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[ANXA2 and NF-κB positive feedback loop promotes high glucose-induced pyroptosis in renal tubular epithelial cells]. [ANXA2和NF-κB正反馈回路促进高糖诱导的肾小管上皮细胞焦亡]。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.250305
Jiayi Yang, Yang Luo, Zixuan Zhu, Wenbin Tang
<p><strong>Objectives: </strong>Pyroptosis plays a critical role in tubulointerstitial lesions of diabetic kidney disease (DKD). Annexin A2 (ANXA2) is involved in cell proliferation, apoptosis, and adhesion and may be closely related to DKD, but its specific mechanism remains unclear. This study aims to investigate the role and molecular mechanism of <i>ANXA2</i> in high glucose-induced pyroptosis of renal tubular epithelial cells, providing new targets for DKD prevention and treatment.</p><p><strong>Methods: </strong>Human renal tubular epithelial HK-2 cells were divided into a normal glucose group (5.5 mmol/L), a high glucose group (30.0 mmol/L), and a osmotic control group (24.5 mmol/L mannitol+5.5 mmol/L glucose). <i>ANXA2</i> expression was modulated by overexpression of plasmids and small interfering RNA (siRNA). Cell proliferation was measured by 5-ethynyl-2'-deoxyuridine (EdU) assay, apoptosis by flow cytometry, and <i>ANXA2</i>, p50, and p65 subcellular localization by immunofluorescence. Western blotting was employed to detect α-smooth muscle actin (α-SMA), fibronectin (FN), and collagen type IV (Col-IV). Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting were used to analyze nuclear factor-κB (NF-κB) subunits p50/p65 and the pyroptosis pathway factors NLR family Pyrin domain containing 3 (<i>NLRP3</i>), <i>caspase-1</i>, inferleukin (<i>IL</i>)<i>-1β</i>, and <i>IL-18</i>. Protein interactions between <i>ANXA2</i> and p50/p65 were examined by co-immunoprecipitation, while chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were used to examine NF-κB binding to the ANXA2 promoter.</p><p><strong>Results: </strong>High glucose upregulated <i>ANXA2</i> expression and promoted its nuclear translocation (<i>P</i><0.01). High glucose reduced cell proliferation, increased apoptosis, and elevated α-SMA, FN, and Col-IV expression (all <i>P</i><0.05); <i>ANXA2</i> overexpression aggravated these effects (all <i>P</i><0.05), while <i>ANXA2</i> knockdown reversed them (all <i>P</i><0.05). High glucose activated NF-κB and increased <i>NLRP3</i>, <i>caspase-1</i>, <i>L-1β</i>, and <i>IL-18</i> mRNA and protein expression (all <i>P</i><0.05); ANXA2 overexpression further enhanced this, whereas knockdown suppressed NF-κB activation and downstream factors (all <i>P</i><0.05). Co-immunoprecipitation confirmed <i>ANXA2</i> directly binds the NF-κB subunit p65. ChIP assays revealed p65 binds specifically to <i>ANXA2</i> promoter regions (ChIP-2, ChIP-4, and ChIP-6), and luciferase activity in corresponding mutant constructs (M2, M4, and M6) was significantly increased versus controls (all <i>P</i><0.05), confirming positive transcriptional regulation of <i>ANXA2</i> by p65.</p><p><strong>Conclusions: </strong><i>ANXA2</i> and NF-κB form a positive feedback loop that sustains <i>NLRP3</i> inflammasome activation, promotes pyroptosis pathway activation, and aggravates high glucose-induced renal tubular epithelial cell
目的:焦亡在糖尿病肾病(DKD)的小管间质病变中起关键作用。Annexin A2 (ANXA2)参与细胞增殖、凋亡和粘附,可能与DKD密切相关,但其具体机制尚不清楚。本研究旨在探讨ANXA2在高糖诱导的肾小管上皮细胞焦亡中的作用及分子机制,为DKD的防治提供新的靶点。方法:将人肾小管上皮HK-2细胞分为正常葡萄糖组(5.5 mmol/L)、高糖组(30.0 mmol/L)和渗透对照组(24.5 mmol/L甘露醇+5.5 mmol/L葡萄糖)。通过过表达质粒和小干扰RNA (siRNA)调节ANXA2的表达。用5-乙基-2′-脱氧尿苷(EdU)法检测细胞增殖,用流式细胞术检测细胞凋亡,用免疫荧光法检测ANXA2、p50和p65亚细胞定位。Western blotting检测α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)、ⅳ型胶原蛋白(Col-IV)。采用实时荧光定量PCR (RT-qPCR)和Western blot检测核因子-κB (NF-κB)亚基p50/p65和焦亡途径因子NLR家族Pyrin domain containing 3 (NLRP3)、caspase-1、白细胞介素(IL)-1β和IL-18。采用共免疫沉淀法检测ANXA2与p50/p65之间的蛋白相互作用,采用染色质免疫沉淀法(ChIP)和双荧光素酶报告基因法检测NF-κB与ANXA2启动子的结合。结果:高糖上调了ANXA2的表达,促进了其核易位(PPANXA2过表达加重了这些作用)(所有PANXA2敲低均逆转了这些作用(所有PNLRP3、caspase-1、L-1β和IL-18 mRNA和蛋白的表达)(所有PPANXA2直接结合NF-κB亚基p65)。ChIP实验显示p65特异性结合到ANXA2启动子区域(ChIP-2、ChIP-4和ChIP-6),相应突变体(M2、M4和M6)的荧光素酶活性与对照组(所有PANXA2)相比显著增加。结论:ANXA2与NF-κB形成正反馈回路,维持NLRP3炎性小体激活,促进焦亡通路激活,加重高糖诱导的肾小管上皮细胞损伤。靶向ANXA2或阻断其与p65的相互作用可能是减缓DKD进展的新策略。
{"title":"[<i>ANXA2</i> and NF<b>-</b>κB positive feedback loop promotes high glucose<b>-</b>induced pyroptosis in renal tubular epithelial cells].","authors":"Jiayi Yang, Yang Luo, Zixuan Zhu, Wenbin Tang","doi":"10.11817/j.issn.1672-7347.2025.250305","DOIUrl":"10.11817/j.issn.1672-7347.2025.250305","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Objectives: &lt;/strong&gt;Pyroptosis plays a critical role in tubulointerstitial lesions of diabetic kidney disease (DKD). Annexin A2 (ANXA2) is involved in cell proliferation, apoptosis, and adhesion and may be closely related to DKD, but its specific mechanism remains unclear. This study aims to investigate the role and molecular mechanism of &lt;i&gt;ANXA2&lt;/i&gt; in high glucose-induced pyroptosis of renal tubular epithelial cells, providing new targets for DKD prevention and treatment.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Human renal tubular epithelial HK-2 cells were divided into a normal glucose group (5.5 mmol/L), a high glucose group (30.0 mmol/L), and a osmotic control group (24.5 mmol/L mannitol+5.5 mmol/L glucose). &lt;i&gt;ANXA2&lt;/i&gt; expression was modulated by overexpression of plasmids and small interfering RNA (siRNA). Cell proliferation was measured by 5-ethynyl-2'-deoxyuridine (EdU) assay, apoptosis by flow cytometry, and &lt;i&gt;ANXA2&lt;/i&gt;, p50, and p65 subcellular localization by immunofluorescence. Western blotting was employed to detect α-smooth muscle actin (α-SMA), fibronectin (FN), and collagen type IV (Col-IV). Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting were used to analyze nuclear factor-κB (NF-κB) subunits p50/p65 and the pyroptosis pathway factors NLR family Pyrin domain containing 3 (&lt;i&gt;NLRP3&lt;/i&gt;), &lt;i&gt;caspase-1&lt;/i&gt;, inferleukin (&lt;i&gt;IL&lt;/i&gt;)&lt;i&gt;-1β&lt;/i&gt;, and &lt;i&gt;IL-18&lt;/i&gt;. Protein interactions between &lt;i&gt;ANXA2&lt;/i&gt; and p50/p65 were examined by co-immunoprecipitation, while chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were used to examine NF-κB binding to the ANXA2 promoter.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;High glucose upregulated &lt;i&gt;ANXA2&lt;/i&gt; expression and promoted its nuclear translocation (&lt;i&gt;P&lt;/i&gt;&lt;0.01). High glucose reduced cell proliferation, increased apoptosis, and elevated α-SMA, FN, and Col-IV expression (all &lt;i&gt;P&lt;/i&gt;&lt;0.05); &lt;i&gt;ANXA2&lt;/i&gt; overexpression aggravated these effects (all &lt;i&gt;P&lt;/i&gt;&lt;0.05), while &lt;i&gt;ANXA2&lt;/i&gt; knockdown reversed them (all &lt;i&gt;P&lt;/i&gt;&lt;0.05). High glucose activated NF-κB and increased &lt;i&gt;NLRP3&lt;/i&gt;, &lt;i&gt;caspase-1&lt;/i&gt;, &lt;i&gt;L-1β&lt;/i&gt;, and &lt;i&gt;IL-18&lt;/i&gt; mRNA and protein expression (all &lt;i&gt;P&lt;/i&gt;&lt;0.05); ANXA2 overexpression further enhanced this, whereas knockdown suppressed NF-κB activation and downstream factors (all &lt;i&gt;P&lt;/i&gt;&lt;0.05). Co-immunoprecipitation confirmed &lt;i&gt;ANXA2&lt;/i&gt; directly binds the NF-κB subunit p65. ChIP assays revealed p65 binds specifically to &lt;i&gt;ANXA2&lt;/i&gt; promoter regions (ChIP-2, ChIP-4, and ChIP-6), and luciferase activity in corresponding mutant constructs (M2, M4, and M6) was significantly increased versus controls (all &lt;i&gt;P&lt;/i&gt;&lt;0.05), confirming positive transcriptional regulation of &lt;i&gt;ANXA2&lt;/i&gt; by p65.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;&lt;i&gt;ANXA2&lt;/i&gt; and NF-κB form a positive feedback loop that sustains &lt;i&gt;NLRP3&lt;/i&gt; inflammasome activation, promotes pyroptosis pathway activation, and aggravates high glucose-induced renal tubular epithelial cell","PeriodicalId":39801,"journal":{"name":"中南大学学报(医学版)","volume":"50 6","pages":"940-954"},"PeriodicalIF":0.0,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12464933/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Successful pregnancies following individualized treatment for diffuse uterine leiomyomatosis: A report of 5 cases]. 弥漫性子宫平滑肌瘤病个体化治疗后成功妊娠5例报告
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.240583
Shudan Chen, Sili He, Ruizhen Li, Chunxia Cheng

