1. Determination of the related substances in the etoricoxib tablets by HPLC
Journal of International Pharmaceutical Research 2019;46(10):795-799
Objective: To establish and validate a high performance liquid chromatography(HPLC)method for the determination of related substances in etoricoxib tablets. Methods: The related substances in the etoricoxib tablets were determined by HPLC. The Waters C18 column(4.6 mm×150 mm,3.5 μm)was used. The mobile phase consisted of the aqueous 0.02 mol/L sodium dihydrogen phosphate solution and acetonitrile(71:29,V/V)and the flow rate was 0.6 ml/min. The detection wavelength was 235 nm. The column temperature was 40℃ and the injection volume was 10 μl. Results: The etoricoxib related substances and other degraded substances in the forced degradation test of etoricoxib were com- pletely separated under the HPLC conditions. The quantification limit and the detection limit were 0.5 ng and 1.0 ng both for etoricoxib and its N-oxide,respectively. The calibration curves showed a good linearity within the range of 61.14- 152.85 μg/ml for etoricoxib and 61.50-153.75 μg/ml for the N-oxide. The recoveries of etoricoxib and its N-oxide were both within the range of 98%-102%. The solution used for determining the related substances were kept stable within 12 h at room temperature. Conclusion: The established HPLC method showed good specificity,sensitivity and accuracy,which could be used for the determination of related substances in the etoricoxib tablets.
2.Effects of STI571 combined with As₂O₃ on proliferation, apoptosis and caspase 3, Bcl-xL expression of K562 cells.
Nai-Yao CHEN ; Jing WANG ; Xue-Ming WANG ; Hai-Xia ZHANG ; Zhen-Yu YAN ; Ying-Man WANG ; Song ZHANG ; Bing YAN
Journal of Experimental Hematology 2010;18(4):882-886
This study was aimed to explore the effects of STI571 alone or with As₂O₃ on proliferation, apoptosis and caspase 3, bcl-xL mRNA expression of K562 cells, and the molecular mechanism of As₂O₃ enhancing the anti-leukemia effect of STI571 so as to provide the scientific basis for clinical treatment of chronic myeloid leukemia. The effect of drugs on proliferation of K562 cells was assayed by MTT method, the apoptosis rate of K562 cells was detected by flow cytometry with Annexin V/PI double staining, the caspase 3, bcl-xL mRNA expressions of K562 cells were determined by real time quantitative PCR. The results showed that STI571 alone or with As₂O₃ both could inhibit the proliferation of K562 cells. OD value in test groups reduced along with prolonging of action times, the OD values between different time points were significantly different (p < 0.05), furthermore the OD values at 72 hours in test groups were lowest, while as compared with control group, OD values at same time points in test groups all gradually decreased, among which decrease of OD value in test 5 group was most significant. The flow cytometric detection indicated that along with time prolonging, the apoptotic rate in control group not obviously changed, but the apoptotic rate in test groups gradually increased, the difference between time points was significant (p < 0.05), moreover apoptotic rate increased most obviously at 72 hours, while as compared with control group, apoptotic rate at same time points in test groups was gradually enhanced (p < 0.05), among which the apoptotic rate in test 5 group was highest. The real time qPCR assay revealed that as compared with control group, the bcl-xL mRNA expression in test groups reduced with decrease of 2-ΔΔCT value, furthermore the decrease of expression level in test 3 group was higher than that in test 2 group (p < 0.05), while the caspase 3 mRNA expression in test groups was enhanced with increase of 2-ΔΔCT value, moreover the increase of expression level in test 3 group was higher than that in test 2 group (p < 0.05). It is concluded that the STI571 can inhibit the proliferation of K562 cells, accelerate the apoptosis of K562 cells. The STI571 combined with As₂O₃ can enhance these two effects, increase the expression of caspase-3 mRNA and decrease the expression of bcl-xL mRNA. Therefore, the effect of STI571 combined with As₂O₃ on expression of caspase 3 and bcl-xL mRNA may be one of molecular mechanisms underlying their synergic antileukemia efficacy.
