This investigation is to explore the feasibility of applying reverse docking method to the selectivity studies of protein kinase inhibitors. Firstly, a database that consists of 422 protein kinase structures was established through collecting the reported crystal structures or homology modeling. Then a reverse docking based method of protein kinase target screening was established, followed by the optimization of related parameters and scoring functions. Finally, seven typical selective kinase inhibitors were used to test the established method. The results show that the selective targets of these inhibitors have relatively high scoring function values (ranking in the first 35% of the tested kinase targets according to the scoring function values). This implies that the reverse docking method can be applied to the virtual screening of kinase targets and further to the selectivity studies of protein kinase inhibitors.
Alternative Splicing ; Drug Delivery Systems ; Drug Evaluation, Preclinical ; methods ; Gene Targeting ; Models, Molecular ; Protein Binding ; Protein Kinase Inhibitors ; chemistry

OBJECTIVETo study the efficacy of erythropoietinin (EPO) in the treatment of moderate or severe hypoxic-ischemic encephalopathy (HIE) in neonates.

METHODSSeventy neonates with moderate or severe HIE were randomly assigned to two groups: EPO treatment and control (n=35 each). The EPO treatment group included 22 cases of moderate HIE and 13 cases of severe HIE. The control group included 24 cases of moderate HIE and 11 cases of severe HIE. Thirty-five healthy full-term infants served as normal group. The control group received a conventional treatment. Beside the conventional treatment, the EPO treatment group was intravenously injected with EPO of 200 IU/kg•d, 3 times weekly. Routine blood test was performed every 6 days. EPO dose was adjusted based on the results of the routine blood test. The course of EPO treatment was 2 to 4 weeks. Neonatal Behavioral Neurological Assessment (NBNA) was performed at age of 28 days. The infant development test of Child Development Centre of China (CDCC) was performed at ages of 3 months and 6 months.

RESULTSThe percentage of normal NBNA scores in the EPO treatment group was significantly higher than that in the control group at age of 28 days (P<0.05), but was significantly lower than that in the normal group (P<0.01). The CDCC test including physical development index (PDI) and physical development index (MDI) showed the percentage of normal results in the EPO treatment group was significantly higher than in the control group at age of 3 months (P<0.05), but was significantly lower than in the normal group (P<0.01). The CDCC test including PDI and MDI showed that the percentage of normal results in the EPO treatment group was significantly higher than in the control group at age of 6 months. The MDI test results in the EPO treatment group were not significantly different from those in the normal group at age of 6 months, but the percentage of normal results in the PDI test in the EPO treatment group was still significantly lower than that in the normal group (P<0.05).

CONCLUSIONSEPO treatment has neuroprotective effects against moderate or severe HIE and improves long-term behavioral neurological developments in neonates.

Child Development ; Erythropoietin ; therapeutic use ; Female ; Humans ; Hypoxia-Ischemia, Brain ; drug therapy ; psychology ; Infant Behavior ; Infant, Newborn ; Male

OBJECTIVETo investigate the clinical and pathological features of progressive muscular dystrophy (PMD) in children and to provide help for the early and accurate diagnosis of PMD.

METHODSRetrospective analysis was performed on the clinical data of 99 hospitalized children with PMD, including clinical manifestations, age of onset, family history, creatase, electromyogram (EMG) and pathological changes of muscles.

RESULTSOf the 99 children with PMD, the age of onset was 0.5-14.5 (4.7 ± 3.1) years. Eleven cases (11%) had a family history of PMD. Twenty-six (26%) were misdiagnosed as other diseases. All patients presented with muscle weakness when seeing the doctor, and 66 (67%) of them had muscle atrophy and/or hypertrophy. All patients had elevated creatine kinase (CK) levels. The 2-7-year-old group (n=51) had a mean CK level of 9965 ± 8876 U/L, and the 7-15-year-old group (n=48) had a mean CK level of 5110 ± 4498 U/L, with a significant difference between the two groups (P<0.01). The EMG examination performed on 66 patients showed that 54 cases (82%) had myogenic damage and 10 cases (15%) had neurogenic damage. Light microscopy revealed coexistence of atrophy and hypertrophy of muscle fibers, hyaline degeneration and granular degeneration. Electron microscopy showed that muscle fibers were different in thickness, some atrophic or hypertrophic; muscle cell nuclei moved inwardly, myofilaments dissolved and disappeared mildly under the sarcolemma, there were scattered melting lesions within muscle fibers, the numbers of glycogen granules and mitochondria increased, mild hyperplasia and expansion of sarcoplasmic reticulum were seen, and a small number of muscle fibers had necrosis.

