1.Clinical efficacy analysis of seven pediatric patients with Acute myeloid leukemia and the t(16;21)(p11;q22) FUS::ERG fusion gene.
Lihuan SHI ; Shan HUANG ; Xing XIE ; Pengkai FAN ; Haili GAO ; Yanna MAO
Chinese Journal of Medical Genetics 2026;43(2):90-95
OBJECTIVE:
To analyze the clinical characteristics, treatment, and prognosis of seven pediatric patients with Acute myeloid leukemia (AML) positive for the t(16;21)(p11;q22) FUS::ERG fusion gene.
METHODS:
A retrospective analysis was carried out on the clinical data, treatment, and prognosis of seven AML patients with t(16;21)(p11;q22) FUS::ERG fusion gene admitted to Henan Children's Hospital between June 2015 and November 2024. Relevant literature was also reviewed. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.: 2024-102-001).
RESULTS:
Among 297 pediatric patients with AML, 7 cases (2.36%) were positive for the t(16;21)(p11;q22) FUS::ERG fusion gene, including 3 males and 4 females, with a median age of 11 years (range: 3 ~ 12 years). According to the FAB classification, these included 1 case of M2, 3 cases of M5, and 3 cases of AML-not otherwise specified (non-M3). All 7 patients were found to harbor the t(16;21)(p11;q22) translocation, with 3 cases showing additional chromosomal abnormalities. Immunophenotyping revealed universal expression of CD13, CD33, CD34, and CD117, with partial expression of CD56, CD4, CD64, CD123, CD15, CD38, CD11b, HLA-DR, cMPO, and CD16. One patient achieved complete remission (CR) after the first course of DAE (cytarabine + daunorubicin + etoposide) induction chemotherapy but relapsed and discontinued the treatment. Six patients received DAH (cytarabine + daunorubicin + homoharringtonine) induction therapy, of whom 2 achieved CR after two courses and underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT), resulting in an overall CR rate of 42.86%. Five children did not receive allo-HSCT and had a median overall survival of 9 months (range: 6 ~ 18 months). Two children who underwent transplantation achieved bone marrow morphological and molecular biological relapse at 6 and 9 months post-transplantation, respectively. After receiving combined chemotherapy and donor lymphocyte infusion, one child failed to achieve remission and died at 22 months post-transplantation, while the other has been followed up to date with positive fusion gene status. Their overall survival was 25 months and 30 months, respectively.
CONCLUSION
The t(16;21)(p11;q22) FUS::ERG fusion gene is rare in pediatric AML and associated with poor prognosis. Allo-HSCT may mitigate the adverse prognostic impact of the FUS::ERG fusion gene and contribute to prolonged survival.
Humans
;
Male
;
Child
;
Female
;
Leukemia, Myeloid, Acute/drug therapy*
;
Oncogene Proteins, Fusion/genetics*
;
Translocation, Genetic
;
Retrospective Studies
;
RNA-Binding Protein FUS/genetics*
;
Chromosomes, Human, Pair 16/genetics*
;
Adolescent
;
Child, Preschool
;
Chromosomes, Human, Pair 21/genetics*
;
Prognosis
;
Treatment Outcome
2.Effects of different electromagnetic stimulation protocols on hand function and activities of daily living for stroke patients: a network meta-analysis
Shan LIU ; Kunyao XU ; Zhimeng ZHOU ; Lingyun ZHOU ; Mao ZENG ; Sha HE ; Luoyang DAI ; Ting QIN ; Qinglin HU ; Jiahai YANG ; Rui LI
Chinese Journal of Rehabilitation Theory and Practice 2026;32(8):916-926
ObjectiveTo systematically compare the effects of diverse electromagnetic stimulation protocols on hand function and activities of daily living (ADL) in stroke patients using network meta-analysis (NMA), and screen the optimal stimulation strategy for post-stroke hand dysfunction rehabilitation. MethodsNMA was performed in accordance with the PRISMA-NMA statement. Databases of CNKI, Wanfang data, VIP, CBM, PubMed, Web of Science and Embase were searched from database inception to April, 2026 for eligible randomized controlled trials (RCTs) comparing different electromagnetic stimulation protocols for post-stroke hand impairment. The Cochrane Risk of Bias tool and PEDro scale were used to assess methodological quality of included studies, while the GRADE