Apoptosis
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drug effects
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Arsenicals
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pharmacology
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Benzamides
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Caspase 3
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metabolism
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Cell Proliferation
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drug effects
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Gene Expression Regulation, Leukemic
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Humans
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Imatinib Mesylate
;
K562 Cells
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Oxides
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pharmacology
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Piperazines
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pharmacology
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Proto-Oncogene Proteins c-bcl-2
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metabolism
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Pyrimidines
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pharmacology
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bcl-X Protein
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metabolism
3.Effectiveness of dynamic contrast-enhanced magnetic resonance imaging in evaluating clinical responses to neoadjuvant chemotherapy in breast cancer.
Yin-Hua LIU ; Jing-Ming YE ; Ling XU ; Qing-Yun HUANG ; Jian-Xin ZHAO ; Xue-Ning DUAN ; Nai-Shan QIN ; Xiao-Ying WANG
Chinese Medical Journal 2011;124(2):194-198
BACKGROUNDUse of neoadjuvant chemotherapy necessitates assessment of response to cytotoxic drugs. The aim of this research was to investigate the effectiveness of dynamic contrast-enhanced magnetic resonance imaging (MRI) for evaluating clinical responses to neoadjuvant chemotherapy in breast cancer patients.
METHODSWe examined patients receiving neoadjuvant chemotherapy for primary breast cancer between October 2007 and September 2008. Dynamic contrast-enhanced MRI was used to examine breast tumors prior to and after neoadjuvant chemotherapy. The MRI examination assessed tumors using Response Evaluation Criteria in Solid Tumors (RECIST). The Miller-Payne grading system was used as a histopathological examination to assess the effect of the treatment. We examined the relationship between the results of RECIST and histopathological criteria. In addition, we used time-signal intensity curves (MRI T-SI) to further evaluate the effects of neoadjuvant chemotherapy on tumor response.
RESULTSMRI examination of patients completing four three-week anthracycline-taxanes chemotherapy treatment revealed that no patients had complete responses (CR), 58 patients had partial responses (PR), 29 patients had stable disease (SD), and four with progressive disease (PD). The effectiveness of neoadjuvant chemotherapy (CR + PR) was 63.7% (58/91). The postoperative histopathological evaluations revealed the following: seven G5 (pCR) cases (7.7%), 39 G4 cases (42.9%), 16 G3 cases (17.6%), 23 G2 cases (25.3%), and six G1 cases (6.6%). The effectiveness (G5 + G4 + G3) was 68.1% (62/91). MRI T-SI standards classified 53 responding cases, 29 stable cases, and nine progressing cases. These results indicated that the treatment was 58.2% effective (53/91) overall.
CONCLUSIONSDynamic contrast-enhanced MRI and histopathological standards were highly correlated. Importantly, MRI T-SI evaluation was found to be useful in assessing the clinical effectiveness of neoadjuvant chemotherapy.
Adult ; Aged ; Anthracyclines ; therapeutic use ; Antineoplastic Agents ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; Contrast Media ; chemistry ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Middle Aged ; Neoadjuvant Therapy ; Taxoids ; therapeutic use
4.Effect of coronary bifurcation angle on clinical outcomes in Chinese patients treated with crush stenting: a subgroup analysis from DKCRUSH-1 bifurcation study.
Shao-liang CHEN ; Jun-jie ZHANG ; Fei YE ; Yun-dai CHEN ; Wei-yi FANG ; Meng WEI ; Ben HE ; Xue-wen SUN ; Song YANG ; Jin-guo CHEN ; Shou-jie SHAN ; Nai-liang TIAN ; Xiao-bo LI ; Zhi-zhong LIU ; Jing KAN ; Lee MICHAEL ; Kwan-tak W
Chinese Medical Journal 2009;122(4):396-402
BACKGROUNDBifurcation angles may have an impact on the clinical outcomes of crush stenting. We sought to compare high (> or = 60 degrees ) with low (< 60 degrees ) bifurcation angle in patients who underwent either classical or double kissing (DK) crush stenting for bifurcation lesions from the DKCRUSH-1 data base.
METHODSThere were 212 patients with 220 lesions, some with low-angle (n = 138) and some with high-angle (n = 74). Angiography was indexed at 8-month after procedure. Primary endpoint was the occurrence of major adverse cardiac events (MACEs), defined as cardiac death, myocardial infarction and target lesion revascularization (TLR). Secondary endpoint included late lumen loss, the rate of restenosis, and final kissing balloon inflation (FKBI).
