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Objective To evaluate the effect of melatonin pretreament on cell apoptosis and autophagy during lung ischemia-reperfusion (I/R) in rats.Methods Thirty-six adult male Sprague-Dawley rats,weighing 230-280 g,were divided into 3 groups (n=12 each) using a random number table method:sham operation group (group Sham),pulmonary I/R group (LIR group) and melatonin pretreatment group (MLT group).Lung I/R injury model was established by clamping the left hilum of lung for 60 min followed by 2 h reperfusion in LIR and MLT groups.Melatonin 1 mg/100 g was intraperitoneally injected at 15 min before clamping the left hilum of lung in group MLT.The rats were sacrificed at the end of reperfusion,the left lung was lavaged and the broncho-alveolar lavage fluid (BALF) was colleted for determination of protein concentrations.Lungs were removed for microscopic examination of the pathological changes (with a light microscope) which were scored and for determination of wet to dry weight ratio (W/D ratio),cell apoptosis (by TUNEL) and expression of Bcl-2,Bax,microtubule-related protein 1 light chain 3B Ⅰ (LC3B Ⅰ),LC3B Ⅱ,Beclin-1 and phosphorylated mammal rapamycin target protein receptor (p-mTOR) (using Western blot).The apoptosis index,Bcl-2/Bax ratio and LC3B Ⅱ/LC3B Ⅰ ratio were calculated.Results Compared with Sham group,the protein concentration in BALF,W/D ratio of lung tissues,apoptosis index,LC3B Ⅱ/LC3B Ⅰ ratio and Beclin-1 expression were significantly increased,and Bcl-2/Bax ratio and p-mTOR expression were decreased in LIR and MLT groups (P＜0.05).Compared with LIR group,the protein concentration in BALF,W/D ratio of lung tissues,apoptosis index,LC3B Ⅱ/LC3B Ⅰ ratio and Beclin-1 expression were significantly decreased,and Bcl-2/Bax ratio and p-mTOR expression were increased in MLT group (P＜0.05).Conclusion The mechanism by which melatonin pretreatment mitigates lung I/R injury may be related to inhibiting cell apoptosis and autophagy in rats.
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Objective To explore the correlation of epicardial adipose tissue (EAT) volume with atrial fibrillation (AF) and its recurrence after radiofrequency ablation (RA).Methods Eighty-five AF patients (AF group) and 90 non-AF patients (control group) were chosen between January 2014 and May 2016.Their EAT volumes were measured by CT scanning.Patients in AF group after RA were followed up 6-18 months,and the recurrence of AF was recorded.The recurrence of AF within 3 months of RA was defined as blanking recurrence group (n =27) and non-blanking recurrence group (n =58),and that after 3 months of RA was defined as the 1 ong-term recurrence.Results The total EAT volume and left atrial EAT volume were significantly larger in blanking recurrence group than in non-blanking recurrence group [(118.71 ±28.94) cn3 vs (97.73 ±24.86)cm3,(29.98 ±8.09)cm3 vs (23.11 ±8.30)cm3,t =6.219,4.451,P ＜ 0.01].Multivariate logistic resgression analysis showed that total EAT volume and left atrial EAT volume were the independent risk factors for AF.All showed that total EAT volume and left atrial EAT volume were the independent risk factors for blanking recurrence.The incidence of long-term recurrence was significantly higher in blanking recurrence group [40.7％ (11/27)] than in non-blanking recurrence group [15.5 ％ (9/58)] (x2 =7.142,P ＜ 0.05).Conclusions The incidence of AF is higher in patients with a Iarger total EAT volume and a larger left atrial EAT volume and AF is easier to recur even though after RA.
