Inflammatory reaction dominated by defense response will arise against infection and trauma. As an important proinflammatory cytokine, high mobility group box 1 (HMGB1) is widely expressed in all nuclear cells to mediate the inflammatory response. However, the biological functions of HMGB1 in inflammation vary depending on the type of HMGB1 protein modification and the localization in the cell. HMGB1 protein will be modified as acetylation of lysine residues, methylation of lysine residues, oxidation of cysteine residues, phosphorylation of serine residues, glycosylation of asparagine residues, adenosine diphosphate-ribosylation and lactylation of the protein in the nucleus, migrate from the nucleus to the cytoplasm, and release into the extracellular compartment. Extracellular HMGB1 can bind to receptors for advanced glycation end products (RAGE) and Toll-like receptors, activate cells and regulate inflammatory responses. The authors review the research progress in regulatory mechanism of HMGB1 in inflammation response from aspects of its post-translational modifications, releases, biological roles and binding receptors, hoping to provide theoretical basis for finding the targets of inflammation intervention.

The body is in a very complex pathophysiological state under trauma, including ischemia and hypoxia, inflammation caused by infection and tissue necrosis and accumulation of metabolic waste. Metastasis associated lung adenocarcinoma transcript 1 (MALAT1) is involved in the regulation of a variety of cell behaviors, such as proliferation, apoptosis, differentiation, migration, epithelial-mesenchymal transition, autophagy and morphological maintenance. Under trauma, the expression of MALAT1 is significantly increased. In different injury models, the role of MALAT1 is slightly different, and the specific mechanism is unknown. The authors summarize the regulatory effects of MALAT1 on the body under traumatic conditions from the biological characteristics of MALAT1 and its role in different injury models, so as to provide references for clinical control of inflammation development and improvement of disease prognosis.

目的 模拟临床创伤休克并发脓毒性休克的情况,建立"二次休克"打击动物模型,连续观察小鼠血压和生存情况,为筛选抗休克药物提供实验动物模型.方法 采用双后肢股骨骨折加眼球放血45％建立创伤休克(TH)模型;于TH术后8h进行盲肠结扎穿孔术(CLP)构建"二次打击"模型.将70只6～8周龄成年雄性C57BL/6小鼠随机分为:创伤休克组(TH+生理盐水),模型组(TH+CLP+生理盐水),假手术组(TH+开腹腔+生理盐水),各组持续观察小鼠平均动脉血压(MPa)以及生存情况;于TH后0 h,2 h,32 h,各组分别处死5只小鼠(处死小鼠前持续观察小鼠生命状态),收集血液检测血清肌酐值.结果 各组TH后血压急剧下降,在第二次打击CLP后血压持续降低,模型组血压下降较假手术组与创伤休克组剧烈(p<0.05).与假手术组、 创伤休克组相比,模型组死亡率明显增加(p<0.05),血清肌酐值显著增高(p<0.05),经CLP后小鼠的呼吸频率增加,渐出现体温冰冷,蜷缩拥挤.结论 小鼠TH术后8 h并发脓毒性休克(即"二次休克"模型)较假手术组与创伤休克组的死亡率明显增加,血压持续下降,肾功能损害,符合临床创伤性休克并发脓毒性休克的病理生理改变,动物模型成功构建.

