This study was aimed to investigate the effect of STI571, an inhibitor of tyrosine kinase, on the proliferation and apoptosis of chronic myelogenous leukemia (CML) cells as well as expression of anti-apoptotic protein Mcl-1, and to explore the possible role of Mcl-1 in apoptosis-inducing mechanism. K562 cell line was used to observe the effect of STI571 on CML cells. Proliferation and cytotoxicity were analyzed by MTT assay. The apoptotic cells were labelled with Annexin V-FITC and PI and then analyzed by flow cytometry. The expression of apoptotic-related proteins in K562 cells was determined by Western blot with specific antibodies. The results showed that STI571 significantly inhibited the proliferation and induced apoptosis of K562 cells in a dose-and time-dependent manner. Coincidently, the protein phosphorylation on tyrosine residues was reduced and the expressions of anti-apoptotic protein Mcl-1 and Bcl-xl were down-regulated after exposure to STI571. It is concluded that STI571 induces the apoptosis of CML cells by down-regulating the expressions of Mcl-1 and Bcl-xl, which suggests that Mcl-1 and Bcl-xl may play an important role in anti-apoptotic process of CML cells.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Benzamides ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Imatinib Mesylate ; K562 Cells ; Myeloid Cell Leukemia Sequence 1 Protein ; Phosphorylation ; Piperazines ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrimidines ; pharmacology ; bcl-X Protein ; metabolism

Une new flavonoids named as notabilisin K (1), together with four known compounds, morusin (2), mulberrofuran A (3), neocyclomorusin (4) and mornigrol F (5) are separated from 95% ethanol extracts of the twigs of Morus notabilis. Compounds 2-5 are separated from this plant for the first time. Notabilisin I, notabilisin J exhibits certain effect against cells of HCT-116, HepG2 and A2780 with IC50 values ranging from 1.47 μmol x L(-1) to 5.46 μmol x L(-1). Morusin exhibits strong effect against five kinds of human cancer cells (BGC823, A2780, HCT-116, HepG2 and NCI-H1650) with IC50 values ranging from 0.74 μmol x L(-1) to 1.58 μmol x L(-1).
Antineoplastic Agents, Phytogenic ; chemistry ; Benzofurans ; chemistry ; Flavonoids ; chemistry ; Hep G2 Cells ; Humans ; Inhibitory Concentration 50 ; Morus ; chemistry ; Plant Extracts ; chemistry ; Terpenes ; chemistry

24 strains obtained from root nodules of Psoralea corylifolia, Pueraria lobata, and Campylotropis macrocarpa of Yunnan province were studied with numerical taxonomy and 16S rDNA PCR-RFLP. Results of numerical taxonomy indicated that all strains included 10 reference strains were divided into 3 groups at 84% similarity. Group III is an unknown group with no reference strains. Group I is slow-growing kind, and group II fast and middle-slow-grower. The dendrogram derived from 16S rDNA PCR-RFLP showed that all strains divided into five phylogenetic branches at the similarity of 70%. They are branches I and V with no reference strains, Agrobacterium-Sinorhizobium-Rhizobium, Mesorhizobium and Bradyrhizobium. Not all results of numerical taxonomy are accord with 16S rDNA PCR-RFLP, and 2 strains at the same group with A. tumefaciens IAM13129T.

Objective Mutated inbred animal model is introduced to the practical course of genetic diagnosis in the hope that medical students are able to apply what they have learned to clinical cases, based on a deep understanding of principle and technology on gene mutation detection. Methods We integrated DNA extraction, polymerase chain reaction, agarose gel electrophoresis, and gel imaging analysis into a comprehensive experiment and arranged 4-year-programme undergraduates majoring in preclinical medical sciences to conduct it with the purpose of investigating the internal relations between phenotype and genotype in a hairless Uncv mouse model. Subsequently, the questionnaire aimed at evaluating learning effect on the part of students was handed out and their feedbacks were analyzed. Results More than 90% of respondents are satisfied with the general learning effect. Especially, 98. 7% of students support the enhancing effect of the new teaching mode on their research skills and 96% consider the practical course helpful to their problem-solving ability. Conclusions The introduction of mutated inbred animal model to the practical system of molecular diagnostics proves beneficial to boost students' learning effect and scientific research quality. Our practice also provokes thoughts on the further utilization of animal models in teaching system of medical sciences.