CONCLUSIONSWeakness of both lower extremities remains the main reason for PMD patients seeing the doctor. CK is the main laboratory indicator for diagnosis of PMD. PMD is mainly manifested as myogenic damage in the early stage and may be accompanied by neurogenic damage in the late stage, according to the EMG examination. With a high misdiagnosis rate, PMD may be misdiagnosed as many other diseases. Pathological examination under light microscope and electron microscope is the main means for confirming a PMD diagnosis.

Adolescent ; Child ; Child, Preschool ; Creatine Kinase ; blood ; Electromyography ; Female ; Humans ; Male ; Muscle, Skeletal ; pathology ; Muscular Dystrophies ; pathology ; physiopathology ; Retrospective Studies

OBJECTIVETo preliminarily investigate the mechanism of small interfering RNA (siRNA) induced apoptosis in glioma U251 cells by silencing basic fibroblast growth factor (bFGF).

METHODSU251 cells were divided into the normal control group, the mock group and experiment group, the mock and experiment group were transfected with mock vector (Ad-null) and the recombinant adenovirus carrying bFGF-siRNA (Ad-bFGF-siRNA) respectively at a multiplicity of infection (MOI) of 100. After 72 hours, the expression of related proteins was revealed by the method of Western blot. Mitochondrial transmembrane potential (ΔΨm) was measured with flow cytometry and confocal microscopy, Groups were compared using single factor analysis of variance (One-way ANOVA).

RESULTSAfter U251 cells were transfected with bFGF-siRNA, the results of Western blot showed that after 72 hours of transfection the bFGF protein in the experiment group decreased obviously, meanwhile Cytochrome C, Caspase-3 and Bax showed increased expression while in the Bcl-xl and Bcl-2 proteins decreased expression. The proportion of high mitochondrial membrane potential of cells by flow cytometry, the experimental group was 74.4% ± 4.7% decreased significantly compared with the control group 92.1% ± 2.5%, the mock group 90.9% ± 1.8% (F = 28.805, P < 0.05); laser scanning confocal microscopy results showed that the red fluorescence and green fluorescence ratio of the experimental group was 0.83 ± 0.12 decreased significantly compared with 1.36 ± 0.40 of the control group and 1.32 ± 0.35 of the mock group(F = 7.920, P < 0.05).

CONCLUSIONsiRNA targeting bFGF induced U251 cell apoptosis may be achieved through the mitochondrial pathway.

Adenoviridae ; genetics ; Apoptosis ; Cell Line, Tumor ; Fibroblast Growth Factor 2 ; genetics ; Glioma ; pathology ; Humans ; RNA Interference ; RNA, Small Interfering ; Transfection

OBJECTIVETo study the anti-glioma effect of recombinant adenovirus mediated combined gene therapy of bFGF-siRNA and HIV1-Vpr in vivo.

METHODSMouse glioma model was established by injecting 5 × 10(6) LN229 cells into BALB/c-nu nude mice. 30 nude mice were randomly divided into 5 groups: the negative control group, mock group, bFGF-siRNA group, Vpr group and combined therapy group, which at regular intervals were injected with PBS, rAd5-null, rAd5-bFGF-siRNA, rAd5-Vpr, rAd5-bFGF-siRNA plus rAd5-Vpr, respectively. The tumor volume was recorded every third day to draw a growth curve. After four weeks treatment, the mice were killed and specimens were taken. HE, immunohistochemical and TUNEL staining were performed to observe the cell morphology, detect the changes of relevant target proteins and cell apoptosis, respectively. Also the ultrastructural changes were observed by electron microscopy.

