Objective To study the inhibitory effects of total flavonoids of scutellaria baicalensis georgi(TFSB) on S180,Hep-A-22 and Bcap-37 tumor cell proliferation in vitro and on S180,Hep-A-22 in mice bearing tumor in vivo.Methods In vitro,S180,Hep-A-22 and Bcap-37 cells were divided into control group and TFSB groups(12.5,25.0,50.0,100.0 mg?L-1).The inhibitory effects of TFSB on proliferation of S180 and Hep-A-22 were measured by XTT colorimetric assay,and Bcap-37 cells were measured by MTT colorimetric assay.In vivo,the mice bearing tumor were divided into control group,CTX group(30 mg?kg-1),high,middle,low doses TFSB groups(200,100,50 mg?kg-1).After the mice bearing S180 and Hep-A-22 tumor cells were treated with TFSB for 15 d,the tumor weights were measured,the inhibitory rates of S180 and Hep-A-22 were calculated and survival of Hep-A22 was measured after administration of TFSB for 10 d.Results TFSB inhibited the proliferation of S180,Hep-A-22 and Bcap-37 cells,IC50 values were 16.04,17.74 and 9.05 mg?L-1,respectively.The tumor weight of mice bearing S180 and Hep-A-22 cells in TFSB groups(200,100,50mg?kg-1) were lowered than that in control(P

Objective To investigate the spiral CT findings in hemopathic patients with druginduced pulmonary injury.Methods CT images obtained in 11patients with drug-induced pulmonary injury were retrospectively analyzed.Six patients had antineoplastic agent-induced pulmonary injury and 5 patients had non-neoplastic agent-induced pulmonary injury (immunosuppressor in 2 patients,antifungal in 2 patients,antineoplastic immunomodulators in 1 patient).CT findings were reviewed by a chest radiologist.Results All 11patients had parenchymal abnormalities on MSCT scans,including ground-glass opacities( n =8 ),consolidation( n =5 ),interlobular septal thickening( n =3 ) and focal fibrosis ( n =2 ).The abnormalities were bilateral and asymmetric in all patients.They were mainly in the peripheral lung regions in 6 patients,in the central lung regions in four,and irregularly located in one.The abnormalities involved mainly the lower lung zones in six patients,the upper lung zones in two,and all lung zones homogeneously in three.One patient had fluid in bilateral pleural cavities.Three patients were given the same agent once more after the imaging turned to normal,and they presented with same clinical symptoms and similar but more serious imaging findings.Conclusions Drug-induced pulmonary injury usually manifests as areas of ground-glass opacity and consolidation,which most commonly involves the peripheral lungs and lower lung zones.Drug-induced pulmonary injury shows reproducible but more serious lesions when the patient is given the same agent once more.

The variability of peak current of L-type calcium channel (I(Ca,L)) shows an increase in cardiomyocytes after 6 h of preservation when the acutely isolated cardiomyocytes are preserved in a small volume buffer solution. The mechanism of the increased variability of I(Ca,L) is not clear. In order to obtain more accurately and stably experimental data of I(Ca,L), the aim of this study was to observe the pH changes of preservation buffer solution with acutely isolated rat cardiomyocytes, and the effects of pH changes on the shape of cardiomyocytes, the function of mitochondria and the gating property of L-type calcium channel. The results indicated that the pH was kept stable in 100 mL buffer solution, but was decreased from 7.20 to 6.95 in 20 mL buffer solution during 10 h of cardiomyocyte preservation. Therefore, 100 mL or 20 mL preservation solution was used as a normal control or acidotic group, respectively. The ratio of abnormal to normal rod-shaped cardiomyocytes increased in the acidotic group after 6 h of preservation. The acidosis induced a reduction in mitochondrial membrane potential indicated by JC-1 fluorescent probe after 8 h of cardiomyocyte preservation. The acidosis also shifted the autofluorescence of NADPH from blue to green after 8 h of cardiomyocyte preservation. The above changes in mitochondrial function induced a significant decrease in the peak I(Ca,L) and a shift in the clamped voltage at peak I(Ca,L) from +10 mV to 0 mV, after 10 h of cardiomyocyte preservation. These results suggest that the best way to preserve acutely isolated cardiomyocytes is to use a larger volume buffer system. In order to get stable peak I(Ca,L), we need to not only select a normal shape of cardiomyocyte at a bright field but also a blue fluorescent myocyte at an ultraviolet excitation.
Animals ; Buffers ; Calcium Channels, L-Type ; physiology ; Cells, Cultured ; Membrane Potential, Mitochondrial ; Myocytes, Cardiac ; physiology ; Preservation, Biological ; Rats

OBJECTIVETo To establish a method of phosphoprotein affinity profiling for identifying phosphoproteomic differences between Thp-1 cells with or without lipopolysaccharide (LPS) stimulation, aiming to screen potential regulators involved in LPS pathway.

