To reveal the underlying mechanism of doxorubicin induced hepatotoxicity, an NMR-based metabolomic approach combined with multivariate statistical analysis was used to observe its metabolic alternations of rat liver. Sixteen differential metabolites between model rats and normal rats were characterized as potential pathological biomarkers related to doxorubicin induced hepatotoxicity. Six pathways, including phenylalanine, tyrosine and tryptophan biosynthesis, valine, leucine and isoleucine biosynthesis, phenylalanine metabolism, glycine, serine and threonine metabolism, alanine, aspartate and glutamate metabolism, and tyrosine metabolism were regarded as the targeted metabolic pathways according to Metabolic Pathway Analysis (MetPA). The results suggested that the metabolic perturbations in rats with doxorubicin induced hepatotoxicity were mainly involved in amino acid metabolism, lipid pathways, purine metabolism, energy metabolism, dysfunction of biotransformation and oxidative stress. The investigation revealed the effects of doxorubicin on liver in a holistic metabolic way, which laid a foundation for further studies on its toxicity mechanism.
Animals ; Biomarkers ; metabolism ; Doxorubicin ; toxicity ; Energy Metabolism ; Liver ; drug effects ; metabolism ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Metabolic Networks and Pathways ; Metabolomics ; Multivariate Analysis ; Oxidative Stress ; Rats

Objective To analyze the relations between medical students' burnout and some related factors for providing some suggestions for educational practice. Methods A total of 80 medical students were investigated and assessed by organizational justice questionnaire and Maslach Burnout Inventory-General Survey(MBI-GS). Results Some subjects were found to experience job burnout.Means of total distributive justice were significantly lower than medians.Means of participation and complaint mechanism were not discrete to medians.Means of motional exhaustion and cynicism were significantly lower than medians.Means of reduced professional efficacy were significantly higher than medians.Conclusion Job burnout existed among interns from non-211 and 211 Project universities.Medical students should establish scientific view of profession through psychological lessons,lectures,simulated diagnosis,clinical practice and self-sentience.

The global regulatory gene, nsdA, negatively regulates antibiotics production in Streptomyces coelicolor. Southern blot experiment, using an nsdA fragment of S. coelicolor as probe, indicated that nsdA gene existed in many Streptomyces. Primers were designed based on the published sequences of S. coelicolor and S. avermitilis. PCR amplification and sequencing showed that nsdA in Streptomyces was conservative and that of S. lividans ZX64 has a 100% identity in the nucleotide sequence comparing with that of S. coelicolor A3 (2). The nsdA disrupted mutant of S. lividans was constructed named as WQ2. WQ2 was able to produce actinorhodin but the wild-type strain ZX64 did not, which has a silent gene cluster contributing to the biosynthesis of actinorhodin. However, the ability was lost when another copy of the wild nsdA gene was introduced into WQ2. All the results above indicate that nsdA homologous gene is wildly existent and conserved in Streptomyces. And it plays a role in negatively regulating the actinorhodin synthesis in S. lividans and disruption of it can activate the silent gene cluster.
Anti-Bacterial Agents ; biosynthesis ; Blotting, Southern ; Genes, Bacterial ; physiology ; Genes, Regulator ; physiology ; Multigene Family ; Streptomyces lividans ; genetics