OBJECTIVETo study the clinical features, distribution of pathogens, drug susceptibility, and treatment effectiveness in neonates with urinary tract infection (UTI) and admitted to the neonatal intensive care unit (NICU).

METHODSThe clinical data of 229 neonates who developed UTI during their stay in the NICU were retrospectively studied.

RESULTSThe main clinical manifestations of these children included fever/irregular body temperature, refusing to milk feeding, jaundice, vomiting, diarrhea, poor weight gain, and lethargy. The top three pathogens were Escherichia coli, Enterococcus feces, and Klebsiella pneumoniae. Escherichia coli and Klebsiella pneumoniae were highly resistant to ampicillin and most cephalosporins (≥ 85%), and were highly sensitive to imipenem (100%), meropenem (100%), cefoperazone/sulbactam and piperacillin/tazobactam (>90%). Enterococcus feces were highly resistant to penicillin (100%), rifampicin (84%) and gentamicin (79%), but were sensitive to vancomycin.

CONCLUSIONSThe clinical manifestations of neonatal UTI are often atypical and manifested as systemic symptoms. The main pathogenic bacterium is Escherichia coli, and the isolation rate of enterococci can also be high. Most pathogenic bacteria are resistant to penicillin and cephalosporins, and therefore decision-making on drug administration must be based on the results of drug sensitivity tests.

Drug Resistance, Bacterial ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; Microbial Sensitivity Tests ; Retrospective Studies ; Urinary Tract Infections ; drug therapy ; microbiology

OBJECTIVETo observe the ventricular and vascular remodeling reversal effect of Erigeron breviscapus injection (EBI), a protein kinase C inhibitor, in spontaneous hypertension rats (SHR).

METHODSTwenty-four SHR were divided into 4 groups, they were treated respectively with EBI, Fosinopril, Enalapril and normal saline 10 mg/kg per day by intraperitoneal injection for 8 weeks. Systolic blood pressure (SBP), ventricular weight index (VWI) and protein kinase C (PKC) activity in myocardial cells were determined, ultrastructural changes of heart and vessel were observed by polaroscope and transmission electron microscope, and the area and content of myocardial interstitial collagen (MIC) were determined by image analyzer system.

RESULTSThe left ventricular hypertrophy was regressed to certain degree after EBI, Fosinopril and Enalapril treatment, but no significant change in heart rate and right VWI was found. Fosinopril and Enalapril were superior to EBI in lowering SBP and left VWI, and EBI was more obvious in improving myocardial ultrastructure such as hypertrophy and degeneration. All the 3 drugs could improve the MIC and vascular remodeling, the MIC area, content and collagen volume fraction in the EBI group were lowered after treatment, as compared with those in the control group, but comparison between the three groups showed no significant difference. The 3 drugs could reduce the PKC activity in myocardial cell membrane, and EBI showed the effect more significant than that of the other two (P < 0.05).

CONCLUSIONEBI could reverse the myocardial, interstitial and vascular remodeling, improve the rigidness of cardiac muscle, thus, has protective effect on heart.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Heart ; anatomy & histology ; Hypertension ; pathology ; physiopathology ; Injections, Intraperitoneal ; Male ; Myocardium ; ultrastructure ; Organ Size ; Protein Kinase C ; metabolism ; Rats ; Rats, Inbred SHR ; Ventricular Remodeling ; drug effects

BACKGROUNDCollapsin response mediator protein-2 (CRMP2), a multifunctional cytosolic protein highly expressed in the brain, is degraded by calpain following traumatic brain injury (TBI), possibly inhibiting posttraumatic neurite regeneration. Lipid peroxidation (LP) is involved in triggering postinjury CRMP2 proteolysis. We examined the hypothesis that propofol could attenuate LP, calpain-induced CRMP2 degradation, and brain injury after TBI.