Objectives: Hysterectomy remains the only definitively effective treatment for diffuse uterine leiomyomatosis (DUL). However, no standardized management strategy exists for DUL patients wishing to preserve fertility. This study summarizes and analyzes 5 cases of individualized treatment in DUL patients desiring fertility preservation, aiming to provide a clinical reference for personalized management of similar patients.

Methods: We retrospectively analyzed the clinical data of 5 DUL patients with fertility intentions admitted to the Department of Obstetrics and Gynecology at Third Xiangya Hospital of Central South University. To preserve fertility, individualized treatment plans were selected based on clinical manifestations and fibroid distribution. One patient received high-intensity focused ultrasound (HIFU); one underwent hysteroscopic myomectomy (HM) combined with laparoscopic myomectomy (LRM); one underwent HIFU combined with HM and LRM; one received drug therapy combined with staged HM; and one underwent HIFU combined with staged HM and drug therapy. Treatment outcomes and pregnancy results were analyzed.

Results: After treatment, all 5 patients showed marked improvement in menstrual volume or dysmenorrhea symptoms and significant reduction in uterine volume; mild intrauterine adhesions occurred in 3 cases. All 5 patients achieved successful pregnancy. One patient with chronic hypertension developed severe preeclampsia at 34 weeks and underwent cesarean section, while the remaining 4 delivered at term by cesarean section. Three cases of placenta accreta and 2 cases of postpartum hemorrhage occurred. During long-term follow-up, one patient underwent hysterectomy 2 years postpartum due to increased menstrual volume, while the other 4 remained stable.

Conclusions: Individualized treatment tailored to DUL patients' conditions can preserve fertility, support successful pregnancy, and achieve favorable pregnancy outcomes.

目的:子宫切除术仍然是唯一明确有效的治疗弥漫性子宫平滑肌瘤病(DUL)。然而,对于希望保持生育能力的DUL患者,没有标准化的管理策略。本研究总结分析5例希望保留生育能力的DUL患者的个体化治疗,旨在为同类患者的个体化治疗提供临床参考。方法:回顾性分析中南大学湘雅第三医院妇产科收治的5例有生育意向的DUL患者的临床资料。为了保持生育能力,根据临床表现和肌瘤分布选择个性化的治疗方案。1例患者接受高强度聚焦超声(HIFU);1例行宫腔镜子宫肌瘤切除术(HM)联合腹腔镜子宫肌瘤切除术(LRM);1例HIFU联合HM和LRM;1例接受药物治疗联合分期HM;1例接受HIFU联合分阶段HM和药物治疗。分析治疗结果和妊娠结果。结果:5例患者经治疗后月经量或痛经症状均明显改善,子宫体积明显缩小;轻度宫内粘连3例。5例患者均成功妊娠。1例慢性高血压患者在34周时出现严重子痫前期,行剖宫产,其余4例足月分娩时行剖宫产。发生胎盘增生3例,产后出血2例。在长期随访中,1例患者因月经量增加于产后2年行子宫切除术,其余4例保持稳定。结论:针对DUL患者的个体化治疗可以保持生育能力,支持成功妊娠,并获得良好的妊娠结局。
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引用次数: 0
[Research progress in the role of caspase-3 in regulating pyroptosis and apoptosis in non-alcoholic fatty liver disease]. [caspase-3在非酒精性脂肪性肝病中调节焦亡和凋亡的研究进展]。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.240688
Saiying Cao, Yi Long, Lina Yang

Non-alcoholic fatty liver disease (NAFLD), including non-alcoholic fatty liver (NAFL), non-alcoholic steatohepatitis (NASH), and advanced fibrosis, is a leading cause of chronic liver disease worldwide, progressing to cirrhosis and ultimately hepatocellular carcinoma (HCC). Excessive accumulation of fatty acids in the liver triggers multiple forms of hepatocyte death and exacerbates NAFLD progression, with pyroptosis and apoptosis considered key events. Recent studies show that cysteine aspartic acid specific protease-3 (caspase-3) is a central regulator of both pyroptosis and apoptosis in NAFLD. Activated caspase-3 not only directly induces apoptosis but also cleaves the N-terminal domain of gasdermin E (GSDME), disrupts cell membranes, releases inflammatory factors, and thereby mediates pyroptosis. Inhibiting caspase-3 expression in NAFLD can alleviate hepatocyte injury (such as ballooning degeneration), dampen pro-inflammatory signaling, and reduce apoptosis. Caspase-3 acts as a key node coordinating pyroptosis and apoptosis and may serve as a novel therapeutic target for the prevention and treatment of NAFLD.