The aim of the paper is to prepare stable antisense oligodeoxynucleotides-loaded cationic liposomes and evaluate the transfection efficiency of asODN to MCF-7 oophoroma cells and study their distribution to different tissues in mice. Antisense oligodeoxynucleotides (asODN)-loaded cationic liposomes were prepared by a thin film-adsorption-lyophilization method which is simple and can overcome crucial pharmaceutical defects (e.g. instability) of liposomes during storage. The morphology was investigated by transmission electron microscope. The size and surface charge of the liposomes were determined by laser particle analyter. The dissociated ligodeoxynucleotides were separated from the liposomes by sephadex column and the entrapment efficiency was determined by using an ultraviolet photometer. Trehalose, mannitol, and glycine were suitable for lyophilization especially trehalose. The resulting liposomes were global microcapsule in a narrow particle size with a mean diameter of 175 nm and 320 nm before and after lyophilization, and a high zeta potentials of +32 mV. The dissociated asODN were separated from the liposomes by sephadex G-50 column and the entrapment coefficient of asODN was 88.4% pre and 83.2% post-lyophilization separately for trehalose. The growth of MCF-7 oophoroma cells were inhibited in vitro obviously (P < 0.05) and transfection efficiency of asODN was 18%, 26%, 44% after 2 h, 4 h and 8 h, respectively. The formulation and method can be used to prepare stable cationic liposomes which can effectively inhibit the growth of MCF-7 oophoroma cells and obtain a high transfection efficiency. This system can improve distribution amount of asODN to tissues especially tumors in mice.
Animals ; Breast Neoplasms ; metabolism ; pathology ; Cations ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Carriers ; Female ; Freeze Drying ; Humans ; Liposomes ; chemistry ; pharmacokinetics ; pharmacology ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Oligodeoxyribonucleotides, Antisense ; chemistry ; genetics ; Particle Size ; Transfection

This study aimed to explore the outcomes of progestin-primed ovarian stimulation protocol (PPOS) in aged infertile women who failed to get pregnant in the first IVF/ICSI-ET cycles with GnRH-a long protocol.A self-controlled study was conducted to retrospectively investigate the clinical outcomes of 104 aged infertile patients who didn't get pregnant in the first IVF/ICSI-ET treatment by stimulating with GnRH-a long protocol (non-PPOS group),and underwent PPOS protocol (PPOS group) in the second cycle between January 2016 and December 2016 in the Center for Reproductive Medicine,Renmin Hospital of Wuhan University.The primary outcomes included clinical pregnancy rate of frozen-thawed embryos transfer (FET) in PPOS group,and good-quality embryo rate in both groups.The secondary outcomes were fertilization rate,egg utilization rate and cycle cancellation rate.The results showed that there were no significant differences in basal follicle stimulating hormone (bFSH),antral follicle count (AFC),duration and total dosage of gonadotropin (Gn),number of oocytes retrieved,intracytoplasmic sperm injection (ICSI) rate,fertilization rate,and cycle cancellation rate between the two groups (P＞0.05).However,the oocyte utilization rate and good-quality embryo rate in PPOS group were significantly higher than those in non-PPOS group (P＜0.05).By the end of April 2017,62 FET cycles were conducted in PPOS group.The clinical pregnancy rate and embryo implantation rate were 22.58％ and 12.70％,respectively.In conclusion,PPOS protocol may provide better clinical outcomes by improving the oocyte utilization rate and good-quality embryo rate for aged infertile patients who failed to get pregnant in the first IVF/ICSI-ET cycles.

OBJECTIVETo investigate the possible relationship between variation of coxsackievirus B3 (CoxB3) VP1 sequence from cerebrospinal fluid of children with severe and mild central nervous system (CNS) infection and damage to CNS in children from Shandong province.

METHODSThe enteroviruses were detected using VP1 typing and sequencing primer for enteroviruses from 73 enterovirus-infected cases confirmed by detection of cerebrospinal fluid by enteroviruses common primer. VP1 sequences (450 nucleotides) were determined and analyzed for 21 CoxB3 enteroviruses strains isolated in Qingdao and Binzhou, and were compared with that of BLAST search procedures from GeneBank in NCBI. The variation of VP1 gene and amino acids sequence of CoxB3 enteroviruses was analyzed for severe and mild CNS infection.

RESULTSThe nucleotide homogeneity of these CoxB3 appeared to be 97% - 99%, however, the homogeneity among different genotypes were 83% - 76%. Replacement of glutamine by histidine at amino acid locus 856 of VP1 CoxB3 was found in 4 cases with severe encephalitis. There were different variation in VP1 nucleotide sequence of CoxB3 in 3 cases with mild encephalitis and 14 cases with meningitis, but amino acids sequences had no regular variation. The modified Glasgow's coma score was below 7 in all the 4 cases with severe encephalitis. Of these 4 cases, 3 had consciousness disturbance for less than 3 days. Lethargy, restlessness and psychiatric symptoms were major manifestations, of whom 3 also had dysphagia, 1 had encephalatrophy obviously, Glasgow's coma score was 3, deep coma lasted for 9 days, and had concomitant fatal epileptic attacks. Of these 4 cases, 2 completely recovered, 1 had high muscle tone, 1 remained under anti-epileptic drug treatment at follow-up 6 months later.

