AIM: To explore the mutual effect of co-culture of wild-type astrocytes (ASC) and motor neurons VSC4.1 (VSC) on the respective ability to produce reactive oxygen species (ROS). METHODS: The inhibition rates of cell growth in ASC and VSC in co-culture or independent culture was detected after exposed to excitatory stimulus by MTT method. Real-time observation of ROS production by ASC and VSC labeled with Hoechst 33342 was detected by confocal microscopy under the conditions of co-culture or independent culture. RESULTS: Higher concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in ASC alone, while lower concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in VSC only. Real-time observation by confocal microscopy showed that ROS production by VSC under the condition of co-culture, which showed a notable increase at 15 min, was significant less than that in independent culture, which peaked at 5 min and was gradually decreased. ROS production by ASC in co-culture began to increase significantly at 10 min. CONCLUSION: Compared to independent culture, ASC reduces the resting ROS production by co-cultured with VSC, while ASC prolongs the duration of ROS production by VSC after exposed to excitatory stimulus.

Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

OBJECTIVETo predict part of medicinal properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces by using the traditional Chinese medicinal property data prediction platform, in order to establish the relationship between properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces.

METHODThe properties of traditional Chinese medicine components were predicted by using the medicinal property data prediction platform based on the pharmacological effects of the components.

RESULTThe total sum of identical or similar results of the prediction for the properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces accounted for over 75%.

CONCLUSIONThe self-similarity exists between properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces, which reflects the inheritance, additivity and emergence among different properties of traditional Chinese medicines.

Drug Combinations ; Drug Prescriptions ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional

OBJECTIVE Toprepareapolyclonalantibodyforspindlin1protein,anovelcancer related protein,and to provide the data for a better understanding of its functions and screening tu mor. METHODS Purifiedspindlin1proteinwasinjectedintorabbitstoproducethepolyclonalantiserumafter removing glutathione S-transferase (GST)from the fusion protein spindlin 1-GST that was expressed in Escherichia coli..The antiserum was purified through the Hitrap Protein A system,and the titer of spin-dlin 1 polyclonal antibody was detected by ELISA.The specificity of the polyclonal antibody was deter-minedbyWesternblottingandimmunohistochemistry.RESULTS Thetiterofspindlin1polyclonalanti-body was 1∶2000.Western blotting detection demonstrated that the spindlin 1 polyclonal antibody recog-nized myc-spindlin 1 reco mbinant fusion protein in HeLa cells transfected with pAdeasy-myc-spindlin 1 , which also corresponded with Myc.antibody.The HeLa cells were transfected with enhanced green fluo-rescence protein (EGFP)and spindlin 1 vector(pEGFP-C3-spindlin 1 ),which was confirmed by the in-dependent GFP fluorescence assay.The results of immunohistochemistry detection with the spindlin 1 polyclonal antibody suggested that spindlin 1 was mainly expressed in the nuclei of HeLa cells.More i m-portantly,in i mmunohistoche mical assays,the spindlin 1 antibody recognized nuclear spindlin 1 expres-sioninclinicalovariancancertissues.CONCLUSION Thespecificspindlin1polyclonalantibodyispre-pared,which may be used to detect cancer-related protein spindlin 1 in HeLa cells and ovarian cancer tissues.

