OBJCETIVETo investigate the method of separation of culture of bone microvascular endothelial cells (BMECs) of human femoral head in vitro.

METHODSFrom October 2013 to January 2014,15 femoral heads without pathologic change from patients resected during hip replacement were selected involving 2 males and 13 females with a mean age of 71.2 years old ranging from 38 to 92. Cancellous bone in femoral head was bited into broken bone grain and transfered into medium in aseptic contidion. Cells were isolated by the methods of enzymic digestion and density gradient centrifugation,purified by differiential attachment. The characteristics of cells was observed by inverted microscope. vWF and CD31 immunofluorescence analysis was applied for identification of cells.

RESULTSThe number of cells was positively correlated with patients' age after 24 hours in primary culture. The older patients had the less cells numbered. After 4 to 5 days' culture, primary cells appeared short spindle,polygon shaped and cobblestone-like morphology. After 7 to 10 days' culture, primary cells proliferated densely, became fusion, arranged in swirl, and contact inhibition appeared significantly. Immunofluorescence staining revealed the cells were 100% positive for vWF and CD31, and it showed that the cultured cells were BMECs.

CONCLUSIONIt was a simple, steady, effective method with good reproducibility, by which highly purified human BMECs can be obtained.

Adult ; Aged ; Aged, 80 and over ; Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Femur Head ; blood supply ; Humans ; Male ; Microvessels ; cytology ; Middle Aged

Objective To explore the therapeutic effects and dose of Topiramate(TPM)in children with Tourette syndrome(TS).Me-thods Seventy-nine children with TS were given oral TPM,which were treated with Topiramate [0.5-3.0 mg/(kg?d),twice a day].The therapeutic effects were assessed using Yale Global Tic Severity Scale(YGTSS)before and 3 months after treatment and the side-effects of the drugs were observed.Results The differences of YGTSS scores before and after treatment,motertic score(19.63?3.09 vs 5.05?1.74),vocaltic score[(18.95?2.56)vs(4.82?1.94)],global scole score[(24.21?5.89)vs(10.42?3.69)],severity score[(62.21?5.81)vs(22.26?4.81)],there were significant differences of every score of YGTSS between before and after treatment(Pa

Objective To promote comprehension of pulmonary sequestration by investigating the diagnosis and surgical treatment methods of this disease.Methods A retrospective analysis of the clinical records of 27 patients with pulmonary sequestration was made,15 male patients and 12 female patients,with the age ranging from 4 to 56 years old.The patients were examined by chest radiography,computerized tomography (CT),computerized tomography angiography (CTA),magnetic resonance imaging (MRI) before the surgery.Eleven patients were confirmed preoperatively as pulmonary sequestration by the contrast enhanced CT and 64slice spiral CT examinations.Sequestrectomy was performed on patients with extralobar sequest ration (ELS) and lobectomy was performed on patients with intralobar sequestration (ILS).There were 21 cases of left lower lobectomy,5 cases of right lowerlobectomy,and 1 case of right sequestrectomy.Results All recovered uneventfully.No significant complications occurred in the perioperative period.All patients were cured and discharged from hospital.No relapse was reported during follow up period.Conclusion Diagnosis mainly depended on the chest radiography,CT,MRI and CTA.Surgical operation should be the best choice of treatment.

Objective To study the conventional MRI and dittusion weighted imaging(DWI) features of Japanese encephalitis(JE)in children.Methods Sixteen patients with JE were included. Conventional MRI and DWI sequences were performed in all patients.Seven patients received MRI within 10 days of onset and 9 after 10 days.The appearances on DWI and T_2 WI were recorded.The ADC values of lesions were calculated,and then were correlated with the corresponding time interval between onset of neurological symptoms and MRI scanning.Results The lesions of JE mainly showed long T_1 and long T_2 signal intensity on MRI.The thalami were the most frequently involved areas,and 15 out of 16 showed thalamic involvement and 6 patients only showed thalamic abnormalities without other lesions.Seven patients within 10 days of onset showed lesions with high signal intensity on both DWI and T_2WI,but whole or partial lesions showed clearer on DWI than on T_2WI,and 2 patients showed extra lesions that were invisible on T_2WI.As for the other 9 patients after 10 days of onset,the lesions showed clearer on T_2WI than on DWI. There was a direct correlations between thalamic ADC values and the disease duration (r=0.84,P

Animals ; Astrocytes ; immunology ; metabolism ; Male ; Microglia ; immunology ; metabolism ; Neurons ; metabolism ; Oxidopamine ; metabolism ; Parkinson Disease ; immunology ; Rats ; Rats, Sprague-Dawley

Animals ; Dynorphins ; metabolism ; Parkinson Disease ; drug therapy ; metabolism ; Peptides ; pharmacology ; Rats ; Scorpion Venoms ; pharmacology

BACKGROUNDAppropriate expression and regulation of the transcriptome, which mainly comprise of mRNAs and lncRNAs, are important for all biological and cellular processes including the physiological activities of bone microvascular endothelial cells (BMECs). Through an intricate intracellular signaling systems, the transcriptome regulates the pharmacological response of the cells. Although studies have elucidated the impact of glucocorticoids (GCs) cell-specific gene expression signatures, it remains necessary to comprehensively characterize the impact of lncRNAs to transcriptional changes.

METHODSBMECs were divided into two groups. One was treated with GCs and the other left untreated as a paired control. Differential expression was analyzed with GeneSpring software V12.0 (Agilent, Santa Clara, CA, USA) and hierarchical clustering was conducted using Cluster 3.0 software. The Gene Ontology (GO) analysis was performed with Molecular Annotation System provided by CapitalBio Corporation.