METHODSThp-1 cells were stimulated with 100 ng/ml PMA for 48 h to induce differentiation into mature macrophages, which, after culture for another 48 h in the absence of PMA, were either treated with 100 ng/ml LPS for 30 min or left untreated. After desalting procedure with ultrafiltration, the phosphoproteins enriched by phosphoprotein metal affinity column (PMAC) of both groups were run on 2-D electrophoresis to find the spots with different phosphorylation status. Finally, some of these spots were identified by mass spectrometry (MS) and subsequent bioinformatic analysis.

RESULTSCompared to untreated Thp-1 cells, LPS stimulated Thp-1 cells showed 29 spots with reproducible alterations on the 2-D map, including 8 representing up-regulated spots, 7 new spots, 10 down-regulated spots, and 4 absent spots. The newly emerged and absent protein spots were subjected to MS analysis, and 4 of them were identified to be involved in various cellular processes such as proteolysis, signal transduction and protein folding. Among these, phosphorylation of proteasome C2 subunit was dramatically up-regulated in LPS-stimulated cells, as was consistent with previous reports; the phosphorylation of Z-DNA-binding protein 1 has not been reported so far and needs further confirmation.

CONCLUSIONPhosphoprotein affinity profiling is an attractive method for screening novel regulators involved in LPS signaling pathways and can be widely used in systemic study of signal transduction.

Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; drug effects ; metabolism ; Mass Spectrometry ; Phosphoproteins ; analysis ; metabolism ; Phosphorylation ; Proteomics ; methods ; Signal Transduction

OBJECTIVETo observe whether the community-based management for patients with hypertension can reduce the incidence of stroke.

METHODSSample of this study included 36 863 people aged 35 years or more who came from a cohort consisting three communities from Tiantan Hospital, Puren Hospital and the Gymnasium Road Hospital in Beijing, based on the surveys on the Integrated Community Intervention Measures of Cerebro-vascular Diseases. Some patients with hypertension in this cohort were followed up and under management. First-ever stroke was considered as the end-point event.

RESULTSIn both groups diagnosed as borderline hypertension or definite hypertension group, the rates of management and control showed an annual increase. The management rate for women was higher, but the control rate was lower (P < 0.05) than that for men. In the third year of this study, the control rate was nearly 18%. With the qualification of control rate, the risk factors of overall stroke, ischemic stroke or hemorrhagic stroke reduced gradually, and the qualification of control rate showed more effects on hemorrhagic stroke. The qualification of control rate in the three years could cause the risk factors of total stroke, ischemic stroke or hemorrhagic stroke to reduce by 25.7%, 19.1%, 27.4%, respectively. When comparing with blood pressure level at < 160/95 mm Hg (1 mm Hg = 0.133 kPa), the level of < 140/90 mm Hg could reduce the risk factors as: 12.3% to total stroke, 12.8% to ischemic stroke and 14.9% to hemorrhagic stroke.

CONCLUSIONPrograms as long-term followed-up and management for patients with hypertension, and control the blood pressure at low level etc. could significantly reduce the incidence of stroke.

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Humans ; Hypertension ; complications ; epidemiology ; Male ; Middle Aged ; Stroke ; epidemiology ; etiology

OBJECTIVETo clone the coding gene of the stage-specific antigen cC1 from Cysticercus cellulosae and express high levels of soluble cC1 in E.coli.

METHODSThe cC1 gene was amplified from Cysticercus cellulosae by RT-PCR and cloned into pMD18-T vector, followed by subcloning into the prokaryotic expression plasmid pET28a. The recombinant plasmid was transformed into E.coli BL21(DE3) and the expression conditions were optimized. The expressed product was purified by Ni(+)-affinity chromatography, analyzed by high-performance liquid chromatography (HPLC), and identified with SDS-PAGE and Western blotting.

