Objective:This study aimed to identify the morphology of mast cells by using a modified toluidine blue staining scheme,so as to provide a powerful reference for the experimental basis research of mast cells.Methods:Bone marrow-derived mast cells were induced in vitro.After 4 weeks,the cells were collected,fixed,and stained.Mast cells were fixed at different temperature during different time.The optimum condition was determined by comparing the effects of toluidine blue staining.Results:Bone marrow cells were induced to differentiate into mast cells by SCF and IL-3 in vitro.When mast cells were stained with modified toluidine blue staining,the staining effect was better.Mast cells were round or oval and the cell membrane was complete and the cytoplasm was filled with a large number of purple particles.Conclusion:In this study,we successfully applied a modified toluidine blue staining method to mast cells cultured in vitro.The results showed that the condition at 37 ℃ full fixation with staining could reduce the degeneration of mast cells.This method was easy to operate with good stability.It was suitable for the morphological observation of mast cells cultured in vitro.

Objective To determine the etiology and clinical characteristics of children with dilated cardiomyopathy.Methods 112 children with DCM from January 2002 to December 2011 were studied.The age,cause and the clinical manifestations were analyzed.According to the children with heart function,they were divided into four groups,the incidence and type of arrhythmia were compared.Results In 112 cases of DCM in children,＜ 1 year old diagnosed 52 cases (46.42％),107 cases (95.53％) with heart failure,25 cases (22.32％) with chear causes,96 cases(85.71％) children with a variety of arrhythmias.According to cardiac function in Ⅰ,Ⅱ and Ⅲ and Ⅳ level,they were divided into four groups.The incidence of one kind of arrhythrnia were 20.00％,31.25％,36.17％ and 42.85％.The incidence with two or more kinds of arrhythmia was 0,18.75％,38.29％ and 50.00％ respectively.The incidence of malignant arrhythmia was 0,12.50％,40.42％and 67.85％ in different groups.There was significant differences among the groups of Ⅱ and Ⅲ and Ⅳ level (P ＜ 0 05).Conclusion Children with dilated cardiomyopathy disease younger than I year old had the highest incidence.The major causes were myocarditis,hereditary and congenital metabolic diseases.The incidence of arrhythmia in DCM were high and diverse,often exist for a variety of arrhythmia,and arrhythmia have closely relation with heart function.The embolism in children with DCM was very rare.

OBJECTIVETo study the clinical effect of development theory based acupuncture on early cerebral palsy (CP) infants with parafunctional sitting position.

METHODSTotally 120 early CP infants were randomly assigned to two groups equally, the treatment group and the control group. All received acupuncture combined with training rehabilitation. Patients in the treatment group adopted acupuncture based on infants development theory, while those in the control group were treated by head acupuncture. Sitting functional points in Gross motor function measure (GMFM) 88 were observed in different groups and infant patients of various types before and after treatment. Root mean square (RMS) signals of sitting correlated muscles (latissimus dorsi, erector spinae, rectus abdominis) were recorded by surface electromyography (sEMG). The effective rate was evaluated by Nimodipine method.

RESULTSCompared with before treatment, sitting functional points were significantly improved in the two groups (P<0.01). After treatment, it was higher in the treatment group than in the control group (P<0.01). The advance amplitude was higher in CP infants of the spastic type and the hypotonic type than other types (P<0.01). Along with sitting process, latissimus dorsi RMS signals were gradually tapered, erector spinae RMS signals were gradually enhanced, and rectus abdominis RMS signals were slightly weakened. Compared with the control group, latissimus dorsi RMS signals obviously decreased, and erector spinae RMS signals obviously increased in the treatment group after treatment (all P<0.01). The total effective rate was higher in the treatment group than in the control group (89.29% vs. 77.78%, P<0.05).

CONCLUSIONInfants development theory based acupuncture could effectively elevate dorsi-extensor muscles force, improve sitting position of 8 months to 1 year old CP infants with parafunctional sitting position.

