Objective To explore the role of apoptosis of neutrophils and vascular endothelial cells and changes of relative cytokines and thrombotic factors in the pathogenesis of traumatic sepsis at the advanced stage.Methods The venous blood was collected from the patients with traumatic sepsis at advanced stage and traumatic patients without sepsis and healthy subjects.The peripheral hlood mononu- clear cells(PBMC)and neutrophils were isolated and cultured.The apoptosis of neutrophils and vascular endothelial cells was assayed,and the level of IL-4,IL-10 in PBMC culture supernatants were deter- mined,and the tissue factor(TF)and vW factor(vWF)of peripheral plasma were measured.Results The percentages of apoptosis of neutrophils and numbers of apoptotic circulating vascular endothelial cells were higher than that of traumatic patients without sepsis and healthy controls significantly.And the levels of anti-inflammatory cytokines IL-4,IL-10 and thrombotic factors TF,vWF in traumatic sepsis were elevated than that of traumatic patients without sepsis and controls too.Conclusion Immune suppres- sion and abnormal thrombotic state may be one characteristic of traumatic sepsis at advanced stage,which perhaps involveds in the pathogenesis of traumatic sepsis at the advanced stage and multiple organ dys- function syndrome.

Objective:To construct nursing-sensitive indicators system of perioperative pulmonary rehabilitation for patients in thoracicsurgery, so as to provide scientific monitoring standards for nursing quality of pulmonary rehabilitation.Methods:Based on the theory of enhanced recovery after surgery (ERAS) , evidence synthesis was conducted based on Johns Hopkins evidence-based nursing method for laying the foundation for nursing-sensitive quality indicators system of perioperative pulmonary rehabilitation.Based on the analysis results of the quality of nursing in the past two years, and though group discussion, appropriate indicators feasible to application were determined, and nursing-sensitive quality indicators, calculation formulas were preliminarily decided. Through two rounds of experts consultation, the nursing-sensitive quality indicators system of pulmonary rehabilitation was improved and established.Results:After two rounds of expert consultations, the constructed nursing-sensitive quality indicators system of pulmonary rehabilitation included 3 first-class indicators, 6 second-class indicators, and 31 third-class indicators. Positive coefficients of two rounds of expert consultation were both 100%, authority coefficients were 0.827 and 0.861, respectively, and the coordination coefficients were 0.309 and 0.372.Conclusion:The nursing-sensitive quality indicators system was scientific and practical and it was beneficial to regulate the behavior of nurses and improve the nursing quality of pulmonary rehabilitation.

OBJECTIVETo observe the effect of electromagnetic pulse (EMP) exposure on cerebral micro vascular permeability in rats.

METHODSThe whole-body of male Sprague-Dawley rats were exposed or sham exposed to 200 pulses or 400 pulses (1 Hz) of EMP at 200 kV/m. At 0.5, 1, 3, 6, and 12 h after EMP exposure, the permeability of cerebral micro vascular was detected by transmission electron microscopy and immunohistochemistry using lanthanum nitrate and endogenous albumin as vascular tracers, respectively.

RESULTSThe lanthanum nitrate tracer was limited to the micro vascular lumen with no lanthanum nitrate or albumin tracer extravasation in control rat brain. After EMP exposure, the lanthanum nitrate ions reached the tight junction, basal lamina and pericapillary tissue. Similarly, the albumin immunopositive staining was identified in pericapillary tissue. The changes in brain micro vascular permeability were transient, the leakage of micro vascular vessels appeared at 1 h, and reached its peak at 3 h, and nearly recovered at 12 h, after EMP exposure. In addition, the leakage of micro vascular was more obvious after exposure of EMP at 400 pulses than after exposure of EMP at 200 pulses.

CONCLUSIONExposure to 200 and 400 pulses (1 Hz) of EMP at 200 kV/m can increase cerebral micro vascular permeability in rats, which is recoverable.