Objective To analyze the features of bone minimal density and bone quantitative ultrasound in men with different osteoporotic risk graded by osteoporosis self-assessment tool for Asian (OSTA).Methods After exclude the secondary osteoporosis,724 subjects over 50 years old were involved.The parameters of hight,weight,quantitative ultrasound index (QUI),QUS-T score were examined.The bone density (BMD) were measured by dual-energy X-ray absorptiometry (DXA)in 120 elderly men.All subjects were grouped into low (osteoporotic) risk group,moderate risk group and high risk group by OSTA index.120 subjects measured BMD were grouped into normal bone mass group,osteopenia group and osteoporosis group by WHO standard.The differences and correlation analysis in BMD,QUST,and QUI between these groups were analysed.Results The percents of low risk people,moderate risk people and high risk people were 56.4％ (408 cases),28.2％ (204cases),15.5％ (112 cases),respectively.There were 30.0％ (36 cases) normal bone mass people,58.3％ (70 cases) osteopenia people and 11.7％ (14 cases) osteoporosis people in groups measured BMD.QUS-T score,QUI were gradually decreased in groups of low risk,moderate risk and high risk (-0.56±1.09,-0.88±-1.28,-1.21±1.40; 98.47±19.04,92.62±22.49,87.68±24.43; all P ＜0.05) and had statistical significant differences between low risk and moderate risk,high risk groups,while had no differences between moderate risk and high risk groups.The femoral neck BMD and total BMD were gradually decreased in all the three groups (0.89±0.12,0.85±0.10,0.77± 0.10; 1.0±0.15,0.93 ± 0.11,0.83±0.1; all P＜0.01).Osteoporosis in the three groups were 3.4％ (2 cases),13.0％ (6 cases),37.5％ (6 cases),respectively and osteoporosis percents in moderate risk group and high risk group were higher compared with low risk group (x2=11.77,P＜0.01).QUS-T score and QUI decreased gradually in groups of normal mass,osteopenia and osteoporosis (0.99±0.08,-0.70±1.07,-1.96±0.73; 109.26±17.05,96.15±18.20,72.54±10.00; F=10.47,11.73,all P＜ 0.01).Except for lumbar BMD,a positive linear correlation emerged between OSTA and QUS-T score,QUI,hip BMD(all P＜0.01).The values of R with femoral neck BMD,torch BMD and total hip BMD were 0.45,0.38,0.39,respectively.And the same value with QUS-T score and QUI was 0.23.Conclusions With the decreasing of OSTA index,risk of osteoporosis is increased and QUS-T score,QUI and BMD are decreased gradually.There are positive linear correlation between OSTA index and QUS-T score,QUI,hip BMD.

This article summarizes the current studies on the pathological mechanism of Parkinson's disease as well as the advance in studies traditional Chinese medicine (TCM) experiments in prevention and treatment of Parkinson's disease in the latest decade in terms of prevention and treatment of PD by TCM, inhibition of oxidative stress, improving mitochondrial energy metabolism, inhibition of neural immune and inflammatory responses, reduction of neural toxicity, inhibition of apoptosis and abnormal protein aggregation.
Animals ; Apoptosis ; drug effects ; Humans ; Medicine, Chinese Traditional ; Oxidative Stress ; drug effects ; Parkinson Disease ; drug therapy ; etiology ; pathology ; prevention & control

OBJECTIVETo detect the loss of heterozygosity (LOH) in esophageal squamous cell carcinoma and adjacent high-grade squamous dysplasia, and to evaluate possible tumor suppressor genes in the development and progression of invasive malignancy.

METHODSLOH was detected in normal esophageal mucosa, high grade squamous dysplasia and esophageal squamous cell carcinoma using microdissection and polymerase chain reaction technology. The changes of LOH at seven microsatellite markers and the relationship between LOH rate and clinicopathologic parameters were analyzed.

RESULTSIn high grade squamous dysplasia, LOH was detected at D13S802 (40%), D13S267 (32%), D13S221 (31%), D9S942 (30%), D17S520 (24%) and D9S171 (33%). However, D17S1798 LOH was not detected. In invasive squamous cell carcinoma, LOH was detected as follows: D13S267 (71%), D13S802 (58%), D17S520 (55%), D13S221 (45%), D9S942 (43%), D9S171 (33%) and D17S1798 (11%). The frequency of LOH in the seven microsatellite markers, the pathologic grade, clinical stage and occurrence of lymph node metastasis did not show any statistically significant correlation (P > 0.05).