RESULTSAt 8 months, clinical follow-up was 100%; angiographic follow-up was 75% in the low-angle group and 83.3% in the high-angle group. There were no significant differences in the FKBI between the high-angle group (91.43%) and the low-angle group (82.39%). In the high angle group, there was a significant difference in contrast volume used (P = 0.005) but no significant difference in acute gain, minimum lumen diameter (MLD), late loss and diameter stenosis in the pre-bifurcation segment, post-bifurcation segment or side branch. When lesions were assigned into with-(n = 133) and without-FKBI (n = 42), significant side-branch late loss was seen in the group without-FKBI ((0.65 +/- 0.49) mm vs (0.47 +/- 0.62) mm, P = 0.02), with a resultant greater restenosis rate (37.68% vs 18.32%, P = 0.001). No difference was detected in the MACE free survival rate between the high and low angle groups (82.39% vs 82.36%, P = 0.84). The rate of stent thrombosis tended to be higher in the lower-angle group although there was no significant difference (P = 0.38). The TLR free survival rate was 87.2% in the with-FKBI group vs 73.5% in the without-FKBI group (P = 0.001). Cox regression analysis showed that the independent predictors for target vessel revascularization were the side branch stent MLD post stenting (hazard ratios (HR) 1.028, 95% CI 2.357 - 16.233, P = 0.002), lack of FKBI (HR 4.910, 95% CI 4.706 - 8.459, P = 0.001) and unsatisfactory kissing (HR 3.120, 95% CI 2.975 - 5.431, P = 0.001).
CONCLUSIONSBifurcation angles do not influence the clinical outcome of crush stenting. Successful final kissing balloon inflation, regardless of bifurcation angles, can predict TLR.
Aged ; Angioplasty, Balloon, Coronary ; methods ; Asian Continental Ancestry Group ; ethnology ; Coronary Angiography ; methods ; Coronary Stenosis ; ethnology ; pathology ; therapy ; Drug-Eluting Stents ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; ethnology ; pathology ; therapy ; Stents ; Treatment Outcome
5.Intravenous injection of human umbilical cord blood mesenchymal stem cells affecting the level of nerve growth factors in rats with traumatic brain injury
Ya-Chen AN ; Yu-Xun WANG ; Nai-Yao CHEN ; Na SHEN ; Da-Li WANG ; Xiao-Jing ZHAO ; Jiang ZHANG ; Chun-Ling LI ; Yan-Bo PENG ; Su-Ling GAO ; Li-Sha CHANG ; Li ZHANG ; Xin-Hong XUE ; Rui-Ying CHEN
Chinese Journal of Neuromedicine 2010;09(12):1193-1196,1200
Objective To investigate the effect of transplantation of human umbilical cord blood mesenchymal stem cells (CB-MSCs) on the levels of NGF and BDNF in rats with traumatic brain injury (TBI) and explore its possible mechanism of cerebral protection. Methods Ninety healthy male SD rats were equally randomized into sham-operated group, injury control group and treatment group.TBI models in the injury control group and treatment group were induced by the improved device of Feeney weight-dropping; the rats in the treatment group were injected Brdu-labeled 3×106 CB-MSCs solved in 1ml PBS by rat-tail vein, while the rats in the sham-operated group and the injury control group were injected the equal volume of PBS solution. No immunosuppressive agents were used in all the rats.HE staining, immunohistochemistry, in-situ hybridization were employed, respectively, to detect the morphological changes, Brdu positive cells, expressions of BDNF and NGF on the 3rd, 7th, 14th, 21st and 28th d of injection. Results Only a very small number of nerve cells were BDNF and/or NGF positive in the sham-operated group. Substantial BDNF and/or NGF positive cells in the injury control group were noted in the surrounding brain damaged area following traumatic brain injury, which peaked at their levels on about 14 d of injection (the A value of NGF=8.35±1.07, that of BDNF=9.01±1.74), following by a gradual decline; however, significant difference was still noted as compared with that in the sham-operated group (P<0.05). BDNF and/or NGF-positive cells significantly increased in the treatment group, especially in the surrounding brain injured areas; their levels peaked on the 14th d of injection,following by a gradual decline on the 21st and 28th d, but they were still higher than those in the injury control group and sham-operated group at each time points (P<0.05). Conclusion Transplantation of CB-MSCs can increase the secretion of BDNF and NGF in rats with TBI, improve the local micro-damage environment and promote the repair of neurons.
6.5'-flanking regulatory sequence methylation of the Boule gene in the testis tissue of infertile men with Sertoli cell-only syndrome.
Hong-Xing LI ; Yuan-Xue JING ; Liang SUN ; Nai-Hui WANG ; De-Xiao SONG ; Yi JIN ; Le-Wei WANG ; Shi-Long XUE
National Journal of Andrology 2016;22(6):516-519
ObjectiveTo investigate the 5'-flanking regulatory sequence methylation status of the Boule gene in the testis tissue of infertile men with Sertoli cell-only syndrome (SCOS).
METHODSWe collected biopsy samples of the testis tissue from 12 men with obstructive azoospermia (the control group) and 15 cases of SCOS, all without varicocele, cryptorchidism, or infectious disease. We extracted genomic DNA from the testis tissue of the SCOS patients, analyzed the characteristics of the 5'-flanking regulatory sequence of the Boule gene using the bioinformatics method, and detected the methylation status of the Boule gene by sodium bisulfite sequencing.
RESULTSA CpG island was observed in the 5'-flanking regulation region of the Boule gene. The methylation level of the Boule gene was remarkably higher in the SCOS group than in the obstructive azoospermia controls (61.4% vs 21.7%, P<0.01), with significant differences in the methylation levels of 14 CpG sites, namely, -58 bp, -50 bp, -48 bp, -38 bp, -28 bp, -24 bp, -20 bp, -15 bp, -1 bp, +5 bp, +8 bp, +15 bp, +29 bp, and +58 bp.
CONCLUSIONSThe methylation level of the Boule gene is significantly higher in the SCOS patients than in the obstructive azoospermia males, which suggests that the changes in Boule methylation may be associated with spermatogenic dysfunction.
Case-Control Studies ; DNA Methylation ; Humans ; Male ; RNA-Binding Proteins ; genetics ; Sertoli Cell-Only Syndrome ; genetics ; Spermatogenesis ; Testis ; metabolism
7.Obstructive Sleep Apnea Affecting Platelet Reactivity in Patients Undergoing Percutaneous Coronary Intervention.
Xiao-Min JIANG ; Xue-Song QIAN ; Xiao-Fei GAO ; Zhen GE ; Nai-Liang TIAN ; Jing KAN ; Jun-Jie ZHANG
Chinese Medical Journal 2018;131(9):1023-1029
BackgroundThe relationship between obstructive sleep apnea (OSA) and platelet reactivity in patients undergoing percutaneous coronary intervention (PCI) has not been defined. The present prospective, single-center study explored the relationship between platelet reactivity and OSA in patients with PCI.
MethodsA total of 242 patients were finally included in the study. OSA was screened overnight by polysomnography. Platelet reactivity was assessed with a sequential platelet counting method, and the platelet maximum aggregation ratio (MAR) and average aggregation ratio were calculated. All patients were assigned per apnea-hypopnea index (AHI) to non-OSA (n = 128) and OSA (n = 114) groups. The receiver operating characteristic curve analysis was used to evaluate the accuracy of AHI for high platelet reactivity (HPR) on aspirin and clopidogrel, and multivariable logistic regression was used to determine the independent predictors of HPR on aspirin and clopidogrel.
ResultsMedian AHI was significantly higher in the OSA group than in the non-OSA group (34.5 events/h vs. 8.1 events/h, Z = -13.422, P < 0.001). Likewise, median arachidonic acid- and adenosine diphosphate-induced maximum aggregation rate (MAR) in the OSA group was significantly higher than those in the non-OSA group (21.1% vs. 17.7%, Z = -3.525, P < 0.001 and 45.8% vs. 32.2%, Z = -5.708, P < 0.001, respectively). Multivariable logistic regression showed that OSA was the only independent predictor for HPR on aspirin (odds ratio [OR]: 1.055, 95% confidence interval [CI]: 1.033-1.077, P < 0.001) and clopidogrel (OR: 1.036, 95% CI: 1.017-1.056, P < 0.001). The cutoff value of AHI for HPR on aspirin was 45.2 events/h (sensitivity 47.1% and specificity 91.3%), whereas cutoff value of AHI for HPR on clopidogrel was 21.3 events/h (sensitivity 68.3% and specificity 67.7%).
ConclusionPlatelet reactivity appeared to be higher in OSA patients with PCI despite having received a loading dose of aspirin and clopidogrel, and OSA might be an independent predictor of HPR on aspirin and clopidogrel.
Adult ; Aged ; Aged, 80 and over ; Blood Platelets ; physiology ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Percutaneous Coronary Intervention ; Prospective Studies ; Sleep Apnea, Obstructive ; physiopathology ; surgery
8.Effect of thermophilic bacterium HB27 manganese superoxide dismutase in a rat model of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS).
Nai-Wen CHEN ; Jing JIN ; Hong XU ; Xue-Cheng WEI ; Ling-Feng WU ; Wen-Hua XIE ; Yu-Xiang CHENG ; Yi HE ; Jin-Lai GAO
Asian Journal of Andrology 2022;24(3):323-331
We investigated the therapeutic effects of superoxide dismutase (SOD) from thermophilic bacterium HB27 on chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and its underlying mechanisms. A Sprague-Dawley rat model of CP/CPPS was prepared and then administered saline or Thermus thermophilic (Tt)-SOD intragastrically for 4 weeks. Prostate inflammation and fibrosis were analyzed by hematoxylin and eosin staining, and Masson staining. Alanine transaminase (ALT), aspartate transaminase (AST), serum creatinine (CR), and blood urea nitrogen (BUN) levels were assayed for all animals. Enzyme-linked immunosorbent assays (ELISA) were performed to analyze serum cytokine concentrations and tissue levels of malondialdehyde, nitric oxide, SOD, catalase, and glutathione peroxidase. Reactive oxygen species levels were detected using dichlorofluorescein diacetate. The messenger ribonucleic acid (mRNA) expression of tissue cytokines was analyzed by reverse transcription polymerase chain reaction (RT-PCR), and infiltrating inflammatory cells were examined using immunohistochemistry. Nuclear factor-κB (NF-κB) P65, P38, and inhibitor of nuclear factor-κBα (I-κBα) protein levels were determined using western blot. Tt-SOD significantly improved histopathological changes in CP/CPPS, reduced inflammatory cell infiltration and fibrosis, increased pain threshold, and reduced the prostate index. Tt-SOD treatment showed no significant effect on ALT, AST, CR, or BUN levels. Furthermore, Tt-SOD reduced inflammatory cytokine expression in prostate tissue and increased antioxidant capacity. This anti-inflammatory activity correlated with decreases in the abundance of cluster of differentiation 3 (CD3), cluster of differentiation 45 (CD45), and macrophage inflammatory protein 1α (MIP1α) cells. Tt-SOD alleviated inflammation and oxidative stress by reducing NF-κB P65 and P38 protein levels and increasing I-κBα protein levels. These findings support Tt-SOD as a potential drug for CP/CPPS.
Animals
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Chronic Pain
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Cytokines/metabolism*
;
Fibrosis
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Humans
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Inflammation/metabolism*
;
Male
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NF-kappa B/metabolism*
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Pelvic Pain/pathology*
;
Prostatitis/metabolism*
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Rats
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Rats, Sprague-Dawley
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Superoxide Dismutase
;
Syndrome
9.CD7 expression and its prognostic significance in acute myeloid leukemia patients with wild-type or mutant CEBPA.
Ming Yu ZHU ; Ying ZHU ; Rong Rong CHEN ; Li Xia ZHU ; Jing Jing ZHU ; Xue Ying LI ; De ZHOU ; Xiu Di YANG ; Yan Long ZHENG ; Mi Xue XIE ; Jia Nai SUN ; Xian Bo HUANG ; Li LI ; Wan Zhuo XIE ; Xiu Jin YE
Chinese Journal of Hematology 2020;41(2):100-105
Objective: To analyze the prognostic value of CD7 expression in newly diagnosed acute myeloid leukemia (AML) patients, and to further explore the correlation between CD7 expression and CEBPA mutation, and to clarify the prognostic value of CD7(+) in AML patients with wild-type (WT) or mutant-type (MT) CEBPA. Methods: The clinical data of 298 newly diagnosed non-M(3) AML patients between January 2010 and December 2016 were analyzed retrospectively. The clinical characteristics and prognosis of CD7(+) and CD7(-) patients were respectively compared in all patients, and in patients with WT and MT CEBPA. The relationship between CD7 expression and CEBPA mutation was determined by chi-square, and the effects of CEBPA mutation on survival and prognosis in CD7(+) group by Kaplan-Meier method. Results: In CD7(+) group, the frequencies of CEBPA mutation were 10.1% (single site) and 33.9% (double site) , significantly higher than those of the CD7(-) group (5.3% and 4.2%) (P=0.000) . Subgroup prognostic analysis showed a lower CR rate (P=0.001) and a higher RR (P=0.023) in CD7(+) group comparing to those of CD7(-) group in AML patients with wild type CEBPA. There were no statistical difference between CD7(+) group and CD7(-) group in overall survival (OS) and disease free survival (P>0.05) , while in the CEBPA mutant group the CD7(+) group has higher OS (P=0.019) and DFS (P=0.010) . Based on the CD7 expression and CEBPA mutation, 298 cases were divided into 3 subgroups, named as CD7(+)-CEBPA MT group, CD7(-) and CD7(+)-CEBPA WT group. The 3-year OS of the 3 groups were 80.2%, 48.0% and 30.6%, respectively (P<0.001) , and the 3-year DFS were 74.1%, 37.4% and 22.2%, respectively (P<0.001) . Conclusion: The CEBPA mutation rate was higher in CD7(+) AML patients then that of CD7(-) patients. CD7 expression has opposite prognostic significance in AML patients carrying the wild-type or mutant-type CEBPA. Based on CD7 expression and CEBPA mutation, a new risk stratification model can be established, which is helpful to guide the clinical individualized treatment for AML patients.
CCAAT-Enhancer-Binding Proteins/genetics*
;
Disease-Free Survival
;
Humans
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Leukemia, Myeloid, Acute/genetics*
;
Mutation
;
Prognosis
;
Retrospective Studies
10.Efficacy of Moxifloxacin against in Zebrafish Model .
Wen Juan NIE ; Zhong Yao XIE ; Shan GAO ; Tian Lu TENG ; Wen Qiang ZHOU ; Yuan Yuan SHANG ; Wei JING ; Wen Hui SHI ; Qing Feng WANG ; Xue Rui HUANG ; Bao Yun CAI ; Jun WANG ; Jing WANG ; Ru GUO ; Qi Ping GE ; Li Hui NIE ; Xi Qin HAN ; Ya Dong DU ; Nai Hui CHU
Biomedical and Environmental Sciences 2020;33(5):350-358
Objective:
Moxifloxacin (MFX) shows good activity against and can be a possible antibiotic therapy to treat infection; however, other studies have shown a lower or no activity. We aimed to evaluate MFX activity against using zebrafish (ZF) model .
Methods:
A formulation of labeled with CM-Dil was micro-injected into ZF. Survival curves were determined by recording dead ZF every day. ZF were lysed, and colony-forming units (CFUs) were enumerated. Bacteria dissemination and fluorescence intensity in ZF were analyzed. Inhibition rates of MFX and azithromycin (AZM, positive control) were determined and compared.
Results:
Significantly increased survival rate was observed with different AZM concentrations. However, increasing MFX concentration did not result in a significant decrease in ZF survival curve. No significant differences in bacterial burdens by CFU loads were observed between AZM and MFX groups at various concentrations. Bacterial fluorescence intensity in ZF was significantly correlated with AZM concentration. However, with increasing MFX concentration, fluorescence intensity decreased slightly when observed under fluorescence microscope. Transferring rates at various concentrations were comparable between the MFX and AZM groups, with no significant difference.
Conclusion
MFX showed limited efficacy against using ZF model. Its activity needs to be confirmed.
Animals
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Anti-Bacterial Agents
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pharmacology
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Disease Models, Animal
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Moxifloxacin
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pharmacology
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Mycobacterium Infections, Nontuberculous
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drug therapy
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Mycobacterium abscessus
;
drug effects
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Zebrafish