system was adopted to grade evidence quality of primary outcomes of Fugl-Meyer Assessment-Upper Extremities (FMA-UE) and modified Barthel Index (MBI). After literature screening, data extraction and bias evaluation, RevMan 5.4 and Stata 17.0 were used for meta-analysis and NMA. ResultsA total of 22 RCT involving 17 electromagnetic stimulation protocols were finally included. The PEDro scores ranged from six to ten. The top five protocols for improving FMA-UE score were repetitive peripheral magnetic stimulation (rPMS) + repetitive transcranial magnetic stimulation (rTMS), intermittent theta-burst stimulation (iTBS) + rPMS, iTBS + transcranial direct current stimulation (tDCS), transcutaneous electrical acupoint stimulation (TEAS) and contralaterally controlled functional electrical stimulation (CCFES). tDCS was the most effective on MBI, followed by TEAS and iTBS+rPMS. Evidence quality for FMA-UE and MBI was moderate. Funnel plots suggested low risk of publication bias, and consistency tests verified good coherence between direct and indirect comparisons in network models. ConclusionElectromagnetic stimulation significantly facilitates upper limb and hand functional recovery, and improves ADL in stroke patients. The combined central-peripheral magnetic stimulation protocol may be the most effective on motor rehabilitation, whereas tDCS on ADL.
3.Dexmedetomidine Improves the Therapeutic Efficacy Against Mammary Tumors in Stress-exposed Mice by Inhibiting the TLR4/NF-κB Pathway
Chuyi ZHANG ; Shan MENG ; Xin XU ; Peng FAN ; Xunyuan MAO ; Yuhui HUANG ; Sanqing JIN
Journal of Sun Yat-sen University(Medical Sciences) 2026;47(4):601-612
ObjectiveThis study aimed to investigate whether dexmedetomidine (Dex) alleviates stress-induced mammary tumor progression in mice by inhibiting the TLR4/NF‑κB signaling pathway, and to examine its effects on anxiety‑depression‑like behaviors and stress hormone levels. MethodsAn orthotopic EMT6 mammary tumor model was established in mice, and chronic mild stress (CMS) was applied to induce anxiety‑depression‑like states. The mice were divided into six groups: naive, tumor, CMS, CMS+tumor, CMS+tumor+Dex, and CMS+tumor+Vehicle groups. Behavioral changes were evaluated using the open field test and tail suspension test. Plasma stress hormone levels were measured by high‑performance liquid chromatography. Western blot and RT‑qPCR were performed to detect the expression of TLR4, p‑NF‑κB p65, p‑IκBα, and inflammatory cytokines (IL‑6, IL‑1β, TNF‑α) in tumor tissues. In vitro, EMT6 cells were treated with cortisol to mimic the stress-related microenvironment, and the effects of Dex on the TLR4/NF‑κB pathway were assessed by CCK‑8, Western blot, and RT-qPCR. ResultsTumor‑bearing mice exhibited significant anxiety‑depression‑like behaviors (P<0.05), along with markedly elevated plasma levels of norepinephrine, epinephrine, and cortisol (P<0.05). Chronic stress further aggravated these behavioral abnormalities and promoted tumor growth (P<0.05). Dexmedetomidine intervention significantly improved anxiety‑depression‑like behaviors (P<0.05) and lowered peripheral stress hormone levels (P<0.05) in stress‑exposed tumor‑bearing mice. In vitro experiments demonstrated that cortisol treatment activated the TLR4/NF‑κB pathway (P<0.001) and upregulated the expression of inflammatory cytokines (P<0.05 in EMT6 cells, whereas dexmedetomidine effectively suppressed this activation (P<0.001). In the in vivo model, dexmedetomidine treatment downregulated the expression of TLR4, p‑NF‑κB p65, p‑IκBα, and pro‑inflammatory cytokines in tumor tissues (P<0.001) and significantly inhibited tumor growth (P<0.05). ConclusionDexmedetomidine inhibits the TLR4/NF‑κB signaling pathway, alleviates stress‑induced inflammatory responses, improves anxiety- and depression‑ like behaviors, reduces stress hormone levels, and suppresses mammary tumor progression. These findings provide an experimental basis for understanding anxiety- and depression- associated breast tumor progression and suggest that Dexmedetomidine and the TLR4/NF-κB pathway may serve as potential therapeutic targets.
