MS-275 potentiates the effect of YM-155 in lung adenocarcinoma via survivin downregulation induced by miR-138 and miR-195.

BACKGROUND: YM-155 has been proven to be an efficient antitumor suppressor in non-small cell lung cancer (NSCLC) cells. However, the suppressive effect of YM-155 on the expression of survivin is not sufficient and has a short half-life. MS-275, a histone deacetylase inhibitor, has significant antitumor capacity with a relatively long half-life. Our study explored whether MS-275 could enhance the inhibitory effect of YM-155 on LUAD proliferation. METHODS: To investigate the synergistic effect of MS-275 and YM-155, we employed methyl thiazolyl tetrazolium and colony formation assays to access the inhibition effect of MS-275, YM-155, or a combination in A549 and HCC827 cell lines. We then detected the effect of MS-275 and YM-155 on the expression of survivin and pro-apoptotic proteins by Western blot and miR-138 or miR-195 expression by quantitative PCR. We also analyzed the methylation level of microRNAs (miRNAs) using methylation-sensitive quantitative PCR. Finally, we investigated the interaction between miRNAs and survivin by luciferase reporter assay. RESULTS: MS-275 facilitated an inhibitory effect of YM-155 on lung adenocarcinoma cell proliferation. MS-275 can upregulate the level of acetylated H3, promote the degradation of DNA methyltransferases, and inhibit the methylation of miR-138 and miR-195 genes to elevate the expression of miR-138 and miR-195. Moreover, miR-138 and miR-195 showed a synergistic effect with YM-155 by directly binding to the 3 untranslated region of survivin to attenuate its expression. CONCLUSION: For the first time, we report the synergistic effective of MS-275 and YM-155 and suggest a new direction for the future application of YM-155.

Tumor necrosis factor-like weak inducer of apoptosis regulates quadriceps muscle atrophy and fiber-type alteration in a rat model of chronic obstructive pulmonary disease.

BACKGROUND: In chronic obstructive pulmonary disease (COPD), weakness and muscle mass loss of the quadriceps muscle has been demonstrated to predict survival and mortality rates of patients. Tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK), as a member of the TNF superfamily, has recently been identified as a key regulator of skeletal muscle wasting and metabolic dysfunction. So our aim was to study the role of TWEAK during quadriceps muscle atrophy and fiber-type transformation in COPD model rats and its possible pathway. METHODS: Forty-four healthy male adult Wistar rats were randomly divided into two groups: A normal control group (n = 16) and a COPD model group (n = 28). The COPD group was exposed to cigarette smoke for 90 d and injected with porcine pancreatic elastase on day 15, whereas the control group was injected with saline alone. Following treatment, weights of the quadriceps muscles were measured and hematoxylin and eosin staining was performed to identify structural changes in lung and quadriceps muscle tissue. Immunohistochemical staining was also conducted to determine the localization of TWEAK, nuclear factor (NF)-κB, muscle ring finger (MuRF)-1 and proliferator-activated coactivator (PGC)-1a proteins in the quadriceps muscle, and western blotting was used to assess the level of protein expression. RESULTS: Compared with controls, COPD model rats exhibited significantly lower quadriceps muscle weight (P < 0.05) accompanied by fiber atrophy and disordered fiber arrangement, a wide gap between adjacent muscle fibers, a significant reduction in nuclear number (P < 0.05) and an uneven size distribution. The proportion of fiber types was also significantly altered (P < 0.05). In addition, TWEAK expression in the quadriceps muscle of COPD model rats was significantly higher than that in control rats (P < 0.05), and was significantly associated with quadriceps atrophy and fiber-type alteration (P < 0.05). Levels of NF-κB, MuRF1 and PGC-1α expression also significantly differed between the two groups (P < 0.05). CONCLUSIONS: Collectively these data suggest that increased levels of TWEAK may lead to skeletal muscle atrophy and fiber-type alteration, which in turn may be associated with activation of the ubiquitin-proteasome pathway, involving NF-κB, MuRF1 and PGC-1α as potential regulatory factors. These preliminary results in rats suggest that TWEAK may be a therapeutic target for the treatment of muscle atrophy in COPD.

A randomized controlled clinical trial of levofloxacin 750 mg versus 500 mg intravenous infusion in the treatment of community-acquired pneumonia.

