老年免疫缺陷病合并败血症患者病原菌分布及药物敏感性分析

目的探究老年免疫缺陷病合并败血症患者病原菌分布及对药物的敏感性。方法回顾性分析2018年1月到2019年10月我院收治的74例老年免疫缺陷病(AIDS)合并败血症患者,血培养细菌检出情况及药物敏感性实验结果。结果74例老年AIDS合并败血症患者共检出真菌50株(67.57%),包括马尼尔菲青霉菌31株、新生隐球菌12株、克鲁维酵母菌6株和葡萄牙假丝酵母菌1株;革兰阴性菌16株(21.62%),包括大肠埃希菌5株、铜绿假单胞菌5株、猪霍乱沙门菌4株、人苍白杆菌1株和肺炎克雷伯菌1株;革兰阳性菌8株(10.81%),包括金黄色葡萄球菌3株、屎肠球菌2株、表皮葡萄球菌2株和溶血葡萄球菌1株。真菌对氟康唑、5-氟胞嘧啶耐药率均为2.00%,对两性霉素B、伊曲康唑耐药率均为100%;革兰阴性菌对阿莫西林、头孢噻吩等抗菌药物耐药率分别为56.25%、43.75%;革兰阳性菌对四环素、青霉素、红霉素等抗菌药物耐药率分别为87.50%、75.00%、50.00%;参照药敏结果给予抗真菌药物和抗生素治疗,因败血症死亡10例(13.51%),其中4例感染大肠埃希菌,3例铜绿假单胞菌,2例感染金黄色葡萄球菌,1例马尼尔菲青霉菌。结论本地区老年免疫缺陷病合并败血症患者感染菌以真菌为主,对大多数抗真菌药物敏感。     

脓毒症患者血清降钙素原、C-反应蛋白与凝血功能、病情评分的关系及预后的影响因素分析

摘要 目的：探讨脓毒症患者血清降钙素原（PCT）、C-反应蛋白（CRP）与凝血功能、病情评分的关系及预后的影响因素。方法：选择2017年8月至2019年10月我院诊治的150例脓毒症患者作为研究对象,根据脓毒症组患者的生存状况将其分为存活组（n=118）和死亡组（n=32）。检测两组患者血清PCT、CRP水平,凝血酶原时间（PT）和部分凝血活酶时间（APTT）。采用急性生理功能和慢性健康状况评分系统Ⅱ(APACHEⅡ)和序贯器官衰竭估计(SOFA)对两组患者病情进行评估。比较两组患者临床资料。相关性分析采用Pearson检验。患者预后的影响因素采用多因素Logistic回归分析。结果：与存活组相比,死亡组血清PCT、CRP,PT、APTT,APACHE II和SOFA评分均明显升高（P<0.05）。血清PCT>7.32 ng/mL、CRP>86.73 ng/mL、PT>15.48s、APTT>36.74s、APACHE Ⅱ评分>15.88分和SOFA评分>6.48分均是脓毒症患者死亡的危险因素（OR=2.099、1.747、2.333、1.596、1.916、2.208,P<0.05）。血清PCT和CRP均与APACHE Ⅱ评分、SOFA评分、PT和APTT呈正相关（P<0.05）。结论：血清PCT和CRP水平升高与脓毒症患者死亡相关,两者与脓毒症患者APACHE Ⅱ评分、SOFA评分、PT和APTT存在正相关,并且是患者不良预后的影响因素,在脓毒症患者预后的评估中具有一定临床价值。     

脓毒症大鼠肝肾功能、凝血功能和炎症指标的变化及意义

摘要 目的：研究脓毒症大鼠肝肾功能、凝血功能和炎症指标的变化及意义。方法：选取SPF级雄性SD大鼠20只,以随机数字表法将其分成实验组与假手术组,每组各10只。其中实验组通过盲肠穿刺结扎制备脓毒症大鼠模型,假手术组大鼠仅翻动盲肠,不作结扎处理。分别采集所有大鼠术后0 h、6 h、12 h、24 h时的尾静脉血3 mL,检测并比较上述时间点两组大鼠的肝肾功能、凝血功能和炎症指标水平的差异。结果：实验组术后6 h、12 h、24 h的谷丙转氨酶（ALT）、尿素氮（BUN）以及肌酐（Cr）水平均高于假手术组（P＜0.05）。实验组术后24 h的活化部分凝血活酶时间（APTT）、凝血酶原时间（PT）、纤维蛋白原（FIB）水平均高于假手术组（P＜0.05）。实验组大鼠术后6 h、12 h、24 h的血清白细胞计数（WBC）、白细胞介素-6（IL-6）、降钙素原（PCT）水平均高于假手术组（P＜0.05）。结论：脓毒症大鼠肝肾功能存在明显的受损,凝血功能障碍明显,且机体内会释放大量的炎症因子。     

The diagnostic and prognostic role of MiRNA 15b and MiRNA 378a in neonatal sepsis