4.Latent tuberculosis infection among enterprise employees in Wenzhou City
ZENG Zhen ; ZHAO Lina ; SHAN Zhili ; MAO Jing ; LIN Hante ; MAO Lingqiong ; LI Jun
Journal of Preventive Medicine 2026;38(4):334-337
Objective:
To investigate the status and influencing factors of latent tuberculosis infection (LTBI) among enterprise employees in Wenzhou City, Zhejiang Province, so as to provide evidence for optimizing tuberculosis prevention and control strategies in enterprises.
Methods:
From August 2024 to September 2025, a convenience sampling method was used to select employees from 26 enterprises in Wenzhou City as the study participants. Demographic information, enterprise scale, and industry type were collected by reviewing enterprise employee management files. Interferon-gamma release assays were used to detect Mycobacterium tuberculosis infection and screen for LTBI. Factors affecting LTBI among enterprise employees were identified using a multivariable logistic regression model.
Results:
A total of 6 476 participants were surveyed, including 3 674 males (56.73%) and 2 802 females (43.27%). The median age was 37.00 (interquartile range, 20.00) years. A total of 994 cases of LTBI were detected, with a detection rate of 15.35%. Multivariable logistic regression analysis showed that male (OR=1.397, 95%CI: 1.211-1.612), age ≥45 years (45-<60 years, OR=1.919, 95%CI: 1.649-2.234; ≥60 years, OR=4.425, 95%CI: 3.170-6.177), household registration outside Zhejiang Province (OR=1.928, 95%CI: 1.577-2.357), employment in small or medium-sized enterprises (medium-sized, OR=1.663, 95%CI: 1.392-1.988; small-sized, OR=1.665, 95%CI: 1.358-2.040), and working in the manufacturing industry (OR=1.799, 95%CI: 1.147-2.822) were associated with a higher risk of LTBI among enterprise employees.
Conclusions
The detection rate of LTBI among enterprise employees in Wenzhou City was relatively low. Male employees, those aged ≥45 years, those with household registration outside Zhejiang Province, those working in small or medium-sized enterprises, and those in the manufacturing industry were identified as high-risk populations for LTBI.
5.Latent tuberculosis infection among HIV/AIDS cases in Wenzhou City
ZHAO Lina ; ZENG Zhen ; LIN Hante ; MAO Jing ; ZHU Tingting ; SHAN Zhili ; MAO Lingqiong ; LI Jun
Journal of Preventive Medicine 2026;38(6):550-554
Objective:
To investigate the status of latent tuberculosis infection (LTBI) and tuberculosis preventive treatment (TPT) among HIV/AIDS cases in Wenzhou City, Zhejiang Province, and to analyze the influencing factors of LTBI, so as to provide the evidence for optimizing active tuberculosis screening and early intervention strategies.
Methods:
Study subjects were HIV/AIDS cases who received antiretroviral therapy and CD4+T lymphocyte count testing at designated HIV treatment hospitals from October 2024 to August 2025. The interferon-gamma release assay was used for LTBI screening. Data on HIV/AIDS cases, including gender, age, and other variables, were collected through the Surveillance System of China Information System for Disease Control and Prevention. A multivariable logistic regression model was used to analyze the influencing factors for LTBI among HIV/AIDS cases.
Results:
A total of 3 719 HIV/AIDS cases were included, including 2 983 males (80.21%) and 736 females (19.79%). The study population was predominantly aged 40-<65 years, with 1 948 individuals accounting for 52.38%. LTBI was detected in 399 cases, with a detection rate of 10.73%. Among LTBI cases, 103 received TPT, representing a TPT rate of 25.81%. Multivariable logistic regression analysis showed that HIV/AIDS cases who were male (OR=1.796, 95%CI: 1.326-2.431), aged ≥40 years (40-<65 years, OR=1.895, 95%CI: 1.424-2.523; ≥65 years, OR=2.365, 95%CI: 1.575-3.551), non-Wenzhou household registration (OR=1.462, 95%CI: 1.150-1.857), with education level below junior college (primary school and below, OR=1.578, 95%CI: 1.023-2.394; junior high school, OR=1.725, 95%CI: 1.161-2.562; high school/technical secondary school, OR=1.639, 95%CI: 1.066-2.518), CD4+T lymphocyte count ≥500 cells/μL (OR=2.642, 95%CI: 1.420-4.915), and at the HIV infection stage (OR=1.322, 95%CI: 1.022-1.709) had a higher risk of LTBI.