The objective of this study was to compare the efficacy and safety of levofloxacin 750 mg for 5 days versus 500 mg for 7-14 days intravenous (IV) in the treatment of community-acquired pneumonia (CAP). This clinical trial was the first of its kind conducted in Chinese people and also in Asian population. A total of 241 were enrolled and randomized to 750 mg group (n = 121) or 500 mg (n = 120) group from 10 study centers. The median treatment duration was 5.0 days in 750 mg and 9.0 days in 500 mg group. The median total dose was 3750 mg in 750 mg and 4500 mg in 500 mg group. The bacterial eradication rate was 100% in both groups. The overall efficacy rate in 750 mg group was 86.2% (94/109), and 84.7% (94/111), in 500 mg group of full analysis set visit 4, 95% confidence interval of 1.6% (-7.8-10.9%); the statistical results showed that 750 mg group was non-inferior to 500 mg group. The most common clinical adverse drug reactions were injection site adverse reactions in both 750 mg group and 500 mg group; the other common adverse drug reactions were insomnia, nausea, skin rash, etc. The most common drug-related laboratory abnormalities were neutrophil percentage decreased, decreased white blood cell count, alanine aminotransferase, and aspartate aminotransferase elevation in both 750 mg group and 500 mg group. Most of adverse drug reactions were mild in severity and well-tolerated. In summary, the regimen of levofloxacin 750 mg IV for 5 days was at least as effective and well tolerated as 500 mg IV for 7-14 days for the treatment of CAP.

[The influence of indomethacin on TNFα and skeletal muscle protein catabolism in chronic obstructive pulmonary disease rat model].

OBJECTIVE: To observe the influence of tumor necrosis factor-alpha (TNFα) on skeletal muscle protein catabolism in rats with chronic obstructive pulmonary disease (COPD) and the effects of indomethacin (IND) on it. METHODS: Duplicated COPD model rats were divided into two groups: the malnutrition group and the normal nutrition group. The malnutrition group were further divided by randomized block design into four groups. Isotonic physiologic saline was administered to group A, the control and the normal nutrition group, and different doses of oral IND were administered to groups B, C, and D daily (group B: 0.5 mg×kg(-1)×d(-1); group C: 1 mg×kg(-1)×d(-1); group D: 2 mg×kg(-1)×d(-1)). The body weight, concentrations of TNFα, contents of 3-methyl-histidine (3-MH) and tyrosine (Tyr)in the diaphragm and extensor digitorum longus muscle homogenates were measured before and after the intervention. RESULTS: Before the intervention, the concentrations of TNFα in the serum of malnutrition groups were all significantly higher than those of normal nutrition group and the control group.After the intervention: (1) The concentrations of TNFα in the serum of the rats of group B, C and D were significantly lower than the group A, especially in group C. The levels of TNFα in serum and body weight of model group rats were negatively correlated (r = -0.846, P < 0.01), as well as the diaphragm and extensor digitorum longus muscle weights (r = -0.778, P < 0.01; r = -0.772, P < 0.01). (2) The levels of 3-methyl-histidine in the diaphragm and extensor digitorum longus muscles of the intervention group C was lower than the COPD normal nutrition group, as well as the intervention groups B and D. The contents of tyrosine in the diaphragm and extensor digitorum longus muscles of the intervention group C was lower than that of the COPD normal nutrition group, as well as the groups B and D. The body weight growth value of the intervention group B were slightly higher than the group A, without significant difference (P > 0.05), while the group C was significantly higher than the group A (P < 0.01). CONCLUSIONS: TNFα is involved in the occurrence of COPD malnutrition and skeletal muscle amyotrophy. IND can reduce the TNFα levels in the serum and the catabolic rates of the skeletal muscle proteins in malnutrition rats with COPD, so as to improve partly the skeletal muscle atrophy.

[Clinical value of CYFRA 21-1, HER-2/neu and NSE in differential diagnosis of pleural effusion].