Animals ; Brain ; blood supply ; Capillary Permeability ; physiology ; Electromagnetic Fields ; adverse effects ; Electrophysiology ; Male ; Rats ; Rats, Sprague-Dawley

The purpose of the present study was to investigate the effects of cornel iridoid glycoside (CIG) on the activity of cholinesterases in vitro, and to investigate the mechanism of CIG's treating Alzheimer's disease (AD). The sources of cholinesterases were prepared from human blood cells, rat brain homogenate and human blood plasma, respectively. The biochemical methods were used to detect the activity of acetylcholine esterase (AChE) and butyryl cholinesterase (BuChE) to investigate the influence of CIG on cholinesterases. The results showed that CIG inhibited the activity of AChE of human blood cells and rat brain homogenate, with the 50% inhibition rate (IC50) of 1.6 g . L-1 and 3.3 g . L-1, respectively; and the inhibition of AChE of CIG is reversible. CIG also inhibited the activity of BuChE of human blood plasma, with the IC50 of 2.9 g . L-1. In conclusion, CIG can inhibit the activity of AChE and BuChE in vitro, which may be one of the mechanisms of CIG to treat AD.
Acetylcholinesterase ; metabolism ; Animals ; Brain ; drug effects ; metabolism ; Cholinesterase Inhibitors ; pharmacology ; Cholinesterases ; metabolism ; Humans ; Iridoid Glycosides ; pharmacology ; Plasma ; enzymology ; Rats

Toxoplasma gondii is an obligate intracellular parasite that stimulates production of high levels of proinflammatory cytokines, which are important for innate immunity. NLRs, i.e., nucleotide-binding oligomerization domain (NOD)-like receptors, play a crucial role as innate immune sensors and form multiprotein complexes called inflammasomes, which mediate caspase-1-dependent processing of pro-IL-1β. To elucidate the role of inflammasome components in T. gondii-infected THP-1 macrophages, we examined inflammasome-related gene expression and mechanisms of inflammasome-regulated cytokine IL-1β secretion. The results revealed a significant upregulation of IL-1β after T. gondii infection. T. gondii infection also upregulated the expression of inflammasome sensors, including NLRP1, NLRP3, NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and NAIP, in a time-dependent manner. The infection also upregulated inflammasome adaptor protein ASC and caspase-1 mRNA levels. From this study, we newly found that T. gondii infection regulates NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and neuronal apoptosis inhibitor protein (NAIP) gene expressions in THP-1 macrophages and that the role of the inflammasome-related genes may be critical for mediating the innate immune responses to T. gondii infection.
Apoptosis ; Cytokines ; Gene Expression ; Immunity, Innate ; Inflammasomes* ; Macrophages* ; Multiprotein Complexes ; Negotiating ; Neurons ; Parasites ; RNA, Messenger ; Toxoplasma* ; Up-Regulation

[Objective]To investigate the effect of KIF23 gene expression on the proliferation,migration and invasion of human hepatocellular carcinoma HepG2 cells in vitro,and to explore the possible mechanism.[Methods]The KIF23 siRNA was transfected into HepG2 cells by lipofectamine 3000.The expression of KIF23 mRNA and protein in HepG2 cells was de-tected by qRT-PCR and Western blot.The effect of silencing KIF23 on the proliferation of HepG2 cells was studied by CCK-8 assay and plate clone formation assay.The tumor cell abilities of migration and invasion after transfection were measured by scratch assay and Transwell assay.The expression of protein kinase B(PKB/Akt)and phosphorylated Akt(p-Akt)protein in HepG2 cells transfected with KIF23-siRNA2 was detected by Western blot.[Results]KIF23-siRNA could effectively si-lence the expression of KIF23 mRNA and protein in HepG2 cells(P<0.01).The results of CCK-8 assay,plate clone forma-tion assay,scratch assay and Transwell assay demonstrated that the cell proliferation,migration and invasion ability of the KIF23-siRNA2 interference group were significantly inhibited,compared to the negative control group and the blank control group(P<0.05).The expression level of total Akt protein in HepG2 cells was not changed,but the expression level of phos-phorylated Akt protein was down-regulated(P<0.05).[Conclusions]KIF23 may promote the proliferation,migration and in-vasion of human hepatocellular carcinoma cells by activating Akt signal transduction pathway.KIF23 is expected to be a new target for gene therapy of hepatocellular carcinoma.