Conclusions
The detection rate of LTBI and the TPT rate among LTBI cases are relatively low among HIV/AIDS cases in Wenzhou City. Gender, age, household registration, education level, CD4+T lymphocyte count, and disease stage are influencing factors for LTBI in HIV/AIDS cases.
6.Relationship between social function and family support in young and middle-aged patients undergoing peritoneal dialysis
Shan LYU ; Chunyan YI ; Shuchao LU ; Xiao YANG ; Haiping MAO ; Wei CHEN ; Jianxiong LIN
Chinese Journal of Nephrology 2025;41(11):811-816
Objective:To analyze the relationship between social function and family support in young and middle-aged patients undergoing peritoneal dialysis (PD).Methods:In this cross-sectional study, the patients undergoing maintenance PD therapy for more than 3 months at the Department of Nephrology, the First Affiliated Hospital of Sun Yat-sen University, from May to October 2023 were recruited retrospectively. The social dysfunction screening scale and family support self-rating scale were used to evaluate the social dysfunction and family support of PD patients, respectively. The social demographic and clinical data of the patients were collected. Logistic regression analysis model was used to identify associated factors of social dysfunction in PD patients.Results:A total of 359 PD patients were recruited with age of (42.6±9.5) years old. Among them, 197 patients (54.9%) were males, and 33 patients (9.2%) were complicated with diabetes. The dialysis age was 28.8 (13.5, 56.3) months. The score of social function was 2 (1, 4), and the score of family support was (10.5±2.2). There were 199 patients (55.4%) having social dysfunction. There were 224 patients (62.4%) employed after PD. Compared with the normal social function group, the social dysfunction group had significantly lower score in family support ( Z=-2.613, P=0.009), serum potassium ( t=-2.725, P=0.007), urea clearance index ( Z=-2.346, P=0.019) and proportions of married status ( χ2=6.847, P=0.009), pre-dialysis employment ( χ2=3.996, P=0.046) and post-dialysis employment ( χ2=8.331, P=0.004), and higher serum creatinine ( Z=2.175, P=0.030), and proportions of annual household income < 100 000 yuan ( χ2=6.270, P=0.012) and diabetes mellitus ( χ2=4.400, P=0.036). Multivariate logistic regression analysis revealed that family support ( OR=0.828, 95% CI 0.733-0.935, P=0.002), diabetes mellitus ( OR=3.551, 95% CI 1.456-8.658, P=0.005) and serum potassium ( OR=0.559, 95% CI 0.374-0.835, P=0.005) were independent correlated factors of social dysfunction in young and middle-aged PD patients. Conclusions:The prevalence of social dysfunction is 55.4%, and the employment rate is 62.4% in young and middle-aged PD patients. Poor family support, diabetes mellitus and decreased serum potassium level are independently associated with social dysfunction in young and middle-aged PD patients.