RESULTSThe tumor growth inhibition rates were 36.9%, 37.2% and 58.6% in the bFGF-siRNA group, Vpr group and combined therapy group, respectively, and the combined therapy group showed the most significant effect (P < 0.05). Also the results of HE, immunohistochemical and TUNEL staining revealed that the combined therapy group had the best effects on proliferation inhibition and apoptosis induced in glioma cells (P < 0.05). The most significant ultrastructural changes were observed in the combined therapy group.

CONCLUSIONThe combined gene therapy of bFGF-siRNA with Vpr shows a prominent and synergistic anti-glioma effect compared with that of mono-gene therapy in nude mice.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Brain Neoplasms ; metabolism ; pathology ; therapy ; Cell Line, Tumor ; Cell Proliferation ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Gene Products, vpr ; genetics ; metabolism ; Genetic Therapy ; Glioma ; metabolism ; pathology ; therapy ; HIV-1 ; genetics ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; RNA, Small Interfering ; genetics ; Random Allocation ; Recombinant Proteins ; genetics ; metabolism

Objective:To explore clinical application effect of cardiac remote real -time monitoring system.Methods:Cardiac remote real -time monitoring warning instrument(iHolter)was applied in 210 patients.The iHolter gave a-lerts automatically in case of ECG abnormalities for corresponding treatment in time,and it offered corresponding reports on monitoring results.According to age,patients were divided into youth group(<45 years,n=59),mid-dle-aged group(45160 years,n=56)and aged group(≥60 years,n=95).Characteristics of arrhythmias among different age and sex were analyzed.Results:ECG abnormalities were found in 191 cases(90.95%)out of 210 pa-tients,including eight cases with malignant arrhythmias,in which five cases were treated and survived after timely intervention via alerts.Compared with youth group and middle -aged group,there were significant rise in percenta-ges of paroxysmal atrial tachycardia(16.9%,26.8% vs.54.7%)and atrial fibrillation(0%,0% vs.8.4%)in aged group,P< 0.05 or <0.01;abnormal percentages of heart rate variability of middle -aged group and aged group were significantly higher than that of youth group(25.0%,36.8% vs.5.1%,P<0.01 both).Male percent-age of paroxysmal ventricular tachycardia was significantly higher than that of female(81.3% vs.18.8%,P=0.013).Conclusion:Cardiac remote real-time monitoring system can offer timely alerts for cardiovascular events such as arrhythmia and myocardial ischemia etc.,fight for more time for rescuing patients,maximally protect cardi-ac function and improve accuracy of clinical diagnosis.

Bardet-Biedl Syndrome ; diagnosis ; pathology ; Child ; Humans ; Male

OBJECTIVETo investigate the effects of recombinant human growth hormone (rhGH) on the morphology and function of the left cardiac ventricle in young rats with dilated cardiomyopathy (DCM), and to evaluate the efficacy and safety of rhGH in the treatment of DCM.

METHODSSixty male Sprague-Dawley rats were randomly and equally assigned to control group, DCM group, and rhGH group. Furazolidone (0.25 mg/g) was given by gavage for 12 weeks to prepare the DCM model. Rats in the rhGH group received an intraperitoneal injection of rhGH (0.15 U/kg) once per day for 12 weeks, while rats in the DCM group received an equal volume of normal saline instead. Rats in the control group did not receive any treatment. Cardiac indices, serum biochemical parameters, hemodynamic indices, cardiac histopathological changes, and levels of myocardial collagen fibrils in each group were determined using Doppler echocardiography, enzyme-linked immunosorbent assay, multi-channel physiological recorder, light and electron microscopy, and picrosirius red staining plus polarization microscopy, respectively.

