Liver toxicity of macrolide antibiotics in zebrafish.

Macrolide antibiotics (macrolides) are among the most commonly prescribed antibiotics worldwide and are used for a wide range of infections, but macrolides also expose people to the risk of adverse events include hepatotoxicity. Here, we report the liver toxicity of macrolides with different structures in zebrafish. The absorption, distribution, metabolism, excretion and toxicology (ADMET) parameters of macrolide compounds were predicted and contrasted by utilizing in silico analysis. Fluorescence imaging and Oil Red O stain assays showed all the tested macrolide drugs induced liver degeneration, changed liver size and liver steatosis in larval zebrafish. Through RNA-seq analysis, we found seven co-regulated differentially expressed genes (co-DEGs) associated with metabolism, apoptosis and immune system biological processes, and two co-regulated significant pathways including amino sugar and nucleotide sugar metabolism and apoptosis signaling pathway. We found that only fosab of seven co-DEGs was in the two co-regulated significant pathways. fosab encoded proto-oncogene c-Fos, which was closely associated with liver diseases. The whole-mount in situ hybridization showed high transcription of c-Fos induced by macrolide compounds mainly in the liver region of zebrafish larvae. Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) leakage assays revealed that macrolides exerts significant cytotoxic effects on L02 cells. qRT-PCR and western blot analysis demonstrated macrolides also promoted human c-Fos expression in L02 cells. The c-Fos overexpression significantly reduced cell viability by using CCK-8 assay. These data indicate that hepatotoxicity induced by macrolides may be correlated with c-Fos expression activated by these compounds. This study may provide a biomarker for the further investigations on the mechanism of hepatotoxicity induced by macrolide drugs with different structures, and extend our understanding for improving rational clinical application of macrolides.

Cardiotoxicity evaluation of anthracyclines in zebrafish (Danio rerio).

Drug-induced cardiotoxicity is a leading factor for drug withdrawals, and limits drug efficacy and clinical use. Therefore, new alternative animal models and methods for drug safety evaluation have been given great attention. Anthracyclines (ANTs) are widely prescribed anticancer agents that have a cumulative dose relationship with cardiotoxicity. We performed experiments to study the toxicity of ANTs in early developing zebrafish embryos, especially their effects on the heart. LC50 values for daunorubicin, pirarubicin, doxorubicin (DOX), epirubicin and DOX-liposome at 72 h post-fertilization were 122.7 µM, 111.9 µM, 31.2 µM, 108.3 µM and 55.8 µM, respectively. At the same time, zebrafish embryos were exposed to ANTs in three exposure stages and induced incomplete looping of the heart tube, pericardia edema and bradycardia in a dose-dependent manner, eventually leading to death. DOX caused the greatest heart defects in the treatment stages and its liposome reduced the effects on the heart, while daunorubicin produced the least toxicity. Genes and proteins related to heart development were also identified to be sensitive to ANT exposure and downregulated by ANTs. It revealed ANTs could disturb the heart formation and development. ANTs induced cardiotoxicity in zebrafish has similar effects in mammalian models, indicating that zebrafish may have a potential value for assessment of drug-induced developmental cardiotoxicity.

Establishment of an HPLC identification system for detection of counterfeit steroidal drugs.

A set of simple HPLC methods employing UV detection were developed for detection of counterfeit drugs by the qualitative and quantitative analysis of nine steroidal drugs, ethinylestradiol, diethylstilbestrol, norethisterone, norgestrel, methyltestosterone, medroxyprogesterone acetate, progesterone, testosterone propionate and nilestriol. The methods were based on studies of the relationships between the retention factors (k) of the nine compounds and the percentages of water to methanol in the mobile phases on a reverse phase Alltima C(18) column giving reliable separation of the compounds under three sets of chromatographic conditions. The methods were validated using statistical tests and were used on nine commercial samples for detection of possible counterfeit drugs.

[Antimicrobial resistance of Escherichia coli isolates collected from inpatients and outpatients].