非酒精性脂肪性肝病(NAFLD),包括非酒精性脂肪性肝(NAFL)、非酒精性脂肪性肝炎(NASH)和晚期纤维化,是世界范围内慢性肝病的主要原因,可发展为肝硬化并最终发展为肝细胞癌(HCC)。肝脏中脂肪酸的过度积累会引发多种形式的肝细胞死亡,并加剧NAFLD的进展,其中焦亡和凋亡被认为是关键事件。最近的研究表明,半胱氨酸天冬氨酸特异性蛋白酶-3 (caspase-3)是NAFLD中焦亡和凋亡的中心调节因子。激活的caspase-3不仅可以直接诱导细胞凋亡,还可以裂解气皮蛋白E (GSDME)的n端结构域,破坏细胞膜,释放炎症因子,从而介导细胞焦亡。抑制caspase-3在NAFLD中的表达可以减轻肝细胞损伤(如球囊变性),抑制促炎信号,减少细胞凋亡。Caspase-3作为协调细胞焦亡和凋亡的关键节点,可能成为预防和治疗NAFLD的新的治疗靶点。
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引用次数: 0
[Itaconic acid alleviates macrophage PANoptosis in sepsis-associated acute lung injury via inhibiting ninjurin-1-mediated plasma membrane rupture]. 衣康酸通过抑制忍素-1介导的质膜破裂减轻脓毒症相关急性肺损伤的巨噬细胞PANoptosis。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.250283
Mengrui Chen, Xiaohua Tan, Wenjing Zhong, Hanxi Sha, Liying Liang, Shaokun Liu
<p><strong>Objectives: </strong>Sepsis-associated acute lung injury (S-ALI) is one of the major causes of death in intensive care unit (ICU) patients, yet its mechanisms remain incompletely understood and effective therapies are lacking. Lytic cell death of macrophages is a key driver of the inflammatory cascade in S-ALI. PANoptosis, a newly recognized form of lytic cell death characterized by PANoptosome assembly and activation, involves plasma membrane rupture (PMR) mediated by ninjurin-1 (NINJ1), a recently identified pore-forming protein. Itaconic acid is known for its anti-inflammatory effects, but its role in macrophage PANoptosis during S-ALI is unclear. This study aims to investigate the protective effect of itaconic acid on macrophage PANoptosis in S-ALI to provide new therapeutic insights.</p><p><strong>Methods: </strong>Male specific-pathogen-free C57BL/6J mice (6-8 weeks, 18-20 g) received intraperitoneal lipopolysaccharide (LPS) to establish a classical S-ALI model. Western blotting was used to assess PANoptosome-related proteins and enzymes involved in the itaconic acid metabolic pathway, while real-time reverse transcription polymerase chain reaction and metabolomics quantified itaconic acid levels. Primary peritoneal macrophages (PMs) were pretreated with the itaconate derivative 4-octyl itaconate (4-OI) and then exposed to tumor necrosis factor alpha (TNF-α) plus interferon gamma (IFN-γ) to induce PANoptosis. Cell viability was evaluated by cell counting kit-8 (CCK-8) assay. Western blotting was employed to quantify enzymes of the itaconate-metabolic pathway in PANoptotic macrophages, to evaluate the impact of 4-OI on PANoptosome-associated proteins, and to determine NINJ1 abundance in lung tissues from S-ALI mice and in PANoptotic macrophages. Fluorescent dye FM<sub>4-64</sub> was used to visualize 4-OI-mediated changes in PMR, whereas immunofluorescence staining mapped the effect of 4-OI on both the expression level and membrane localization of NINJ1 in PANoptotic macrophages. The effect of 4-OI on lactate dehydrogenase (LDH) release in culture supernatants and peripheal blood serum was assessed using a LDH assay kit, and non-denataring polyacylamide gel electrophoresis was used to assess the expression of NINJ1 in S-ALI mouse lung tissues and the impact of 4-OI on the expression of PANoptosis-associated NINJ1 multimeric reflected protein in macropahges.</p><p><strong>Results: </strong>In S-ALI mouse lungs, PANoptosome components [NOD-like receptor thermal protein domain associated protein 3 (NLRP3), Gasdermin D (GSDMD), Caspase-1, Z-DNA binding protein (ZBP1), and Caspase-3] and phosphorylated mixed lineage kinase domain-like protein (MLKL) S345 were significantly upregulated (all <i>P</i><0.05), while metabolomics showed compensatory increases in itaconic acid and its key enzymes [aconitate decarboxylase 1 (ACOD1)/immunoresponsive gene 1 (IRG1)]. In macrophages, 4-OI obviously suppressed PANoptosome protein expression, reduce
目的:脓毒症相关急性肺损伤(S-ALI)是重症监护病房(ICU)患者死亡的主要原因之一,但其机制尚不完全清楚,缺乏有效的治疗方法。巨噬细胞的溶解性死亡是S-ALI炎症级联的关键驱动因素。PANoptosis是一种新发现的以PANoptosome组装和激活为特征的溶解性细胞死亡形式,涉及由最近发现的成孔蛋白ninjurin-1 (NINJ1)介导的质膜破裂(PMR)。衣康酸以其抗炎作用而闻名,但其在S-ALI期间巨噬细胞PANoptosis中的作用尚不清楚。本研究旨在探讨衣康酸对S-ALI巨噬细胞PANoptosis的保护作用,为治疗提供新的思路。方法:雄性无特异性病原体C57BL/6J小鼠(6-8周,18-20 g)腹腔注射脂多糖(LPS),建立经典S-ALI模型。Western blotting检测参与衣康酸代谢途径的panoptosome相关蛋白和酶,real-time逆转录聚合酶链反应和代谢组学定量衣康酸水平。用衣康酸衍生物4-辛酯衣康酸(4-OI)预处理原代腹膜巨噬细胞(pm),然后暴露于肿瘤坏死因子α (TNF-α)和干扰素γ (IFN-γ)诱导PANoptosis。采用细胞计数试剂盒-8 (CCK-8)法测定细胞活力。采用Western blotting定量PANoptotic巨噬细胞中itaconate代谢途径的酶,评估4-OI对panoptosome相关蛋白的影响,并测定S-ALI小鼠肺组织和panopto巨噬细胞中NINJ1的丰度。荧光染料FM4-64用于观察4-OI介导的PMR变化,免疫荧光染色显示4-OI对PANoptotic巨噬细胞中NINJ1表达水平和膜定位的影响。采用乳酸脱氢酶(LDH)检测试剂盒检测4-OI对培养上清和外周血血清乳酸脱氢酶(LDH)释放的影响,采用非变性聚丙烯酰胺凝胶电泳检测S-ALI小鼠肺组织中NINJ1的表达及4-OI对巨噬细胞panoposis -associated NINJ1多聚体反射蛋白表达的影响。结果:在S-ALI小鼠肺中,PANoptosome组分[nod样受体热蛋白结构域相关蛋白3 (NLRP3)、Gasdermin D (GSDMD)、Caspase-1、Z-DNA结合蛋白(ZBP1)和Caspase-3]和磷酸化的混合谱系激酶结构域样蛋白(MLKL) S345]显著上调(所有ppp)。结论:衣康酸可能通过抑制ninj1介导的质膜破裂来减轻S-ALI中巨噬细胞PANoptosis。靶向NINJ1或增强衣康酸通路可能为S-ALI提供新的治疗策略。
{"title":"[Itaconic acid alleviates macrophage PANoptosis in sepsis<b>-</b>associated acute lung injury via inhibiting ninjurin<b>-</b>1<b>-</b>mediated plasma membrane rupture].","authors":"Mengrui Chen, Xiaohua Tan, Wenjing Zhong, Hanxi Sha, Liying Liang, Shaokun Liu","doi":"10.11817/j.issn.1672-7347.2025.250283","DOIUrl":"10.11817/j.issn.1672-7347.2025.250283","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Objectives: &lt;/strong&gt;Sepsis-associated acute lung injury (S-ALI) is one of the major causes of death in intensive care unit (ICU) patients, yet its mechanisms remain incompletely understood and effective therapies are lacking. Lytic cell death of macrophages is a key driver of the inflammatory cascade in S-ALI. PANoptosis, a newly recognized form of lytic cell death characterized by PANoptosome assembly and activation, involves plasma membrane rupture (PMR) mediated by ninjurin-1 (NINJ1), a recently identified pore-forming protein. Itaconic acid is known for its anti-inflammatory effects, but its role in macrophage PANoptosis during S-ALI is unclear. This study aims to investigate the protective effect of itaconic acid on macrophage PANoptosis in S-ALI to provide new therapeutic insights.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Male specific-pathogen-free C57BL/6J mice (6-8 weeks, 18-20 g) received intraperitoneal lipopolysaccharide (LPS) to establish a classical S-ALI model. Western blotting was used to assess PANoptosome-related proteins and enzymes involved in the itaconic acid metabolic pathway, while real-time reverse transcription polymerase chain reaction and metabolomics quantified itaconic acid levels. Primary peritoneal macrophages (PMs) were pretreated with the itaconate derivative 4-octyl itaconate (4-OI) and then exposed to tumor necrosis factor alpha (TNF-α) plus interferon gamma (IFN-γ) to induce PANoptosis. Cell viability was evaluated by cell counting kit-8 (CCK-8) assay. Western blotting was employed to quantify enzymes of the itaconate-metabolic pathway in PANoptotic macrophages, to evaluate the impact of 4-OI on PANoptosome-associated proteins, and to determine NINJ1 abundance in lung tissues from S-ALI mice and in PANoptotic macrophages. Fluorescent dye FM&lt;sub&gt;4-64&lt;/sub&gt; was used to visualize 4-OI-mediated changes in PMR, whereas immunofluorescence staining mapped the effect of 4-OI on both the expression level and membrane localization of NINJ1 in PANoptotic macrophages. The effect of 4-OI on lactate dehydrogenase (LDH) release in culture supernatants and peripheal blood serum was assessed using a LDH assay kit, and non-denataring polyacylamide gel electrophoresis was used to assess the expression of NINJ1 in S-ALI mouse lung tissues and the impact of 4-OI on the expression of PANoptosis-associated NINJ1 multimeric reflected protein in macropahges.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;In S-ALI mouse lungs, PANoptosome components [NOD-like receptor thermal protein domain associated protein 3 (NLRP3), Gasdermin D (GSDMD), Caspase-1, Z-DNA binding protein (ZBP1), and Caspase-3] and phosphorylated mixed lineage kinase domain-like protein (MLKL) S345 were significantly upregulated (all &lt;i&gt;P&lt;/i&gt;&lt;0.05), while metabolomics showed compensatory increases in itaconic acid and its key enzymes [aconitate decarboxylase 1 (ACOD1)/immunoresponsive gene 1 (IRG1)]. In macrophages, 4-OI obviously suppressed PANoptosome protein expression, reduce","PeriodicalId":39801,"journal":{"name":"中南大学学报(医学版)","volume":"50 6","pages":"970-985"},"PeriodicalIF":0.0,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12464917/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Ameliorative effects and mechanisms of an integrated endoplasmic reticulum stress inhibitor on lipopolysaccharide-induced cognitive impairment in mice]. [综合内质网应激抑制剂对脂多糖诱导的小鼠认知障碍的改善作用和机制]。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.240262
Dandan Liu, Wenjia Liu, Lihua Xie, Xiaofan Xu, Xiaolin Zhong, Wenyu Cao, Yang Xu, Ling Chen