7.Chemical constituents from ethyl acetate fraction of Balanophora harlandii and their tyrosinase inhibitory activity
Zhang-xian CHEN ; Hai-ming WANG ; Yun-tao ZHANG ; Mao-xin DENG ; Kui-lin ZHU ; Jin-lian ZOU ; Jian WANG ; Shan-shan WEI ; Hong-ping HE ; Fa-wu DONG
Chinese Traditional Patent Medicine 2025;47(10):3290-3297
AIM To study the chemical constituents from ethyl acetate fraction of Balanophora harlandii Hook.f.and their tyrosinase inhibitory activity.METHODS Separation and purification were performed using silica gel,MCI,ODS,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The monophenolase inhibitory activity was determined by the tyrosinase-catalyzed oxidation of L-tyrosine.RESULTS Twenty-four compounds were isolated and identified as sesamin(1),methyl caffeate(2),quercetin(3),5,7-dihydroxychromanone(4),methyl 3,4-dihydroxybenzoate(5),esculetin(6),kaempferol(7),naringenin(8),pyrogallic acid(9),pinosylvin(10),methyl propionate(11),caffeic acid(12),saccharinol(13),ferulic acid(14),trans-p-hydroxycinnamic acid(15),cinnamic acid(16),vanillic acid(17),vanillin(18),4-hydroxyacetophenone(19),4-hydroxybenzaldehyde(20),apigenin(21),(-)-isolariciresinol(22),(-)-secoisolariciresinol(23)and meso-2,3-di(3′,4′-methylenedioxybenzyl)butane-1,4-diol(24).The IC50 values of compounds 3,5,7,8,19,and 20 ranged from(0.246 5±0.028 3)to(1.278 2±0.021 3)mmol/L.CONCLUSION Compounds 1-9、11、15、17-21、24 are isolated from this plant for the first time,and 1,6,9,17-19,24 are first isolated from genus Balanophora.Compounds 3、5、7、8、19 and 20 have tyrosinase inhibitory activity.
8.PI-RADS v2.1 score combined with PSA density for diagnosis of clinically significant prostate cancer in the PSA grey zone by MRI-TRUS cognitivefusion-guided transperineal targeted prostate biopsy.
Yue LI ; Shan ZHOU ; Jing CHEN ; Fei MAO ; Xiao-Bing NIU ; Li SUN ; Ming XU ; Jin-Tao LIU
National Journal of Andrology 2025;31(1):50-54
OBJECTIVE:
To assess the value of the Prostate Imaging Reporting and Data System version 2.1 (PI-RADS v2.1) score combined with PSA density (PSAD) in the diagnosis of clinically significant prostate cancer (CSPCa) in the PSA grey zone by MRI-TRUS cognitive fusion-guided transperineal targeted prostate biopsy.
METHODS:
This retrospective study included 327 male patients with total PSA (tPSA) levels of 4-10 μg/L undergoing MRI-TRUS cognitive fusion-guided transperineal targeted prostate biopsy in our hospital between January 2021 and December 2023. According to the pathological results, we divided the patients into a CSPCa (n = 44) and a non-CSPCa group (n = 283), collected their clinical and imaging data, and subjected them to statistical analysis.
RESULTS:
The age, tPSA level, PSAD and PI-RADS score were significantly higher, while the free PSA (fPSA) level, f/tPSA ratio and prostate volume remarkably lower in the CSPCa than in the non-CSPCa group (P<0.05). The areas under the curve (AUCs) of PSAD, PI-RADS score and their combination were 0.772, 0.730 and 0.801, with sensitivities of 63.63%, 70.45% and 72.73%, and specificities of 84.10%, 75.62% and 83.75%, respectively (P<0.01). With PSAD 0.2 μg/(ml·cm3) as the best cut-off value and based on the PI-RADS scores, the patients were divided into two groups for analysis. In the patients with PI-RADS scores 2 and 5, the AUCs were 0.534 and 0.643, with sensitivities of 16.67% and 63.64%, and specificities of 85.14% and 64.29%, with no statistically significant differences (P= 0.784, P= 0.228), and in those with PI-RADS scores 3 and 4, the AUCs were 0.794 and 0.843, with sensitivities of 57.14% and 80.00%, and specificities of 87.14% and 81.82%, with statistically significant differences (P= 0.009, P<0.001).
CONCLUSION
PI-RADS v2.1 score combined with PSAD can effectively improve the diagnostic efficiency of CSPCa in the PSA grey zone by MRI-TRUS cognitive fusion-guided transperineal targeted prostate biopsy and serve as a guide for selection of prostate biopsy.
Humans
;
Male
;
Prostatic Neoplasms/diagnostic imaging*
;
Retrospective Studies
;
Prostate-Specific Antigen
;
Magnetic Resonance Imaging
;
Image-Guided Biopsy
;
Prostate/pathology*
;
Aged
;
Middle Aged
9.Dexmedetomidine attenuates heat stress-induced oncosis in human skeletal muscle cells by activating the Nrf2/Ho-1 pathway.
