Pretreatment with anti-flagellin serum delays acute lung injury in rats with sepsis.

OBJECTIVE: This study examined the reduction of sepsis-induced ALI by inhibition of flagellin-stimulated TLR5 signaling. METHODS: Rats were randomly divided into three groups: one group served as the sham-operated group (control group), and the other two groups received the induction of sepsis (sepsis and treatment groups). The treatment group was injected with anti-flagellin serum before induction of sepsis. At 2, 4, 6, 12, 24, and 48 h following induction of sepsis (six time-point subgroups, n = 10 per subgroup), arterial PaO(2), wet/dry (W/D) lung weight ratios, levels of serum and BALF flagellin and TNF-α, pulmonary pathological alterations, and TLR5 mRNA expression in the lungs were examined. RESULTS: Compared to sham-operated rats, septic rats had: increased levels of serum and BALF flagellin at 6, 12, 24, and 48 h; reduced arterial PaO(2); elevated W/D lung weight ratio; increased serum and BALF TNF-α levels; and up-regulated TLR5 mRNA expression at 12, 24, and 48 h (P < 0.01). Pretreatment with anti-flagellin serum, however, significantly inhibited sepsis-associated declines in arterial PaO(2), increased W/D lung weight ratios, elevated serum and BALF TNF-α levels, and up-regulated TLR5 mRNA expression at 24 and 48 h (P < 0.01). CONCLUSION: Neutralizing the actions of circulating flagellin with anti-flagellin serum delayed the development of ALI in rats with sepsis.

[The effect of seawater on expression of protease-activated receptor 2 in adenocarcinoma of lung cell line A549 cells].

OBJECTIVE: To investigate the effect of seawater on expression of protease-activated receptor 2 (PAR-2) in adenocarcinoma of lung cell line A549 cells, and the inflammatory injury on A549 cells induced by seawater. METHODS: A549 cells were randomly divided into four groups: control group, 2 hours group, 4 hours group, 8 hours group, in which cells were treated with seawater for 2, 4 and 8 hours respectively. After seawater treatment, cells were collected for real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis to determine the expression of PAR-2 mRNA and its protein. Supernatant were collected for tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) which were determined by enzyme-linked immunosorbent essay (ELISA). RESULTS: The expression of PAR-2 mRNA and protein of A549 cells increased in a time-dependent manner after seawater treatment, significantly so after 2 hours in all groups (both P<0.05), and peaked at 4 hours after seawater treatment (1.8-fold and 2.2-fold respectively, both P<0.01), followed by a decrease though still higher than those of control group significantly (both P<0.01). TNF-alpha and IL-8 in supernatant increased significantly after seawater treatment, peaking at 2 hours after seawater treatment [(214.35+/-20.85) ng/L, (55.86+/- 5.65) ng/L ] and then followed by a slight decrease though still significantly higher than those of control group [(25.86+/-3.85) ng/L, (6.97+/-1.77) ng/L, all P<0.01]. CONCLUSION: Seawater can induce significant inflammation of A549 cells and up-regulate the expression of PAR-2 on A549 cells.

Effectiveness of varenicline and counselling for smoking cessation in an observational cohort study in China.

OBJECTIVES: To evaluate the effectiveness of varenicline for smoking cessation in Chinese smokers in a real world cessation clinic practice. DESIGN: A prospective observational study. SETTING: Beijing, China. PARTICIPANTS: A total of 924 smokers (883 men and 41 women) who attended a smoking cessation clinic of a large general hospital were assessed with data from structured questionnaires at baseline and follow-up at 1, 3 and 6 months. Trained physician counsellors provided free individual counselling for all subjects and follow-up interviews with brief counselling. 332 subjects additionally prescribed varenicline according to their own choice were compared with those without varenicline. MAIN OUTCOME MEASURES: Primary outcomes were self-reported 7-day point prevalence abstinence rate and 3-month continuous abstinence rate at 6-month follow-up. Secondary outcomes were 7-day point prevalence abstinence rates at 1 and 3-month follow-up, and 1-month continuous abstinence rate at 3-month follow-up. RESULTS: By intention-to-treat, the 7-day point prevalence abstinence rate with varenicline and counselling at 6 months was significantly higher than counselling only (37.0% vs 23.1%; OR, 1.75; 95% CI 1.46 to 2.62; p=0.001). The 3-month continuous abstinence rate at 6 months was higher with varenicline (33.1% vs 18.4%; OR, 2.04; 95% CI 1.61 to 2.99; p<0.001). Varenicline also showed better secondary outcomes. CONCLUSIONS: Varenicline prescription in the smoking cessation clinic appeared to be effective with doubling of quit rates in Chinese smokers in a real world cessation clinic practice. CLINICAL TRIAL REGISTRATION: NCT01935505; Results.