Objectives: The integrated endoplasmic reticulum stress inhibitor (ISRIB) is a selective inhibitor of the protein kinase R-like endoplasmic reticulum kinase (PERK) signaling pathway within endoplasmic reticulum stress (ERS) and can improve spatial and working memory in aged mice. Although ERS and oxidative stress are tightly interconnected, it remains unclear whether ISRIB alleviates cognitive impairment by restoring the balance between ERS and oxidative stress. This study aims to investigate the effects and mechanisms of ISRIB on lipopolysaccharide (LPS)-induced cognitive impairment in mice.

Methods: Eight-week-old male ICR mice were randomly divided into 3 groups: Normal saline (NS) group, LPS group, and ISRIB+LPS group. NS and LPS groups received daily intraperitoneal injections of normal saline for 7 days; on day 7, LPS group mice received intraperitoneal LPS (0.83 mg/kg) to establish a cognitive impairment model. ISRIB+LPS group received ISRIB (0.25 mg/kg) intraperitoneally for 7 days, with LPS injected 30 minutes after ISRIB on day 7. Cognitive ability was evaluated by the novel place recognition test (NPRT). Real-time fluorogenic quantitative PCR (RT-qPCR) was used to detect changes in nitric oxide synthase (NOS), superoxide dismutase-1 (SOD-1), and catalase (CAT) gene expression in the hippocampus and prefrontal cortex. Oxidative stress markers malondialdehyde (MDA), glutathione (GSH), and oxidized glutathione (GSSG), were measured in hippocampal and prefrontal cortex tissues.

