血必净对脓毒症早期大鼠急性肺损伤的作用及可能机制

目的:探讨血必净注射液对脓毒症早期肺组织损伤的影响及其可能作用机制,为临床脓毒症时急性肺损伤的治疗提供新思路。方法:选用健康雄性S-D大鼠30只,随机分3组,即假手术组(Sham组)、盐水组(NS组)和血必净组(XBJ组)。应用盲肠结扎穿孔法(CLP)建立脓毒症大鼠模型,术后6 h处死动物,通过HE染色光镜观察组织形态学改变及透射电镜观察肺组织超微结构,测定各组肺组织湿干重比,采用RT-PCR检测肺组织内皮素-1(ET-1)、诱导型一氧化氮合酶(iNOS)、基质金属蛋白酶-9(MMP-9)、组织金属蛋白酶抑制物-1(TIMP-1)mRNA的表达。结果:XBJ组肺泡及肺间质水肿和内皮细胞超微结构改变均较NS组明显减轻。术后6 h,NS组大鼠肺组织湿干重比(W/D)显著高于Sham组(5.37±0.12 vs 4.33±0.06,P0.01),XBJ组肺组织W/D较NS组显著降低(4.67±0.09 vs 5.37±0.12,P0.05)。此外,XBJ组大鼠肺组织ET-1(0.511±0.111 vs 0.705±0.122,P0.01)、iNOS(0.456±0.075 vs 0.548±0.098,P0.05),MMP-9(0.617±0.079 vs 0.732±0.131,P0.05)、TIMP-1(0.438±0.043vs 0.515±0.049,P0.01)mRNA的表达均较NS组明显降低,而NS组大鼠肺组织ET-1(0.705±0.122 vs 0.400±0.033,P0.01)、iNOS(0.548±0.098 vs 0.334±0.027,P0.01)、MMP-9(0.732±0.131 vs 0.352±0.061,P0.01)、TIMP-1mRNA(0.515±0.049 vs 0.365±0.068,P0.01)水平均较Sham组明显升高。结论:血必净治疗对脓毒症大鼠的肺组织损伤有一定的保护作用,可能与其降低肺组织ET-1、iNOS、MMP-9、TIMP-1的表达有关。     

Characterization of SVEP1, KIAA, and SRPX2 in an in vitro cell culture model of endotoxemia

To assess the influence of unknown factors in endotoxemia, a conditioned medium, achieved by the stimulation of THP1 monocytes with lipopolysaccharide (LPS) [4 h], was used for the stimulation of human umbilical vein endothelial cells (HUVECs) [16 h]. SVEP1, KIAA0247, and SRPX2 were selected after microarray analysis. To study their possible functions, siRNAs of SVEP1, KIAA0247, or SRPX2 were used for the transfection of HUVECs and cells were stimulated with conditioned medium [16 h]. Inhibition of SVEP1 expression resulted in an increase of soluble intercellular adhesion molecule (sICAM) 1 (10%) and soluble E-selectin (sE-selectin) (19%). Inhibition of SRPX2 led to an increase of sICAM (11%) and sE-selectin (14%). KIAA0247 negative HUVECs showed a decrease in monocyte chemoattractant protein (MCP) 1 of 16%. SVEP1 and SRPX2 seemed to act as regulators of ICAM1 and E-selectin shedding and influence the expression of membrane bound adhesion molecules.     