Background and objectivesSepsis is one of the major factors for both term and preterm babies with morbidity and mortality. On the basis of recent clinical trials, altered circulating micro-RNAs (miRNAs) may serve as possible biomarkers in sepsis for diagnosis and prognosis. The aim of this research is to assess the diagnostic and prognostic biomarkers of miRNA 15b and miRNA 378a for neonatal sepsis.Subjects & methodsThis study was carried out 25 neonates with sepsis admitted to neonatal ICU of Menoufia University Hospital and 25 healthy controls from February 2019 to May 2020. The relative quantification (RQ) of miRNA-15b and miRNA-378a expression was assessed using real time PCR technique. Results: Our results demonstrated that patients with sepsis had significantly higher level of MiRNA-15b than the healthy volunteers. On the other hand, patients with sepsis had significantly lower level of MiRNA-378a than the healthy volunteers. The ROC curve showed that the serum MiRNA-15b was a significant discriminator of sepsis with a combined sensitivity and specificity of 76% and 88% with cutoff point of 3.24. In addition, serum MiRNA-378a was a significant discriminator of sepsis with a combined sensitivity and specificity of 60% and 88% with cutoff point of 0.361. The miRNA-15b expression significantly correlated positive with respiratory rate (r =0.415,p =0.039), WBCs (r = 0.408, p =0.043), and CRP (r =0.407, p=0.043). Likewise, miRNA-378a expression significantly correlated negative with respiratory rate (r =-0.415p =0.024), WBCs (r =- 0.442, p =0.027), and CRP (r =- 0.459, p=0.021).ConclusionBoth MiRNA 15b and 378a are promising biomarker for neonatal sepsis.     

Therapeutic potential of plasma gelsolin administration in a rat model of sepsis

BackgroundGelsolin is an actin-binding protein found in the cytoplasm and in extracellular fluids including blood plasma. Plasma gelsolin concentration decreases after a wide range of injuries. We hypothesized that the repletion of gelsolin would limit inflammation and tissue injury in a rat model of sepsis using cecal ligation and double puncture (2CLP).MethodsHuman plasma gelsolin (pGSN, 10 mg in 1 ml saline) was administered once immediately following surgery, and control 2CLP (2CLP Alb) and sham animals were injected with 1 ml saline containing equimolar albumin. Treatments were administered intraperitoneally (IP), intravenously (IV), or subcutaneously (SC).ResultsGelsolin levels in the 2CLP Alb group were lower than in sham animals. Administration of pGSN increased levels when administered IV and SC, but not IP. Morbidity scores were significantly less severe in the 2CLP pGSN group than in the 2CLP Alb group when pGSN was administered IV and SC, but not IP. Furthermore, enzymatic activity indicative of tissue damage (lactate dehydrogenase and alanine transaminase) was significantly lower in 2CLP pGSN group when treated SC compared to 2CLP Alb group.ConclusionThese data provide further evidence that exogenous gelsolin can reduce morbidity from sepsis.     

Protective effects of finasteride on the pulmonary immune response in a combined model of trauma-hemorrhage and polymicrobial sepsis in mice

Literature supports findings about a gender specific outcome following multiple trauma. Male sex hormones such as dihydrotestosterone (DHT) exert deleterious effects on the posttraumatic immune response whereas increased estradiol concentrations are correlated with improved outcome. Pretreatment with the 5α-reductase inhibitor finasteride resulted in an improved outcome following trauma-hemorrhage (TH) in mice. The present study tested the hypothesis that finasteride exerts beneficial effects on the posttraumatic immune response also in a combined setting of TH and sepsis when administered during the resuscitation process.

Material and Methods

Male C57BL/6N-mice were subjected to TH (blood pressure, 35 mm Hg, 60 min) followed by finasteride application and fluid resuscitation. Thereafter, finasteride was administered every 12 h. 24 h after TH, sepsis was induced by cecal ligation and puncture (CLP) or sham operation was performed. Plasma cytokines (MIP-1α, MIP-1β, TNF-α, MCP-1, IL-6), productive capacity by alveolar macrophages (AM) and systemic estradiol levels were determined 4 h thereafter. The expression of pro-inflammatory mediators in lung tissue was evaluated by PCR. Pulmonary infiltration of PMN was determined by immunohistochemical staining.

Results

Finasteride treatment resulted in a reduced posttraumatic cytokine secretion of AM as well as in a decreased concentration of MCP-1 and MIP-1β in lung tissue. Systemic estradiol levels were increased following finasteride treatment.

Conclusion

Finasteride mediates salutary effects on the pulmonary immune response using a therapeutical approach following TH–CLP in mice. Thus, finasteride might represent a relevant therapeutic substance following major trauma also in the clinical setting.     