Results: Compared with the NS group, mice in LPS group showed a significant reduction in novel place recognition ratio, upregulation of hippocampal NOS-1 and NOS-2 mRNA, downregulation of SOD-1 and CAT mRNA, increased MDA and GSSG, decreased GSH, and reduced GSH/GSSG ratio (all P<0.05). Compared with the LPS group, mice in ISRIB+LPS group exhibited significantly improved novel place recognition, downregulated NOS-1 and NOS-2 mRNA, upregulated SOD-1 and CAT mRNA, decreased MDA and GSSG, increased GSH, and an elevated GSH/GSSG ratio in the hippocampus (all P<0.05). No significant changes were observed in the prefrontal cortex.

Conclusions: ISRIB improves LPS-induced cognitive impairment in mice by restoring the oxidative/antioxidant balance in the hippocampus.

目的:综合内质网应激抑制剂(ISRIB)是内质网应激(ERS)中蛋白激酶r样内质网激酶(PERK)信号通路的选择性抑制剂,可改善老年小鼠的空间记忆和工作记忆。尽管ERS和氧化应激紧密相连,但ISRIB是否通过恢复ERS和氧化应激之间的平衡来减轻认知障碍尚不清楚。本研究旨在探讨ISRIB对脂多糖(LPS)诱导的小鼠认知功能障碍的影响及其机制。方法:将8周龄雄性ICR小鼠随机分为生理盐水(NS)组、LPS组、ISRIB+LPS组。NS组和LPS组每天腹腔注射生理盐水,连续7 d;第7天,LPS组小鼠腹腔注射LPS (0.83 mg/kg),建立认知功能障碍模型。ISRIB+LPS组腹腔注射ISRIB (0.25 mg/kg) 7 d,第7天ISRIB后30分钟注射LPS。采用新颖位置识别测验(NPRT)评价受试者的认知能力。采用实时荧光定量PCR (RT-qPCR)检测海马和前额叶皮层一氧化氮合酶(NOS)、超氧化物歧化酶-1 (SOD-1)、过氧化氢酶(CAT)基因表达的变化。在海马和前额叶皮层组织中测定氧化应激标志物丙二醛(MDA)、谷胱甘肽(GSH)和氧化谷胱甘肽(GSSG)。结果:与NS组比较,LPS组小鼠新位置识别率显著降低,海马NOS-1和NOS-2 mRNA表达上调,SOD-1和CAT mRNA表达下调,MDA和GSSG升高,GSH降低,GSH/GSSG比值降低(均为PNOS-1和NOS-2 mRNA表达上调,SOD-1和CAT mRNA表达上调,MDA和GSSG降低,GSH升高,海马GSH/GSSG比值升高(均为pp)。ISRIB通过恢复海马的氧化/抗氧化平衡,改善lps诱导的小鼠认知障碍。
{"title":"[Ameliorative effects and mechanisms of an integrated endoplasmic reticulum stress inhibitor on lipopolysaccharide-induced cognitive impairment in mice].","authors":"Dandan Liu, Wenjia Liu, Lihua Xie, Xiaofan Xu, Xiaolin Zhong, Wenyu Cao, Yang Xu, Ling Chen","doi":"10.11817/j.issn.1672-7347.2025.240262","DOIUrl":"10.11817/j.issn.1672-7347.2025.240262","url":null,"abstract":"<p><strong>Objectives: </strong>The integrated endoplasmic reticulum stress inhibitor (ISRIB) is a selective inhibitor of the protein kinase R-like endoplasmic reticulum kinase (PERK) signaling pathway within endoplasmic reticulum stress (ERS) and can improve spatial and working memory in aged mice. Although ERS and oxidative stress are tightly interconnected, it remains unclear whether ISRIB alleviates cognitive impairment by restoring the balance between ERS and oxidative stress. This study aims to investigate the effects and mechanisms of ISRIB on lipopolysaccharide (LPS)-induced cognitive impairment in mice.</p><p><strong>Methods: </strong>Eight-week-old male ICR mice were randomly divided into 3 groups: Normal saline (NS) group, LPS group, and ISRIB+LPS group. NS and LPS groups received daily intraperitoneal injections of normal saline for 7 days; on day 7, LPS group mice received intraperitoneal LPS (0.83 mg/kg) to establish a cognitive impairment model. ISRIB+LPS group received ISRIB (0.25 mg/kg) intraperitoneally for 7 days, with LPS injected 30 minutes after ISRIB on day 7. Cognitive ability was evaluated by the novel place recognition test (NPRT). Real-time fluorogenic quantitative PCR (RT-qPCR) was used to detect changes in nitric oxide synthase (<i>NOS</i>), superoxide dismutase-1 (<i>SOD-1</i>), and catalase (<i>CAT</i>) gene expression in the hippocampus and prefrontal cortex. Oxidative stress markers malondialdehyde (<i>MDA</i>), glutathione (<i>GSH</i>), and oxidized glutathione (<i>GSSG</i>), were measured in hippocampal and prefrontal cortex tissues.</p><p><strong>Results: </strong>Compared with the NS group, mice in LPS group showed a significant reduction in novel place recognition ratio, upregulation of hippocampal <i>NOS-1</i> and <i>NOS-2</i> mRNA, downregulation of <i>SOD-1</i> and <i>CAT</i> mRNA, increased <i>MDA</i> and <i>GSSG</i>, decreased <i>GSH</i>, and reduced <i>GSH/GSSG</i> ratio (all <i>P</i><0.05). Compared with the LPS group, mice in ISRIB+LPS group exhibited significantly improved novel place recognition, downregulated <i>NOS-1</i> and <i>NOS-2</i> mRNA, upregulated <i>SOD-1</i> and <i>CAT</i> mRNA, decreased <i>MDA</i> and <i>GSSG</i>, increased <i>GSH</i>, and an elevated <i>GSH/GSSG</i> ratio in the hippocampus (all <i>P</i><0.05). No significant changes were observed in the prefrontal cortex.</p><p><strong>Conclusions: </strong>ISRIB improves LPS-induced cognitive impairment in mice by restoring the oxidative/antioxidant balance in the hippocampus.</p>","PeriodicalId":39801,"journal":{"name":"中南大学学报(医学版)","volume":"50 6","pages":"986-994"},"PeriodicalIF":0.0,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12464928/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Construction and phenotypic analysis of p2rx2 knockout zebrafish lines. p2rx2基因敲除斑马鱼系的构建及表型分析。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.240659
Yong Zhang, Qingying Shi, Hao Xie, Binling Xie, Lihua Li, Weijing Wu, Huaping Xie, Zi'an Xiao, Dinghua Xie, Ruosha Lai
<p><strong>Objectives: </strong>The purinergic receptor P2X2 (<i>P2RX2</i>) encodes an ATP-gated ion channel permeable to Na<sup>+</sup>, K<sup>+</sup>, and especially Ca²⁺. Loss-of-function mutations in <i>P2RX2</i> are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a <i>p2rx2</i> knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of <i>p2rx2</i> deficiency on the auditory system, providing a basis for understanding <i>P2RX2</i>-related hearing loss and developing gene therapy strategies.</p><p><strong>Methods: </strong>Two CRISPR targets (<i>sgRNA1</i> and <i>sgRNA2</i>) spaced 47 bp apart were designed within the zebrafish <i>p2rx2</i> gene. Synthesized <i>sgRNAs</i> and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group.</p><p><strong>Results: </strong>A stable <i>p2rx2</i> knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all <i>P</i><0.001), indicating reduced hearing sensitivity.</p><p><strong>Conclusions: </strong>We successfully generated a <i>p2rx2</i> loss-of-function zebrafish model using CRISPR/Cas9 technology. <i>p2rx2</i> deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotyp
目的:嘌呤能受体P2X2 (P2RX2)编码一个atp门控离子通道,该通道可通过Na+、K+,尤其是Ca 2 +。已知P2RX2的功能缺失突变可导致常染色体显性非综合征性耳聋41 (DFNA41),其表现为高频听力损失、加速老年性耳聋和对噪声诱发损伤的易感性增加。斑马鱼具有体积小、发育快、繁殖力强、胚胎透明、与人类基因高度保守等特点,为研究人类疾病和发育机制提供了理想的模型。本研究旨在利用CRISPR/Cas9基因编辑系统构建p2rx2敲除斑马鱼模型,研究p2rx2缺失对听觉系统的影响,为理解p2rx2相关性听力损失和制定基因治疗策略提供依据。方法:在斑马鱼p2rx2基因中设计两个相距47 bp的CRISPR靶点(sgRNA1和sgRNA2)。将合成的sgRNAs和Cas9蛋白微注射到单细胞期t宾根(TU)株斑马鱼胚胎中。PCR和凝胶电泳验证了受精后36小时(hpf)的编辑效率。将存活的胚胎饲养至成年(F0),剪去尾巴,进行基因分型,筛选阳性嵌合。异种杂交获得F1杂合子,杂交获得F2纯合突变体。聚合酶链反应(PCR)结合测序验证突变类型和遗传力。受精后5天,采用YO-PRO-1染色检测侧线神经突和耳石区毛细胞形态和计数。在每组9只4月龄野生型和突变型斑马鱼中测量600、800、1 000和2 000 Hz的听觉诱发电位阈值。结果:成功建立了稳定的p2rx2基因敲除斑马鱼系。测序显示,在第一个目标位点插入66 bp,引入过早终止密码子(TAA),导致蛋白质翻译的早期终止和功能丧失。胚胎发育正常,无明显畸形。在5 dpf时,突变体与野生型相比,耳石区域的毛细胞密度显著降低,但侧线神经鞘未受影响。AEP测试显示,与野生型相比,纯合突变体在所有4个频率上的听觉阈值都显著升高(均为pp)。结论:我们利用CRISPR/Cas9技术成功构建了p2rx2功能丧失斑马鱼模型。P2rx2缺乏导致耳石区毛细胞缺陷,各频率的听觉阈值升高,表明其在维持斑马鱼听觉毛细胞功能和听觉感知中起关键作用。该表型对耳石区域的限制表明p2rx2在感觉器官中的组织特异性作用。该模型为阐明p2rx2相关听力损失的分子机制、筛选耳保护药物和开发基因治疗提供了有价值的工具。
{"title":"Construction and phenotypic analysis of <i>p2rx2</i> knockout zebrafish lines.","authors":"Yong Zhang, Qingying Shi, Hao Xie, Binling Xie, Lihua Li, Weijing Wu, Huaping Xie, Zi'an Xiao, Dinghua Xie, Ruosha Lai","doi":"10.11817/j.issn.1672-7347.2025.240659","DOIUrl":"10.11817/j.issn.1672-7347.2025.240659","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Objectives: &lt;/strong&gt;The purinergic receptor P2X2 (&lt;i&gt;P2RX2&lt;/i&gt;) encodes an ATP-gated ion channel permeable to Na&lt;sup&gt;+&lt;/sup&gt;, K&lt;sup&gt;+&lt;/sup&gt;, and especially Ca²⁺. Loss-of-function mutations in &lt;i&gt;P2RX2&lt;/i&gt; are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a &lt;i&gt;p2rx2&lt;/i&gt; knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of &lt;i&gt;p2rx2&lt;/i&gt; deficiency on the auditory system, providing a basis for understanding &lt;i&gt;P2RX2&lt;/i&gt;-related hearing loss and developing gene therapy strategies.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Two CRISPR targets (&lt;i&gt;sgRNA1&lt;/i&gt; and &lt;i&gt;sgRNA2&lt;/i&gt;) spaced 47 bp apart were designed within the zebrafish &lt;i&gt;p2rx2&lt;/i&gt; gene. Synthesized &lt;i&gt;sgRNAs&lt;/i&gt; and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;A stable &lt;i&gt;p2rx2&lt;/i&gt; knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all &lt;i&gt;P&lt;/i&gt;&lt;0.001), indicating reduced hearing sensitivity.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;We successfully generated a &lt;i&gt;p2rx2&lt;/i&gt; loss-of-function zebrafish model using CRISPR/Cas9 technology. &lt;i&gt;p2rx2&lt;/i&gt; deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotyp","PeriodicalId":39801,"journal":{"name":"中南大学学报(医学版)","volume":"50 6","pages":"919-930"},"PeriodicalIF":0.0,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12464923/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Effect of morphine pump in prepontine cistern via lumbar approach for intractable head and neck cancer pain]. [腰路入路吗啡泵对顽固性头颈癌疼痛的影响]。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.250049
Wenjie Zhang, Bohua Yin, Xinning Li, Jiaxin Lei, Yanying Xiao, Yaping Wang, Dingquan Zou