METHODSA unilateral moderate controlled cortical impact injury was induced in adult male Sprague-Dawley rats. The animals were randomly divided into seven groups: Sham control group, TBI group, TBI + propofol groups (including propofol 1 h, 2 h, and 4 h groups), TBI + U83836E group and TBI + fat emulsion group. The LP inhibitor U83836E was used as a control to identify that antioxidation partially accounts for the potential neuroprotective effects of propofol. The solvent of propofol, fat emulsion, was used as the vehicle control. Ipsilateral cortex tissues were harvested at 24 h post-TBI. Immunofluorescent staining, Western blot analysis, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling were used to evaluate LP, calpain activity, CRMP2 proteolysis and programmed cell death. The data were statistically analyzed using one-way analysis of variance and a paired t-test.

RESULTSPropofol and U83836E significantly ameliorated the CRMP2 proteolysis. In addition, both propofol and U83836E significantly decreased the ratio of 145-kDa αII-spectrin breakdown products to intact 270-kDa spectrin, the 4-hydroxynonenal expression and programmed cell death in the pericontusional cortex at 24 h after TBI. There was no difference between the TBI group and the fat emulsion group.

CONCLUSIONSThese results demonstrate that propofol postconditioning alleviates calpain-mediated CRMP2 proteolysis and provides neuroprotective effects following moderate TBI potentially by counteracting LP and reducing calpain activation.

Animals ; Blotting, Western ; Brain Injuries ; drug therapy ; metabolism ; Calpain ; metabolism ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Nerve Tissue Proteins ; metabolism ; Propofol ; therapeutic use ; Proteolysis ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the relationship between hepatic lipase activity and fatty degeneration of hepatocytes and explore the effects of tea polyphennols (TP) on the changes of hepatic lipase (HL) activity in rabbits with fatty liver.

METHODSAccording to serum cholesterol and triglyceride (TC) levels, 19 rabbits were divided into fatty liver group (FL, n=6) fed with high cholesterol diet, TP group (n=7) fed with high cholesterol diet and 20mug/g/d tea polyphennols everyday orally, control group (n=6) fed with normal diet. After 8 weeks, the levels of serum TC, HL activity, HL activity and malondildehyde (MDA) in hepatic tissue were detected, and the pathomorphology of hepatic tissue were determined in all rabbits.

RESULTSThe fatty degeneration of hepatocyts in FL group was more severe than that in TP and control group. The serum TC level in TP group (16.87 6.58) mmol/L was higher than that (1.11 0.82) mmol/L in control group (t=5.786, p<0.05), but lower than that (28.49 5.99) mmol/L in FL group (t=3.968, p<0.05). The serum low density lipoprotein-cholesterol level in Tp group (5.10 4.19) mmol/L also higher than that (0.71 1.14) mmol/L in control group (t=3.763, p<0.05), but lower than that (12.15 1.95) mmol/L in FL group (t=2.478, p<0.05). The number of positive dots presenting HL activity level in 100 square micron, hepatic tissue in TP group (3.24 0.17) was higher than that (1.76 0.10) in FL group (t=-3.153, p<0.05), but lower than that (4.14 0.05) in control group (t=-2.902, p<0.05). The levels of MDA in hepatic tissue in TP group (44.66 26.18) nmol/mg was significantly lower than that (75.58 29.88) nmol/mg in FL group (t=2.261, p<0.05), but no evidently different from that (43.64 16.95) nmol/mg in control group. The plasma HL activity was no difference among the three groups.

CONCLUSIONThe HL activity in hepatic tissue with fatty degeneration of hepatocytes was lower than that in normal liver. Tea polyphennols can increase HL activity in hepatic tissue and protect hepatocytes from fatty degeneration.