RESULTSThe fragment length of the amplification product by RT-PCR was 1056 bp. Comparison of the amplified gene sequence with the cC1 gene in Genbank identified a samesense point mutation at 423 position in the gene cloned into the expression plasmids. After a 6-h induction with 0.05 mmol/L IPTG at 37 degrees celsius;, the expression of the 40 kd soluble fusion protein exceeded 60% of the total bacterial protein, and the fusion protein was recognized by Cysticercus-infected human sera. The purity of the fusion protein was about 94% after purification by affinity chromatography.

CONCLUSIONThe stage-specific antigen cC1 from Cysticercus cellulosae has been successfully cloned and the soluble protein efficiently expressed in E.coli, which provides the basis for its further study and application.

Animals ; Antigens, Helminth ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Cysticercus ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Solubility ; Swine ; Taenia solium ; immunology

OBJECTIVE: To detect pathogenic variant in a child featuring Usher syndrome type II. METHODS: Peripheral blood samples of the child and his parents were collected for the analysis of variants of hearing impairment-related genes. The findings were verified in 100 individuals with normal hearing. RESULTS: The child was found to harbor compound heterozygous variants of the USH2A gene, namely c.8224-1G>C in intron 41 and c.5678C>G(p.Ser1893X) in exon 28, which were inherited respectively from his mother and father. Based on the American College of Medical Genetics and Genomics standards and guidelines, both c.8224-1G>C and c.5678C>G(p.Ser1893X) variants of USH2A gene were predicted to be pathogenic(PVS1+PM2+PM3). CONCLUSION The compound heterozygous variants c.8224-1G>C and c.5678C>G of the USH2A gene probably underlay the disease in this child. Above finding has enriched the spectrum of USH2A gene variants.
Child ; Exons ; Extracellular Matrix Proteins/genetics* ; Family ; Humans ; Introns ; United States ; Usher Syndromes/genetics*

Objective To analyze the property of the new antibacterial agent (N, N-di-n-decyl-N, N-dimethyl-ammonium 5-oxopyrrolidine-2-carboxylate, DAPC) to prevent Streptococcus mutans' adhesion on tooth surface.Method determine the minimal inhibitory concentration and minimal bactericidal concentration of DAPC and Chlorhexidine gluconate by liquid dilution method.Set the Chlorhexidine gluconate as the positive control, while PBS as the negative control.Use the crystal violet staining to measure the quantity of biofilm.Re s ult The MIC of DAPC on Streptococcus mutans was 0.0031250％ and MBC was 0.0062500％. The MIC of Chlorhexidine gluconate was 0.0015625％, while the MBC was 0.0031250％.When concentration of the two antibacterial agents was MIC, the quantity of biofilm have no significance among three groups.However, when concentration of Chlorhexidine gluconate and DAPC were 0.12％ and 0.24％ respectively, biofilm of experimental group was lower than PBS, and there is no significance between Chlorhexidine gluconate group and DAPC group after 24 hours incubation.Conclus ion New antibacterial agent DAPC have significant effect in inhibiting of Streptococcus mutans.And the residual of DAPC on teeth can maintain a long time of antibacterial effect to inhibit Streptococcus mutans' adhesion.

Objective To study the influence on the tumor after percutaneous intra-tumor injection of ~(32)P-GMS in liver cancer as well as its suitable dose.Methods 24 New Zealand rabbits were used to establish the animal model of VX-2 liver cancer,and divided into A,B,C and D groups with individually 37,74,111 and 148 MBq of ~(32)P-GMS being injected,respectively;and then pathological changes of tumor were observed by light and electron microscope respectively.Result The dose of ~(32)P-GMS was obviously correlated with the radioactivity damage of tumor cells.In the A and B groups,the tumor cells were not observed to disappear completely after injection of ~(32)P-GMS,but in C group,tumor cells were almost completely disappeared and surrounded by a lot of connective tissue.Although the tumor cells were found to disappear completely in D group,normal liver tissues were also involved.Conclusion Percutaneous intra-tumor injection of ~(32)P-GMS with suitable dose that may induce the tumor tissue to be maximally damaged and may also provide some significances to prevent the tumor metastasis.