目的制备转录因子Musculin（MSC）敲除（knockout,KO）-叉状头转录因子家族成员3（forkhead box P3,Foxp3）-绿色荧光蛋白（green fluorescent protein,GFP）敲入（knockin,KI）小鼠模型,并初步应用于严重创伤介导调节性T细胞（regulatory T cell,Treg）-Th17失衡研究。方法通过MSCKO纯合子（homozygote,-/-）小鼠和Foxp3-GFPKI纯合子（homozygote,+/+）小鼠杂交,获取MSCKO（heterozygote,+/-）-Foxp3-GFPKI（+/-）杂合子小鼠,再按同笼兄妹近亲繁殖方法进行杂合子配对,直到获取MSCKO-Foxp3-GFPKI纯合子小鼠。将获得MSCKO-Foxp3-GFPKI小鼠连续交配5代以上,观察各代小鼠的基因型、表现型和遗传稳定性。对获取的MSCKO-Foxp3-GFPKI小鼠和Foxp3-GFPKI小鼠实施失血/骨折,3 h后取周围淋巴结（lymph nodes,LN）、肠系膜淋巴结（mesenteric lymph nodes,MLN）、肠道集合淋巴结（Peyer’s patches,PP）、脾（spleen,SP）、骨髓（bone marrow,BM）和胸腺（thymus,Thy）。流式细胞术检测机体免疫器官和组织Treg的分布变化,qPCR检测MSC、Foxp3、孤核受体-γt（orphan nuclear receptor-γt,ROR-γt）的表达水平,进一步验证MSCKO-Foxp3-GFPKI小鼠的实用性,并初步探讨MSC对严重创伤介导Treg-Th17失衡的影响。结果杂交获得的各代MSCKO-Foxp3-GFPKI纯合子小鼠均能表达各分子标志物,活力、体质量和繁育能力正常。流式细胞术检测结果显示,各免疫器官和组织中均存在不同强度的GFP信号;严重创伤3 h后,MSCKO-Foxp3-GFPKI小鼠Treg在LN、MLN、PP和SP中的百分比分别为5.81%、4.81%、1.41%和3.1%,较创伤前差异有统计学意义。qPCR检测结果显示,相对正常组而言,野生型创伤组MSC在外周淋巴器官MLN（6.154 9）和PP（7.789 7）中的相对表达量显著升高（P＜0.05,P＜0.01）;Foxp3在MSCKO创伤组MLN（2.591 2）和PP（1.506 4）中的相对表达量上升（P＜0.01）,表明MSC缺失可使创伤后Treg-Th17平衡左移;而ROR-γt在同组MLN（0.5395）和PP（0.544 9）中的相对表达量下降（P＜0.01）,同样证实了Treg-Th17平衡发生了偏移。结论成功制备MSCKO-Foxp3-GFPKI小鼠,遗传性状稳定,分子标志物和表现型明确。严重创伤后MSC可以影响Treg的分布,并使Treg-Th17平衡发生偏移,提示MSC可能作为严重创伤后恢复机体内环境免疫稳态的潜在调节靶点。

Field surgery training is an important part of professional education in medical staff of war wound/trauma care.The aim of this study is to suggest the improved strategy in training course of field surgery in the light of the curriculum demands of trainees.The investigation of trainees' demands held in the Institute of Surgery Research,Daping Hospital of Third Military Medical University from 2005 to 2016 was analyzed and related strategy was put forward.The results showed that there were several disadvantages in current training,such as inadequate diversification and practicality,poor popularity and unitary evaluation system.Therefore,the corresponding strategy was suggested in the aspects of idea,mode and assessment so as to improve the professional education system of field surgery.

目的 探讨吴茱萸碱(evodiamine,EVO)对酵母多糖(zymosan,ZYM)诱导的小鼠腹膜炎中炎症反应及中性粒细胞(polymorphonuelear neutrophil,PMN)浸润的影响.方法 将54只成年雄性C57BL/6小鼠按随机数字表法分为正常对照组、模型组和EVO治疗组.模型组腹腔注射1 mg/mLZYM溶液(1 mL)建立急性腹膜炎小鼠模型,EVO治疗组则同时腹腔注射10 mg/kg EVO及1 mg/mLZYM溶液,正常对照组腹腔注射等量磷酸盐缓冲液.各组处理后2、6、12 h处死6只小鼠,收集眼眶血及腹腔灌洗液.采用酶联免疫吸附法检测血清及腹腔灌洗液中白介素6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、角化细胞来源趋化因子(keratinocyte-derived chemokine,KC)及巨噬细胞炎性蛋白2(macrophage inflammatory protein 2,MIP-2)的含量;通过血细胞计数板计数腹腔灌洗液中混合细胞总数;利用流式细胞仪检测腹腔灌洗液中PMN比例及PMN中活性氧(reactive oxygen species,ROS)水平;采用Western blot检测6h时腹腔灌洗液混合细胞中核转录因子-kB(nuclear factorκB,NF-κB) p65入核情况.结果 建模后2、6、12 h,模型组血清及腹腔灌洗液中IL-6、TNF-α、KC及MIP-2含量均高于正常对照组(P＜0.05),EVO治疗组上述指标均较模型组明显降低(P＜0.05),其中,血清中IL-6、KC及MIP-2均于2h达高峰,随后逐渐下降,而TNF-α在2h逐渐上升,6h达到高峰,随后逐渐下降;腹腔灌洗液中TNF-α、KC及MIP-2均于2h达高峰,随后逐渐下降,而IL-6在2h逐渐上升,6h达到高峰,随后逐渐下降.制模后6h,EVO治疗组可显著降低腹腔混合细胞数与PMN数及其百分比(P＜0.05),且PMN中ROS显著降低(P＜0.05).另EVO治疗可显著降低腹腔混合细胞中细胞核的NF-κB p65表达(P＜0.05).结论 EVO可抑制ZYM诱导急性小鼠腹膜炎模型中血清及腹腔灌洗液中炎症因子及趋化因子的分泌,减轻腹腔PMN的浸润,并抑制PMN中ROS的产生,该作用可能是通过抑制NF-kB p65入核实现的.