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Objective To study the relation of epicardial adipose tissue (EAT) thickness with 1eftventricular remodeling and dysfunction in dilated cardiomyopathy (DCM) patients.Methods One hundred and twenty DCM patients who received treatment from Febuary 2012 to February 2016 were served as DCM group,and 76 healthy subjects undergoing physical examination were served as control group.Their left ventricular end-diastolic diameter (LVEDD),left ventricular end systolic diameter (LVESD),left ventricular end diastolic volume (LVESV),left ventricular end systolic volume (LVEDV),left ventricular end diastolic volume index (LVEDVI),left ventricular end systolic volume index (LVESVI),sphericity index (SI),leftventricular ejection fraction (LVEF),and EAT thickness were measured by routine cardiac ultrasonography and compared between two groups.Results The levels of LVESD,LVEDD,LVESV,LVEDV,LVESVI,LVEDVI and EAT thickness in DCM group were significantly higher,and the levels of LVEF,SIS and SID in DCM group were significantly lower (P＜0.05).The EAT thickness in DCM group with NYHA class Ⅱ,Ⅲ,Ⅳ was (8.1 ± 1.8),(7.8 ± 2.0),(7.9 ± 1.7) mm,and there was significant difference (F=1.973,P＞ 0.05) Linear correlation analysis showed that the EAT thickness was positively related with the LVESD,LVEDD,LVESV,LVEDV,LVEDVI,LVESVI,SISand SID (r =0.247,0.231,0.256,0.267,0.293,0.281,0.261,0.237,P＜0.05).There was no relationship between EAT thickness and LVEF (r =0.132,P＞ 0.05).Logistic multifactor regression analysis showed that EAT thickness was an independent risk factor for left ventricular remodeling in DCM patients (OR =0.793,95％CI:0.431-1.734,P =0.039).Conclusions The EAT thickness is significantly related with the left ventricular remodeling and can be used as an independent risk factor for predicting left ventricular remodeling in DCM patients.
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<p><b>BACKGROUND</b>Protectin D1 (PD1), derived from docosahexaenoic acid, has been shown to control and resolve inflammation in some experimental models of inflammatory disorders. We investigated the protective roles of protectin D1 in pulmonary inflammation and lung injury induced by lipopolysaccharide (LPS).</p><p><b>METHODS</b>Mice were randomly assigned to six groups (n = 6 per group): sham-vehicle group, sham-PD1 group, sham-zVAD-fmk group, LPS-vehicle group, LPS-PD1 group, and LPS-PD1-zVAD-fmk group. Mice were injected intratracheally with 3 mg/kg LPS or saline, followed 24 hours later by intravenous injection of 200 µg/mouse PD1 or vehicle. At the same time, some mice were also injected intraperitoneally with the pan-caspase inhibitor zVAD-fmk. Seventy-two hours after LPS challenge, samples of pulmonary tissue and bronchoalveolar lavage fluid were collected. Optical microscopy was used to examine pathological changes in lungs. Cellularity and protein concentration in bronchoalveolar lavage fluid were analyzed. Lung wet/dry ratios and myeloperoxidase activity were measured. Apoptosis of neutrophils in bronchoalveolar lavage fluid (BALF) was also evaluated by flow cytometry.</p><p><b>RESULTS</b>Intratracheal instillation of LPS increased neutrophil counts, protein concentration in bronchoalveolar lavage fluid and myeloperoxidase activity, it induced lung histological injury and edema, and also suppressed apoptosis of neutrophils in BALF. Posttreatment with PD1 inhibited LPS-evoked changes in BALF neutrophil counts and protein concentration and lung myeloperoxidase activity, with the outcome of decreased pulmonary edema and histological injury. In addition, PD1 promoted apoptosis of neutrophils in BALF. The beneficial effects of PD1 were blocked by zVAD-fmk.</p><p><b>CONCLUSION</b>Posttreatment with PD1 enhances resolution of lung inflammation during LPS-induced acute lung injury by enhancing apoptosis in emigrated neutrophils, which is, at least in part, caspase-dependent.</p>
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Animals , Male , Mice , Acute Lung Injury , Drug Therapy , Allergy and Immunology , Apoptosis , Docosahexaenoic Acids , Therapeutic Uses , Inflammation , Drug Therapy , Lipopolysaccharides , Toxicity , Mice, Inbred BALB C , Neutrophils , Cell Biology , Peroxidase , MetabolismABSTRACT