AIM: To explore the clinical value of CYFRA 21-1, HER-2/neu and NSE as tumor markers for differential diagnosis of malignant and benign pleural effusion. METHODS: Radioimmunoassay and ELISA were used to detect the serum and effusion levels of CYFRA 21-1, HER-2/neu and NSE in 62 patients with malignat pleural effusion and 48 patients with benign pleural effusion. The clinical properties were analyzed and the cut off values were determined through receiver operating characteristic(ROC) curve. RESULTS: The levels of CYFRA 21-1, HER-2/neu and NSE in serum and pleural effusion in malignant groups were significant higher than those in benign group (P < 0.01).The levels of CYFRA 21-1, HER-2/neu and NSE in pleural effusion were obviously higher than those in serum (P < 0.01 or 0.05). The cut off value of NSE in serum was 11.40 µg/L, While the cut off value of NSE in pleural effusion was 16.47 µg/L. The cut off values of CYFRA 21-1 in serum and in pleural effusion were 4.94 µg/L and 7.70 µg/L respectively. The cut off values of HER-2/neu in serum and in pleural effusion were 2.50 µg/L and 3.50 µg/L, respectively. Independent examination of the levels of CYFRA 21-1, HER-2/neu and NSE in serum and pleural effusion showed that the area under ROC curve was (0.852, 0.932), (0.867, 0.887) and (0.773, 0.846), respectively. Double-combined detection enlarged the area under ROC curve, while three-combined detection caused an increase in the area under ROC curve up to 0.999. CONCLUSION: The detection of CYFRA 21-1, HER-2/neu and NSE in pleural effusion has certain clinical significance in differential diagnosis of malignant and benign effusions. ROC curve can be used to determine the cut off values. Given the limitation of single index, comprehensive application of multiple indexes may increase the accuracy of diagnosis.

Population pharmacokinetics of oral levofloxacin 500 mg once-daily dosage in community-acquired lower respiratory tract infections: results of a prospective multicenter study in China.

This study aimed to explore the pharmacokinetic features of levofloxacin (LVFX) in Chinese patients with infections and to confirm oral LVFX 500 mg once daily as an optimal treatment regimen based on pharmacokinetic-pharmacodynamic (PK-PD) analysis. A total of 1052 plasma samples from 164 Chinese adult patients with community acquired lower respiratory tract infections (CALRTIs) and 18 healthy volunteers were used for population PK analysis. LVFX 500-mg tablets were given once daily. A nonlinear mixed effects model (NONMEM) program was used for population PK model-building and a two-compartment model with first-order absorption process was established. Creatinine clearance (CL(cr)) and body weight were identified as intrinsic factors which significantly affected oral clearance (CL(t)/F) and the apparent volume of distribution of the central compartment (V1/F), respectively. The final model is described as follows: CL(t)/F (l/h) = (8.97 + 0.917 x (CL(cr) (ml/min)-100.92) x 60/1000) x exp (eta(CLt/F)). V1/F (l) = (85.3 + 1.22 x (weight (kg)-60.75)) x exp (eta(V1/F)). Q/F (l/h) = 0.351. V2/F (l) = 6.81. k(a) (h(-1)) = 1.44 x exp(eta(ka)). Based on the population PK model, mean C(max) and AUC(0-24h) in CALRTI patients were estimated as 5.13 microg/ml and 58.98 microg.h/ml, respectively. A subgroup analysis showed that patients with mild renal dysfunction (50 ml/min < or = CL(cr) < 80 ml/min) had 34% higher AUC(0-24h) values compared to patients with normal renal function (CL(cr) > or = 80 ml/min). Postmodeling simulation using final population PK estimates also showed that C(max) and AUC(0-24h) increased markedly in patients with severe renal dysfunction. The results indicate that LVFX dosage adjustment should be individualized on the basis of the CL(cr), especially in those with CL(cr) less than 50 ml/min. None of the PK parameters had any correlation with the occurrence of adverse events. PK-PD analysis indicated that, in patients treated with LVFX 500 mg once daily, the AUC(0-24h)/MIC ratio exceeded the target for those major CALRTI pathogens isolated. In addition, the C(max)/MIC ratio reached 5 for Streptococcus pneumoniae, indicating that the emergence of LVFX-resistant S. pneumoniae could be prevented during the therapy with this dosage regimen. These results demonstrate that oral LVFX 500 mg once daily has favorable PK parameters and PK-PD features in patients with CALRTIs, and the results strongly support this dosage regimen for the treatment of CALRTI.

Clinical evaluation of oral levofloxacin 500 mg once-daily dosage for treatment of lower respiratory tract infections and urinary tract infections: a prospective multicenter study in China.