T淋巴细胞在适应性免疫中发挥细胞免疫功能.调节性T细胞(regulatoryT cell,Treg)是重要的T淋巴细胞亚群,可有效维持免疫耐受和机体内环境稳定.深入挖掘能够影响Treg生物学特征的关键因子有助于干预Treg相关免疫性疾病和炎症反应的进程,对于控制炎症发展、改善疾病预后具有潜在的临床应用价值.新近发现,芳香烃受体(aryl hydrocarbon receptor,AhR)与机体免疫性疾病和炎症反应密切相关,而且活化的AhR可以影响Treg的生物学特征.该文就AhR对Treg的分布、增殖、分化、凋亡、分泌的影响以及相关调控机制作一综述.

This study was performed to establish a murine macrophage RAW264.7 cell line stably overexpressing AhR for evaluating the regulatory effects of AhR in inflammation induced by LPS stimulation.A lentiviral vector containing AhR-FIag-EGFP gene was constructed and then packaged in HEK293T cells;RAW264.7 cells were infected with collected supernatant and screened by puromycin.The gene and protein expression of AhR were analyzed by qRT-PCR and Western blotting,respectively;the expression of inflammatory cytokines were detected by qRT-PCR and ELISA.Data showed that a RAW264.7 cell strain with stable expression of AhR-Flag-EGFP was successfully constructed.Fluorescent microscopy and flow cytometry demonstrated that the expression rates of EGFP reached to 100％;the qRT-PCR and Western blot showed that the gene and protein expression of AhR were significantly upregulated in RAW/AhR group compared to RAW/Vector group.The expression of IL-6 and IL-1β in LPS-induced RAW/AhR group were much lower than that in LPS-induced RAW/Vector group.In contrast,the level of IL-10 in LPS-induced RAW/AhR group was much higher than that in LPS-induced RAW/Vector group.In conclusion,a RAW264.7 cell line stably expressing AhR has constructed,and AhR negatively regulates the LPS-induced inflammatory response in macrophages,which lay a foundation for further study AhR function.

