Objective:To observe whether nitric oxide (NO) could enhance the damaging effect of cyclophosphamide on L1210 cells cultured in vitro, and to investigate the mechanism of this action. Methods:L1210 cells were co-cultured with 3T3 cells in DMEM medium supplemented with cyclophosphamide (400 ?g/ml). The L1210 cells were divided into three groups based on different transfected 3T3 cells: pcDNA3.0-iNOS plasmid transfected 3T3 cells (Group 1), pcDNA3.0 plasmid transfected 3T3 cells (Group 2), pcDNA3.0-iNOS plasmid transfected 3T3 cells plus DEVD-CHO(Group 3). The viability and apoptosis rate of L1210 cells at different culture periods were determined by trypan blue exclusion and TUNNEL method, respectively. And the cell cycles at G_1 and S phase were detected by flow cytometry. Results:①After cyclophosphamide treatment, the viability of L1210 cells was significantly lower in Group 1 than that in Group 2 during 12-72 h (P

AIM: To explore the immunomodulatory effect of paecilomyces cicadidae polysaccharides (PCPS).METHODS: Subcutaneous injection with 50, l00, 200 mg/kg of PCPS were given in the back of the rats everyday for l5 days. The number of white blood cells (WBC) was counted. The activities of acid phosphatase (ACP) and lactase dehydrogenase (LDH)in liver, kidney, spleen and thymus were detected by automatic biochemistry analyzer. The ability of devouring neutral red and activity of ACP, LDH, arginase in alveolar macrophages were also detected. The body weight of the rat everyday during experiment and weight of the spleen and thymus after the rats were killed were measured and wet weight index was calculated.RESULTS: The wet weight index of spleen and thymus, the activity of ACP, LDH, arginase and ability of devouring neutral red in alveolar macrophages in the test group treated with PCPS were significantly higher than those in control group. CONCLUSION: PCPS shows a significant immunomodulatory effect with the increasing counts of WBC and activation of alveolar macrophages in a dose-dependent manner.

BACKGROUNDAlpha 2A adrenergic receptor (AR) is a subtype of α2 AR belonging to G protein-coupled receptors, and exerts a variety of biological effects. Recent studies have demonstrated that the α2A AR activation was closely related with inflammatory reaction. The present study aimed to investigate the influence of α2A AR antagonist, yohimbine, on the severity of endotoxin-induced acute lung injury in rats.

METHODSA total of 72 male Sprague-Dawley rats were randomly divided into three groups: control group, lipopolysaccharide (LPS) group and LPS + yohimbine group. Rats were intratracheally administrated with normal saline or LPS (300 µg), and the rats in the LPS + yohimbine group were treated with additional yohimbine (2 mg/kg, i.p) soon after LPS administration. Six, 24 and 48 hours after treatment, arterial blood gas analysis was carried out, and optical microscopy was performed to evaluate pathological changes in the lung, and lung injury score was assessed. The count of white blood cells in bronchoalveolar lavage fluid (BALF) was determined. The levels of norepinephrine, tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in BALF were measured with enzyme-linked immunosorbent assay. Immunocytochemistry was performed for the detection of α2A AR on inflammatory cells in BALF.

RESULTSWhen compared with the control group, the oxygenation index in the LPS group was significantly decreased, and white blood cell count, the lung histopathological scores, levels of norepinephrine and IL-6 as well as α2A AR expression on inflammatory cells in the BALF were dramatically increased at different time points, and the concentrations of TNF-α and IL-1β were also increased except at 48 hours after LPS administration. The oxygenation index decreased while white blood cell count in BALF and the lung histopathological scores were obviously increased in the LPS + yohimbine group. The level of norepinephrine in BALF was increased at each time interval in the LPS + yohimbine group, and so did the levels of TNF-α, IL-1β and IL-6 at 6 and 48 hours after LPS administration respectively. When compared with the LPS group, the oxygenation index, white blood cell count, the lung histopathological scores and the level of IL-6 in the LPS + yohimbine group were significantly improved at each time interval, and the concentrations of TNF-α and IL-1β were also lower at 24 hours of LPS administration (all P < 0.05). Correlation analysis indicated the level of norepinephrine was related to the levels of TNF-α, IL-1β and IL-6 in the BALF and the lung histopathological scores (r = 0.703, r = 0.595, r = 0.487 and r = 0.688, respectively, P < 0.001) and the intensity scores of immunoreactivity to α2A AR on inflammatory cells were also associated with the levels of TNF-α, IL-1β and IL-6 as well as the lung histopathologial scores (r = 0.803, r = 0.978, r = 0.716 and r = 0.808, respectively, P < 0.001).