Acupuncture Therapy ; methods ; Cerebral Palsy ; therapy ; Female ; Humans ; Infant ; Male ; Medicine ; Posture ; Research ; Spine

Using the weight-average molecular weight 50 000 polylactic acid (PLA) as a carrier, and a certain proportion of erythromycin (EM) and prednisone acetate (PNA) to mixed prepare the compound erythromycin sustained release preparation (sustained-release tablets). Using ultraviolet spectrophotometry and high performance liquid chromatography (HPLC) to detect separately the release amount of EM and PNA in vitro medium. The sustained-release tablets release for about 21 days, the average content of EM is 99.7 mg/table, RSD = 0.82%; and the average content of PNA is 10.03 mg/table, RSD = 0.93%. Within 21 days, the cumulative releases of EM and PNA are 86.1% and 78.3%, respectively. The drug release is steady and slow after 5 days, the burst release phenomenon in early stage is more significant. The results showed that the sustained-release tablet preparation method is feasible, the release performance is good and the clinical efficacy is significant.
Chromatography, High Pressure Liquid ; Delayed-Action Preparations ; administration & dosage ; chemistry ; therapeutic use ; Drug Carriers ; Drug Combinations ; Erythromycin ; administration & dosage ; chemistry ; therapeutic use ; Humans ; Lactic Acid ; administration & dosage ; Polyesters ; Polymers ; administration & dosage ; Prednisone ; administration & dosage ; chemistry ; therapeutic use ; Sinusitis ; drug therapy ; Spectrophotometry, Ultraviolet ; Tablets

OBJECTIVETo investigate changes in serum insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-binding protein-3 (IGFBP-3) and their significance in children with left-to-right shunt congenital heart disease (CHD) associated with heart failure (HF).

METHODSTwenty healthy children (control group), 20 children with HF, without basic heart disease (HF group), 20 children with left-to-right shunt CHD, without HF (CHD group), and 30 children with left-to-right shunt CHD associated with HF (CHD+HF group) were included in the study. These groups were compared in terms of serum IGF-1 and IGFBP-3 levels. According to the New York Heart Association (NYHA) Functional Classification, the CHD+HF group was further divided into NYHA-II, NYHA-III and NYHA-IV subgroups and the subgroups were compared in terms of serum IGF-1, IGFBP-3, and cardiac troponin I (cTnI) levels. The correlation of serum IGF-1 and IGFBP-3 levels with serum cTnI level in the CHD+HF group was analyzed.

RESULTSThe CHD group showed decreased serum IGF-1 and IGFBP-3 levels compared with the control group (P<0.01). The CHD+HF group showed a significantly decreased serum IGF-1 level compared with the control group (P<0.01) and CHD group (P<0.05). The HF group had significantly increased serum IGF-1 and IGFBP-3 levels compared with other groups (P<0.01). The NYHA-II subgroup had the highest serum IGF-1 level and the NYHA-IV subgroup had the lowest serum IGF-1 level (P<0.01). In the CHD+HF group, serum IGF-1 and IGFBP-3 levels were negatively correlated with serum cTnI level (r=-0.692, P<0.05; r=-0.530, P<0.05).

CONCLUSIONSSerum IGF-1 level can be used as an objective condition evaluation indicator for CHD, and low serum IGF-1 level is a risk factor for HF. This also provides a clinical basis for treatment of HF using exogenous IGF-1.