RESULTSCompared with the control group, rats in the DCM group had significantly increased cardiac chamber size, significantly reduced ventricular wall thickness, and significantly decreased fractional shortening (FS) and ejection fraction (EF) (P<0.05). Rats in the rhGH group had significantly improved cardiac chamber size, ventricular wall thickness, FS, and EF compared with the DCM group (P<0.05). Those indices in the rhGH group were similar to those in the control group (P>0.05). There were significant differences in serum biochemical parameters and hemodynamic indices between the DCM and control groups (P<0.05). Compared with the DCM group, the rhGH group had significantly improved serum biochemical parameters and hemodynamic indices (P<0.05). Those indices in the rhGH group were similar to those in the control group (P>0.05), except for the levels of insulin-like growth factor-1 and insulin-like growth factor-binding protein-3. The DCM group had a significantly higher collagen type I/collagen type III (Col I/Col III) ratio in the myocardium than the control group (P<0.05), and there was no significant difference in the Col I/Col III ratio between the control and rhGH groups (P>0.05).

CONCLUSIONSrhGH plays a certain role in improvement in the morphology and function of the left cardiac ventricle in young rats with DCM.

Animals ; Cardiomyopathy, Dilated ; drug therapy ; pathology ; Collagen Type III ; analysis ; Echocardiography ; Hemodynamics ; drug effects ; Human Growth Hormone ; therapeutic use ; Male ; Myocardium ; pathology ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To study the association between clinical phenotypes and genotypes in children with Becker muscular dystrophy (BMD)/Duchenne muscular dystrophy (DMD) so as to provide a theoretical basis for disease management, gene therapy, and prenatal diagnosis. METHODS: A retrospective analysis was performed for the clinical data and gene detection results of 52 children with BMD/DMD. Multiplex ligation-dependent probe amplification (MLPA) was used to detect the DMD gene. The children with negative results of MLPA were further screened by exon chip capture combined with next-generation sequencing (NGS). The mothers of 20 probands were validated by sequencing. RESULTS: The pathogenic genes for BMD/DMD were detected in 50 children by MLPA and NGS, with a detection rate of 96%. Among the 52 children, 36 (69%) had gene deletion, 7 (13%) had duplication, and 7 (13%) had micromutation. Among the 43 children with deletion/duplication, 32 had DMD and 11 had BMD; 37 children (86%) met the reading frame rule, among whom 27 (96%) had DMD and 10 (67%) had BMD. All 7 children with micromutation had DMD. CONCLUSIONS The reading frame rule has an extremely high predictive value for DMD but a limited predictive value for BMD.
Child ; Dystrophin ; Female ; Gene Deletion ; Genotype ; Humans ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; Mutation ; Phenotype ; Pregnancy ; Retrospective Studies

OBJECTIVETo evaluate the correlation of 64-multidetector-row CT (64MDCT) perfusion imaging with microvessel density(MVD) and vascular endothelial growth factor(VEGF) in colorectal carcinoma.

METHODS64MDCT perfusion imaging was performed in 33 patients with pathologically verified colorectal carcinoma. Time-density curves (TDC) were created from the region of interest (ROI) drawn over the tumor, target artery and vein by 64MDCT with perfusion functional software. The individual perfusion maps generated were for blood flow (BF), blood volume (BV), mean transit time (MTT) and permeability-surface area product (PS). MVD and VEGF expression of surgical specimens were examined by immunohistochemical staining with anti-CD34, anti-VEGF monoclonal antibody. MVD and VEGF were compared among the different types of TDC in colorectal carcinoma. The correlation of CT perfusion parameters with MVD and VEGF was also examined.

RESULTSTDC of colorectal carcinoma was divided into five types according to their shapes. MVD in the colorectal carcinoma was 22.61+/-9.01. VEGF staining was found in 25 of 29 tumors (86.2%). The score of VEGF expression was 4.15+/-1.09. No significant differences of MVD and VEGF expression among TDC types were found (F=2.59, 1.11, P>0.05). There were also no correlations of MVD and VEGF expression with any dynamic CT parameters (P>0.05).

CONCLUSION64MDCT perfusion imaging, MVD and VEGF may reflect angiogenic activity, but no significant correlations are found among them.

Adult ; Aged ; Colorectal Neoplasms ; blood supply ; diagnostic imaging ; Female ; Humans ; Male ; Microvessels ; Middle Aged ; Neovascularization, Pathologic ; Tomography, Spiral Computed ; methods ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult