Klebsiella pneumoniae is a major pathogen responsible for severe pulmonary infections, yet the early mechanisms of infection remain incompletely understood. This study investigates the role of exosomes derived from K. pneumoniae in polarizing macrophages to the M1 phenotype, thereby facilitating early lung infections. Utilizing single-cell Raman spectroscopy, we rapidly detected K. pneumoniae within host cells and observed significant lipid expression changes. Metabolomic analysis of exosomes from infected epithelial cells uncovered an elevation of phosphatidylcholine, which disrupted endothelial tight junctions and promoted M1 macrophage recruitment and polarization. This process activated the NF-κB signaling pathway, increasing inflammatory responses and attracting neutrophils. Our findings, validated in infected tissue models, suggest that these exosomal mechanisms significantly contribute to the early stages of pulmonary infection by K. pneumoniae. This study offers crucial insights into potential therapeutic targets for controlling K. pneumoniae infections.

Exosomes are released from a variety of immune cells and nonimmune cells, the phospholipid vesicle bilayer membrane structure actively secreted into tissues. Recently, exosomes were demonstrated to be effectively delivered proteins, cholesterol, lipids, and amounts of DNA, mRNA, and noncoding RNAs to a target cell or tissue from a host cell. These can be detected in blood, urine, exhaled breath condensates, bronchoalveolar lavage fluid (BALF), ascites, and cerebrospinal fluid. BALF is a clinical examination method for obtaining alveolar cells and biochemical components, reflecting changes in the lungs, so it is also called liquid biopsy. Exosomes from BALF become a new method for intercellular communication and well‐documented in various pulmonary diseases. In chronic obstructive pulmonary disease (COPD), BALF exosomes can predict the degree of COPD damage and serve as an effective monitoring indicator for airflow limitation and airway remodeling. It also mediates antigen presentation in the airways to the adaptive immune system as well as costimulatory effects. Furthermore, BALF exosomes from acute lung injury and infective diseases are closely related to various infections and lack of oxygen status. BALF exosomes play an important role in the diagnosis and prognosis of lung cancer. The effect of immunomodulatory role for BALF exosomes in adaptive and innate immune responses has been studied in sarcoidosis. The intercellular communication in the microenvironment of BALF exosomes in pulmonary fibrosis and lung remodeling have been studied. In this review, we summarize the novel findings of exosomes in BALF, executed function by protein, miRNA, DNA cytokine, and so on in several pulmonary diseases.

Recently, there is increasing evidence that microRNAs are related to the development, diagnosis, treatment, and prognosis of glioblastoma. microRNA-210 (miR-210) had been identified in many human cancers, but the specific function of miR-210 remains unclear in glioblastoma. The present study mainly focused on exploring its biological role and potential molecular mechanisms in glioblastoma. We found that miR-210 expression was decreased in glioblastoma, and downregulation of miR-210 was related to worse prognosis in glioblastoma patients. In addition, miR-210 overexpression inhibited the migration and invasion of human glioblastoma cells. At the same time, we found that miR-210 directly targets the brain-derived neurotrophic factor (BDNF) and reduces BDNF expression level. Consistently, BDNF silencing had the same effects as miR-210 overexpression in glioblastoma, and upregulation of BDNF counteracted the inhibitory effect of miR-210 in glioblastoma. In conclusion, miR-210 suppressed the migration and invasion of glioblastoma cells by targeting BDNF.