Levofloxacin (LVFX), a fluoroquinolone agent, has a broad spectrum that covers Gram-positive and -negative bacteria and atypical pathogens. It demonstrates good clinical efficacy in the treatment of various infections, including lower respiratory tract infections (LRTIs) and urinary tract infections (UTIs). To evaluate the efficacy and safety of oral LVFX 500 mg once daily, a large open-label clinical trial was conducted in 1266 patients (899 with LRTIs and 367 with UTIs) at 32 centers in China. In the per-protocol population, the clinical efficacy rate (cure or improvement) at 7 to 14 days after the end of treatment was 96.4% (666/691) for LRTIs and 95.7% (267/279) for UTIs. In 53 patients diagnosed with atypical pneumonia the treatment was effective. The bacteriological efficacy rate was 96.6% (256/265) for LRTIs and 93.3% (126/135) for UTIs. The eradication rate of the causative pathogens was 100% (33/33) for Haemophilus influenzae and 96.0% (24/25) for Streptococcus pneumoniae in LRTIs, and 94.1% (80/85) for Escherichia coli in UTIs. The overall efficacy rates were 89.3% (617/691) for LRTIs and 87.8% (245/279) for UTIs. The incidence of drug-related adverse events (ADRs) was 17.3% (215/1245), and the incidence of drug-related laboratory abnormalities was 15.7% (191/1213). Common ADRs were dizziness, nausea, and insomnia. Common laboratory abnormalities included "WBC decreased", "alanine aminotransferase (ALT) increased", "aspartate aminotransferase (AST) increased", and "lactate dehydrogenase (LDH) increased". All of these events were mentioned in the package inserts of fluoroquinolones including LVFX, and most events were mild and transient. Thirty-four patients (2.7%) were withdrawn from the study because of the ADRs. No new ADRs were found. This study concluded that the dosage regimen of LVFX 500 mg once daily was effective and tolerable for the treatment of LRTIs and UTIs.

[Effects of tumor necrosis factor alpha on proteolysis of respiratory muscles in rats with chronic obstructive pulmonary disease].

OBJECTIVE: To investigate the impact of tumor necrosis factor alpha (TNF-alpha) on proteolysis of respiratory muscles in rats with chronic obstructive pulmonary disease (COPD). METHODS: Ninety healthy male adult Wistar rats were randomly divided into two groups: a normal control group (n = 20) and a model group (n = 70). The COPD rat model was established by intratracheal instillation of porcine pancreatic elastase and exposure to cigarette smoke for 60 days. Malnutrition was defined as the weight of the rats in the model group lower than 90% of the mean body weight of the control group. Two weeks after the establishment of the COPD model, 10 rats were randomly chosen in the malnutrition group, and received 4 days' therapy of intravenous injection of TNF-alpha monoclonal antibody (McAb) 0.1 mg/kg. The concentrations of TNF-alpha in the homogenates of respiratory muscles were measured by ELISA, and the contents of 3-methylhistidine, tyrosine in homogenates of respiratory muscle were measured by high performance liquid chromatography. RESULTS: The levels of TNF-alpha in the homogenates of diaphragmatic muscle of the malnutrition group [(125 +/- 11) pg/g] were significantly higher than that of the control group [(64 +/- 5) pg/g]; The levels of TNF-alpha in the homogenates of internal intercostal muscle of the malnutrition group [(119 +/- 11) pg/g] were significantly higher than that of the control group [(59 +/- 5) pg/g]. The contents of 3-methylhistidine in homogenates of diaphragmatic muscle [(7.1 +/- 0.6) nmol/g] and internal intercostal muscle [(7.4 +/- 0.6) nmol/g] of the malnutrition group were significantly higher than that of the control group [(4.0 +/- 0.5) nmol/g]. The contents of tyrosine in homogenates of diaphragmatic muscle [(639 +/- 24) nmol/g] and internal intercostal muscle [(660 +/- 25) nmol/g] of the malnutrition group were significantly higher than that of the control group [(579 +/- 26) nmol/g]. TNF-alpha in the respiratory muscle showed a strong positive correlation with proteolysis of respiratory muscle (r = 0.854, P < 0.01). CONCLUSION: An increase of proteolysis of respiratory muscles was found in COPD rats, more significant in the malnutrition rats. TNF-alpha is one of the causes for the increase of proteolysis of respiratory muscles.