Child, Preschool ; Female ; Heart Defects, Congenital ; blood ; Heart Failure ; blood ; Humans ; Infant ; Insulin-Like Growth Factor Binding Protein 3 ; blood ; Insulin-Like Growth Factor I ; analysis ; Male ; Troponin I ; blood

This study was designed to evaluate the effects of drilling through the growth plate and using adipose-derived stem cells (ADSCs) and bone morphogenetic protein-2 (BMP-2) to treat femoral head epiphyseal ischemic necrosis,which can be done in juvenile rabbits.Passage-four bromodeoxyuridine (BrdU)-labeled ADSCs were cultured,assayed with MTT to determine their viability and stained with alizarin red dye to determine their osteogenic ability.Two-month-old,healthy male rabbits (1.2 to 1.4 kg,n=45) underwent ischemic induction and were randomly divided into five groups (group A:animal model control;group B:drilling;group C:drilling & ADSCs;group D:drilling & BMP-2;and group E:drilling & ADSCs & BMP-2).Magnetic resonance imaging (MRI),X-ray imaging,hematoxylin and eosin staining and BrdU immunofluorescence detection were applied 4,6 and 10 weeks after treatment.Approximately 90％ of the ADSCs were labeled with BrdU and showed good viability and osteogenic ability.Similar results were observed in the rabbits in groups C and E at weeks 6 and 10.The animals of groups C and E demonstrated normal hip structure and improved femoral epiphyseal quotients and trabecular areas compared with those of the groups A and B (P＜0.01).Group D demonstrated improved femoral epiphyseal quotients and trabecular areas compared with those of groups A and B (P＜0.05).In summary,drilling through the growth plate combined with ADSC and BMP-2 treatments induced new bone formation and protected the femoral head epiphysis from collapsing in a juvenile rabbit model of femoral head epiphyseal ischemic necrosis.

Objective:To explore the risk factors,preventive measures and therapeutic methods for bronchopleural fistula (BPF) after lung resections.Methods:A restrospective analysis for 11 patients with BPF after pneumonectomy from April 2012 to June 2016 in Department of Thoracic Surgery,Xiangya Hospital,Central South University was performed.Their clinical characteristics,treatment and prognosis were analyzed,and the risk factors and effective therapeutic strategies were summarized.Results:Among the 11 patients with BPF,10 cases were cured finally,and 1 case with conservative treatment was dead.The total mortality rate was 9.09％.The 10 patients treated with positive measures were all cured,including 5 cases with pulmonary lobectomy and pneumonectomy,4 cases with amplatzer and covered stent,and 1 case with fibrin glue.One case with conservative treatment was dead because of respiratory failure.Conclusion:It is important to intervene BPF as early as possible.Fibrin glue via bronchoscope for tiny BPF after lung resection is preferred to be considered.We recommend to take early positive operation (pulmonary lobectomy and pneumonectomy) after pulmonary resection if the BPF cannot be cured via bronchoscope whereas the patients' condition is allowed.The amplatzer or covered stent should be considered first for the patient with BPF after pneumonectomy.

OBJECTIVETo screen and identify the proteins that interact with connexin 26 (CX26) and to analyze the expressions of these proteins in cochlea so as to explore the proteins that relate to the trafficking, assembly, localizing and gap junction functions of CX26.

METHODSThe whole coding region of GJB2 (CX26) gene was amplified from normal human genomic DNA by polymerase chain reaction (PCR) and then directionally subcloned into the vector pGBKT7 plasmid of the Match Maker Ga14 Two-Hybrid System 3 as a target to screen the interactive proteins of CX26 from the human fetal brain cDNA library by the yeast two hybrid technique. The false positive clones were discarded from the preys by repeated yeast two hybrid method between CX26 and everyone of the preys respectively. The DNAs of the insert of the identified positive clone were sequenced and BLAST analyzed against the GenBank. Lastly, the mRNA of the gene encoding the identified protein was analyzed in the murine inner ear by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe insert of one positive clone contained 867 bp with the former 525 bp being coding region. The DNA sequence and the open reading frame of the insert were identical to the 525 bp before the stop codes (including the stop codes) and the 238 bp after the stop codes of RTN4 gene which encoded Nogo protein. And the 174 amino acid residues encoded by the insert were those of the C-terminal of Nogo protein: Nogo-A, Nogo-B and Nogo-C. RTN4 mRNA expressed in the murine inner ear was confirmed by RT-PCR method.