Non-toxigenic Clostridium sordellii: clinical and microbiological features of a case of cholangitis-associated bacteremia

Toxigenic Clostridium sordellii strains are increasingly recognized to cause highly lethal infections in humans that are typified by a toxic shock syndrome (TSS). Two glucosylating toxins, lethal toxin (TcsL) and hemorrhagic toxin (TcsH) are believed to be important in the pathogenesis of TSS. While non-toxigenic strains of C. sordellii demonstrate reduced cytotoxicity in vitro and lower virulence in animal models of infection, there are few data regarding their behavior in humans. Here we report a non-TSS C. sordellii infection in the context of a polymicrobial bacterial cholangitis. The C. sordellii strain associated with this infection did not carry either the TcsL-encoding tcsL gene or the tcsH gene for TcsH. In addition, the strain was neither cytotoxic in vitro nor lethal in a murine sepsis model. These results provide additional correlative evidence that TcsL and TcsH increase the risk of mortality during C. sordellii infections.     

脓毒症患者胃肠功能障碍的中医药治疗进展

脓毒症是内外科危重病患者常见的并发症,近年来全球发病率明显增高,且以每年1.5%-8%的速度增加,成为危重病患者的主要死亡原因之一,随着其发病机制的进一步研究,胃肠道的屏障功能越来越受到重视,传统的治疗措施未能明显改善病死率,且大量以抗炎症介质的实验以失败告终。而近年来中医药在危重症患者中的作用越来越受到重视,中西医结合治疗的病死率远远低于目前国际病死率,本文对中医药治疗脓毒症患者胃肠功能进展及现状做如下综述,并简单分析了目前存在的问题和解决办法。     

Phagocytosis and digestion of pH-sensitive fluorescent dye (Eos-FP) transfected E. coli in whole blood assays from patients with severe sepsis and septic shock

The function of phagocytic and antigen presenting cells is of crucial importance to sustain immune competence against infectious agents as well as malignancies. We here describe a reproducible procedure for the quantification of phagocytosis by leukocytes in whole blood. For this, a pH-sensitive green-fluorescent protein- (GFP) like dye (Eos-FP) is transfected into infectious microroganisms. After UV-irradiation, the transfected bacteria emit green (≈5160 nm) and red (≈581 nm) fluorescent light at 490 nm excitation. Since the red fluorescent light is sensitive to acidic pH, the phagocytosed bacteria stop emitting red fluorescent light as soon as the phagosomes fuse with lysosomes. The green fluorescence is maintained in the phagolysosome until pathogen degradation is completed. Fluorescence emission can be followed by flow cytometry with filter settings documenting fluorescence 1 (FL 1, FITC) and fluorescence 2 (FL 2, phycoerythrin, PE). Eos-FP transfected bacteria can also be traced within phagocytes using microscopical techniques. A standardized assay has been developed which is suitable for clinical studies by providing clinicians with syringes pre-filled with fixed and appropriately UV-irradiated Eos-FP E. coli (TruCulture™). After adding blood or body fluids to these containers and starting the incubation at 37°C, phagocytosis by granulocytes proceeds over time. Cultures can be terminated at a given time by lysing red blood cells followed by flow cytometry. A pilot study demonstrated that Eos-FP E. coli phagocytosis and digestion was up-regulated in the majority of patients with either severe sepsis or septic shock as compared to healthy donors (p < 0.0001 after o/n incubation). Following treatment with recombinant human granulocyte colony-stimulating factor (rhG-CSF) in selected patients with sepsis, phagolysosome fusion appeared to be accelerated.     

Global Shapes of F-actin Depolymerization-competent Minimal Gelsolins: INSIGHT INTO THE ROLE OF g2-g3 LINKER IN pH/Ca2+ INSENSITIVITY OF THE FIRST HALF*

Because of its ability to rapidly depolymerize F-actin, plasma gelsolin has emerged as a therapeutic molecule in different disease conditions. High amounts of exogenous gelsolin are, however, required to treat animal models of different diseases. Knowing that the F-actin depolymerizing property of gelsolin resides in its N terminus, we made several truncated versions of plasma gelsolin. The smaller versions, particularly the one composed of the first 28–161 residues, depolymerized the F-actin much faster than the native gelsolin and other truncates at the same molar ratios. Although G1-G3 loses its dependence on Ca²⁺ or low pH for the actin depolymerization function, interestingly, G1-G2 and its smaller versions were found to regain this requirement. Small angle x-ray scattering-based shape reconstructions revealed that G1-G3 adopts an open shape in both the presence and the absence of Ca²⁺ as well as low pH, whereas G1-G2 and residues 28–161 prefer collapsed states in Ca²⁺-free conditions at pH 8. The mutations in the g2-g3 linker resulted in the calcium sensitivity of the mutant G1-G3 for F-actin depolymerization activity, although the F-actin-binding sites remained exposed in the mutant G1-G3 as well as in the smaller truncates even in the Ca²⁺-free conditions at pH 8. Furthermore, unlike wild type G1-G3, calcium-sensitive mutants of G1-G3 acquired closed shapes in the absence of free calcium, implying a role of g2-g3 linker in determining the open F-actin depolymerizing-competent shape of G1-G3 in this condition. We demonstrate that the mobility of the G1 domain, essential for F-actin depolymerization, is indirectly regulated by the gelsolin-like sequence of g2-g3 linker.