OBJECTIVE: To investigate the antimicrobial resistance of Escherichia coli (E. coli) isolates collected from the inpatients in the departments of medicine, surgery, and pediatrics, and intensive care unit (ICU), and from the outpatients. METHODS: Disc diffusion test was used to study the antimicrobial resistance of 3909 strains of E. coli collected from the inpatients in the departments of medicine, surgery, and pediatrics, and Intensive care unit (ICU), and from the outpatients, mostly isolated from urine, sputum, blood, and different secreta in the year 2001. WHONET 5 software was used for analysis of the antimicrobial resistance; and significant differences were tested by chi(2) to compare the resistance rates to antibiotics. RESULTS: The incidences of extended-spectrum beta-lactamases producing strains were 11.2% (195/1737), 14.3% (141/983), 17.7% (28/158), 19.7% (24/122) and 8.4% (76/909) in the strains of E. coli isolated from the inpatients in the departments of medicine, surgery, and pediatrics, and ICU and from the outpatients respectively, with a detectable rate among the outpatients significantly lower than that among the inpatients (P < 0.005), and a detectable rate among the inpatients in the department of medicine significantly lower than those among the inpatients in the department of pediatrics and ICU (both P < 0.05). The resistance rates to cefazolin, cefotaxime, gentamicin and aztreonam of the isolates from the outpatients were significantly lower than those of the inpatients (all P < 0.05). The resistance rates to amoxicillin/clavulanic acid, ceftazidime, cefepime and amikacin of the isolates from the outpatients were significantly lower than those of the inpatients in the department of surgery and ICU (all P < 0.05); The resistance rates to ciprofloxacin and trimethoprim/sulfamethoxazole of the isolates from the outpatients were significantly lower than those from the ICU patients (both P < 0.05). The resistance rates to cefazolin and cefotaxime of E. coli isolates collected from the inpatients in the department of medicine were significantly lower than those of the isolates from the department of surgery and ICU (all P < 0.01); the resistance rates to gentamicin of the isolates from the department was significantly lower than that of the isolates from the department of surgery (P < 0.05). The resistance rates to amoxicillin/clavulanic acid and aztreonam of the isolates from the department of medicine were significantly lower than those of the isolates from the ICU (both P < 0.05). The resistance rate to ciprofloxacin of isolates from the inpatients in the department of pediatrics was significantly lower than that of the other isolates (all P < 0.01). CONCLUSIONS: It is of guiding significance for empirical use of antimicrobial agents in clinic to study on the resistant rates of the strains of E. coli isolated from different departments in hospital.

[Antimicrobial resistance of Staphylococcus aureus isolates from inpatients of departments of internal medicine, surgery, and pediatrics and intensive care unit].

OBJECTIVE: To investigate the antimicrobial resistance of Staphylococcus aureus isolates obtained from inpatients of departments of internal medicine, surgery, and pediatrics and intensive care unit (ICU), and study on the differences of resistant rates among the clinical isolates. METHODS: The strains of Staphylococcus aureus were cultured and their antimicrobial resistance was assayed by disc diffusion test (K-B method) and the data was analyzed by WHO NET 5 software. chi(2) test was made to identify the significance of difference. RESULTS: 2 625 strains of Staphylococcus aureus were obtained from the clinical departments of 60 hospitals all over China, among which 1 669 strains (63.3%) were obtained from the inpatients of departments of internal medicine, surgery, and pediatrics and ICU from 1 January to 31 December 2001. Most of the strains were isolated from sputum (34.3%, 572/1 669), secretion (13.2%, 221/1 669), pus (11.7%, 195/1 669), blood (10.2%, 171/1 669), and wound (5.6%, 94/1 669). The resistant rates of Staphylococcus aureus isolates from the inpatients in department pediatrics of to oxacillin, gentamicin, clindamycin, ciprofloxacin, levofloxacin, and chloramphenicol were 23.3%, 16.1%, 29.3%, 11.2%, 4.0% and 14.4% respectively, all significantly lower than those of the isolates from the inpatients of the departments of internal medicine and surgery, and ICU (all P < 0.001). The resistant rates of Staphylococcus aureus isolates from the inpatients of departments of internal medicine and surgery to oxacillin, gentamicin, clindamycin, ciprofloxacin, and levofloxacin were significantly lower than those of isolates from the patients of ICU (all P < 0.001). CONCLUSION: A high isolation rate of multi-drug resistant Staphylococcus aureus among the inpatients of departments of internal medicine, surgery, and pediatrics and ICU. It is important to select appropriate antimicrobial agents based on the origin of bacterial strains and avoid blindness before the results of drug sensitivity are obtained.