Animals ; Fatty Liver ; blood ; enzymology ; pathology ; Flavonoids ; Lipase ; metabolism ; Lipids ; blood ; Liver ; enzymology ; metabolism ; pathology ; Male ; Malondialdehyde ; analysis ; Phenols ; pharmacology ; Polymers ; pharmacology ; Polyphenols ; Rabbits ; Tea

OBJECTIVETo investigate the expression of Rab5a and APPL1 in breast cancer and fibroma, and analyze their correlation with HER-2 expression, metastasis and development of breast cancer.

METHODSRab5a and APPL1 in paraffin embedded tissues of 74 breast carcinomas and 40 breast fibromas were detected by immunohistochemistry. Their relationship with metastasis, pathological grade, and HER-2 expression in breast cancer was determined by statistical analysis.

RESULTSThere was no expression or low expression of Rab5a and APPL1 in the breast fibroma, but the positive expression rate of Rab5a and APPL1 in the breast carcinomas were 91.9% and 83.8%, respectively. No significant difference in expression of Rab5a and APPL1 was found between metastatic and non-metastatic groups, and pathological grade I/II and grade III groups. But Rab5a was overexpressed in HER-2-positive group compared with that in the HER-2-negative group.

CONCLUSIONSRab5a and APPL1 are overexpressed in breast cancer, and are positively correlated with the HER-2 expression. These proteins may influence the growth and proliferation of breast cancer cells by HER-2 signal transduction.

Adaptor Proteins, Signal Transducing ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Fibroma ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Grading ; Paraffin Embedding ; Receptor, ErbB-2 ; metabolism ; Young Adult ; rab5 GTP-Binding Proteins ; metabolism

OBJECTIVETo observe the laryngopharynx manifestation and electromyography characteristics of myasthenia gravis (MG) patients.

METHODSThirty cases of MG were included in this study, their laryngopharynx symptoms and signs, voice acoustic assessment, laryngeal electromyography (LEMG) behaviors and repetitive nerve stimulation test(RNS) were analyzed, and the data was compared with that of normal subjects.

RESULTSAbout 36.7% of MG patients (11/30) had the symptoms of hoarseness, voice fatigue, dysphonia and dysphagia. The vocal folds movements of 16.7% of MG patients(5/30) appeared weaker than normal, and their vocal glottic couldn't close completely, while with a seam during phonation. Voice amplitude (68.3 +/- 14.6) dB (x +/- s, same at below), and maximum phonation time (15.1 +/- 4.0) s, were greatly lower than normal; shimmer(2. 43 +/- 1.19)%, and normalized noise energy (-9.6 +/- 3.3) dB, were greatly higher than normal. The amplitudes of interference patterns in MG patients' LEMG markedly decreased, except introarytenoid muscle, during low, normal and high pitch phonation, the amplitudes of thyoiarytenoid muscle were (215 +/- 69) microV, (298 +/- 113) microV and (380 +/- 153) microV, those of cricoarytenoid muscle were (253 +/- 92) microV, (361 +/- 116) microV and (486 +/- 155) RV. The turns increased but had no statistical difference. In the RNS test, 83.3% MG patients (25/30) showed masculine response. There were about 2.20 +/- 1.32 pieces of laryngeal muscles involved, and the reduction rate in amplitude of the compound muscle action potential for RNS was about (27.9 +/- 19.2)%.

CONCLUSIONSOnly parts of MG patients had laryngopharyngeal symptoms, but the laryngeal muscles of most of them were involved, appearing as the masculine response for RNS, the decreased synchronization of the laryngeal muscles' interference patterns, the decreased capacity of phonation. MG must be differentiated when a patient has the symptoms of voice weakness, hoarseness and dysphonia. Laryngeal RNS test should be used in the early diagnosis of MG.

Adolescent ; Adult ; Aged ; Articulation Disorders ; Case-Control Studies ; Electromyography ; Female ; Humans ; Hypopharynx ; physiopathology ; Laryngeal Muscles ; physiopathology ; Male ; Middle Aged ; Myasthenia Gravis ; physiopathology ; Young Adult