CONCLUSIONThere were a small epidemic of CoxB3 CNS infection in children in 2005 in this area. The amino acid variation of CoxB3 VP1 possibly caused increased viral virulence and caused damage to CNS.

Amino Acid Sequence ; Base Sequence ; Capsid Proteins ; cerebrospinal fluid ; genetics ; Central Nervous System ; pathology ; virology ; Child ; Coxsackievirus Infections ; cerebrospinal fluid ; epidemiology ; virology ; Encephalitis ; virology ; Enterovirus B, Human ; genetics ; pathogenicity ; Female ; Humans ; Male ; Molecular Sequence Data ; RNA, Viral ; genetics ; Virulence

OBJECTIVEThe aim of this study was to explore the possibility of creating a toxin, C-CPE-ETA', by fusing C-terminal high affinity binding domain of CPE (C-CPE) with a truncated form of Pseudomonas aeruginosa exotoxin A (ETA') and to examine whether C-CPE-ETA' could specifically target CLDN-3, 4 molecule and the targeted toxin was cytotoxic against CLDN-3,4-overexpressing ovarian cancer.

METHODSCLDN-3 and CLDN-4 expressions were analyzed at the mRNA level in three ovarian cancer cell lines and epithelial ovarian cancer tissues from 20 patients. After transforming an expression plasmid of C-CPE-ETA' into E. coli BL21 (DE3) plysS strain, the recombinant protein was purified using His-Bind resin chromatography column and analyzed by Western blot and Coomassie blue staining. The specific binding, proapoptotic and cytolytic activities were evaluated by flow cytometry, fluorescence microscopy with the JC-1 probe and MTT assay in CLDN-3,4-overexpressing ovarian cancer cells.

RESULTSQuantitive RT-PCR results showed there existed high levels of CLDN-3 and CLDN-4 in ovarian cancer cells, CAOV3, OVCAR3 and SKOV3. Moreover, high expressions of CLDN-3 and CLDN-4 were observed in 90.0% (18/20) and 60.0% (12/20) of ovarian cancer tissues, with an expression level 10-fold higher than that in the normal ovarian tissue. A 58 000 recombinant protein C-CPE-ETA' was demonstrated by Western blot and Coomassie blue staining. Purified and recombinant C-CPE-ETA' was bound with high affinity to CLDN-3,4-overexpressing ovarian cancer cells, CAOV3, OVCAR3 and SKOV3 cells. C-CPE-ETA' was strongly proapoptotic and cytotoxic towards the CLDN-3,4-overexpressing ovarian cancer cells. The concentration of IC(50) was 7.364 ng/ml for CAOV3 cells, 8.110 ng/ml for OVCAR3 cells and 22.340 ng/ml for SKOV3 cells, respectively. However, control CLDN-3,4-deficient cell line HUVEC was not susceptible to the recombinant C-CPE-ETA' at a concentration up to 10 µg/ml.

CONCLUSIONSThe C-CPE-ETA' protein exhibits remarkably specific cytotoxicity for CLDN-3,4-overexpressing ovarian cancer cells. Its therapeutic potential warrants further development for ovarian cancer molecular targeted therapy.

ADP Ribose Transferases ; metabolism ; physiology ; Apoptosis ; Bacterial Toxins ; metabolism ; Cell Line, Tumor ; Claudin-3 ; Claudin-4 ; Claudins ; genetics ; metabolism ; Enterotoxins ; metabolism ; physiology ; Exotoxins ; metabolism ; physiology ; Female ; Humans ; Immunotoxins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Recombinant Fusion Proteins ; metabolism ; physiology ; Virulence Factors ; metabolism ; physiology

OBJECTIVETo analyze the clinical features, renal damage and prognosis of primary vesicoureteric reflux (VUR) in children, hoping to give more attention to early recognition and treatment of the disease.

