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Objective To examine the effects of ethyl pyruvate (EP) on mitochondrial dynamics and cell apoptosis in lipopolysaccharide (LPS)-induced human kidney-2 (HK-2) cells. Methods HK-2 cells were divided into three groups: HK-2 cells were challenged with LPS (800 μg/L) for 24 hours as LPS group, or LPS mixed with EP (0.25 mmol/L) for 24 hours as EP group. Cells were incubated with normal saline for 24 hours as control group. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and intracellular adenosine triphosphate (ATP) were detected by enzyme linked immunosorbent assay (ELISA). JC-1 staining and Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) assays were used to evaluate mitochondrial membrane potential and cell apoptosis, respectively. Western Blot was used to evaluate the protein expressions of mitochondrial dynamics, including death-associated protein kinase 2 (DAPK-2), mitofusin (Mfn-1 and Mfn-2), and apoptotic associated biomarkers, including caspase-3, caspase-9, Bcl-2, Bcl-xL, cytochrome C (Cyt C), and DNA repair enzyme poly ADP-ribose polymerase (PARP). Results Compared with the NC group, MDA, IL-6, TNF-α of LPS group were significantly increased, the expression of SOD, mitochondrial membrane potential and ATP level were significantly decreased, the expression of mitochondrial fission protein DAPK-2 was significantly increased, and mitochondrial fusion proteins Mfn-1 and Mfn-2 were significantly decreased, cell apoptosis and apoptotic protein caspase-3, caspase-9 and Cyt C were increased, and anti-apoptotic protein Bcl-2, Bcl-xL, PARP were significantly decreased. Compared with the LPS group, the oxidative activities and inflammatory factors above were inhibited in EP group [MDA (μmol/L):12.35±2.21 vs. 45.95±1.76, SOD (kU/L): 54.68±1.42 vs. 40.73±1.60, IL-6 (ng/L): 67.87±2.61 vs. 338.92±20.91, TNF-α (ng/L): 19.23±1.80 vs. 180.69±6.51], mitochondrial membrane potential and ATP level were significantly increased [mitochondrial membrane potential: (99.43±0.25)％ vs. (69.40±0.75)％, ATP (×106 RLU): 0.19±0.01 vs. 0.12±0.05], the expression of mitochondrial fission protein was significantly decreased (DAPK-2/β-actin:0.03±0.01 vs. 0.61±0.02), mitochondrial fusion proteins were significantly increased (Mfn-1/β-actin: 0.43±0.04 vs. 0.17±0.01, Mfn-2/β-actin: 0.201±0.004 vs. 0.001±0.001), percentage of cell apoptosis was significantly decreased [(5.25±0.17)％ vs. (34.42±0.64)％], the expressions of apoptotic proteins were significantly decreased (caspase-3/β-actin: 0.25±0.15 vs. 1.76±0.01, caspase-9/β-actin: 0.09±0.02 vs. 1.52±0.12, Cyt C/β-actin: 0.001± 0.001 vs. 0.350±0.030), and the expressions of anti-apoptotic proteins and PARP were significantly increased (Bcl-2/β-actin: 0.500±0.010 vs. 0.009±0.004, Bcl-xL/β-actin: 0.550±0.010 vs. 0.009±0.001, PARP/β-actin:0.94±0.01 vs. 0.16±0.13), with statistically significant differences (all P < 0.05). Conclusions There are enhanced mitochondrial fission and diminished mitochondrial fusion in LPS-induced HK-2 cells. EP can protect mitochondria functions by regulate mitochondrial dynamics, and reducethe apoptosis of LPS-induced HK-2 cells.
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Objective@#To examine the effects of ethyl pyruvate (EP) on mitochondrial dynamics and cell apoptosis in lipopolysaccharide (LPS)-induced human kidney-2 (HK-2) cells.@*Methods@#HK-2 cells were divided into three groups: HK-2 cells were challenged with LPS (800 μg/L) for 24 hours as LPS group, or LPS mixed with EP (0.25 mmol/L) for 24 hours as EP group. Cells were incubated with normal saline for 24 hours as control group. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and intracellular adenosine triphosphate (ATP) were detected by enzyme linked immunosorbent assay (ELISA). JC-1 staining and Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) assays were used to evaluate mitochondrial membrane potential and cell apoptosis, respectively. Western Blot was used to evaluate the protein expressions of mitochondrial dynamics, including death-associated protein kinase 2 (DAPK-2), mitofusin (Mfn-1 and Mfn-2), and apoptotic associated biomarkers, including caspase-3, caspase-9, Bcl-2, Bcl-xL, cytochrome C (Cyt C), and DNA repair enzyme poly ADP-ribose polymerase (PARP).@*Results@#Compared with the NC group, MDA, IL-6, TNF-α of LPS group were significantly increased, the expression of SOD, mitochondrial membrane potential and ATP level were significantly decreased, the expression of mitochondrial fission protein DAPK-2 was significantly increased, and mitochondrial fusion proteins Mfn-1 and Mfn-2 were significantly decreased, cell apoptosis and apoptotic protein caspase-3, caspase-9 and Cyt C were increased, and anti-apoptotic protein Bcl-2, Bcl-xL, PARP were significantly decreased. Compared with the LPS group, the oxidative activities and inflammatory factors above were inhibited in EP group [MDA (μmol/L): 12.35±2.21 vs. 45.95±1.76, SOD (kU/L): 54.68±1.42 vs. 40.73±1.60, IL-6 (ng/L): 67.87±2.61 vs. 338.92±20.91, TNF-α (ng/L): 19.23±1.80 vs. 180.69±6.51], mitochondrial membrane potential and ATP level were significantly increased [mitochondrial membrane potential: (99.43±0.25)% vs. (69.40±0.75)%, ATP (×106 RLU): 0.19±0.01 vs. 0.12±0.05], the expression of mitochondrial fission protein was significantly decreased (DAPK-2/β-actin: 0.03±0.01 vs. 0.61±0.02), mitochondrial fusion proteins were significantly increased (Mfn-1/β-actin: 0.43±0.04 vs. 0.17±0.01, Mfn-2/β-actin: 0.201±0.004 vs. 0.001±0.001), percentage of cell apoptosis was significantly decreased [(5.25±0.17)% vs. (34.42±0.64)%], the expressions of apoptotic proteins were significantly decreased (caspase-3/β-actin: 0.25±0.15 vs. 1.76±0.01, caspase-9/β-actin: 0.09±0.02 vs. 1.52±0.12, Cyt C/β-actin: 0.001±0.001 vs. 0.350±0.030), and the expressions of anti-apoptotic proteins and PARP were significantly increased (Bcl-2/β-actin: 0.500±0.010 vs. 0.009±0.004, Bcl-xL/β-actin: 0.550±0.010 vs. 0.009±0.001, PARP/β-actin: 0.94±0.01 vs. 0.16±0.13), with statistically significant differences (all P < 0.05).@*Conclusions@#There are enhanced mitochondrial fission and diminished mitochondrial fusion in LPS-induced HK-2 cells. EP can protect mitochondria functions by regulate mitochondrial dynamics, and reducethe apoptosis of LPS-induced HK-2 cells.
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Objective To analyze clinical effectiveness of tigecycline for complicated intra abdominal infections (cIAIs) in surgical intensive care units.Methods The clinical data of patients diagnosed as cIAIs from Nov 2011 to Aug 2014 were retrospectively collected.Data of sex, age, severity of disease, bacterial strains and drug resistance, prior antibiotics, dosage of tigecycline were included.Results 1 862 patients were admitted into surgical intensive care unit from Nov 2011 to Aug 2014.54 patients were finally treated by tigecycline among 304 patients diagnosed as cIAIs.Acinetobacter baumannii (23.1％), Klebiella pneumonia (18.5％), Escherichia coli (16.9％) were the top three pathogenic bacteria.41/50 were of multiple bacterial infection.Previously adopted antibiotics were miscellaneous, the number of used antibiotics was up to 13 for one patient.Coverage of tigecycline averaged at 8.9 days.Tigecycline effective rate was 62％, 38％ (19/50) cIAIs were completely controlled and cured,24％ (12/50) patients showed effectiveness of tigercycline that procalcitonin decreased 50％ within 72 h.Logistic regression analysis showed that severity of disease and tigecycline dose contribute to the effectiveness within 72 h.Conclusions Tigecycline is effective alterative for patients diagnosed as complicated intra abdominal infections in surgical intensive care units.
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Objective To observe the changes in serum vancomycin trough concentration,and explore its optimal dosage in critical patients.Methods A retrospective study was conducted.Data of 66 patients who used vancomycin during July 2010 to May 2012 in surgical intensive care unit (SICU) of the First Affiliated Hospital of Sun Yat-Sen University were collected and analyzed.According to the endogenous creatinine clearance rate (CCr),the patients were divided into two groups:CCr normal group (≥ 70 mL/min) and CCr lowered group (＜70 mL/min).The distribution of vancomycin serum trough concentration between two groups,relationship between vancomycin serum trough concentrations and CCr,and the influence of vancomycin serum trough concentrations on the prognosis was analyzed.The difference between actual dosage and the recommended dosage in guideline was compared between two groups.Results 119 times of vancomycin serum trough concentration in 66 patients were enrolled,and it was found that only 20.17％ (24/119) reached the target concentration (15-20 mg/L),45.38％ (54/119)＜ 15 mg/L and 34.45％ (41/1 19) ＞20 mg/L.Vancomycin serum trough concentration in CCr normal group (55 cases) was (13.11 ± 6.84) mg/L,among them 65.5％ (36/55) attained lower trough concentrations (＜ 15 mg/L).In the subgroup with 15-20 mg/L trough serum concentrations,vancomycin doses were significantly lower than that of recommendation (g/d:1.95 ± 0.61 vs.2.73 ± 0.32,F=1.739,P=0.001).Vancomycin serum trough concentration in CCr lowered group (64 cases) was (20.49 ± 8.12) mg/L,with 51.5％ (33/64) of them showed higher trough concentrations (＞20 rag/L).In the subgroup with 15-20 mg/L vancomycin trough serum concentration,vancomycin doses were higher than that of recommendation (g/d:1.08 ±0.49 vs.0.78 ±0.19,F=11.294,P=0.062).There was no significant difference in 28-day mortality between patients with targeting trough serum concentrations and those without [22.2％ (4/18) vs.18.8％ (9/48),x2=0.009,P=0.924].Serum creatinine [odds ratio (OR)=1.001,95％ confidence interval (95％CI):0.990-1.012,P=0.000],vancomycin doses (OR=0.600,95％CI:0.251-1.434,P=0.003),age (OR=0.985,95％CI:0.955-1.015,P=0.015) and body mass index (OR=1.013,95％CI:0.967-1.062,P=0.022) were found to be correlated to serum trough concentrations by multiple linear regression analysis.Conclusions The rate of vancomycin serum trough concentrations reaching the standard is low in critical patients,so constant monitoring is necessary.Creatinine,vancomycin dosage,age and body mass index show a relatively significant influence on the serum trough concentrations,and they should be taken into consideration in dosage to be given.
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OBJECTIVE@#To evaluate the relation of EB virus latent membrane protein-1 (LMP-1) and the phenotype of epithelial-mesenchymal transition (EMT) and cervical lymph node metastasis in nasopharyngeal carcinoma.@*METHOD@#Based on histopathology and MRI imaging, nasopharyngeal biopsy tissues from 88 patients with nasopharyngeal carcinoma were divided into 3 groups: pathologic metastasis (18), MRI metastasis(40) and without metastasis (30). The expressions of LMP-1, STAT3, Twist, E-Cadherin and Vimentin were examined immunohistochemically in biopsy tissues.@*RESULT@#LMP-1 expression was found in 35 of 88 biopsy tissues with a positive rate of 38.7%. The positive rates of LMP-1 in groups of pathologic metastasis, MRI metastasis and without metastasis were 38.9% (7/18), 47.5% (19/40) and 30.0% (9/30), respectively, and significant difference were not found among three groups. The expression of LMP-1 was positively correlated to both expressions of Twist and Vimentin (r = 0.276 and 0.282, are P < 0.01), but not to both expressions of STAT3 and E-Cadherin. The positive expressions or abnormal expression of STAT3, Twist, Vimentin and E-Cadherin were found in 57 of 88 (64.8%), 48 of 88 (54.5%), 22 of 88 (20.0%)and 53 of 88 (60.2%), respectively. Significant differences in the expression of STAT3, Twist, Vimentin and E-Cadherin were all found among groups of pathologic metastasis, MRI metastasis and without metastasis, respectively (are P < 0.05). The expression of STAT3 was positively correlated to both expressions of Twist and Vimentin (r = 0.712 and 0.316, P < 0.01).@*CONCLUSION@#EMT plays important role in cervical lymph node metastasis in nasopharyngeal carcinoma. LMP-1 may be only as one of upstream factors associated with the EMT, but not the decisive factor for cervical lymph node metastasis.