Adolescent ; Adult ; Bundle of His ; pathology ; physiopathology ; Child ; Female ; Humans ; Male ; Purkinje Fibers ; pathology ; physiopathology ; Tachycardia, Ventricular ; pathology ; physiopathology

Objective To establish a method of SYBR Green Ⅰ FQ-PCR for detecting the expression of miR-202 in peripheral blood mononuclear cells ( PBMC ) and analyze the expression of miR-202 and its clinical significance in MM.Methods Reverse transcription was performed with specific stemloop primer for miR-202,and then FQ-PCR was used to detected the expression of miR-202 in 21 MM patients and 20 healthy people.Data was presented as mean ± standard deviation ( (x) ± s ).Non-parametric Mann-Whitney test was used to analyze the difference between MM group and control group.In addition,1∶ 125 dilution of one test sample was detected by repeated 5 times,and the same sample was tested one time a day for 3 days,repeated 5 each time.The assessment of repeatability of measurements was done by calculating standard deviation and variation coefficient from threshold cycle (Ct).Results FQ-PCR detection of miR-202 in PBMC was amplified by the standard S-curve,with a single melting curve peak and no complex peak,which showed good specificity.The assay showed good reproducibility (intra-assay coefficient 1.2％ and inter-assay coefficient 3.2％ ) and high sensitivity ( 12.8 pmol/μ1).The expression of miR-202 in MM was 0.014 ( 0.007 - 0.221 ) and 1.844 ( 0.162 - 3.966 ) in normal controls.The expression level of miR-202 was significantly higher in MM than in normal controls (U =48.000,P ＜0.01 ).Conclusions FQ-PCR provide us a rapid,sensitive and specific method for detection of miR-202.The expression level of miR-202 is higher in MM than in normal controls.It is possible to play a role in MM progress and may be a useful marker to evaluate the development and treatment of MM.

Aorta ; pathology ; surgery ; Aortopulmonary Septal Defect ; therapy ; Balloon Occlusion ; methods ; Catheterization, Swan-Ganz ; methods ; Child ; Female ; Humans ; Pulmonary Artery ; pathology ; surgery ; Treatment Outcome

Taking α-asarone as model drug, mono methoxy polyethylene glycol-polylactic acid copolymer (mPEG-PLA) as the drug carrier material to prepare drug-loading nanoparticles by premix membrane emulsification for nasal administration. The prepared nanoparticles were spherical with smooth surface and average particle size of 360 nm. Polydispersity index (PDI) was 0. 030, average drug loading of (11.5 ± 0.045) % (n = 3), and the encapsulation efficiency of (86.34 ± 0.11) % (n = 3). X-ray diffraction and differential scanning calorimetry results showed that, α-asarone existed in mPEG-PLA carrier in amorphous or molecular state, different from simple physical mixture. In the in vitro release test in simulated human nasal cavity, α-asarone apis can be released quickly at close to 94% at 102 h, in line with the first-order kinetics (R² = 0.981 9). mPEG-PLA drug-loading nanoparticles release only 54%, with slow release effect, in line with Riger-Peppas model (R² = 0.967 9, n = 0.630 2), for non-fick diffusion, released by the spread of drugs and skeleton dissolution dual control. This provided the foundation for nasal drug delivery in vivo pharmacokinetic study.
Administration, Intranasal ; Anisoles ; chemistry ; Calorimetry, Differential Scanning ; Nanoparticles ; chemistry ; Polyesters ; chemistry ; Polyethylene Glycols ; chemistry ; Solubility ; X-Ray Diffraction

Hepatitis B ; Humans ; Th17 Cells

Animals ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cell Separation ; Cells, Cultured ; Interferon-alpha ; pharmacology ; Liver ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Rats ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; bcl-2-Associated X Protein ; fas Receptor ; biosynthesis ; genetics

OBJECTIVETo explore the utility of Principal Factor Analysis (PFA) in chromatographic data for quality control.