METHODSBetween June 1995 and December 2006, 974 patient were admitted in our hospital because of urinary tract infection (UTI), 139 primary VUR children were enrolled in the retrospective study. VUR grades, renal scar development, renal ultrasound and urinalysis were evaluated.

RESULTSThe incidence of VUR in UTI children was 14.3% (139/974), however, the incidence in infants and toddlers (younger than two years of age) was 17.2% (79/458), which was greatly higher than that in school children. Of the 139 children, 79(56.8%) were younger than two years and the number of boys was much larger than that of girls (P = 0.001). Bilateral reflux was found in 69 cases, unilateral in 70 cases; the percentages of mild reflux (grade I-II) was 19.7% (41/208), moderate reflux (grade III) was 35.6% (74/208), severe reflux (grade IV-V) was 44.7% (93/208). Dimercaptosuccinic acid (DMSA) scanning was performed for 135 patients, and renal scar was found in 37% (50/135), and the rate of scar in infants was the highest (42.4%), and 30 (60%) patients with renal scar were younger than two years of age. Among them five patients were found to have renal scar during the follow up period, four of them were younger than two years. The rate of renal scar in moderate and severe reflux was much higher than that in mild reflux cases, the risk of occurring renal scar would increase with its severity of reflux (P < 0.001). The sensitivity and specificity of renal ultrasound in suggesting VUR were 24.8% and 94.3%, respectively. The positive outcome of urinary N-acetyl-beta-glucosaminidase was a bit high, but there were no associations between renal scar and urinary microprotein (P > 0.05). Thirty-one cases were followed up for long time (> 1 year); and 90% of urinary infection was controlled and in 44.4% of patients VUR disappeared with medical treatment. No child had decreasing renal function in follow-up period.

CONCLUSIONChildren with primary VUR need early diagnosis and treatment. Insisting on normal therapy and long-term follow-up will protect renal function effectively.

Child ; Child, Preschool ; Cicatrix ; Female ; Humans ; Infant ; Kidney ; pathology ; Male ; Retrospective Studies ; Ultrasonography ; Urinary Tract Infections ; epidemiology ; Vesico-Ureteral Reflux ; diagnostic imaging ; epidemiology ; pathology

The present study is to determine the flavonoid glycosides, terpene lactones, biflavones, gingko acid and procyanidins of ginkgo pollen. UPLC-TQ-MS technology was used for the determination of 24 kinds of resource chemical composition in ginkgo pollen qualitatively and quantitatively. The results shows that the contents of rutin, quercetion 3-O-[4-O-(α-L-rhamnosyl )-β-D-glucoside] and kaempferolis were 120.9, 114.0, 222.1 μg x g(-1). In this paper, the contents of 24 kinds of chemical components of ginkgo pollen were determinated by UPLC-TQ-MS for the first time. This method is simple and quick, which will be benefit for recycling utilization of ginkgo pollen.
Chromatography, High Pressure Liquid ; methods ; Flavonoids ; analysis ; Ginkgo biloba ; chemistry ; Mass Spectrometry ; Pollen ; chemistry ; Proanthocyanidins ; analysis ; Rutin ; analysis ; Terpenes ; analysis

Objective: To investigate the effects of hyaluroni dase (HAase) and hyaluronan (HA) on proliferation of vascular endothelial cells and its mechanism. Methods: The cultured bovine aortic endothel ial cells (BAEC) were treated with HAase or HA. Cell proliferation rate was dete cted by MTT assay. The expression of CD44 and DNA content of the cells were meas ured by flow cytometry (FCM). Results: HAase (50 μg/ml) stimula ted cell proliferation ［(50.10±1.23）% vs control, P<0.01］, incre ased S phase cell rate and induced the expression of CD44, but HA (100 μg/ml) i nhibited cell proliferation and the expression of CD44. Conclusion: HAase may degrade antiangiogenic HA of extracellular matrix, which may stim ulate proliferation of endothelial cells and enhance the curative effect of grow th factors to myocardial ischemia.