Influence of Per3 genotypes on circadian rhythmicity in flight cadets after militarized management.

OBJECTIVE: The purpose of this study was to explore the effect of PERIOD3 (PER3) genotypes on circadian rhythmicity in flight cadets after militarized management. METHODS: We performed a preliminary study in 146 newly enrolled male flight cadets. Venous blood samples were collected, and genotyping of PER3 (4/5) was determined by using PCR. The morningness-eveningness questionnaire (MEQ) survey was given to flight cadets upon enrollment and after militarized management for 24 months respectively. Comparison of frequency distribution of PER3 genotypes between cases and controls (120 well-matched civilians) was performed using the X(2) test. We also compared the circadian rhythmicity upon enrollment and 24 months after enrollment in flight cadets, and analyzed the connection of changes in circadian clock with PER3 genotypes. RESULTS: The frequency distribution of PER3 genotypes in flight cadets was not significantly different from that in controls subjects. MEQ survey results showed chronotype within flight cadet group varied widely at the two time-points: the moderately morning type (50%) and the neither type (41.1%) upon enrollment; the neither type (76.7%) and the moderately morning type (21.2%) 24 months after enrollment. The circadian rhythm of individuals with the PER3 (5/5) genotype showed no significant difference before and after 24 months of militarized management, whereas notable changes were found in individuals with the PER3 (4/4) genotype (n=116, X(2) =37.26, P < 0.001). CONCLUSION: In conclusion, we provide some evidence that circadian rhythm of flight cadets with the PER3 (5) allele are less likely to be affected compared to those with the PER3 (4) allele.

Changes in hepatic blood flow during transcatheter arterial infusion with heated saline in hepatic VX2 tumor.

PURPOSE: This study evaluates the influence of transcatheter arterial infusion with heated saline on hepatic arterial and portal venous blood flows to tumor and normal hepatic tissues in a rabbit VX2 tumor model. METHODS: All animal experiments were approved by the institutional animal care and use committee. Twenty rabbits with VX2 liver tumors were divided into the following two groups: (a) the treated group (n = 10), which received a 60 mL transarterial injection of 60 °C saline via the hepatic artery; (b) the control group (n = 10), which received a 60 mL injection of 37 °C saline via the hepatic artery. Using ultrasonography, the blood flows in both the portal vein and hepatic artery were measured, and the changes in the hemodynamic indices were recorded before and immediately after the injection. The changes in the tumor and normal liver tissues of the two groups were histopathologically examined by hematoxylin and eosin staining after the injection. RESULTS: After the transcatheter arterial heated infusion, there was a decrease in the hepatic arterial blood flow to the tumor tissue, a significant decrease in the hepatic artery mean velocity (P < 0.05), and a significant increase in the resistance index (P < 0.05). On hematoxylin and eosin staining, there were no obvious signs of tissue destruction in the normal liver tissue or the tumor tissue after heated perfusion, and coagulated blood plasma was observed in the cavities of intratumoral blood vessels in the treated group. CONCLUSIONS: The changes in tumor blood flow in the rabbit VX2 tumor model were presumably caused by microthrombi in the tumor vessels, and the portal vein likely mediated the heat loss in normal liver tissue during the transarterial heated infusion.

The expression of V-ATPase is associated with drug resistance and pathology of non-small-cell lung cancer.

OBJECTIVE: This article aims to investigate the expression of vacuolar-H + -ATPase (V-ATPase) in non-small cell lung cancer (NSCLC) and its variations with pathological type and grade. Furthermore, to evaluate the chemotherapy drug sensitivity of different cancer tissues as well as its correlation with V-ATPase expression in NSCLC. METHODS: V-ATPase expression was examined in 92 NSCLC tissue samples using the immunohistochemical Envision method and immunofluorescence assay. The location of V-ATPase expression was observed by confocal laser scanning microscopy and the difference of its expression rate was evaluated. The sensitivity of cancer tissues to chemotherapy drug was examined using MTT assay and its correlation with the V-ATPase expression was tested in NSCLC by Spearman rank correlation analysis. RESULTS: V-ATPase expression was mainly localized in the cell membrane and cytoplasm. The expression rate of V-ATPase was 71.43% in squamous cell lung cancer, significantly lower than that of the lung adenocarcinoma (83.72%, P = 0.000). In different pathological grades of squamous cell lung cancer, the expression rate of V-ATPase was 58.33% in grade II, significantly lower than that of the grade III (84.00%, P = 0.014). The expression rate of V-ATPase in grade II lung adenocarcinoma was 76.67%, significantly lower than that of the grade ΙΙΙ adenocarcinoma (100.0%, P = 0.012). Correlation analysis showed that the sensitivity of NSCLC tissues to cyclophosphamide, gemcitabine, doxorubicin, paclitaxel and cisplatin was significantly correlated with the V-ATPase expression rate (P < 0.05). CONCLUSIONS: V-ATPase was overexpressed in NSCLC. The expression of V-ATPase was related to the pathological type and grade of cancer and was likely associated with chemotherapy drug resistance in NSCLC. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/7515811511020000.

