Background:Sepsis-associated acute kidney injury (SA-AKI) is a life-threatening condition with poor prognosis. However, due to significant inter-individual variability in disease progression, clinicians lack reliable tools to stratify patients based on long-term mortality risk. Objective:This study explores how the timing of SA-AKI (early vs. late onset) affects survival, based on the latest ADQI consensus, and aims to build a simple nomogram to predict 1-year mortality. Methods:A retrospective cohort study enrolled 422 SA-AKI patients (from 3273 sepsis patients) admitted to the First Affiliated Hospital of Zhengzhou University between January 2020 and December 2024. Patients were stratified into early-onset SA-AKI(E-SA-AKI, within 48 hours of sepsis diagnosis) and late-onset SA-AKI (L-SA-AKI, 48 hours-7 days post-sepsis) groups, with a maximum 5-year follow-up. Results:The study found that most SA-AKI cases were early-onset (E-SA-AKI vs. L-SA-AKI: 73% vs. 27%). The median age of patients was 65 years, with 72% males. The primary infection sources were abdominal (41.9%) and pulmonary/urinary (36.3%). The cumulative survival rates at 28 days, 90 days, 1 year, and 5 years were 50.5%, 44.8%, 38.9%, and 33.9%, respectively. Both short-term and long-term survival rates were significantly higher in E-SA-AKI patients than L-SA-AKI patients(all P < 0.001). Using LASSO and multivariable Cox regression analyses, a prognostic model was developed, identifying five independent predictors of one-year mortality:L-SA-AKI and septic shock as risk factors, while high albumin level, 28-day renal function recovery, and 48-hour lactate normalization as protective factors. The model's C-statistic (C-index)was 0.79, demonstrating good discrimination and calibration in both the training set [AUC (95% CI): 0.897(0.853-0.931)] and the validation set (AUC(95% CI): 0.909 [0.839-0.951)]. Decision curve analysis indicated favorable clinical utility. Conclusion:The timing of SA-AKI onset is a critical prognostic stratification indicator. A one-year mortality risk nomogram model constructed based on the temporal staging, septic shock, albumin level, 28-day renal function recovery status, and 48-hour lactate normalization provides important reference for clinical decision-making.

The study was to explore the efficacy and safety of sivelestat (SV) in the treatment of severe acute pancreatitis (SAP) with systemic inflammatory response syndrome (SIRS). A total of 102 SAP patients diagnosed and treated in the Emergency Intensive Care Unit of the First Affiliated Hospital of Zhengzhou University from January 2021 to August 2024 were selected. The changes of disease outcome, hospital stays and mortality were compared between the two groups. A total of 102 patients were recruited to control group (n = 56) or SV group (n = 46) according to whether SV was applied or not. There was no significant difference in baseline data at admission between the two groups. After 1 week of treatment, all the indexes in both groups improved. The duration of ventilator use (p = 0.0400) and ICU stays (p = 0.0495) in SV group was shorter than that in control group, but there was no significant difference in mortality between the two groups. Although SV did not reduce the mortality of patients with SAP, it reduced the length of ventilator use and ICU stay.

Antibiotic abuse has led to an increasingly serious risk of antimicrobial resistance, developing alternative antimicrobials to combat this alarming issue is urgently needed. Rhesus theta defensin-1 (RTD-1) is a theta-defensin contributing to broad-spectrum bactericidal activity via the mechanisms of membrane perturbation. Intriguingly, human defensin-6 (HD6), an enteric defensin secreted by Paneth cells without direct bactericidal effect, could self-assembled into fibrous networks to trap enteric pathogens for assistance of innate immunity. The direct bactericidal action of RTD-1 and the bacterial trapping of HD6 inspire a promising antimicrobial paradigm for unique antibacterial strategies. In this study, we utilized the principle of alternating arrangement of D- and L-amino acids in cyclic peptides, which endows them with the potential to self-assemble into nanotubes, mimic the antimicrobial processes of RTD-1 and HD6. We designed and synthesized five cyclic biomimetic peptides (CBPs), among these biomimetics, CBP-4, which possessed a nanotube-like structure, demonstrated the ability to directly and rapidly disrupt the cell membranes of Gram-positive S. aureus and MRSA, while also targeting the surfaces of Gram-negative E. coil using its nanofibrous network to capture bacteria, preventing invasion and migration, and indirectly killing the bacteria. Moreover, CBP-4 eliminated pathogens, inhibited excessive inflammatory responses caused by infections, and maintained immune system homeostasis in septic mice. By fully emulating the antimicrobial mechanisms of both RTD-1 and HD6, CBP-4 showed promising potential for anti-infectious therapies.

重型颅脑损伤是重物撞击或严重的机械作用下导致的脑组织损伤,具有很高的致残率及病死率[1,2],是ICU常见病之一.重型颅脑损伤可使患者脑组织发生严重的病理改变,引起炎症反应,造成脑出血或脑水肿,甚至引起神经功能的缺失,严重影响患者的预后及生活质量.近年来,中药治疗在临床上广泛应用,并逐渐应用于颅脑损伤等脑血管疾病.在常规基础上联合安宫牛黄丸能促进颅脑损伤患者意识恢复,减少高热惊厥等发生率[3].叶恭杰等[4]在颅脑损伤研究中发现安宫牛黄丸能改善患者的炎症指标,提高临床疗效.

Abstract Gram-negative sepsis has become one of major increasing medical burdens globally, which is subjected to growing antibiotic resistance problem and the relatively delayed development of new antibiotics. LL-37, the only type of Cathelicidin identified in humans, has diverse biological activities including direct bactericidal action, regulation of inflammation and LPS-neutralization. The KR-12 peptide is the smallest portion of LL-37 with antibacterial action, which has been shown that could be modified into more effective antimicrobials. Here, we synthesize two myristoylated derivatives of KR-12, Myr-KR-12N and Myr-KR-12C, which can spontaneously form nanoparticles when mixed with deionized water. We show that myristoylated KR-12 derivatives possess a broad-spectrum and more powerful bactericidal activity through interrupting the membranes of bacteria. Myr-KR-12N rescues mice from lethal sepsis induced by E. coli, even more potent rescuing activity than meropenem. We also demonstrate that myristoylated KR-12 nanobiotic can significantly bind with LPS and inhibit the inflammation in vitro and Myr-KR-12N rescue mice from LPS-induced sepsis in vivo, even more potent rescuing activity than polymyxin B. Toxic experiments indicate that neither Myr-KR-12N nor Myr-KR-12C nanobiotics exhibits meaningful hemolytic activity, liver and kidney injury. We thus developed a novel nanobiotic with dual bactericidal and LPS-neutralization properties, which may provide good insights for clinical translation of antimicrobial peptides and the creation of new antibiotics.

Shenfu Injection(SFI) is praised for the high efficacy in the treatment of septic shock. However, the precise role of SFI in the treatment of sepsis-associated lung injury is not fully understood. This study investigated the protective effect of SFI on sepsis-associated lung injury by a clinical trial and an animal experiment focusing on the hypoxia-inducing factor-1α(HIF-1α)-mediated mitochondrial autophagy. For the clinical trial, 70 patients with sepsis-associated lung injury treated in the emergency intensive care unit of the First Affiliated Hospital of Zhengzhou University were included. The levels of interleukin(IL)-6 and tumor necrosis factor(TNF)-α were measured on days 1 and 5 for every patient. Real-time quantitative polymerase chain reaction(RT-qPCR) was performed to determine the mRNA level of hypoxia inducible factor-1α(HIF-1α) in the peripheral blood mononuclear cells(PBMCs). For the animal experiment, 32 SPF-grade male C57BL/6J mice(5-6 weeks old) were randomized into 4 groups: sham group(n=6), SFI+sham group(n=10), SFI+cecal ligation and puncture(CLP) group(n=10), and CLP group(n=6). The body weight, body temperature, wet/dry weight(W/D) ratio of the lung tissue, and the pathological injury score of the lung tissue were recorded for each mouse. RT-qPCR and Western blot were conducted to determine the expression of HIF-1α, mitochondrial DNA(mt-DNA), and autophagy-related proteins in the lung tissue. The results of the clinical trial revealed that the SFI group had lowered levels of inflammatory markers in the blood and alveolar lavage fluid and elevated level of HIF-1α in the PBMCs. The mice in the SFI group showed recovered body temperature and body weight. lowered TNF-α level in the serum, and decreased W/D ratio of the lung tissue. SFI reduced the inflammatory exudation and improved the alveolar integrity in the lung tissue. Moreover, SFI down-regulated the mtDNA expression and up-regulated the protein levels of mitochondrial transcription factor A(mt-TFA), cytochrome c oxidase Ⅳ(COXⅣ), HIF-1α, and autophagy-related proteins in the lung tissue of the model mice. The findings confirmed that SFI could promote mitophagy to improve mitochondrial function by regulating the expression of HIF-1α.