METHODChromatographic fingerprints of processed root pieces of Paeonia lactiflora were determined by HPLC, the PFA was used for data processing.

RESULTThe quantitative differences among different growing areas and different processing batches were found with the method.

CONCLUSIONThe method could be used in quality control for monitoring between-batch products of traditional Chinese pharmaceutical process.

Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Factor Analysis, Statistical ; Hot Temperature ; Pinellia ; chemistry ; Plant Preparations ; analysis ; chemistry ; standards ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Wine

OBJECTIVETo investigate the antitumor efficacy of death receptor 5, its ligand (TRAIL) and DR5mAb in human hepatocellular carcinoma.

METHODSExpression of DR5 in the HCC cell lines HepG2, SMMC 7721 and normal human liver cell line LO2 was measured at mRNA and protein level by semi-quantitative RT-PCR and Western blot, respectively. MTT method was used to measure the cell viability and flow cytometry assay was used to detect apoptosis so as to observe the inhibitory effect of TRAIL and DR5mAb on HCC cells.

RESULTSDeath receptor 5 was highly expressed in the HCC cell lines, but rarely expressed in normal human liver cell line (P < 0.01). With the increase of TRAIL concentration, the cell viability of HCC cells decreased gradually. However, when the concentration of TRAIL was above 1000 ng/ml, HCC cells were resistant to TRAIL, but still sensitive to DR5mAb. After incubation with DR5mAb (1000 ng/ml) for 24 h, the rate of apoptosis in HCC cells reached to 52.45% +/- 0.57%, which was higher than that incubated with TRAIL under the same condition (14.74% +/- 0.48%) (P < 0.05). The cell viability of normal human liver cell line treated with TRAIL tended to decline with the increase of the concentration, which was significantly different from that of matched control group. But DR5mAb had little effect on normal human liver cell line.

CONCLUSIONDeath receptor 5 as a target plays an important role in the course of HCC apoptosis induction. Agonistic monoclonal antibody specific for human DR5 can selectively and effectively kill hepatocellular carcinoma cells in vitro, while is not harmful to normal human hepatocytes. It reveals that DR5mAb might provide a new direction in hepatocellular carcinoma treatment research.

Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Cell Survival ; drug effects ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; genetics ; immunology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis

OBJECTIVETo characterize the structural and the functional feature of a novel gene HSPCSET isolated from human CD34+ hematopoietic stem/progenitor cells (HS/PCs).

METHODSBioinformatic technology was used to identify the structural features of the HSPCSET protein and perform the multiple sequence alignment. Yeast-two-hybrid system was used to identify the proteins interacting with the HSPCSET protein. After sequencing, we selected out the positive clones which had clear functions, and carried out beta-gal experiment and GST pull down assay to confirm the results. The cellular location of the HSPCSET was checked by immunofluorescence assay.

RESULTSThe HSPCSET protein belongs to a SET domain family, which is evolutionarily conserved across species. It implied that HSPCSET may have biologically important function. Using yeast-two-hybrid system, we showed that the protein sequence with SET domain might bind to 13 proteins, which involved in signaling transduction, transcriptional regulation, apoptosis, tumorigenesis, development, etc. And 4 proteins (GADD34, SIVA, DNAJ and PHF1) were confirmed by one-on-one back of the hybrid experiment, beta-gal test and GST pull down assay. When GADD34 and HSPCSET were co-transfected, they co-localized in the nucleus, suggesting a strong interaction.

CONCLUSIONThe novel gene HSPCSET is likely to have biologically important function. This study provides the basis for further studies of its function in hematopoiesis and tumorigenesis.

Amino Acid Sequence ; Animals ; Antigens, Differentiation ; metabolism ; Cell Cycle Proteins ; metabolism ; Computational Biology ; Conserved Sequence ; Hematopoietic Stem Cells ; metabolism ; Humans ; Molecular Sequence Data ; Protein Phosphatase 1 ; Protein Structure, Tertiary ; Proteins ; chemistry ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Two-Hybrid System Techniques