CONCLUSIONThe C-terminal of Nogo protein interacts with CX26. Nogo protein expresses in the inner ear and may take part in the trafficking of CX26 or CX26 gap junction function.

Animals ; Base Sequence ; Connexin 26 ; Connexins ; genetics ; metabolism ; Ear, Inner ; metabolism ; Gene Expression ; Humans ; Mice ; Molecular Sequence Data ; Myelin Proteins ; genetics ; metabolism ; Nogo Proteins ; Protein Binding ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Two-Hybrid System Techniques

This study was purposed to investigate the effect of crocin on the proliferation in vitro and immune function of dendritic cells (DC) derived from the bone marrow of children with acute leukemia. The mononuclear cells were isolated from bone marrow of leukemia children by Ficoll-Hypaque. The experiment was divided into six groups: blank control group (A), crocin 1.25 mg/ml group (B), cytokines (rhGM-CSF 75 ng/ml+rhIL-4 75 ng/ml+rhTNF-α 50 ng/ml) group (C), cytokines+crocin 0.3125, 1.25 or 5.0 mg/ml groups (D, E, F). The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture. The DC of different groups were mixed with T cells just separated from peripheral blood of another children with acute lymphoblastic leukemia, and cultured with rhIL-2 200 U/ml for 5 d. The function of DC was detected by mixed lymphocyte reaction (MLR). The results indicated that the test groups and control group all obtained a certain amount of typical DC, but the DC numbers in test groups were all higher than those in control group (P < 0.01). Cultured for 9 days, the rates of CD1a(+), CD83(+), and HLA-DR(+) in group C, D, E, F were higher than group A (P < 0.01). There was no statistically significant difference between A and B groups (P > 0.05). MLR showed that with the increasing of DC, the stimulation index of T cells in group A and B was not rising (P > 0.05); the stimulated index of T cells in group C and E was significantly rising, there was statistically significant difference between them (P < 0.01). When the number of stimulated cells was the same, the stimulation index of T cell in group E was the highest (P < 0.01). It is concluded that the capability of DC proliferation promoted by crocin alone is lower than that of its combination with rhGM-CSF, rhIL-4 and rhTNF-α, but the crocin can synergically promote the maturity of DC cooperating with rhGM-CSF, rhIL-4 and rhTNF-α. The DC induced by crocin can particularly enhance the proliferation of T cells.
Bone Marrow Cells ; cytology ; drug effects ; Carotenoids ; pharmacology ; Cell Proliferation ; drug effects ; Child ; Dendritic Cells ; cytology ; drug effects ; Humans ; Leukemia ; pathology ; Lymphocyte Culture Test, Mixed ; T-Lymphocytes ; cytology ; Tumor Cells, Cultured

OBJECTIVETo screen all ten genes between D15S971 and D15S1012 in five Chinese families with hereditary spastic paraplegia with thin corpus callosum (HSP-TCC).

METHODSDNA samples from 5 HSP-TCC families were screened for mutations in AK128197, MGC14798, HH114, MEIS2, MGC35118, SPRED1, AK128458, FLJ38426, RASGRP1 and AK093014 on chromosome 15q13-15 between microsatellites D15S971 and D15S1012 by polymerase chain reaction, direct sequencing and cosegreagation analysis.

RESULTSNo disease-causing mutations were found in the 10 genes, but 13 polymorphisms were identified in which two were novel.

CONCLUSIONThis study did not support the ten genes between D15S971 and D15S1012 were the disease-causing genes of the 5 HSP-TCC families.

Adult ; Asian Continental Ancestry Group ; genetics ; Chromosomes, Human, Pair 15 ; Corpus Callosum ; pathology ; Female ; Genes, Recessive ; Humans ; Male ; Paraparesis, Spastic ; genetics ; Spastic Paraplegia, Hereditary ; complications ; genetics