Objectives: Managing patients with refractory head and neck cancer pain is one of the more challenging issues in clinical practice, and traditional intrathecal drug delivery also fails to provide adequate analgesia. There are currently no comprehensive and effective treatment methods. This study aims to observe the efficacy and safety of treating intractable head and neck cancer pain with morphine pump via lumbar approach to the prepontine cistern.

Methods: A total of 18 patients with intractable head and neck cancer pain treated with prepontine cistern morphine pumps were selected from the Department of Pain Management, Second Xiangya Hospital, Central South University between September 2019 and July 2023. Statistical analysis was performed on patients' preoperative and postoperative (1 week, 1 month, and 2 months after surgery), Numerical Rating Scale (NRS) scores, Self-Rating Depression Scale (SDS) scores, daily oral morphine consumption, the number of daily breakthrough pain episodes, and postoperative daily intrathecal morphine dosage.

Results: The NRS scores, SDS scores, daily oral morphine consumption, and the number of daily breakthrough pain episodes of patients at each time point after surgery were significantly lower than before surgery (all P<0.05). With the gradual increase in the dosage of intrathecal morphine, the daily oral morphine consumption of patients at each postoperative time point was significantly reduced compared to preoperative levels (all P<0.05). The complications related to the operation were mild, including nausea in 5 cases (31.3%), headache in 2 cases (12.5%); hypotension, urine retention, hypersomnia and constipation in 1 case (6.3% each), and no serious adverse events occurred. All improved and were discharged after symptomatic treatment.

Conclusions: The implantation of prepontine cistern morphine pump effectively controls intractable head and neck cancer pain, demonstrating characteristics of minimal invasiveness, mild side effects, and low medication dosage under the premise of standardized procedures.