Hypoxia-inducible factor 1 promoter-induced JAB1 overexpression enhances chemotherapeutic sensitivity of lung cancer cell line A549 in an anoxic environment.

The presence of lung cancer cells in anoxic zones is a key cause od chemotherapeutic resistance. Thus, it is necessary to enhance the sensitivity of such lung cancer cells. However, loss of efficient gene therapeutic targeting and inefficient objective gene expression in the anoxic zone in lung cancer are dilemmas. In the present study, a eukaryotic expression plasmid pUC57-HRE-JAB1 driven by a hypoxia response elements promoter was constructed and introduced into lung cancer cell line A549. The cells were then exposed to a chemotherapeutic drug cis-diamminedichloroplatinum (C-DDP). qRT-PCR and western blotting were used to determine the mRNA and protein level and flow cytometry to examine the cell cycle and apoptosis of A549 transfected pUC57-HRE-JAB1. The results showed that JAB1 gene in the A549 was overexpressed after the transfection, cell proliferation being arrested in G1 phase and the apoptosis ratio significantly increased. Importantly, introduction of pUC57-HRE-JAB1 significantly increased the chemotherapeutic sensitivity of A549 in an anoxic environment. In conclusion, JAB1 overexpression might provide a novel strategy to overcome chemotherapeutic resistance in lung cancer.

Transcatheter arterial infusion with heated saline changes the vascular permeability of rabbit hepatic tumors.

RATIONALE AND OBJECTIVES: The vascular permeability of tumors can be changed by transarterial infusion heat, but the mechanisms remain unknown. The aim of this study was to analyze the underlying causes of changes in tumor vascular permeability after heated perfusion via two different modes. MATERIALS AND METHODS: Thirty rabbits with VX2 hepatic tumors were randomly divided into three groups of 10 rabbits each. The hepatic artery was selectively catheterized via a femoral approach, and unheated saline (control group) or heated saline (60°C) was then injected in either a continuous (transcatheter arterial continuous perfusion [TACP]) or a pulsed (transcatheter arterial pulsed perfusion [TAPP]) manner. Changes in vascular permeability in the tumors were assessed using the following markers and methods: (1) qualitative assessment by visual estimation on digital subtraction angiography performed after the heat infusion procedure on live animals and quantitative assessment by spectrophotometry using Evans blue dye extravasation on tumor and liver tissue after animals were sacrificed and (2) kinase domain receptor or vascular endothelial growth factor (VEGF), expressed in vascular endothelial cells, assessed by immunohistochemical staining, Western blot analysis, and reverse transcription polymerase chain reaction. RESULTS: Tumor staining increased in the TAPP group more than in the TACP group, but not in the control group, assessed on digital subtraction angiography. Extracted dye was higher in tumors in the TAPP group than in those in the TACP group; extracted dye in both groups was higher than in the control group. Kinase domain receptor protein and messenger ribonucleic acid expression were both higher in the TAPP group than in the TACP and control groups. VEGF protein expression was lower in the TAPP and TACP groups than in the control group, but VEGF messenger ribonucleic acid expression was higher in the TACP group than in the TAPP and control groups, and VEGF messenger ribonucleic acid expression was lower in the TAPP group than in the control group. CONCLUSIONS: The vascular permeability of rabbit VX2 tumors significantly increased after arterial pulsed heated infusion, and the protein kinase domain receptor may play a key role in this increase of tumor vascular permeability.

[Study design and the preliminary results on the modes of smoking cessation in general hospitals].

To study the intervention programs on smoking cessation in a general hospital and to evaluate its effects of the programs. Four methods including: a) the intervention through specialists in the smoking cessation clinic, b) short-time intervention in the out-patient department, c) free medical intervention, d) group intervention, were adopted for different smokers, with health counseling, psychological intervention and drug treatment. Intervention effect was evaluated by standard methods. During the 20-month period of the project, we treated 690 cases and 402 completed 6-month follow-up. Preliminary results in 402 cases showed that the three methods of smoking cessation interventions could reduce the amount of cigarette smoking and increase the quitting rate. Motivation to quit smoking, intervention methods and intensity of intervention seemed the main factors. The quit rate of 6-month follow-up in the 'specialist intervention' in the smoking cessation clinic (31.6%) and in the group intervention (30.9%) was higher than short-time intervention in free medical events (15.1%). The successful rate of smoking cessation depended on the motivation of quitters, and the attitude, methods and intervention skills of the physicians. Therefore, it is necessary to explore and develop smoking cessation service models suitable to national context and individual intervention methods in China.