目的 探讨血液灌流联合持续性血液滤过在敌草快中毒中的临床应用效果.方法 选取2020年1月至2021年12月郑州大学第一附属医院急诊医学部收治的84例敌草快中毒患者作为研究对象,根据治疗方式分为对照组和试验组.对照组接受血液灌流治疗,试验组给予血液灌流联合持续性血液滤过治疗,对比两组患者的肝肾功能、炎性细胞因子等,比较两组患者脏器功能障碍发生率、病死率及毒物清除时间的差异.结果 试验组患者第 7天谷丙转氨酶(ALT)、谷草转氨酶(AST)、血肌酐(Cr)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平较对照组患者均明显降低,差异均有统计学意义(P＜0.05);而氧分压(PaO2)较对照组明显上升,差异有统计学意义(P＜0.05).两组患者血尿中毒物清除的时间比较,差异有统计学意义(P＜0.05).预后分析比较,试验组多脏器功能障碍综合征(MODS)的发生率(42.2％)与对照组(69.2％)比较,差异有统计学意义(P＜0.05);两组病死率比较[22(48.9％)vs.24(61.5％)],差异无统计学意义(P＞0.05).结论 早期血液灌流联合持续性血液滤过在一定程度上改善了患者的临床症状,降低了多脏器功能损伤发生率,在临床上值得应用.

BackgroundThe present study aimed to identify the module genes and key gene functions and biological pathways of septic shock (SS) through integrated bioinformatics analysis. MethodsIn the study, we performed batch correction and principal component analysis on 282 SS samples and 79 normal control samples in three datasets, GSE26440, GSE95233 and GSE57065, to obtain a combined corrected gene expression matrix containing 21,654 transcripts. Patients with SS were then divided into three molecular subtypes according to sample subtyping analysis. ResultsBy analyzing the demographic characteristics of the different subtypes, we found no statistically significant differences in gender ratio and age composition among the three groups. Then, three subtypes of differentially expressed genes (DEGs) and specific upregulated DEGs (SDEGs) were identified by differential gene expression analysis. We found 7361 DEGs in the type I group, 5594 DEGs in the type II group, and 7159 DEGs in the type III group. There were 1698 SDEGs in the type I group, 2443 in the type II group, and 1831 in the type III group. In addition, we analyzed the correlation between the expression data of 5972 SDEGs in the three subtypes and the gender and age of 227 patients, constructed a weighted gene co-expression network, and identified 11 gene modules, among which the module with the highest correlation with gender ratio was MEgrey. The modules with the highest correlation with age composition were MEgrey60 and MElightyellow. Then, by analyzing the differences in module genes among different subgroups of SS, we obtained the differential expression of 11 module genes in four groups: type I, type II, type III and the control group. Finally, we analyzed the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment of all module DEGs, and the GO function and KEGG pathway enrichment of different module genes were different. ConclusionsOur findings aim to identify the specific genes and intrinsic molecular functional pathways of SS subtypes, as well as further explore the genetic and molecular pathophysiological mechanisms of SS.

Objective:By detecting the expression of collagen triple helix repeat 1 (CTHRC1) in human venous malformation tissues and normal venous tissues, the relationship between CTHRC1 and the invasion and development of venous malformation was explored.Methods:From December 2020 to December 2021, 35 cases of venous malformation tissues and normal venous tissues (> 3 cm from the edge of venous malformation tissues) in the Department of Hemangioma Surgery, the Third Affiliated Hospital of Zhengzhou University were collected. The expression levels of CTHRC1 mRNA and protein in human venous malformation tissues (experimental group) and normal venous tissues (control group) were detected by immunohistochemical staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) and Western blotting, respectively. The results of immunohistochemistry were tested by χ2 tests, and the results of RT-qPCR and Western blotting were tested by independent sample t test. P<0.05 indicated that the difference was statistically significant. Results:The results of immunohistochemical staining showed that the positive expression rate was 74.29% (26/35) in the experimental group and 22.86% (8/35) in the control group, and the difference was statistically significant ( χ2=18.529, P<0.01). RT-qPCR results showed that the expression of CTHRC1 mRNA in the experimental group was significantly higher than that in the control group, and the difference was statistically significant (2.327±0.472 vs. 0.926±0.174, t=16.460, P<0.01). Western blotting results showed that the expression of CTHRC1 protein in the experimental group was significantly higher than that in the control group, and the difference was statistically significant (1.020±0.078 vs. 0.429±0.029, t=42.151, P<0.01). Conclusion:CTHRC1 mRNA and protein are highly expressed in venous malformation tissues than in normal venous tissues, which may be closely related to the local invasion and development of venous malformation diseases.

Objective:To explore the efficacy and safety of sivelestat, a neutrophil elastase (NE) inhibitor, in the treatment of acute lung injury (ALI) in the intensive care unit (ICU).Methods:A retrospective analysis was performed on 171 patients with ALI in the ICU of the First Affiliated Hospital of Zhengzhou University from June 2020 to June 2021, including 77 patients in the sivelestat group and 94 patients in the conventional treatment group. Acute physiology and chronic health evaluation (APACHE) Ⅱ score, Murray lung injury score, oxygenation index (PaO 2/FiO 2 ratio), inflammatory cytokines (IL-6, IL-10, TNF-α), ventilator-free days (VFD), the length of ICU stay, and the 28-day mortality were collected to assess the efficacy of sivelestat. At the same time, adverse reactions and laboratory test results within 30 days after the use of sivelestat were recorded to assess the safety. Results:Compared with conventional treatment, oxygenation index, Murray lung injury scores, IL-6, IL-10, and TNF-α were significantly improved after 7 days of sivelestat treatment. Compared with the conventional treatment group, the VFD was significantly longer ( P = 0.0119) and the length of ICU stay was significantly shorter ( P = 0.0269) in the sivelestat group. The mortality was 14.29% in the sivelestat group and 22.34% in the conventional treatment group and, with no statistically significant. In the meantime, sivelestat did not increase adverse reactions within 30 days after treatment. Conclusions:Sivelestat treatment is safe and more effective than conventional treatment for ALI patients in the ICU.

Aim To evaluate the roles of plasma soluble cluster of differentiation 14 (sCD14) and sCD14 subtype (sCD14-ST) in the diagnosis of chronic obstructive pulmonary disease (COPD) and in the prediction of an acute exacerbation of COPD (AECOPD). Methods We quantified the levels of white blood cell count (WBC), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), interleukin (IL)-6, IL-8, sCD14, and sCD14-ST in patients with COPD and healthy controls. The relationships between sCD14 or sCD14-ST and inflammatory markers were analyzed in each group. We used receiver operating characteristics (ROC) curves to evaluate the potential roles of sCD14 and sCD14-ST in the diagnosis of COPD and in predicting AECOPD. Results A total of 62 subjects were recruited, including 15 controls and 47 COPD patients, with the latter including 32 stable COPD and 15 AECOPD. WBC, IL-8, sCD14, and sCD14-ST were significantly higher in COPD than in the controls (all P < 0.05). WBC, CRP, ESR, IL-6, IL-8, sCD14, and sCD14-ST were higher in AECOPD than in the controls (all P < 0.05). In the COPD group, sCD14 levels were positively correlated with WBC, IL-8, and sCD14-ST ( P < 0.05), and sCD14-ST levels were positively correlated with WBC and IL-8 ( P < 0.05). In the AECOPD group, sCD14 was positively correlated with WBC, CRP, IL-8, and sCD14-ST ( P < 0.05); sCD14-ST was positively correlated with WBC, IL-6, and IL-8 ( P < 0.05). Discrimination between COPD and controls was tested by calculating areas under the ROC curve (AUCs) for sCD14 and sCD14-ST showing scores of 0.765 (95% CI 0.648–0.883) and 0.735 (95% CI 0.537–0.933) respectively. Similarly, discrimination between AECOPD and controls using sCD14 and sCD14-ST showed scores of 0.862 (95% CI 0.714–1.000) and 0.773 (95% CI 0.587–0.960), respectively. Conclusion Our study suggests that the inflammatory markers sCD14 and sCD14-ST might play an important diagnostic role in COPD and help predict AECOPD.

创伤在我国目前已成为青壮年第一位死亡原因[1].有研究报道,创伤是导致患者发生急性呼吸窘迫综合征(ARDS)的常见原因,根据相关统计结果发现临床常见的ARDS发病诱因中,创伤占7％[2],约50％的创伤患者合并有严重的胸部损伤,导致创伤性ARDS的发生[3];另有报道ICU创伤患者的病死率约为10％,其中ARDS是主要诱因[4].笔者报道1例患者被煤矿车挤压胸腹部约10min,出现创伤性湿肺并ARDS,亦合并腹腔活动性出血并意识障碍、面部青紫、间接性眼部病变,并出现心跳呼吸骤停,经心肺复苏、损害控制性剖腹术及ECMO应用成功挽救了患者生命.

目的 分析双水平气道正压通气(BiPAP)无创呼吸机联合经管道雾化吸入布地奈德对慢性阻塞性肺疾病急性加重(AECOPD)合并Ⅱ型呼吸衰竭患者APACHEⅡ评分及机械通气的影响.方法 将我院收治的56例AECOPD合并Ⅱ型呼吸衰竭患者依据治疗方案分为试验组(n=28,BiPAP无创呼吸机联合经管道雾化吸入布地奈德治疗)和参照组(n=28,BiPAP无创呼吸机治疗).比较两组患者的治疗效果.结果 治疗后,试验组的FEV1、FVC均高于治疗前和参照组,APACHEⅡ评分均显著低于治疗前和参照组(P＜0.05).试验组呼吸困难改善时间、机械通气时间及住院时间均显著短于参照组(P＜0.05).结论 AECOPD合并Ⅱ型呼吸衰竭患者采取BiPAP无创呼吸机联合经管道雾化吸入布地奈德治疗,能改善肺功能及APACHEⅡ评分,加快康复进程.

Melanoma is a kind of tumor that originates from melanocytes and is characterized by chemoresistance and distant metastasis. Although the complete pathogenesis of melanoma remains unclear, increasing evidence suggests that circular RNAs (circRNAs) may be involved. In the present study, we identified a circular RNA, circ_0002770, which is produced from the well-known oncogene MDM2, and was sharply increased in melanoma and correlated with a poor prognosis. Knockdown of circ_0002770 suppressed melanoma cell invasion, migration and proliferation. Mechanistically, circ_0002770 acted as a sponge of miR-331-3p and could indirectly regulate DUSP5 and TGFBR1. Inhibition of miR-331-3p reversed the inhibitory effect of si-circ_0002770 on melanoma cell proliferation and invasion. In vivo evidence further confirmed that silencing circ_0002770 inhibited melanoma tumor formation. In conclusion, circ_0002770 facilitated melanoma cell proliferation, invasion and migration by sponging miR-331-3p and modulating DUSP5 and TGFBR1.

Objective:To explore the effect of Hsp22 on the activation of cardiac fibroblasts stimulated by TGFβ1 and its possible molecular mechanism.Methods:Cardiac fibroblasts of adult mice were isolated and cultured, and stimulated with TGFβ1 to induce fibroblast activation. Fibroblasts were incubated with Hsp22 of different concentrations (1, 2, 4, 8, 10 μg/mL) for 24 h, and their activation, proliferation and secretion were observed. CCK8 kit was used to detect cell proliferation. RT-PCR was used to detect the transcription of fibrogenic factor. Immunofluorescence was used to detect the expression of α-SMA protein. Immunoblotting was used to detect the possible signal protein.Results:CCK8 results showed that fibroblast increased significantly after TGFβ1 stimulation ( P<0.05). The expression of α-SMA in fibroblasts and the transcription of fibrosis-related genes increased significantly after TGFβ1 stimulation ( P<0.05). Different concentrations (1, 2, 4, 8, and 10 μg/mL) of Hsp22 all inhibited the proliferation of fibroblasts significantly (( P<0.05). Eight μg/mL and 10 μg/mL Hsp22 inhibited the expression of α-SMA ( P<0.05). and reduced the transcription of fibrosis-related genes ( P<0.05). Immunoblotting results indicated that after induced by TGFβ1, the expression of WNT and β-catenin, the phosphorylation level of GSK3β, and the nuclear translocation of β-catenin increased ( P<0.05). Ten μg/mL Hsp22 inhibited the expression of WNT and β-catenin, and reduced the phosphorylation of GSK3β the nuclear translocation of β-catenin and the phosphorylation of smad2 and smad3( P<0.05). Conclusions:Hsp22 could block TGFβ1-induced fibroblast activation, proliferation and secretion via inhibiting the WNT/β-catenin signaling pathway.

Objective:To investigate the effect of thymosin α1 on the differentiation of T lymphocyte and the secretion of inflammatory factors in septic mice, thus to explore the effect of thymosin α1 on the prognosis of sepsis.Methods:Adult female C57 mice were randomly (random number) divided into 3 groups: blank control group, sepsis group, and thymosin α1 treatment group. T cell counts and the corresponding inflammatory factors in the further differentiation of T lymphocytes as well as plasma and lung tissues were statistically analyzed, and the survival rate of the mice within 96 h was also analyzed. Graphpad 7.0 software was used for statistically analysis of the study results.Results:There was no significant difference in T cell counts among the three groups of mice, but in the further differentiation of T lymphocytes, the expression of Th17 in the thymosin α1 treatment group was significantly lower than that in the sepsis group, and the expression of Treg was significantly increased in the sepsis group. The expression of the inflammatory cytokine IL-10 was significantly increased in plasma and lung tissues of the thymosin α1 treatment group, while the expression of IL-17A in plasma and lung tissues of the thymosin α1 treatment group was significantly lower ( P <0.05). Survival analysis showed that the survival rate of the thymosin α1 treatment group increased significantly at 96 h, and the difference was significant statistically ( P <0.05). Conclusions:Thymosin α1 can enhance the cellular immunity in sepsis, ameliorate the systemic inflammation, and further protect against sepsis.

Background: Cholangiocarcinoma (CCA) is a serious malignant tumor. Long non-coding RNA NNT-AS1 (NNT-AS1) takes crucial roles in several tumors. So, we planned to research the roles and underlying mechanism of NNT-AS1 in CCA. Results: NNT-AS1 overexpression was appeared in CCA tissues and cell lines. Proliferation was promoted by NNT-AS1 overexpression in CCLP1 and TFK1 cells. Besides, NNT-AS1 overexpression reduced E-cadherin level and raised levels of N-cadherin, vimentin, Snail and Slug. However, the opposite trend was occurred by NNT-AS1 knockdown. Further, NNT-AS1 overexpression promoted phosphatidylinositol 3 kinase (PI3K)/AKT and extracellular signal-regulated kinase (ERK)1/2 pathways. MiR-203 was sponged by NNT-AS1 and miR-203 mimic reversed the above promoting effects of NNT-AS1. Additionally, insulin-like growth factor type 1 receptor (IGF1R) and zinc finger E-box binding homeobox 1 (ZEB1) were two potential targets of miR-203. Conclusion: NNT-AS1 promoted proliferation, EMT and PI3K/AKT and ERK1/2 pathways in CCLP1 and TFK1 cells through down-regulating miR-203. Methods: CCLP1 and TFK1 cells were co-transfected with pcDNA-NNT-AS1 and miR-203 mimic. Bromodeoxyuridine (BrdU), flow cytometry, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot were employed to detect roles and mechanism of NNT-AS1. Interaction between NNT-AS1 and miR-203 or miR-203 and target genes was examined through luciferase activity experiment.

Background: Polymyxin B is used as the last treatment resort for multidrug-resistant gram-negative bacterial infections. This study aimed to develop and validate a simple and robust liquid chromatography with tandem mass spectrometry analytical method for therapeutic drug monitoring of plasma and cerebrospinal fluid (CSF) polymyxin B1 and B2. Methods: Plasma and CSF polymyxin B1 and B2 were chromatographically separated on a Thermo Hypersil GOLD aQ C18 column and detected using electrospray ionization(+)mode coupled with multiple reaction monitoring. Blood and CSF samples for pharmacokinetic analysis were collected from 15 polymyxin B-treated patients. Results: The calibration curve showed acceptable linearity over 0.2-10 mcg/mL for polymyxin B1 and 0.05-2.5 mcg/mL for B2 in the plasma and CSF, respectively. After validation, according to the Food and Drug Administration (FDA) method validation guideline, this method was applied for polymyxin B1 and B2 quantification in over 100 samples in a clinical study. Conclusions: A simple and robust method to measure polymyxin B1 and B2 in human CSF was first exploited and validated with good sensitivity and specificity, and successfully applied in polymyxin B pharmacokinetic analysis and therapeutic monitoring in Chinese patients.

MiR-367 was reported to regulate inflammatory response of microglia. CCAAT/enhancer-binding protein α (C/EBPA) could mediate microglia polarization. In this study, we explored the possible roles of miR-367 and CEBPA in intracerebral hemorrhage (ICH). ICH and normal specimens were obtained from the tissue adjacent to and distant from hematoma of ICH patients, respectively. Microglia were isolated and identified by immunofluorescence. The isolated microglia were treated with erythrocyte lysate and randomly divided into 8 groups using different transfection reagents. The transfection efficiency of miR-367 was determined by qRT-PCR. The expressions of M1 and M2 microglia markers were detected by Western blotting. The relationship between CEBPA and miR-367 was confirmed by dual luciferase reporter system. Flow cytometry was performed to determine the level of apoptosis in the cells transfected with miR-367 and CEBPA in erythrocyte lysate–treated microglia. We found that miR-367 expression level was downregulated in ICH specimens. Erythrocyte lysate–treated microglia was successfully established using erythrocyte lysate, as decreased miR-367 expression was observed. Overexpression of miR-367 could significantly decrease the expressions of MHC-ІІ, IL-1β, and Bax, reduced apoptosis rate, and increased the expressions of CD206, Bal-2, and Arg-1 in erythrocyte lysate–treated microglia. CEBPA was proved to be a direct target for miR-367, which could inhibit microglia M2 polarization and increase apoptosis rate. However, in the presence of both CEBPA and miR-367 mimic, the protein and mRNA expressions of CEBPA were decreased, leading to promoted microglia M2 polarization and a decreased apoptosis rate. MiR-367 regulates microglia polarization by targeting CEBPA and is expected to alleviate ICH-induced inflammatory injury.