CONCLUSIONSYohimbine can inhibit TNF-α, IL-1β and IL-6 overproduction and relieve the severity of pulmonary inflammation induced by endotoxin, which is maybe mediated by blockade of α2A AR on inflammatory cells.

Acute Lung Injury ; chemically induced ; drug therapy ; Adrenergic alpha-2 Receptor Antagonists ; therapeutic use ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Immunohistochemistry ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; toxicity ; Male ; Norepinephrine ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, alpha-2 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Yohimbine ; therapeutic use

Objective To observe the hemodynamic changes of acute after cardiac resynchronization therapy(CRT) interruption in patients with chronic heart failure (CHF),and explore the benefit of hemodynamic of the reverse remodeling and non-reverse remodeling CHF patients during CRT.Methods 46 CHF patients who had undergone implantation of CRT device for more than 6 months were enrolled in the study.The reverse remodeling and non-reverse remodeling were identified by reduction of LV end-systolic volumes ≥ 15％.The two groups underwent echocardiography,and mitral regurgitation area (MRA),velocity time integral of aortic valve (VTI-AV),velocity time integral of mitral valve(VTI-MV),maximum rising rate of left intra-ventricular pressure(+ dp/dtmax),LVFT/T were measured in on and off mode of CRT.Hemodynamic indicators of two groups were compared in CRT on and off modes.The rate of changes of above mentioned parameters ΔMRA,ΔVTI-AV,ΔVTI-MV,Δdp/dt and ΔLVFT/T were calculated and compared in the two groups.Results MRA had significant increasing in both group after 10 minutes intereuption of CRT (P ＜0.01).Two groups had significant worsening of VTI-AV,VTI-AV,+ dp/dtmax and LVFT/T after 10 minutes interruption of CRT as compared to CRT-on mode (P ＜0.05).ΔMRA,ΔVTI-MV,Δdp/dt of reverse remodeling group significantly higher than non-reverse remodeling group(P ＜0.05).But there was no difference in ΔVTI-AV and ΔLVFT/T(P ＞0.05).Conclusions The hemodynamic index become worsening after 10 minutes interruption of CRT in both group of medium-and long-term CRT patients especially in reverse remodeling group,suggesting that patients of both groups have obtained hemodynamic benefits continuously during CRT.Reverse remodeling group get more benefits than non reverse remodeling group.

Objective To investigate the relationship between wound age and the expressions of frizzled-2 (Fzd2) mRNA and its protein in rats skeletal muscle after contusion,and to explore its possibility of being an index for wound age estimation.Methods The mRNA and protein expressions of Fzd2 in rats skeletal muscle of the control group and the experimental group within 4-48 h after contusion were detected per 4 h by RT-qPCR and Western blotting.Results The relative expression of Fzd2 mRNA increased at 24h,36h and 40h after contusion,and the expression at 24h was twice as the control group (P＜0.05).The relative expression of Fzd2 protein changed inconspicuously after contusion (P＞0.05).Conclusion The changes of Fzd2 mRNA expression after contusion in a certain time can be a basis to estimate wound age by combined multi-indicators.

OBJECTIVES: To explore the homogeneity level of four different functional mRNA （PUM2, TAB2, Cx45 and CHRNA1） expressions in rats with skeletal muscle contusion. METHODS: The relative expressions of PUM2, TAB2, Cx45 and CHRNA1 mRNAs were detected by quantitative real-time reverse transcription-polymerase chain reaction （RT-qPCR）. The coefficient of variation （CV） of the relative expressions for different individuals in each injury group was calculated. The extreme value of CV, cumulative variability, and CV_CV were compared. RESULTS: A high CV of PUM2 and TAB2 mRNAs appeared on several different time points. However, the CV of Cx45 and CHRNA1 mRNAs was relatively low. The cumulative variability from high to low was PUM2, CHRNA1, TAB2 and Cx45 mRNAs. The relative expression of PUM2 mRNA was significantly higher than that of TAB2, Cx45 and CHRNA1 mRNAs （ P<0.05）. There was no statistical significance （P>0.05） in the CV_CV of the relative expression of TAB2, CHRNA1 and Cx45 mRNAs. CONCLUSIONS As the mRNAs involving in biological process regulation, PUM2 and CHRNA1 mRNAs show a lowest individual homogeneity of the relative expression followed by TAB2 mRNA. As the mRNAs participating in the composition of cellular structure, Cx45 and CHRNA1 mRNAs show a high individual homogeneity of the relative expressions. The functional classification should be considered for the screening of the mRNA indicators used for wound age estimation.
Animals ; Contusions/diagnosis* ; RNA, Messenger ; RNA-Binding Proteins/metabolism* ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Time Factors ; Wound Healing