Objective To investigate the cytolytic activity of extracellular cytolytic toxin rVVC of Vibrio vulnificus on the apoptosis of human ECV304 cells, and to analyze the activities of Caspase-3,-8 and -9. Methods The cytotoxic effect of refolded rVVC on the growth and apoptosis of ECV304 cells was identified by MTT, Hochest33342/PI fluorescent staining, flow cytometry and DNA agarose electrophoresis analysis, respectively. The activities of Caspase-3, -8 and -9 was measured using a colorimetric method. Results The viability of human ECV304 cells exposed to rVVC was inhibited by rVVC after 24 h. 2.0 HU/ml rVVC groups had a better cytotoxic effect to human ECV304 than that of 0.5 HU /ml rVVC groups. The apoptosis of human ECV304 cells in 2.0 HU/ml rVVC+40 μmol/L Z-VAD-FMK groups was relative reduced than that of 2.0 HU/ml of rVVC groups. After 0.5 h treatment with 2.0 HU/ml of rVVC, the Caspase-3 activity in human ECV304 cells increased gradually and reached the peak at 3 h (versus control groups, P<0.01). The activity of Caspase-8 and -9 remained unchanging. Conclusion The rVVC has cytotoxic effect on human ECV304 and the cytolysin is probably correlated with Caspase-3.

Air Pollutants, Occupational ; adverse effects ; analysis ; Dust ; analysis ; Environmental Monitoring ; instrumentation ; methods ; Humans ; Industry ; Inhalation Exposure ; statistics & numerical data ; Maximum Allowable Concentration ; Occupational Exposure ; statistics & numerical data ; Quartz

Stress is a kind of coping mechanism of human and most animals in the face of external survival pressure, which can regulate an individual's physiological, psychological function and behavior. Some studies have shown that there are differences in stress response and cognition of different stress intensity on individuals. In addition, for different individuals, the stress response caused by the same stress source is also varied, which is related to the changes in neurotransmitter and hormone levels, as well as difference in the activation patterns of downstream signa-to individual genetic difference and other factors. In this paper, the cognitive changes from different stress states are summarized from the aspects of neurotransmitters, hormones and their downstream mechanisms, so as to provide ideas for the reasonable application of stress and the prevention and treatment of stress-related diseases.

BACKGROUNDNon-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. Platelet activation may play an important role in pathologic progress in lung cancer. In this study, we aimed to clarify the influence of activated platelets on lung cancer generation and growth, and the relationship among these functional and ultrastructural changes of platelets and the severity of pathogenetic condition in these patients with NSCLC.

METHODSOne hundred and thirty-six cases of patients with pathologically confirmed NSCLC were included in this study. Fifty-four healthy people were enrolled as controls. The change of ultra microstructure and activity of blood platelets were observed under the transmission and scanning electron microscope. Simultaneous determination of plasma granule membrane protein 140 (GMP-140) was made.

RESULTSTransmission electron microscopy showed remarkable changes of ultra microstructure of platelets in patients with NSCLC, including swelling, increase of a-granules, vesicles, and glycogenosome. Scanning electron microscopy showed many more surface processes and wrinkles on platelets in patients with NSCLC. The reference plasma levels of GMP-140 of healthy controls were (18.2 +/- 2.7) microg/L. The plasma levels of GMP-140 in patients with NSCLC were (47.8 +/- 12.3) microg/L, which were much higher than those of the controls. There was a medium positive correlation between plasma levels of GMP-140 and amount of a-granules (r = 0.514, P < 0.01) and a high positive correlation between plasma levels of GMP-140 and area of platelet (r = 0.84, P < 0.01) in patients with NSCLC. The Kaplan-Meier survival curve analysis showed significant shift to the left in patients with NSCLC whose a-granules per platelet were 19 or more compared to those 18 or less (Log rank statistic, chi(2) = 17.38, P < 0.01).

CONCLUSIONSThere are significant activated changes of ultra microstructure and increased activity of blood platelets in patients with NSCLC. These activated platelets may play an important role in the generation and growth of lung cancer. These changes can be used as a diagnostic index of severity, progression, and prognosis of NSCLC.

Adult ; Aged ; Blood Platelets ; ultrastructure ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; mortality ; ultrastructure ; Female ; Humans ; Male ; Microscopy, Electron, Transmission ; Middle Aged ; P-Selectin ; blood ; Survival Analysis

BACKGROUNDAbnormal apoptosis of pulmonary artery smooth muscle cells (PASMCs) is an important pathophysiological process in the pulmonary artery structural remodeling and pulmonary hypertension. We investigated possible effect of endogenous hydrogen sulfide (H2S) on apoptosis of PASMCs during the development of pulmonary hypertension induced by high pulmonary blood flow.