Sepsis is a systemic inflammatory response syndrome, which is mostly induced by infection in the lungs, the abdomen and the urinary tract. The present study is aimed to investigate the mechanisms of sepsis. Expression profile of E‑MTAB‑4421 (including leukocytes isolated from 207 survived and 58 non‑survived patients with sepsis) and E‑MTAB‑4451 (including leukocytes isolated from 56 survived and 50 non‑survived patients with sepsis) were downloaded from the European Bioinformatics Institute database. Based on the E‑MTAB‑4421 expression profile, several differentially expressed genes (DEGs) were identified and performed with hierarchical clustering analysis by the limma and pheatmap packages in R. Using the BioGRID database and Cytoscape software, a protein‑protein interaction (PPI) network was constructed for the DEGs. Furthermore, module division and module annotation separately were conducted by the Mcode and BiNGO plugins in Cytoscape software. Additionally, the support vector machine (SVM) classifier was constructed by the SVM function of e1071 package in R, and then verified using the dataset of E‑MTAB‑4451. A total of 384 DEGs were screened in the survival group. The PPI network was divided into 4 modules (modules A, B, C and D) involving 11 DEGs including microtubule‑associated protein 1 light chain 3 alpha (MAP1LC3A), protein kinase C‑alpha (PRKCA), metastasis associated 1 family member 3 (MTA3), and scribbled planar cell polarity protein (SCRIB). SCRIB and PRKCA in module B, as well as MAP1LC3A and MTA3 in module D, might function in sepsis through PPIs. Functional enrichment demonstrated that MAP1LC3A in module D was enriched in autophagy vacuole assembly. Finally, the SVM classifier could correctly and effectively identify the samples in E‑MTAB‑4451. In conclusion, DEGs such as MAP1LC3A, PRKCA, MTA3 and SCRIB may be implicated in the progression of sepsis, and need further and more thorough confirmation.

Purpose: This study investigated the frequency and absolute numbers of different subsets of peripheral blood cells, including CD4(+) T cells and CD19(+) interleukin (IL) 10(+) B regulatory cells (Breg) and their potential association with clinical laboratory measurements in children diagnosed with simple asthma or asthma plus allergic rhinitis (AR). Method: The frequency and numbers of peripheral blood CD4(+) interferon (IFN) gamma(+) T-helper (Th) type 1, CD4(+) IL-4(+) Th2, CD4(+) IL-17A(+) Th17, CD4(+) CD25(+) Forkhead Box P3(+) regulatory T cells (Treg) and CD19(+) IL-10(+) Bregs in 22 children with asthma, 17 children with asthma and AR, and 25 healthy controls were determined by flow cytometry. The levels of serum cytokines were determined by enzyme-linked immunosorbent assay. Results: In comparison with healthy controls, patients with asthma alone had significantly increased numbers of Th1, Th2, and Th17 cells, and their cytokines but decreased numbers of Tregs and Bregs, and the cytokines IL-10 and transforming growth factor beta 1. This imbalance between effector and regulatory cells and their cytokines further increased in patients with asthma and AR. The ratios of percentage effector T cells (Th1 + Th2 + Th17) to regulatory cells (Treg + Breg) were positively correlated with fractional exhaled nitric oxide but negatively correlated with forced expiratory volume in 1 second in patients with asthma or asthma plus AR. Conclusion: The imbalance of effector T cells and regulatory cells contributed to the pathogenesis of airway inflammation of asthma and AR in children.

Objective To explore the expression of CD1a and CD83 marked tumor infiltrating dendritic cells(TIDC) in breast tissues and its correlation with prognosis. Methods Eighty?five cases of breast cancer tissues were collected and CD1a, CD83 expressions of breast cancer tissues were detected by immunochemical method. The relationship between their expression and clinico?pathological features and prognosis were analyzed. Results There were 77 cases with CD1a positive expression in 85 cases of breast cancer tissues, and most TIDC expressing CD1a were distributed in tumor tissues, and less expressed in tumor stroma and surrounding tissues;there were 44 cases with CD83 positive expression in 85 cases of breast cancer tissues, and TIDC expressing CD83 were ob?served in surrounding and interstitial tissues of tumors, with scattering distribution. There were 35 cases with CD1a and CD83 double positive expression. CD1a, CD83 double positive expression had no correlation with lymph node metastasis, progestrogen receptor( PR) and neutrophil?lymphocyte ratio(NLR)(P>0?05), but correlated with estrogen receptor(ER), HER?2 and TNM staging(P<0?05). The median disease?free survival( DFS) of patients with both CD1a, CD83 double positive expression and non?double positive expres?sion were not reached. Multivariate analysis indicated that non?double positive expression of CD1a, CD83 was an independent factor af?fecting prognosis of patients (HR=2?216,95%CI:1?682?3?431;P<0?05). Conclusion The expression of characteristic markers CD83 and CD1a of TIDC in breast cancer tissues are related with prognosis, and may be associated with immune escape mechanism of tumors, so they can be the reference indicators to detect the prognosis of breast cancer.