目的:治疗难治性头颈癌疼痛是临床实践中较具挑战性的问题之一,传统的鞘内给药也不能提供足够的镇痛。目前还没有全面有效的治疗方法。本研究旨在观察吗啡泵经腰前池入路治疗顽固性头颈癌疼痛的疗效和安全性。方法:选取2019年9月至2023年7月中南大学湘雅第二医院疼痛管理科采用前置池式吗啡泵治疗的顽固性头颈癌疼痛患者18例。统计分析患者术前、术后(术后1周、1个月、2个月)、数值评定量表(NRS)评分、抑郁自评量表(SDS)评分、每日口服吗啡用量、每日突破疼痛发作次数、术后每日鞘内吗啡用量。结果:术后各时间点患者NRS评分、SDS评分、每日口服吗啡用量、每日突破疼痛发作次数均显著低于术前(均为ppp)。结论:在规范操作的前提下,预置池式吗啡泵可有效控制顽固性头颈癌疼痛,具有侵入性小、副作用轻、用药剂量小的特点。
{"title":"[Effect of morphine pump in prepontine cistern via lumbar approach for intractable head and neck cancer pain].","authors":"Wenjie Zhang, Bohua Yin, Xinning Li, Jiaxin Lei, Yanying Xiao, Yaping Wang, Dingquan Zou","doi":"10.11817/j.issn.1672-7347.2025.250049","DOIUrl":"10.11817/j.issn.1672-7347.2025.250049","url":null,"abstract":"<p><strong>Objectives: </strong>Managing patients with refractory head and neck cancer pain is one of the more challenging issues in clinical practice, and traditional intrathecal drug delivery also fails to provide adequate analgesia. There are currently no comprehensive and effective treatment methods. This study aims to observe the efficacy and safety of treating intractable head and neck cancer pain with morphine pump via lumbar approach to the prepontine cistern.</p><p><strong>Methods: </strong>A total of 18 patients with intractable head and neck cancer pain treated with prepontine cistern morphine pumps were selected from the Department of Pain Management, Second Xiangya Hospital, Central South University between September 2019 and July 2023. Statistical analysis was performed on patients' preoperative and postoperative (1 week, 1 month, and 2 months after surgery), Numerical Rating Scale (NRS) scores, Self-Rating Depression Scale (SDS) scores, daily oral morphine consumption, the number of daily breakthrough pain episodes, and postoperative daily intrathecal morphine dosage.</p><p><strong>Results: </strong>The NRS scores, SDS scores, daily oral morphine consumption, and the number of daily breakthrough pain episodes of patients at each time point after surgery were significantly lower than before surgery (all <i>P</i><0.05). With the gradual increase in the dosage of intrathecal morphine, the daily oral morphine consumption of patients at each postoperative time point was significantly reduced compared to preoperative levels (all <i>P</i><0.05). The complications related to the operation were mild, including nausea in 5 cases (31.3%), headache in 2 cases (12.5%); hypotension, urine retention, hypersomnia and constipation in 1 case (6.3% each), and no serious adverse events occurred. All improved and were discharged after symptomatic treatment.</p><p><strong>Conclusions: </strong>The implantation of prepontine cistern morphine pump effectively controls intractable head and neck cancer pain, demonstrating characteristics of minimal invasiveness, mild side effects, and low medication dosage under the premise of standardized procedures.</p>","PeriodicalId":39801,"journal":{"name":"中南大学学报(医学版)","volume":"50 6","pages":"995-1001"},"PeriodicalIF":0.0,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12464918/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Application progress of single-cell RNA sequencing technology in breast development and related diseases]. [单细胞RNA测序技术在乳腺发育及相关疾病中的应用进展]。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.250039
Shiyi Wen, Yang Hu, Xiangyu Chen, Jianda Zhou, Ping Li

The spatio-temporal heterogeneity of breast cell subsets forms the fundamental biological basis for physiological development and pathological progression, including tumorigenesis; however, its complex regulatory mechanisms are not yet fully elucidated. With its high-resolution capabilities, single-cell RNA sequencing (scRNA-seq) technology offers a powerful tool for dissecting this cellular heterogeneity. This technology enables the construction of high-precision breast cell atlases, the accurate identification of distinct cell subsets, and the reconstruction of differentiation trajectories from stem/progenitor cells to functional epithelial cells. By resolving the transcriptional regulatory networks that govern cell fate determination, intercellular communication patterns, and dynamic microenvironmental interactions, scRNA-seq has unveiled the molecular foundations of breast development and provided new perspectives on the pathogenesis of related diseases such as breast cancer and macromastia. Furthermore, scRNA-seq demonstrates significant potential for discovering early molecular markers of disease, deciphering tumor heterogeneity, and elucidating mechanisms of therapeutic resistance. The continued application of scRNA-seq for dissecting breast cell heterogeneity, combined with its integration with multi-modal data such as spatial omics, promises to provide critical evidence and new insights for revealing the molecular mechanisms of breast development-related diseases and for formulating precision therapeutic strategies.

乳腺细胞亚群的时空异质性构成了包括肿瘤发生在内的生理发育和病理进展的基本生物学基础;然而,其复杂的调控机制尚未完全阐明。凭借其高分辨率的能力,单细胞RNA测序(scRNA-seq)技术为解剖这种细胞异质性提供了强大的工具。该技术能够构建高精度的乳腺细胞图谱,准确识别不同的细胞亚群,并重建从干细胞/祖细胞到功能性上皮细胞的分化轨迹。通过解析控制细胞命运决定、细胞间通讯模式和动态微环境相互作用的转录调控网络,scRNA-seq揭示了乳房发育的分子基础,并为乳腺癌和巨乳症等相关疾病的发病机制提供了新的视角。此外,scRNA-seq在发现疾病的早期分子标记、破译肿瘤异质性和阐明治疗耐药机制方面显示出巨大的潜力。scRNA-seq在剖析乳腺细胞异质性方面的持续应用,结合其与空间组学等多模态数据的整合,有望为揭示乳腺发育相关疾病的分子机制和制定精确的治疗策略提供关键证据和新的见解。
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引用次数: 0
[Therapeutic effects of natural products on animal models of chronic obstructive pulmonary disease]. [天然产物对慢性阻塞性肺疾病动物模型的治疗作用]
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.240124
Xinru Fei, Guixian Yang, Junnan Liu, Tong Liu, Wei Gao, Dongkai Zhao

Chronic obstructive pulmonary disease (COPD) currently lacks effective treatments to halt disease progression, making the search for preventive and therapeutic drugs a pressing issue. Natural products, with their accessibility, affordability, and low toxicity, offer promising avenues. Investigating the pharmacological effects and related signaling mechanisms of active components from natural products on COPD animal models induced by various triggers has become an important focus. In animal models induced by cigarette smoke, cigarette smoke combined with lipopolysaccharide (LPS), air pollution, elastase, bacterial or viral infections, the active compounds of natural products, such as flavonoids, terpenoids, and phenolics, can exert anti-inflammatory, antioxidant, mucus-regulating, and airway remodeling-inhibiting effects through key signaling pathways including nuclear factor-erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1), nuclear factor-kappa B (NF-κB), and mitogen-activated protein kinase (MAPK). These findings not only provide a theoretical basis for the clinical diagnosis and treatment of COPD but also point to new directions for future scientific research.