Objective To analyze the immunophenotypic characteristics of the patients with minimal residual disease (MRD) positive acute B lymphocytic leukemia (B-ALL). Methods The leukemia-associated immunophenotype (LAIP) of 106 cases with MRD positive B-ALL from Department of Hematology, Tianjin KingMed Diagnois Center between June 2014 and January 2016 were retrospectively analyzed. CD10, CD13/CD33, CD19, CD38, CD58, CD45 and other antibodies were used to analyze the MRD of B-ALL. Results All the patients were positive for CD19. CD34 was negatively or weakly positive expressed in 27 cases (25.4％). CD10 was negatively or weakly positive expressed in 23 cases (21.7％). CD10 was strongly positive in 24 cases (22.6％). Totally, CD10 was weakly or strongly expressed in 47 cases (44.3％). CD58 was strongly positive in 98 cases (92.5％). CD13/CD33 was positively or weakly positive expressed in 64 cases (60.4％). CD38 was negative or weakly expressed in 33 cases (31.1％). CD45 was negative in 21 cases (19.8％). 15 cases (14.1％) were positive for 6 types of LAIP; 30 (28.3％) cases were positive for 5 types of LAIP; 42 (39.6％) cases were positive for 4 types of LAIP; 13 (12.3％) cases were positive for 3 types of LAIP;5 cases (4.7％) were positive for 2 types of LAIP; only one case (0.9％) was positive for 1 type of LAIP. Conclusion The combination of CD58, CD13/CD33, CD10, CD38 and CD34 antibodies can distinguish the neoplastic blast/immature B lymphocytes from progenitor B cells. This strategy has a high accuracy for the judgement of MRD in B-ALL.

ATG8-binding proteins play a key role in autophagy, selective autophagy or non-autophagy process by interacting between ATG8 and the ATG8-interacting motif (AIM) or the ubiquitin-interacting motif (UIM). There is great progress of ATG8-binding proteins in yeast and mammalian studies. However, the plant domain is still lagging behind. Therefore, the structure characteristics of plant ATG8 binding protein were firstly outlined. Unlike the single copy of ATG8 gene in yeast, many homologous genes have been identified in plant. The LIR/ AIM-docking site (LDS) of ATG8 protein contains W and L pockets and is responsible for binding to AIM. The ATG8 protein binds to UIM-containing proteins via UIM-docking site (UDS) instead of LDS. UDS is in the opposite position to LDS, so the ATG8 can bind both AIM and UIM proteins. Secondly, the structure and function of ATG8-binding proteins, especially the selective autophagy receptors, were systematically described. The protein NBR1 and Joka2, as proteaphagy receptors, guide ubiquitination protein aggregates to autophagosome for degradation by binding to AIM and ATG8 in Arabidopsis and tobacco, respectively. AtNBR1 also promotes plant immunity by binding the capsid protein of cauliflower mosaic virus and silencing suppressor HCpro of turnip mosaic virus, mediating pathogen autophagy. AtNBR1 still degrades chloroplast by microautophagy under photoinjure or chlorophagy during ibiotic stress. And the protein ORM mediates the degradation of plant immune receptor flagellin sensing 2 (FLS2) through AIM binding to ATG8. Interestingly, ATI1 and ATI2 participate in both chlorophagy and ERphagy. Otherwise, ER membrane protein AtSec62, soluble protein AtC53, and ubiquitin-fold modifier1-specific ligase 1 (UFL1) can be directly bound to ATG8 as ER autophagy receptors. As pexophagy receptor, AtPEX6 and AtPEX10 bind to ATG8 via AIM and participate in pexophagy. RPN10, as a 26S proteasome subunit, whose C-terminal UIM1 and UIM2 bind ubiquitin and ATG8, respectively, mediates the selective autophagy degradation of 26S proteasome inactivation when fully ubiquitinated. Plant-specific mitochondrial localization proteins FCS-like zinc finger (FLZ) and friendly (FMT) may also be mitophagy receptors. CLC2 binds to ATG8 via the AIM-LDS docking site and is recruited to autophagy degradation on the Golgi membrane. The tryptophan-rich sensory protein (TSPO) in Arabidopsis was involved in clearing free heme, porphyrin and plasma membrane intrinsic protein 2;7 (PIP2;7) through the combination of AIM and ATG8. The conformation of GSNOR1 changes during anoxia, exposing the interaction between AIM and ATG8, leading to selective degradation of GSNOR1. At last, the ATG8 binding proteins involved in autophagosome closure, transport and synthetic synthesis was summarized. For example, plant-specific FYVE domain protein required for endosomal sorting 1 (FREE1) is involved in the closure of autophagosomes during nutrient deficiency. Therefore, according to the recent research advances, the structure and function of plant ATG8-binding proteins were systematically summarized in this paper, in order to provide new ideas for the study of plant selective autophagy and autophagy.

OBJECTIVETo study the effect of electromagnetic pulse (EMP) on the permeability of blood-brain barrier, tight junction (TJ)-associated protein expression and localization in rats.