Yang LIU ; Yiqing JIA ; Chengcheng LI ; Handing MAO ; Shuyuan LIU ; Yi SHAN
Journal of Southern Medical University 2025;45(3):603-613
OBJECTIVES:
To investigate the protective effects of dexmedetomidine (DEX) against heat stress (HS)-induced oncosis in human skeletal muscle cells (HSKMCs) and its underlying mechanisms.
METHODS:
A HSKMC model of HS-induced oncosis were established by 43 ℃ water bath for 4 h, and the effects of treatments with 30 μmol/L DEX, ML385 (a Nrf2 inhibitor) +DEX, si-Nrf2+HS, and si-Nrf2+DEX prior to modeling on cell viability was assessed using CCK-8 assay. Oncosis characteristics were evaluated using transmission electron microscopy and Annexin V-FITC/PI flow cytometry. The oxidative stress markers (GSH, GSH-Px, MDA, SOD and ROS), mitochondrial membrane potential, energy metabolism, and inflammatory cytokines (TNF-α, IL-6 and IL-1β) in the cells were quantified using standard kits, and the expressions of porimin, caspase-3 and Nrf2 pathway proteins were analyzed using Western blotting and qRT-PCR.
RESULTS:
HS induced typical oncotic features in HSKMCs including organelle swelling and cytoplasmic vacuolization. DEX pretreatment significantly attenuated these changes, reduced Annexin V+/PI+ cell ratio and cellular porimin expression, and lowered the levels of ROS and MDA while restoring GSH and SOD levels. DEX pretreatment also significantly increased the mitochondrial membrane potential and ATP level, upregulated the expressions of Nrf2, p-Nrf2, HO-1 and NQO1, and suppressed the expressions of TNF-α, IL-6 and IL-1β. The protective effects of DEX were obviously attenuated by interventions with ML385 or si-Nrf2.
CONCLUSIONS
DEX mitigates HS-induced HSKMC oncosis by activating the Nrf2/HO-1 pathway to relieve oxidative stress, mitochondrial dysfunction, and inflammatory responses.
Humans
;
Dexmedetomidine/pharmacology*
;
NF-E2-Related Factor 2/metabolism*
;
Oxidative Stress/drug effects*
;
Heat-Shock Response/drug effects*
;
Signal Transduction/drug effects*
;
Membrane Potential, Mitochondrial
;
Muscle, Skeletal/cytology*
;
Heme Oxygenase-1/metabolism*
;
Apoptosis/drug effects*
10.Role and mechanism of microRNA-145-5p in hypoxia-induced pyroptosis of human alveolar epithelial cells.
Runqi YUAN ; Junmiao GUO ; Zhenting LIANG ; Yongxin ZHENG ; Yongbo HUANG ; Yonghao XU ; Pu MAO ; Jinglan SHAN
Chinese Critical Care Medicine 2025;37(4):354-360
OBJECTIVE:
To elucidate the role and mechanism of microRNA-145-5p (miR-145-5p) in hypoxia-induced pyroptosis of human alveolar epithelial cells.
METHODS:
In vitro, human alveolar epithelial cell line BEAS-2B was cultured. Cells in the logarithmic growth phase were cultured to 80% confluence and then used for the experiment. (1) BEAS-2B cells were cultured under 1% O2 hypoxic condition, with a normoxic control group. Western blotting was employed to detect the expressions of pyroptosis marker proteins [NOD-like receptor protein 3 (NLRP3), Gasdermin D N-terminal domain (GSDMD-N), and caspase-1] in cells cultured for 24 hours. Real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of miR-145-5p in cells cultured for 6 hours and 12 hours. (2) Cells were transfected with 30 nmol/L miR-145-5p mimic to overexpress miR-145-5p expression under normoxic condition or 30 nmol/L miR-145-5p inhibitor to suppress miR-145-5p expression under hypoxic condition. Control group and negative control group were respectively set up. After 24 hours of cell culture, Western blotting was used to detect the expressions of pyroptosis marker proteins and nuclear factor-E2-related factor 2 (Nrf2) in cells. Flow cytometry was applied to detect the level of reactive oxygen species (ROS) in cells. The target genes of miR-145-5p were predicted by miR target gene prediction software miRWalk and verified by Western blotting. (3) Under hypoxic condition, cells were transfected with 6.94 ng/μL silent information regulator 5 (Sirt5) overexpression plasmid or pretreated with 12.5 mmol/L N-acetyl-L-cysteine (NAC) as an ROS inhibitor. The empty plasmid group and control group were set up. After 24 hours of cell culture, Western blotting was used to detect the expressions of Sirt5, Nrf2, and pyroptosis marker proteins in cells. Flow cytometry was used to detect the level of ROS in cells.