Objective To investigate the expression of matrix metalloproteinases 1 (TIMP-1)products inhibitor on smooth muscle cell (SMC) proliferation in rat autologous vein graft.Methods Autogenous vein transplantation model was established in 30 Wistar rats. The vein grafts were harvested at different time after grafting. The change of TIMP-1 was detected by using hematoxylin and eosin, immunohistochemistry and in situ hybridization (ISH). Results 1. Changes of histopathology in vein grafts: Intimal hyperplasia (IH) could be seen in 7 - 14 days, the peak at 2 ~ 4 weeks after operation.2. ISH results: TIMP-1mRNA positive cells appeared at 24 hours, increased significantly in 72 hours and reached the peak at 1 ~ 2 week after operation. There was significant difference between day 1 and 2 week.TIMP-I expression was not detected in normal vessels (P <0. 01). 3. Immunohistochemistry results: There was trace TIMP-I expression in normal vessels. The expression of TIMP-1 appeared at 72 hours after vein graft, increased mostly in 1 week, reached the peek at 2 week and reduced later. There was significant difference between day 1 and 2 week (P < 0. 01). Conclusions 1. The activation of TIMP-1 exists in autogenous vein grafts. 2. The intima experienced hyperplasia in spite of increased secretion of endogenous TIMP-1 after autogenous vein grafts.
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Objective BALB/C mice were inoculated subcutaneously with hepatoma 22 (H22) cells, and the treatment effect of thalidomide and intratumor injection of ethanol extract of Scutellaria Barbata (SB) on tumor growth were evaluated. Methods In vitro, mouse H22 liver cells were cultivated, and H22 cells in the logarithmic growth phase were tested. SB was extracted in the ethanol recirculation. A total of 28 BALB/C male mice inoculated subcutaneously with H22 cells were randomly divided into 4 groups, 7 mice per group. Group A:intragastric administration with thalidomide. Group B:SB intratumor injection. Group C:combination of thalidomide and SB with the same administration as Group A and Group B. Group D (control group):1% Ethanol 0.2～0.5 mL intratumor injection. Tumor volumes of four groups were observed and the tumors were weighed. The microvessel densities (MVD) of all transplantation tumors were measured by immunohistochemical staining with anti-CD31 monoclonal antibody. Results There were statistically significant differences between Group C and Group A (P 0.05). The studies showed statistically differences between Group C and Group A (P
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Objective To study the significance of c-met mRNA in axillary drainage after operations for breast cancer. Methods RT-PCR assay was used to examine c-met mRNA in axillary drainage after operations in 52 cases of breast cancer. The relationships between the expression of c-met and the tumor size, metastatic lymph nodes, the expressions of estrogen receptor (ER), progesterone receptor (PR) and c-erbB-2 were analyzed, respectively. In addition, the effect of douching operative field with 5-FU and distilled water on the expression of c-met mRNA was also analyzed. Results ①The proto-oncogene c-met mRNA could be detected in axillary drainage after operations for breast cancer by RT-PCR, and its positive rate was higher than that in routine pathological detection for micrometastasis in the axillary lymph nodes (P
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OBJECTIVE:To establish an HPLC method for determination of major component and related peptide in eptifibatide injection.METHODS:The determination was performed on AichromBond-AQ C18 column with column temperature at 25 ℃ and flow rate of 1.0 mL?min-1.The mobile phase consisted of acetonitrile-water(0.13% trifluoroacetic acid).The detection wavelength was set at 220 nm and injection volume was 20 ?L.RESULTS:The linear range was 75.2～1 203.6 ?g?mL-1(r=0.9999) with mean recovery of 99.73%～99.95% (RSD=0.26%).CONCLUSION:The method is simple and rapid and it is applicable for determination of major component and related peptide in eptifibatide injection.