慢性阻塞性肺疾病(COPD)目前缺乏有效的治疗方法来阻止疾病的进展,因此寻找预防和治疗药物成为一个紧迫的问题。天然产品,以其可及性、可负担性和低毒性,提供了有希望的途径。研究天然产物活性成分对各种诱发因素诱导的COPD动物模型的药理作用及相关信号机制已成为一个重要的研究热点。在香烟烟雾诱导的动物模型中,香烟烟雾与脂多糖(LPS)、空气污染、弹性酶、细菌或病毒感染联合作用,其天然产物黄酮类、萜类、酚类物质等活性化合物可通过核因子-红细胞2相关因子2 (Nrf2)/血红素加氧酶-1 (HO-1)、核因子-κB (NF-κB)等关键信号通路发挥抗炎、抗氧化、调节粘液、抑制气道重塑等作用。和丝裂原活化蛋白激酶(MAPK)。这些发现不仅为慢性阻塞性肺病的临床诊断和治疗提供了理论依据,也为今后的科学研究指明了新的方向。
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引用次数: 0
[Establishment and application of a UPLC-MS/MS method for the determination of tiletamine and its metabolite in biological samples]. [生物样品中替乐胺及其代谢物的UPLC-MS/MS测定方法的建立与应用]。
Q3 Medicine Pub Date : 2025-06-28 DOI: 10.11817/j.issn.1672-7347.2025.240609
Zihao Cai, Wenguang Yan, Jiahao Li, Yanjun Ding, Jiang Ling
<p><strong>Objectives: </strong>Tiletamine, a veterinary anesthetic, has emerged as a novel psychoactive substance and has been abused in many parts of the world, causing great harm to public health. However, the sensitivity of existing detection methods cannot meet the needs of forensic practice. This study aims to establish an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of tiletamine and its metabolite desethyltiletamine in human biological samples, and to verify its applicability in forensic practice.</p><p><strong>Methods: </strong>SKF<sub>525A</sub> was used as the internal standard. Biological samples were extracted with acetonitrile containing 1 ng/mL SKF<sub>525A</sub>, vortexed for 10 min, ultrasonicated for 20 min, centrifuged at 10 000 r/min for 10 min, and 500 μL of the supernatant was filtered through a 0.22 μm membrane. Analyses were performed using an ACQUITY UPLC H-Class PLUS system and an XEVO TQ-S Micro triple quadrupole mass spectrometer. An ACQUITY UPLC<sup>®</sup> BEH C18 (1.7 µm, 2.1 mm×100 mm) column at a flow rate of 0.3 mL/min was used, and four mobile phase systems were tested to optimize separation. Detection used positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode, with quantifier ion transitions of mass to charge 224.043→179.016 for tiletamine and mass to charge 196.08→151.06 for desethyltiletamine. Calibration curves were established over 0.1-200 ng/mL in spiked blood samples. The linear range, limit of detection (LOD), and limit of quantification (LOQ) were determined. Low (5 ng/mL), medium (20 ng/mL), and high (100 ng/mL) concentrations of tiletamine were spiked into blood, liver, and kidney to evaluate precision, accuracy, matrix effect, recovery, and stability. Finally, actual forensic case samples were tested to validate applicability.</p><p><strong>Results: </strong>The established UPLC-MS/MS method achieved simultaneous detection of tiletamine and desethyltiletamine in human biological samples, with retention times of 3.42 min and 2.82 min, respectively. Using mobile phase A (20 mmol/L ammonium acetate and 0.1% formic acid in water) and mobile phase B (acetonitrile) produced the best separation. In blood, tiletamine showed good linearity from 0.1-200 ng/mL (<i>r</i>=0.992, <i>R</i><sup>2</sup>=0.983), LOD 0.03 ng/mL, LOQ 0.1 ng/mL, recovery 92%-107%, and matrix effect 71%-99%. In liver and kidney, recoveries were 91%-98% and 93%-104%, and matrix effects were 69%-96% and 72%-100%, respectively. Intra- and inter-day precision [expressed as relative standard deviation (RSD)] and accuracy [expressed as relative error (RE)] were within 15%, and samples were stable at -20 ℃. Tiletamine was detected in actual case samples at 0.37 μg/mL (blood), 0.15 μg/g (liver), 0.11 μg/g (kidney) in case 1, and 8.75 ng/mL (blood) in case 2; desethyltiletamine was also detected in blood.</p><p><strong>Conclusions: </strong>The UPLC-MS/
目的:替乐胺是一种兽药,是一种新型精神活性物质,在世界许多地方被滥用,对公众健康造成极大危害。然而,现有的检测方法的灵敏度不能满足法医实践的需要。本研究旨在建立一种超高效液相色谱-串联质谱(UPLC-MS/MS)测定人体生物样品中替乐胺及其代谢物去乙基替乐胺的方法,并验证其在法医实践中的适用性。方法:以SKF525A为内标。生物样品用含有1 ng/mL SKF525A的乙腈提取,旋涡10 min,超声20 min, 10000 r/min离心10 min, 0.22 μm膜过滤500 μL上清。使用ACQUITY UPLC H-Class PLUS系统和XEVO TQ-S Micro三重四极杆质谱仪进行分析。采用ACQUITY UPLC®BEH C18(1.7µm, 2.1 mm×100 mm)色谱柱,流速为0.3 mL/min,采用四种流动相体系进行分离优化。检测采用多反应监测(MRM)模式下的正电喷雾电离(ESI+),对替乐胺的定量离子跃迁为质量电荷224.043→179.016,对去乙基替乐胺的定量离子跃迁为质量电荷196.08→151.06。在0.1 ~ 200 ng/mL范围内建立校正曲线。确定了该方法的线性范围、检出限和定量限。将低浓度(5ng /mL)、中浓度(20ng /mL)和高浓度(100ng /mL)的替乐胺加入血液、肝脏和肾脏中,以评估精密度、准确度、基质效应、回收率和稳定性。最后,对实际法医案例样本进行了测试,以验证其适用性。结果:建立的UPLC-MS/MS方法可同时检测人体生物样品中的替乐胺和去乙基替乐胺,保留时间分别为3.42 min和2.82 min。采用流动相A (20 mmol/L乙酸铵和0.1%甲酸水溶液)和流动相B(乙腈)分离效果最佳。在血液中,替乐胺在0.1 ~ 200 ng/mL范围内呈良好的线性关系(r=0.992, R2=0.983),检出限0.03 ng/mL,检出限0.1 ng/mL,回收率92% ~ 107%,基质效应71% ~ 99%。在肝脏和肾脏中,加样回收率分别为91% ~ 98%和93% ~ 104%,基质效应分别为69% ~ 96%和72% ~ 100%。日内、日间精密度[以相对标准偏差(RSD)表示]和准确度[以相对误差(RE)表示]均在15%以内,样品在-20℃下稳定。实际病例标本中tilletamine的检测值分别为0.37 μg/mL(血)、0.15 μg/g(肝)、0.11 μg/g(肾)、8.75 ng/mL(血);血液中也检测到去乙基替乐胺。结论:UPLC-MS/MS方法高效、准确、灵敏,适用于人体生物样品中替乐胺和去乙基替乐胺的检测。
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中南大学学报(医学版)
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