RESULTS:
(1) Compared with the normoxic control group, the expression levels of pyroptosis marker proteins in the 24-hour hypoxia group was significantly increased, indicating that hypoxia could induce pyroptosis in BEAS-2B cells. The expression level of miR-145-5p in cells gradually increased with the extension of hypoxia induction time, indicating that hypoxia could cause the increase of miR-145-5p expression level. (2) The expression levels of pyroptosis marker proteins in cells of miR-145-5p mimic group significantly increased under normoxic condition as compared with the control and negative control groups [NLRP3 protein (NLRP3/β-actin): 1.58±0.07 vs. 1.00±0.01, 0.98±0.07, GSDMD-N protein (GSDMD-N/β-actin): 1.71±0.03 vs. 1.01±0.01, 0.85±0.03, caspase-1 protein (caspase-1/β-actin): 2.33±0.04 vs. 1.01±0.01, 1.05±0.04, all P < 0.05], Nrf2 protein expression level was significantly decreased (Nrf2/β-actin: 0.79±0.03 vs. 1.00±0.01, 1.03±0.04, both P < 0.05), ROS level was significantly up-regulated (fluorescence intensity: 1.74±0.03 vs. 1.00±0.01, 0.92±0.03, both P < 0.05). Under hypoxia condition, compared with control group and negative control group, the expression levels of pyroptosis marker proteins in miR-145-5p inhibitor group were significantly decreased [NLRP3 protein (NLRP3/β-actin): 0.21±0.04 vs. 1.70±0.02, 1.63±0.04; GSDMD-N protein (GSDMD-N/β-actin): 1.32±0.02 vs. 2.51±0.02, 2.72±0.03; caspase-1 protein (caspase-1/β-actin): 0.56±0.01 vs. 2.77±0.02, 3.12±0.03; all P < 0.05], Nrf2 protein expression level was significantly increased (Nrf2/β-actin: 1.57±0.04 vs. 1.22±0.01, 1.28±0.04, both P < 0.05), ROS level was significantly down-regulated (fluorescence intensity: 0.64±0.05 vs. 1.87±0.04, 1.70±0.07, both P < 0.05). The results indicated that miR-145-5p could promote cell pyrodeath. The predictive result of miRWalk showed that the 3' untranslated region (3'UTR) of Sirt5 had complementary base binding sites with miR-145-5p. The expression level of Sirt5 protein in cells of miR-145-5p mimic group was significantly lower than that of control group and negative control group under normoxic condition (Sirt5/β-actin: 0.59±0.03 vs. 1.00±0.01, 1.01±0.03, both P < 0.05), which verified that Sirt5 was the target gene of miR-145-5p. (3) The occurrence of pyrodeath could be partially reversed by transfection with Sirt5 overexpression plasmid or adding ROS inhibitor NAC into cells, and Sirt5 overexpression could also up-regulate Nrf2 expression and eliminate intracellular ROS.
CONCLUSION
In human alveolar epithelial cells, miR-145-5p can down-regulate Nrf2 by targeting Sirt5, thereby increasing ROS expression and inducing pyrodeath.
Humans
;
MicroRNAs
;
Pyroptosis
;
Cell Hypoxia
;
Alveolar Epithelial Cells/cytology*
;
Cell Line
;
NLR Family, Pyrin Domain-Containing 3 Protein
;
Caspase 1/metabolism*
;
Epithelial Cells/metabolism*
;
Gasdermins
;
Phosphate-Binding Proteins


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