目的 探讨胍丁胺对脓毒症小鼠血管内皮功能的影响.方法 将42只C57BL/6小鼠按随机数字表法分为假手术组、模型组、胍丁胺治疗组,每组14只.采用盲肠结扎穿孔术(CLP)复制脓毒症小鼠模型;假手术组只开腹不进行结扎穿孔术,术后腹腔注射磷酸盐缓冲液.胍丁胺组术后腹腔注射400mg/kg胍丁胺;其余两组术后腹腔注射等量磷酸盐缓冲液.于制模后24h处死小鼠取血,采用酶联免疫吸附试验(ELISA)检测血清细胞间黏附分子-1(ICAM-1)、血管间黏附分子-1(VCAM-1)、血管内皮生长因子(VEGF)、血管性血友病因子(vWF)、血管紧张素-Ⅱ(Ang-Ⅱ)、单核细胞趋化因子-1(MCP-1)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-6、IL-10)的含量;通过异硫氰酸荧光素(FITC)-葡聚糖荧光示踪法检测肝、肺组织的血管通透性.结果 造模后24h,模型组小鼠腹部呈浅粉色、体温下降、精神萎靡、活动减少;胍丁胺组小鼠治疗后精神和活动状态均有明显好转.与假手术组比较,模型组血管内皮相关生物标志物的含量均显著升高[ICAM-1(ng/L):9887.12±897.90比4747.12±958.00,VCAM-1(ng/L):2884.09±230.00比1349.43±238.47,vWF(ng/L):48.61±0.69比31.82±2.38,VEGF(ng/L):45.14±1.76比20.65±0.57,Ang-Ⅱ(ng/L):13.26±0.84比8.87±0.80,MCP-1(ng/L):1881.31±106.71比66.21±6.68,均P＜0.01];胍丁胺组上述各指标含量均较模型组明显降低[ICAM-1 (ng/L):6380.45±1017.73比9887.12±897.90,VCAM-1(ng/L):1894.76±235.39比2884.09±230.00,vWF(ng/L):37.77±4.42比48.61±0.69,VEGF(ng/L):32.19±1.96比45.14±1.76,Ang-Ⅱ(ng/L):10.81±0.45比13.26±0.84,MCP-1 (ng/L):553.58±56.19比1881.31±106.71,均P＜0.01].同时,与假手术比较,模型组血清炎症因子水平显著升高[IL-6 (ng/L):5225.61±600.99比49.33±5.20,IL-10 (ng/L):1034.02±139.34比21.86±2.25,TNF-α(ng/L):135.84±18.86比68.64±17.85,均P＜0.01],而胍丁胺治疗后炎性介质的水平均较模型组明显降低[IL-6 (ng/L):1268.63±219.63比5225.61±600.99;IL-10(ng/L):240.84±75.20比1034.02±139.34;TNF-α(ng/L):68.67±20.30比135.84±18.86,均P＜0.01];模型组中肝、肺组织荧光强度较假手术组明显增强[肝:4998.00±616.33比2626.67±103.00;肺:495.33±67.11比255.33±28.67,均P＜0.01],而胍丁胺能显著降低肝、肺组织荧光强度[肝:3472.00±420.33比4998.00±616.33;肺:399.67±11.67比495.33±67.11,均P＜0.01].结论 胍丁胺通过抑制ICAM-1、VEGF及vWF等生物标志物和炎性因子的分泌,降低肝、肺组织血管通透性,从而缓解脓毒症小鼠的血管内皮损伤.

目的 研究胰岛素样生长因子-1受体(insulin-like growth factor 1 receptor,IGF-1R)的选择性抑制剂PQ401在体内外的抗炎效应.方法 体外实验:小鼠RAW264.7细胞株经0.1、1、10 μmol/LPQ401预作用1h后,加入10 μg/mL脂多糖(LPS)作用12h,采用酶联免疫吸附实验(ELISA)检测细胞上清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-1 β、IL-6)的含量.另将小鼠RAW264.7细胞株经10 μmol/L PQ401预作用1h后,加入10 μg/mL LPS作用6、12、24 h后再次检测以上指标的变化.体内实验:将50只成年雄性C57BL/6小鼠按随机数字表法分为对照组、模型组和不同剂量PQ401组,每组10只.腹腔注射LPS 10 mg/kg制备急性炎症反应小鼠模型,对照组腹腔注射等量二甲基亚砜(DMSO),PQ401组分别腹腔注射LPS 10 mg/kg和PQ401 25、50、100 mg/kg.于给药12 h后通过ELISA检测血清和肝组织中TNF-ot、IL-1β、IL-6的含量;用全自动生化分析仪测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及总胆红素(TBil)水平;采用Western blot检测肝细胞核内核转录因子-κB p65(NF-κB p65)的含量.结果 与LPS单独作用组相比,PQ401呈浓度和时间依赖性降低RAW264.7细胞上清中TNF-α、IL-1β、IL-6的水平.在LPS诱导的小鼠急性腹膜炎模型中,小鼠经LPS注射12 h后表现为精神萎靡、活动减少,而PQ401组小鼠的精神状态、活动均好于模型组.PQ401治疗后能有效降低LPS所致急性腹膜炎小鼠血清中TNF-α、IL-1β、IL-6的含量,降低肝组织TNF-α、IL-1β的水平和NF-κB p65的含量.结论 PQ401在体内外对LPS诱导的炎症因子分泌具有明显的抑制效应.