CONCLUSIONSThe progression from normal squamous epithelium to high grade squamous dysplasia and subsequently to invasive squamous cell carcinoma of the esophagus was associated with accumulation of genetic errors. Possible tumor suppressor genes related to the development of esophageal squamous cell carcinoma may exist near D13S802 (13q12.12). Possible tumor suppressor genes near D13S267 (13q13.1), D17S1798 (17p13.3) and D17S520 (17p13.1) may be related to the progression of esophageal squamous cell carcinoma.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; Chromosome Mapping ; Chromosomes, Human, Pair 13 ; Chromosomes, Human, Pair 17 ; Chromosomes, Human, Pair 9 ; Esophageal Neoplasms ; genetics ; Female ; Genes, Retinoblastoma ; Genes, Tumor Suppressor ; Genes, p16 ; Genes, p53 ; Humans ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Middle Aged ; Precancerous Conditions ; genetics

ObjectiveTo investigate the clinical value of color Doppler ultrasound examination in the diagonosis of acute and chronic artery occlusion of the extremities.MethodsA review was made on 129 extremetiy artery occlusion patients at Anzhen Hospital during 2006 -2010. 85 cases were male, and 44 cases were female. Age was from 17 to 94 years (average: 62 ±9 years). We analyzed two-dimensional and color Doppler flow imagings of 39 acute occlusion arteries and 97 chronic occlusion arteries. We compared factors including the echoes of artery lumens, the vessel wall structures, hemodynamic parameters of inlet and outlet at the occlusion, and collaterals between groups.ResultsThe factors of depths of vessel wall,internal diameters of ccclusion arteries, proximal resistant index and collaterals were significantly different between groups ( P ＜ 0. 05 ). The internal diameters of acute occlusion arteries were wider than chronic occlusion arteries. The depths of vessel wall, proximal resistant index and collaterals were thinner, smaller,and less than chronic occlusion arteries. The total accurate rate of differential diagnosis for acute and chronic artery occlusion by color Doppler ultrasound was 95.6％.ConclusionsColor Doppler ultrasound is an effective method for the differential diagnosis of acute and chronic artery occlusion of the extremities.

The transcriptional repressor RE1 silencer transcription factor(NRSF/REST) is an important factor that restricts some neuronal traits in neurons.Since these traits are also present in pancreatic islet cells,NRSF-regulated genes involved in islet function are searched.A NRSE-like motif was analysed in human insulin promoter.The role of NRSE was evaluated by generating a model of insulin-secreting cells that firmly express NRSF.The presence of NRSF led to a decrease in activity of human insulin promoter by stable or transient transfection with human insulin-promoter luciferase.The predicted NRSE-like motif also confers NRSF-dependent transcriptional repression in the context of a surrogate gene promoter.Specific binding activity of NRSF/REST to the NRSE-like motif was confirmed by EMSA.Moreover,the binding activity is competed by consensus NRSE sequence.These data showed that human insulin promoter is regulated by the transcriptional repressor NRSF/REST via the NRSE-like motif.

OBJECTIVETo investigate the sustaining effects of gene-transferring hepatic stellate cell strain CFSC/HGF on the development of hepatocytes.

METHODSA CFSC/HGF strain, expressing HGF steadily and effectively was established by recombined retroviral vector pMSCV-HGF infection. Morphology and ultra structure of hepatocytes, albumin and urea production, as well as ICG uptake and excretion were studied continuously following the hepatocytes cultured on the CFSC/HGF feeder layers. Parallel group of collagen-dependent hepatocytes culturing and hepatocytes culturing on CFSC were also conducted. Semi-quantitative RT-PCR analysis was made to evaluate the expression of HGF receptor c-Met.

RESULTSThe hepatocytes cocultured on CFSC/HGF feeder layers had a higher survival rate, and the functions of albumin secretion and urea syntheses and ICG uptake and excretion, were superior to the other two culture methods. The result of RT-PCR indicated that the c-Met expressed on the CFSC/HGF coculturing hepatocytes was up-regulated 2.23 times.

CONCLUSIONGene-transferring hepatic stellate cell strain CFSC/HGF exhibited a remarkable sustaining effect on the hepatocytes development. The up-regulation of c-Met expressed on the surfaces of the hepatocytes induced by CFSC/HGF might play some part in this function.

Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Colony-Forming Units Assay ; Gene Transfer Techniques ; Hepatocyte Growth Factor ; metabolism ; Hepatocytes ; cytology ; Humans ; Liver ; cytology ; Proto-Oncogene Proteins c-met ; metabolism ; Transfection