脓毒症患者发生凝血功能紊乱的临床特征及预后影响因素分析

摘要 目的：探究脓毒症患者发生凝血功能紊乱的临床特征及预后影响因素。方法：选择2019年9月~2023年1月本院收治的80例脓毒症患者为本次研究对象,对所有患者开展凝血功能检验,并依据检验结果,将患者分为凝血功能异常组（n=60）及正常组（n=20）,分析凝血功能障碍异常及正常患者临床特征、凝血功能障碍异常及正常患者病情严重程度,并依据脓毒症患者预后,将其分为存活组（n=64）及死亡组（n=16）脓毒症患者预后的单因素与多因素Logistic回归分析。结果：凝血功能正常组及异常组平均动脉压、体温、白细胞计数（WBC）、纤维蛋白原（FIB）、凝血酶时间（TT）指标水平无较大差异（P＞0.05）,与凝血功能正常组比较,凝血功能异常组患者呼吸、脉率、降钙素原（PCT）、C反应蛋白（CRP）指标水平相对较高,部分凝血活酶时间（APTT）、凝血酶原时间（PT）指标水平相对较长,PLT指标水平相对较高（P＜0.05）;与凝血功能正常的脓毒症患者相比较,凝血功能异常患者急性肾损伤（AKI）及急性呼吸窘迫综合征（ARDS）发生率、急性生理与慢性健康评分（APACHE Ⅱ）及急性生理功能评分（SOFA）评分更高,住ICU时间均相对较长（P＜0.05）,机械通气及住院时间比较（P＞0.05）;脓毒症患者预后影响因素分析中,结果显示,年龄、体质量指数（BMI）、性别、糖尿病史、高血压病史均未对脓毒症患者预后产生较大影响（P＞0.05）,PT、PCT、APTT、CRP、SOFA评分及APACHE Ⅱ评分对脓毒症预后产生严重影响,死亡组患者各指标水平均高于存活组（P＜0.05）;将影响脓毒症患者预后的单因素予以赋值,纳入Logistic回归分析,以PCT、PT、APTT、CRP、SOFA、APACHE Ⅱ 评分为自变量,结果显示,PCT、PT、CRP、SOFA、APACHE Ⅱ 评分是影响脓毒症患者预后的独立危险因素（P＜0.05）。结论：脓毒症患者发生凝血功能紊乱会对临床相关指标产生影响,从而增加疾病严重程度,依据脓毒症患者预后,研究结果显示,PT、PCT、CRP、SOFA及APACHE Ⅱ评分均会对脓毒症患者预后产生不良影响,检验上述指标水平,可为临床评估脓毒症预后提供一定参考价值。     

下调miR-223表达对脓毒症心肌病小鼠心肌的保护作用及机制研究

摘要 目的：探讨下调miR-223表达对脓毒症心肌病（SCM）小鼠心肌的保护作用及其机制。方法：按随机数字表法将27只8-10 周龄SPF级雄性C57BL/6小鼠分配至SCM模型（0 h,6 h,12 h,18 h,24 h）时相组、Normal组、SCM组、miR-223 antagomir NC组、miR-223 antagomir组,每组3只。腹腔注射脂多糖（lipopolysaccharide, LPS）15 mg/kg构建SCM小鼠模型。miR-223 antagomir NC组与miR-223 antagomir组分别于建模前连续3天鼠尾静脉注射miR-223 antagomir NC、miR-223 antagomir预处理。采用反转录-聚合酶链反应（RT-PCR）研究SCM模型各个时相组小鼠心肌组织miR-223的表达情况。采用苏木素伊红（HE）染色法观察Normal组、SCM组、miR-223 antagomir NC组和miR-223 antagomir组小鼠心肌病理形态变化。采用酶联免疫吸附实验（ELISA）测定Normal组、SCM组、miR-223 antagomir NC组和miR-223 antagomir组小鼠血清cTnI、BNP、CK-MB、IL-6、IL-1β、TNF-α的含量并进行相关性分析。结果：SCM模型时相组小鼠随刺激时间延长,心肌组织miR-223表达水平逐渐升高。与Normal组比较,SCM组、miR-223 antagomir NC组、miR-223 antagomir组小鼠心肌组织出现不同程度损伤;血清心肌损伤标记物cTnI、BNP、CK-MB及炎性因子IL-6、IL-1β、TNF-α的表达水平均上升,差异具有统计学意义（P<0.05）;与SCM组比较,miR-223 antagomir NC组各项指标相差不大,差异均无统计学意义（P>0.05）;miR-223 antagomir组小鼠心肌组织病理损伤程度有所减轻,心肌损伤标记物cTnI、BNP、CK-MB及炎性因子IL-6、IL-1β、TNF-α水平下降,差异具有统计学意义（P<0.05）。相关性分析结果显示小鼠miR-223表达与心肌损伤标记物cTnI、BNP、CK-MB及炎性因子IL-6、IL-1β、TNF-α的表达呈正相关。结论：下调miR-223表达可通过减轻炎症反应对SCM小鼠心肌产生保护作用。     

静注人免疫球蛋白联合万古霉素治疗小儿败血症的疗效及外周血NLR、PCT变化观察

摘要 目的：观察静注人免疫球蛋白联合万古霉素治疗小儿败血症的疗效及外周血中性粒细胞/淋巴细胞比值（NLR）、降钙素原(PCT)变化。方法：选取2011年1月~2020年1月我院收治的败血症患儿80例为研究对象,按数字随机表法分为对照组和观察组各40例,对照组给予万古霉素治疗,观察组在对照组基础上给予静注人免疫球蛋白治疗,比较两组临床疗效、症状改善时间和住院时间、NLR、PCT、超敏C反应蛋白(hs-CRP)、白细胞计数（WBC）、免疫功能及不良反应发生率。结果：观察组治疗有效率高于对照组（87.50%vs65.00%）（P<0.05）。观察组神经系统症状改善时间、体温改善时间、拒奶改善时间和住院时间为（6.22±1.05）d、（3.88±0.25）d、（5.10±0.86）d、（8.71±2.05）d,均短于对照组的（8.76±1.53）d、（6.22±0.64）d、（7.53±1.46）d和（11.24±3.36）d,比较差异有统计学意义（P<0.05）。治疗后观察组外周血NLR、PCT、hs-CRP、WBC水平为（1.35±0.20）、（0.80±0.34）mg/mL、（3.56±0.62）g/L、（9.12±1.80）×10⁹/L,均显著低于对照组的（1.83±0.32）、（2.19±0.73）mg/mL、（9.78±2.64）g/L和（12.26±2.59）×10⁹/L,比较差异有统计学意义（P<0.05）。治疗后观察组CD4⁺、CD3⁺、CD4/CD8为（42.77±11.36）％、（41.27±11.26）％、（1.70±0.33）,均显著高于对照组的（35.80±9.32）％、（35.66±9.40）％和（1.29±0.25）,比较差异有统计学意义（P<0.05）。两组不良反应发生率比较无差异（10.00%vs7.50%）（P>0.05）。结论：静注人免疫球蛋白联合万古霉素治疗小儿败血症的疗效显著,可降低炎症因子,提高免疫功能,且安全性较高。     

Effect of 593C > T GPx1 SNP alone and in synergy with 47C > T SOD2 SNP on the outcome of critically ill patients

During critical illness and sepsis there is severe antioxidant depletion, and this scenario raises the critical ill patient’s mortality risk. Glutathione peroxidase (GPx) is one of the first endogenous antioxidant defense enzymes, and it works cooperatively with superoxide dismutase (SOD) and catalase (CAT) to detoxify free radicals from the cellular environment. Genetic studies are important to understand the complexity of human oxidative stress and how the organism responds to an extreme situation such as critically care conditions. Previous studies with a GPx1 single nucleotide polymorphism (593C > T SNP; rs1050450; protein variant in GPx1: Pro198Leu) showed 593T carriers and 593TT homozygotes present higher risk to develop different diseases. We assessed the relationship of the genotype distribution of GPx1 SNP in critically ill patients with their conditions (organ dysfunction, sepsis, and septic shock) and their outcome. We monitored 626 critically ill patients daily from the ICU (intensive care unit) admission to their discharge from hospital, or death. Our study revealed a significant association between 593TT GPx1 genotype and mortality; the mortality rate was higher in homozygous 593TT GPx1 (N = 94) when compared with the group of subjects with genotypes 593CT or 593CC GPx1 (N = 532) (52% vs. 38%, P = 0.009; OR = 1.79; 95% CI = 1.13–2.85). Evaluating the subgroup of 293 ICU patients with sepsis, a pooled analysis including two genetic variants GPx1 and SOD2 (47C > T SNP, rs4880; protein variant in MnSOD: Ala–9Val) showed a significant difference in relation to progression to septic shock. The frequency of septic shock among septic patients with 593T GPx1 and 47C SOD2 alleles (N = 122) was higher when compared with septic patients carrying other settings of genotypes (N = 174) (78% vs. 66%; P = 0.028; OR = 1.81; 95% CI = 1.03–3.18). Accepting the previously reported functional effects of these two SNPs on GPx1 and SOD2 gene expressions and, consequently, on GPx1 and MnSOD enzyme activities, we believe our results may be considered as an important contribution for the understanding of oxidative imbalance during the critical ill.     

谷氨酰胺对脓毒症患者肠粘膜屏障功能和免疫功能的影响

目的:研究谷氨酰胺对脓毒症患者肠道黏膜屏障功能和免疫功能的影响。方法:将2012年10月至2013年10本院收治的40例脓毒症患者随机分为治疗组和对照组,每组20例。两组患者均给予常规对症治疗,治疗组在此基础上加用谷氨酰胺治疗,对照组接受安慰剂治疗。采用分光光度法监测其血清D-乳酸水平,高效液相色谱法监测尿乳果糖/甘露醇(L/M)值。对比两组肠道黏膜屏障功能和免疫功能指标及ARDS、MODS发生率。结果:治疗后,治疗组ARDS和MODS发生率分别为24.1%,17.2%,均低于对照组的38.9%,33.3%,两组比较差异均有统计学意义(P0.05);治疗14 d后,治疗组D-乳酸及尿L/M水平较对照组明显降低,血清Ig G、Ig M、CD4+和CD4+/CD8+水平均增高,两组比较差异均有统计学意义(P0.05)。结论:谷氨酰胺治疗脓毒性患能明显改善肠道黏膜屏障功能,促进患者免疫功能和营养状态的提高,同时还能够降低患者肠道细菌、内毒素移位。     

Mesenchymal stem cells as a therapeutic tool to treat sepsis

Sepsis is a clinical syndrome caused by a deregulated host response to an infection. Sepsis is the most frequent cause of death in hospitalized patients. Although knowledge of the pathogenesis of sepsis has increased substantially during the last decades, attempts to design effective and specific therapiestargeting components of the derailed host response have failed. Therefore, there is a dramatic need for new and mechanistically alternative therapies to treat this syndrome. Based on their immunomodulatory properties, adult mesenchymal stem or stromal cells(MSCs) can be a novel therapeutic tool to treat sepsis. Indeed, MSCs reduce mortality in experimental models of sepsis by modulating the deregulated inflammatory response against bacteria through the regulation of multiple inflammatory networks, the reprogramming of macrophages and neutrophils towards a more antiinflammatory phenotype and the release of antimicrobial peptides. This report will review the current knowledge on the effects of MSC treatment in preclinical experimental small animal models of sepsis.     

Sepsis Associated Encephalopathy Studied by MRI and Cerebral Spinal Fluid S100B Measurement

The pathogenesis of sepsis associated encephalopathy (SAE) is not yet clear: the blood–brain barrier (BBB) disruption has been indicated among the possible causative mechanisms. S100B, a calcium binding protein, originates in the central nervous system but it can be also produced by extra-cerebral sources; it is passively released from damaged glial cells and neurons; it has limited passage through the BBB. We aimed to demonstrate BBB damage as part of the pathogenesis of SAE by cerebral spinal fluid (CSF) and serum S100B measurements and by magnetic resonance imaging (MRI). This paper describes four septic patients in whom SAE was clinically evident, who underwent MRI and S100B measurement. We have not found any evidence of CSF-S100B increase. Serum S100B increase was found in three out of four patients. MRI did not identify images attributable to BBB disruption but vasogenic edema, probably caused by an alteration of autoregulation, was diagnosed. S100B does not increase in CSF of septic patients; S100B increase in serum may be due to extracerebral sources and does not prove any injury of BBB. MRI can exclude other cerebral pathologies causing brain dysfunction but is not specific of SAE. BBB damage may be numbered among the contributors of SAE, which aetiology is certainly multifactorial: an interplay between the toxic mediators involved in sepsis and the indirect effects of hyperthermia, hypossia and hypoperfusion.