higher than that of male rats(P<0.05).Both male and female CIA rats showed pulmonary interstitial inflammatory cell infiltration and mild fibrosis,but the pulmonary interstitial lesions in females were more severe than those in males.Conclusions Female and male CIA models established in SD rats have arthritis and pulmonary interstitial lesions,but the lesion degree in female CIA rats is more serious.When using CIA models for RA-related research,attention should be focused on the effect of gender differences.
5.Research on Improvement of Health Status and Health Inequalities among Older Adults in Urban and Rural China by Centralized Drug Volume-based Purchasing Policy
Shaoliang TANG ; Yuxin QIAN ; Lei CHEN ; Huiqiu DONG ; Yuli FENG ; Yue GONG ; Wenting SUN
Chinese Health Economics 2024;43(6):28-35
Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P>0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.
6.Factors influencing social participation in schizophrenia patients and research progress
Wenting WU ; Xiuying XU ; He SUN ; Peng ZHANG
Chinese Journal of Practical Nursing 2024;40(23):1830-1835
Introduced the concept of social participation, and reviewed the current status of social participation among patients with schizophrenia, and summarized the assessment tools and influencing factors of social participation in schizophrenia patients, with the aim of providing a reference for related research on social participation in patients with schizophrenia.
7.Immunohistochemical expression of TRPS1 in synovial sarcoma and its significance
Qianqian YANG ; Lingchuan GUO ; Pei PANG ; Hongli YANG ; Xiaojing SUN ; Wenting LIU
Chinese Journal of Clinical and Experimental Pathology 2024;40(8):839-844
Purpose To investigate the expression of TRPS1 in synovial sarcoma(SS)and its significance.Methods Twenty-one cases of synovial sarcoma diagnosed by SS1 8(18q11)(SYT)fluorescence in situ hybridization(FISH)or SS18-SSX immunohistochemistry were collected.The expression of TRPS1 was detected by immunohistochemistry EnVision two-step staining,and the relevant literature was reviewed.Results There were 21 cases of synovial sarcoma,including 9 females and 12 males,aged 17-79 years,with a median age of 50 years and an average age of 47.1 years.The tumors occurred in the lung(3 cases),legs(3 cases),kidney(3 cases),feet(2 cases),psoas major muscle(2 cases),rectum(1 case),tho-racic cavity(1 case),ankle canal(1 case),inguinal area(1 case),knee(1 case),palm(1 case),nasal cavity(1 case),and forearm(1 case).The maximum diameter of the tumor was 1.2-13 cm.Grossly,the tumor section was grayish-white,grayish-yellowish solid,and the lesion with hemorrhage was dark red.Microscopically,there were monophasic fibrous SS(16/21,76.2%),poorly differential SS(small cell)(3/21,14.3%),and biphasic SS(2/21,9.5%).Immunophenotypi-cally,tumor cells expressed SS18-SSX(18/18,100%),TLE1(12/12,100%),BCL2(18/18,100%),partially expressed EMA(10/16,62.5%),CK(12/18,66.7%),CD99(4/10,40.0%),SMA(3/16,18.6%),S-100(3/19,15.8%),and desmin(0/21).The proliferation index of Ki67 ranged from 3%-80%,with an average of 39.9%.FISH test showed that all 11 cases were positive for SS18(18q11)split-ting.All 21 cases of SS expressed TRPS1 to varying degrees and intensities(21/21,100%),of which 19 cases(19/21,90.5%)had a positive range of>50%.There were 19 cases(19/21,90.5%)with a positive intensity of 2+or above.All the 21 patients underwent surgical resection of the tumor,and 20 patients received follow-up visits.3 patients treated with postop-erative combination of radiotherapy and chemotherapy,18 pa-tients treated with postoperative chemotherapy,and 7 patients showed different degrees of recurrence or metastasis,and 9 pa-tients who received follow-up visits died(45%,9/20).Con-clusion Although TRPS1 is regarded as a highly sensitive and specific marker of breast tumor origin,it is also highly expressed in synovial sarcoma,and it is necessary to be vigilant at its pit-falls in the pathological diagnosis.
8.Research on Improvement of Health Status and Health Inequalities among Older Adults in Urban and Rural China by Centralized Drug Volume-based Purchasing Policy
Shaoliang TANG ; Yuxin QIAN ; Lei CHEN ; Huiqiu DONG ; Yuli FENG ; Yue GONG ; Wenting SUN
Chinese Health Economics 2024;43(6):28-35
Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P>0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.
9.Research on Improvement of Health Status and Health Inequalities among Older Adults in Urban and Rural China by Centralized Drug Volume-based Purchasing Policy
Shaoliang TANG ; Yuxin QIAN ; Lei CHEN ; Huiqiu DONG ; Yuli FENG ; Yue GONG ; Wenting SUN
Chinese Health Economics 2024;43(6):28-35
Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P>0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.
10.Research on Improvement of Health Status and Health Inequalities among Older Adults in Urban and Rural China by Centralized Drug Volume-based Purchasing Policy
Shaoliang TANG ; Yuxin QIAN ; Lei CHEN ; Huiqiu DONG ; Yuli FENG ; Yue GONG ; Wenting SUN
Chinese Health Economics 2024;43(6):28-35
Objective:Clarify the correlation between the Centralized Drug Volume-based Purchasing Policy,changes in physical&mental health status,and health inequalities among the older adults,as well as exploring whether this correlation differs between urban and rural areas so as to determine the role played by the total cost of healthcare.Methods:Based on CFPS 2018 and 2020,DID and PSM-DID were used to verify the impact of the Centralized Drug Volume-based Purchasing Policy on physical&mental health status and health inequalities of the elderly in urban and rural China.Three-step method and Sobel test were used to verify the mediating effect of total medical costs.Results:The pilot effect of the Centralized Drug Volume-based Purchasing Policy was first realized among the urban elderly population,promoting their physical and mental health through the intermediary mechanism of reducing the total medical costs,but for the time being did not show a significant correlation with health inequalities within this group.Among the rural elderly population,none of the three dependent variables was significant(P>0.1).Conclusion:The underlying logic of the Centralized Drug Volume-based Purchasing Policy is right,but its future reform should focus on the rural elderly group,and consider the fairness of the effect within the group and between urban and rural areas,so as to improve the health level of the whole society in the longer term.

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