METHODSThirty-nine male Sprague-Dawley rats were randomly assigned to 4-week control, 4-week shunt, 4-week shunt + propargylglycine (PPG), 11-week control, 11-week shunt and 11-week shunt + sodium hydrosulfide (NaHS) groups. Rats in 4-week shunt, 4-week shunt + PPG, 11-week shunt and 11-week shunt + NaHS groups underwent an abdominal aorta-inferior vena cava shunt. Rats in 4-week shunt + PPG group were intraperitoneally injected with PPG, an inhibitor of endogenous H2S production, for 4 weeks. Rats in 11-week shunt + NaHS group were intraperitoneally injected with NaHS, a H2S donor, for 11 weeks. Lung tissue H2S was evaluated by sulfide-sensitive electrode. Apoptosis of PASMCs were detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL). Expressions of Fas, bcl-2 and caspase-3 in the PASMCs were analyzed with immunochemical staining.

RESULTSFour weeks after the shunting operation, the apoptosis of PASMCs and expression of Fas and caspase-3 were significantly decreased (P < 0.01), but expression of bcl-2 increased significantly (P < 0.01). PPG administration further inhibited the apoptosis of PASMCs, downregulated the expression of Fas and caspase-3 (P < 0.01), but increased the expression of bcl-2 (P < 0.01). After 11 weeks of shunting operation, the apoptosis of PASMCs and expression of Fas and caspase-3 were significantly decreased (P < 0.01), but expression of bcl-2 increased obviously (P < 0.01). NaHS administration significantly increased the apoptosis of PASMCs, upregulated the expression of Fas and caspase-3, but inhibited the expression of bcl-2.

CONCLUSIONSH2S induces the apoptosis of PASMCs in the development of high pulmonary blood flow-induced pulmonary hypertension by activating the Fas pathway and inhibiting the bcl-2 pathway.

Alkynes ; pharmacology ; Animals ; Apoptosis ; drug effects ; Blood Flow Velocity ; physiology ; Blotting, Western ; Glycine ; analogs & derivatives ; pharmacology ; Hemodynamics ; drug effects ; Hydrogen Sulfide ; pharmacology ; Hypertension, Pulmonary ; etiology ; physiopathology ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Pulmonary Artery ; cytology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

BACKGROUNDThe mechanism of the neural injury caused by chronic intermittent hypoxia (CIH) that characterizes obstructive sleep apnea syndrome (OSAS) is not clearly known. The purpose of this study was to investigate whether P2X7 receptor (P2X7R) is responsible for the CIH-induced neural injury and the possible pathway it involves.

METHODSEight-week-old male C57BL/6 mice were used. For each exposure time point, eight mice divided in room air (RA) and IH group were assigned to the study of P2X7R expression. Whereas in the 21 days-Brilliant Blue G (BBG, a selective P2X7R antagonist) study, 48 mice were randomly divided into CIH group, BBG-treated CIH group, RA group and BBG-treated RA group. The hippocampus P2X7R expression was determined by Western blotting and real-time polymerase chain reaction (PCR). The spatial learning was analyzed by Morris water maze. The nuclear factor kappa B (NFκB) and NADPH oxidase 2 (NOX2) expressions were analyzed by Western blotting. The expressions of tumor necrosis factor α, interleukin 1β (IL-β), IL-18, and IL-6 were measured by real-time PCR. The malondialdehyde and superoxide dismutase levels were detected by colorimetric method. Cell damage was evaluated by Hematoxylin and Eosin staining and Terminal Transferase dUTP Nick-end Labeling method.

RESULTSThe P2X7R mRNA was elevated and sustained after 3-day IH exposure and the P2X7R protein was elevated and sustained after 7-day IH exposure. In the BBG study, the CIH mice showed severer neuronal cell damage and poorer performance in the behavior test. The increased NFκB and NOX2 expressions along with the inflammation injury and oxidative stress were also observed in the CIH group. BBG alleviated CIH-induced neural injury and consequent functional deficits.

CONCLUSIONSThe P2X7R antagonism attenuates the CIH-induced neuroinflammation, oxidative stress, and spatial deficits, demonstrating that the P2X7R is an important therapeutic target in the cognition deficits accompanied OSAS.

Animals ; Disease Models, Animal ; Hypoxia ; Male ; Metabolic Networks and Pathways ; Mice ; Mice, Inbred C57BL ; Purinergic P2 Receptor Antagonists ; pharmacology ; Receptors, Purinergic P2X7 ; analysis ; physiology ; Rosaniline Dyes ; pharmacology ; Sleep Apnea, Obstructive ; metabolism