RESULTSOur results highlight the involvement of genes implicated in development, differentiation and apoptosis following GC stimulation. Elucidation of differential gene expression emphasizes the importance of regulatory gene networks induced by GCs. We identified 73 up-regulated and 166 down-regulated long noncoding RNAs, the expression of 107 of which significantly correlated with 172 mRNAs induced by hydrocortisone.

CONCLUSIONSTranscriptome analysis of BMECs from human samples was performed to identify specific gene networks induced by GCs. Our results identified complex RNA crosstalk underlying the pathogenesis of steroid-induced necrosis of femoral head.

Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Femur Head ; cytology ; Gene Expression Profiling ; Glucocorticoids ; pharmacology ; Humans ; Oligonucleotide Array Sequence Analysis ; Osteonecrosis ; genetics ; RNA, Messenger ; genetics ; RNA, Untranslated ; genetics ; Transcriptome ; drug effects ; genetics

OBJECTIVE: Visual search is an important attention process that precedes the information processing. Visual search also mediates the relationship between cognition function (attention) and social cognition (such as facial expression identification). However, the association between visual attention and social cognition in patients with schizophrenia remains unknown. The purposes of this study were to examine the differences in visual search performance and facial expression identification between patients with schizophrenia and normal controls, and to explore the relationship between visual search performance and facial expression identification in patients with schizophrenia. METHODS: Fourteen patients with schizophrenia (mean age=46.36+/-6.74) and 15 normal controls (mean age=40.87+/-9.33) participated this study. The visual search task, including feature search and conjunction search, and Japanese and Caucasian Facial Expression of Emotion were administered. RESULTS: Patients with schizophrenia had worse visual search performance both in feature search and conjunction search than normal controls, as well as had worse facial expression identification, especially in surprised and sadness. In addition, there were negative associations between visual search performance and facial expression identification in patients with schizophrenia, especially in surprised and sadness. However, this phenomenon was not showed in normal controls. CONCLUSION: Patients with schizophrenia who had visual search deficits had the impairment on facial expression identification. Increasing ability of visual search and facial expression identification may improve their social function and interpersonal relationship.
Asian Continental Ancestry Group ; Automatic Data Processing ; Cognition ; Facial Expression* ; Humans ; Schizophrenia*

This study was aimed to investigate the anti-angiogenesis of IFN-alpha2b in chronic myeloid leukemia (CML) in vitro by using K562 cell line and human umbilical vein endothelial cells (HUVEC). The levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in culture supernatant of K562 cells were determined by ELISA; the expressions of VEGF and bFGF mRNA after treating K562 cells with 10(3), 10(2) and 10 U/ml IFN-alpha2b for 24, 36, 48 hours were detected by real-time RT-PCR; the effects of K562 cell culture supernatant and IFN-alpha2b on proliferation, migration and differentiation of HUVEC in vitro were assayed by MTT, Transwell chamber and tubule formation assay respectively. The results showed that the K562 cells expressed and secreted VEGF and bFGF. The culture supernatant of K562 cells significantly promoted the proliferation, migration and tubule formation of HUVEC in vitro in a concentration-dependent manner. After treating K562 cells with IFN-alpha2b 10 U/ml for 24, 36 and 48 hours, the expression levels of VEGF and bFGF mRNA were 1.64+/-0.18, 1.49+/-0.14, 1.31+/-0.05 and 1.53+/-0.10, 1.29+/-0.15, 0.79+/-0.13 respectively (p=0.002), but the expression levels of VEGF and bFGF mRNA were not significantly different along with increasing of IFN-alpha2b concentration. It is concluded that the angiogenesis exists in CML. The K562 cell expresses and secrets VEGF and bFGF, which promotes the proliferation, migration and differentiation of HUVEC. The IFN-alpha2b displays anti-angiogenesis by inhibiting the proliferation, migration and tubule formation in vitro of HUVEC and down regulating the expression of VEGF and bFGF mRNA.
Angiogenesis Inhibitors ; pharmacology ; Endothelial Cells ; drug effects ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Interferon-alpha ; pharmacology ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Neovascularization, Pathologic ; Recombinant Proteins ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo explore the alterations of apoptosis factor Bcl-2/Bax in the early Parkinson's disease (PD) rats and the protective effect of scorpion venom derived bioactive peptide.

METHODSHealthy male SD rats (180-220 g) were randomly divided into 4 groups (n = 10): early PD model group, sham operation group, scorpion venom derived bioactive peptide control group, scorpion venom derived bioactive peptide therapy group. 6-hydroxydopamine (6-OHDA) was used to prepare the early PD rat model. The immunohistochemistry was used to detect the expression of Bax and Bcl-2 and further explore the mechanism of anti-apoptosis regarding the neuroprotective effect of scorpion venom derived bioactive peptide.

RESULTSThe results indicated that compared with the control rats, the immunostaining of Bax in the brain increased significantly while that of Bcl-2 decreased significantly in the lesion side of 6-OHDA treated rats. Interestingly, scorpion venom derived bioactive peptide could attenuate the above abnormal changes.

CONCLUSIONUp-regulation of Bax and down-regulation of Bcl-2 could participate in the early stage of PD and the anti-apoptotic mechanism could be involved in the neuroprotective effect exerted by scorpion venom derived activity peptide regarding the dopaminergic neuron in the early stage.

Animals ; Apoptosis ; Disease Models, Animal ; Down-Regulation ; Male ; Neuroprotective Agents ; chemistry ; Oxidopamine ; Parkinson Disease ; metabolism ; Peptides ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Scorpion Venoms ; chemistry ; Up-Regulation ; bcl-2-Associated X Protein ; metabolism