A sensitive LC-MS/MS method for the determination of bruceine D in rat plasma was developed. The analyte and IS were separated on a Luna C18 column (2.1 × 50 mm, 1.7 μm) using a mobile phase of acetonitrile and 0.1% formic acid in water (40:60, v/v) at a flow rate of 0.25 mL/min. The selected reaction monitoring mode was chosen to monitor the precursor-to-product ion transitions of m/z 409.2 → 373.2 for bruceine D and m/z 469.2 → 229.3 for IS using a negative ESI mode. The method was validated over a concentration range of 0.5-2000 ng/mL for bruceine D. Total chromatography time for each run was 3.5 min. The method was successfully applied to a pharmacokinetic study of bruceine D in rats. Copyright © 2016 John Wiley & Sons, Ltd.

Objective: To build oxygen glucose deprivation/reperfusion(OGD/R) model in vitro to mimic ischemia and reperfusion inducing apoptosis of SH-SY5Y cells, to detect differential proteins by employing proteomics technique and to reveal the potential mechanisms of neuronal apoptosis. Methods: SH-SY5Y cells were employed and divided into control group and experimental group in which the cells treated with OGD10h/R24h to set up apoptosis model. After the proteins were extracted from SH-SY5Y cells in control and OGD10h/R24h groups, 2D-DIGE techniques was used to detect differential proteins. Results: By DeCyder software mean (1 800 ± 156) protein spots were displayed on 2D-DIGE image. In OGD10h/R24h group, 30 protein spots were detected to differentially express compared with control group, of which 22 spots were up-regulated and 8 spots were downregulated. Compared with control groups at the same time, the differences were significant (P<0.05 or P<0.01). Conclusion: By employing 2D-DIGE proteomics techniques to analyse OGD/R model, the protein spots are detected to differentially express.

Objective To explore time window of inducing neuronal apoptosis successfully via building and evaluating oxygen glucose deprivation/reperfusion(OGD/R) model in vitro to mimic ischemia and reperfusion inducing apoptosis of SH-SY5Y cells,and to build a basis for revealing the potential mechanisms of neuronal apoptosis.Methods SH-SY5Y cells were employed and divided into control group and experimental group treated with OGD/R for different time.The cell viability was calculated by the method of MTT method.The OGD10h/R groups in which the cell viability was decreased approximately 50% 24 h after reperfusion and control groups were selected for further assay.With light microscope,fluorescence microscope and transmission electron microscope the variation of structure and morphous in cells at different time were observed.The changes in lactate dehydrogenase(LDH) levels,malondialdehyde(MDA) contents,superoxide dismutase(SOD) activities were detected at different time of reperfusion.Flow cytometry was used to analyze the cell cycle distribution and the apoptotic rates at different time of reperfusion.Results The MTT results showed that the cell viabilities were decreased significantly with the prolongation of OGD time.At reperfusion 24 h,the cell viability was(44.91±1.21)% in OGD10h/R groups.The cell viabilities in other different OGD groups had significant differences(P0.05 or P0.01).In OGD10h/R groups the inverted microscope and HE staining results showed that the cell densities were decreased following varied shape and deep blue pycnotic nuclei.The AO/EB fluorescence staining results showed the apoptotic body emerged.Under electron microscope,the cells showed heterochromatin fringe agglutinated in nucleus which were viewed as the appearance of apoptosis and autophagia.The level of LDH in culture fluid and the contents of intracytoplasm MDA at different reperfusion time were increased with the prolonged time and at reperfusion 24 h respectively reached the peak and platform phase,compared with control groups the differences were significant(P0.01).The intracytoplasm SOD activities at different reperfusion time were declined at first and recovered later with the prolonged reperfusion time,compared with control groups the differences were significant(P0.01).By flow cytometry analysis on cell cycle phase at different reperfusion time,in OGD 10h/R groups with the prolonged reperfusion time,the percentage of the cells in G0/G1 phase was increased at first and was decreased later with its peak of(74.09±2.62)% at reperfusion 24 h.The percentage of the cells in G2/M phase reached its peak of(26.85±1.35)% at R 0 h and was declined gradually later.The percentage of the cells in S phase was declined gradually and reached the lowest level of(19.2±1.58)% at reperfusion 24 h.Compared with control groups at the same time,the differences were significant(P0.05 or P0.01).Conclusion OGD10h and reperfusion(OGD10h/R) to mimic cerebral ischemia and reperfusion in vitro is a reliable model that induces SH-SY5Y cells apoptosis,which provide a basis for the futher study on neuronal apoptotic mechanisms.