Objective To study the pathogenicity of Streptococcus suis type 7 in CD1 mice and to explore the feasibility of CD1 mice as an animal infection model of Streptococcus suis type 7. Methods CD1 mice were infected with Streptococcus suis type 7 isolated from Guangdong Province by intraperitoneal inoculation. The pathological model of CD1 mice was evaluated by the observation of clinical symptoms, pathological autopsy, histopathological examination of liver and brain tissue sections, determination of LD₅₀, and identification of bacteria and detection of bacterial load in tissues and organs. Results The pathological incidence of infected CD1 mice showed good regularity and repeatability. The clinical symptoms, pathological changes and tissue damage of infected CD1 mice were basically consistent with the previous experimental results of Streptococcus suis type 2 and 9 infection in mice and were similar to the natural disease of pigs. Live bacteria could be isolated from blood, heart, liver, spleen, lung, kidney and brain of infected mice, and the recovered bacteria were consistent with the original bacteria. The detection of bacterial load in tissues and organs showed that the bacterial load in heart was the highest, followed by liver, kidney, spleen, lung and blood, and the bacterial load in brain was the lowest. Conclusion In this study, we demonstrate that Streptococcus suis type 7 strain can cause sepsis and brain inflammation in CD1 mice, resulting in tissue damage and death. CD1 mice are sensitive to the experimental strain and can be used as an animal model of streptococcus suis type 7 infection.

Objective: To evaluate the efficiency and safety of low intensity conditional regimen for children with Fanconi anemia (FA) receiving allogenic hematopoietic stem cells transplantation (allo-HSCT). Methods: Four patients diagnosed as Fanconi anemia were enrolled in this study. One patient received HLA-identical sibling donor hematopoietic stem cell transplantation, two patients underwent unrelated donor matched (UD) HSCT, and one patient received unrelated cord blood transplantation. The conditional regimen consisted of Busulfan with low dose of cyclophosphamide. Results: All 4 cases succeeded in allo-HSCT. The median time for neutrophils engraftment was 11(9-15) day, median time to platelets (PLT) engraftment was 12 (8-28) day. One case occurred with grade I of aGVHD, 1 case with hemorrhagic cystitis. No patient happened with hepatic veno-occlusive disease (VOD). Conclusion: Low intensity of conditional regimen is efficient and safe which should be recommended for FA patients with HSCT.
Busulfan ; Fanconi Anemia ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Transplantation Conditioning

Objective To study the tumor inhibitory effects and an in-fluence on immune organs of the water extract from stem of Coix Lachry-ma-jobi L.(WESCLL) in mice with liver cancer H22.Methods The experimental subjects were using mice with liver cancer H 22.The mice are randomly divided into 7 groups and were fed 0.9%sodium chloride , cyclophosphamide (0.02 g? kg -1 ), WESCLL(2,4,6,8,10 g? kg -1 ) by oral gavages.After 9 days′administration, the mice were killed. Their subcutaneous sarcoma ,liver,thymus and spleen were separated and weighted.Then the tumor inhibiting rates and liver , thymus and spleen index were calculated.Results The WESCLL has obvious effects on re-straining the liver cancer H22.There was significant difference compared with model group (P<0.05).The effect of WESCLL is more noticeable on liver index and weigh′s increase of the mice ( P<0.05 ) .There was not significant difference on thymus and spleen indexes compared with model group ( P<0.05 ) .Conclusion WESCLL have distinct effect on anti-tumor.

Background:The effects of keto acid (KA) supplements on Chinese patients receiving maintenance hemodialysis (MHD) are unclear.This study aimed to evaluate the effects of KA supplementation on nutritional status,inflammatory markers,and bioelectric impedance analysis (BIA) parameters in a cohort of Chinese patents with MHD without malnutrition.Methods:This was a prospective,randomized,controlled,single-center clinical study conducted in 2011 till 2014.Twenty-nine patients with MHD were randomly assigned to a control (n =14) or a KA (n =15) group.The control group maintained a dietary protein intake of 0.9 g/kg/day.The KA group received additional KA supplement (0.1 g/kg/day).BIA was used to determine the lean tissue mass,adipose tissue mass,and body cell mass.The patients' nutritional status,dialysis adequacy,and biochemical parameters were assessed at the ends of the third and sixth months with t test or Wilcoxon rank-sum test.Results:The daily total energy intake for both groups was about 28 kcal/kg/day.After 6 months,the Kt/V (where K is the dialyzer clearance of urea,t is the dialysis time,and V is the volume of the distribution of urea) was 1.33 ± 0.25 in KA group,and 1.34 ± 0.25in the control group.The median triceps skin-fold thickness in KA group was 12.00 and 9.00 mm in the control group.In addition,the median hand-grip strength in KA group was 21.10 and 25.65 kg in the control group.There were no significant differences between the groups with respect to the anthropometry parameters,dialysis adequacy,serum calcium and phosphorus levels,inflammatory markers,and amino-acid profiles,or in relation to the parameters determined by BIA.Both groups achieved dialysis adequacy and maintained nutritional status during the study.Conclusions:In this cohort of Chinese patients with MHD,the patients in the control group whose dietary protein intake was 0.9 g/ kg/day and total energy intake was 28 kcal/kg/day,maintained well nutritional status during study period.The KA supplement (0.1 g/kg/day) did not improve the essential amino acid/non-essential amino acid ratio,nor did it change the patients' mineral metabolism,inflammatory parameters,or body compositions.