METHODS66 male SD rats, weighing (200 approximately 250) g, were sham or whole-body exposed to EMP at 200 kV/m for 200 pulses. The repetition rate was 1 Hz. The permeability of the blood-brain barrier in rats was assessed by albumin immunohistochemistry. The expression of typical tight junction protein ZO-1 and occludin in both cerebral cortex homogenate and cerebral cortex microvessel homogenate was analyzed by the Western blotting and the distribution of ZO-1 and occludin was examined by immunofluorescence microscopy.

RESULTSIn the sham exposure rats, no brain capillaries showed albumin leakage, at 0.5 h after 200 kV/m EMP exposure for 200 pulses; a few brain capillaries with extravasated serum albumin was found, with the time extended, the number of brain capillaries with extravasated serum albumin increased, and reached the peak at 3 h, then began to recover at 6 h. In addition, no change in the distribution of the occludin was found after EMP exposure. Total occludin expression had no significant change compared with the control. However, the expression level of ZO-1 significantly decreased at 1 h and 3 h after EMP exposure in both cerebral cortex homogenate and cerebral cortex microvessel homogenate. Furthermore, immunofluorescence studies also showed alterations in ZO-1 protein localization in cerebral cortex microvessel.

CONCLUSIONThe EMP exposure (200 kV/m, 200 pulses) could increase blood-brain barrier permeability in rat, and this change is associated with specific alterations in tight junction protein ZO-1.

Animals ; Blood-Brain Barrier ; radiation effects ; Brain ; metabolism ; Capillary Permeability ; radiation effects ; Electromagnetic Fields ; adverse effects ; Male ; Membrane Proteins ; metabolism ; Phosphoproteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Zonula Occludens-1 Protein

OBJECTIVETo study the effects of combination of angiopoietin-1 (ANG-1) and vascular endothelial growth factor₁₆₅ (VEGF₁₆₅) gene transfer mediated by recombinant adeno-associated viral vector on the neovascularization in chronic ischemic porcine myocardium.

METHODSAn ameroid constrictor was implanted around the left circumflex coronary artery (LCX) via endoscopy. Six weeks later, coronary angiography revealed that the myocardial ischemia was established by gradual occlusion of the left circumflex coronary artery (LCX). Sixteen swine with the total occlusion or partial stenosis (> 85 %) of the LCX were divided into 4 groups (4 in each group): group I, group II and group IV (control) received direct myocardium injection of rAAV₂ VEGF₁₆₅, rAAV₂ ANG-1 or PBS alone, respectively; group III received rAAV₂ VEGF₁₆₅ and rAAV₂ ANG-1. Selective coronary angiography and ultrasonography were performed perioperatively to evaluate the cardiac function and the formation of collateral circulation. The expression of VEGF₁₆₅ and ANG-1 proteins were assessed using ELISA or Western blot. The degree of angiogenesis was assessed by use of immunohistochemical analysis.

RESULTAngiography showed that the occlusion of all LCX was completed or exceeded 95% 6 weeks after ameroid constrictor implantation, indicating the successful establishment of animal model. The expression levels of VEGF₁₆₅ in group I and III and ANG-1 in groups II and III began to increase at d7 after transfection and reached the peak at d14; then decreased gradually to the normal level after 3 months. The expression levels of VEGF₁₆₅ in group II and group IV or that of ANG-1 protein in group I and group IV had no markedly changes at different time after transfection. There were significant increase in capillary density and arteriole density and more side branch vessels formed in group III compared with other groups. Echocardiographic measurements showed that the left ventricular systolic function of animals in groups I, II and III increased significantly after gene transfection, especially in group III; but there was no changes in group IV.

CONCLUSIONMyocardial perfusion and the left ventricular systolic function are improved after rAAV₂ VEGF₁₆₅ or rAAV₂ ANG-1 transfection, which is associated with the angiogenesis in porcine model of chronic myocardial ischemia.

Adenoviridae ; genetics ; Angiopoietin-1 ; genetics ; Animals ; Collateral Circulation ; Coronary Vessels ; physiopathology ; Disease Models, Animal ; Genetic Therapy ; Genetic Vectors ; Male ; Myocardial Ischemia ; physiopathology ; therapy ; Neovascularization, Physiologic ; Swine ; Swine, Miniature ; Transfection ; Vascular Endothelial Growth Factor A ; genetics