脑血管病是导致人类死亡的三大病因之一,而其中缺血性脑血管病约占75%～80%,其发病机制涉及多方面复杂因素;蛋白质组学作为新兴学科从整体角度,系统地研究动态变化的蛋白质组成分构成及其活动规律,为探索该病机制开辟了新途径.本文旨在综述蛋白质组学概念和主要研究技术,以及脑缺血个体体液、脑缺血个体脑组织、体外模拟缺血的细胞模型三方面的脑缺血性疾病蛋白质组学研究现状,为进一步开展该病蛋白质水平研究奠定基础.

Objective To explore diagnosis feature of the lateral intraventricular meningiomas and microsurgical approaches for removing it.Method There were 24 patients with 24 tumors,of which,13 tumors on the left trigon,7 tumors on the right trigon,1 tumor on the right body,and 1 tumor strideon both lateral trigons and 2 tumors on the left temporal horn.All the 24 cases were diagnosed by MRI and/or CT and removed by microsurgical operation,of which,2 cases by neuroendoscopy-assisted microsurgical operation,15 cases by paramedian parieto-occipital approach,7 cases by middle temporal gyrus approach and 2 cases by transcallosal approach.Results All the 24 cases were diagnosed in coincidence between imaging and pathology and removed totally,of which,5 cases were removed wholly,19 cases were removed by piecemeal.18 cases were followed-up,10 cases were normal,3 cases were relapsed and removed totally again,2 cases and had 3 cases epilepsy,and had homonymous hemianopia.Conclusion CT and MRI are the most reliable diagnosis measure and microsurgical operation is the best treatment method for lateral intraventricular meningiomas.

我院采用YL-1型颅内血肿粉碎穿刺针[1]治疗多部位老年高血压中等量脑出血取得了良好的疗效.本文对出血量、出血部位以及手术时机的选择对治疗效果的影响做进一步探讨.

OBJECTIVE To investigate the activities' alteration of the proteasome in neurons of cortex and its relation with delayed neuron death after reperfusion following ischemia. METHODS 20 minutes transient global ischemia rat model was used. Following different reperfusion period, all the 50 rats were divided into 5 groups, sham-operation group, 0.5 hour recovery group, 4 hours recovery group, 24 hours recovery group and 72 hours recovery group, 10 rats each group. Suc-llvy-amc was used as substrate for measuring proteasome's activities. Delayed neuron death after reperfusion following ischemia was observed under light microscope by HE staining. Proteasome's distribution was observed under laser scanning confocal microscope after immuno-histo-chemical staining. RESULTS The proteasome activity of sham group was 54 602 +/- 1602, and that of 0.5 h reperfusion following ischemia was 42,036 +/- 1465 (compared with sham group, P < 0.01). Although the proteasome activity temporarily recovered to 47,536 +/- 2532 (P < 0.05) after 4 h reperfusion, it still decreased to 45,450 +/- 649 (P < 0.01) after 24 h reperfusion, and to 43,108 +/- 995 (P < 0.01) after 72 h reperfusion. HE staining showed that parts of neurons in the cortex died after 72 hours reperfusion. Under laser scanning microscope, we could observe that after 24 hours reperfusion, proteasome in both nucleus and cytoplasma significantly decreased; after 72 hours reperfusion, proteasome almost disappeared totally in nucleus and only a small part of proteasome still existed in cytoplasm. CONCLUSIONS Decreasing of the proteasome's activities is an important factor for delayed neuron death after reperfusion following ischemia.

重度原发性脑室出血以中老年人高发,病情多呈快速进展性恶化,既往内科保守治疗和外科常规手术死亡率高,属于比较棘手的一类脑室出血[1].神经内窥镜锁孔手术具有创伤小、疗效确切、恢复快的优点,主要用于治疗脑室内疾病.本文旨在探讨中老年重度原发性脑室出血的特点以及神经内窥镜锁孔技术在其治疗中的应用.