American Medical Association ; Humans ; Liver Failure, Acute ; diagnosis ; therapy ; Liver Transplantation ; Practice Guidelines as Topic ; United States

Europe ; Hepatitis C ; diagnosis ; therapy ; Humans ; Practice Guidelines as Topic ; Societies, Medical

Objective To examine the expression of Toll-like receptor(TLR)4 protein and the transcription of TLR4 mRNA in the peripheral blood mononuclear cells(PBMC)from patients with hepatitis B virus(HBV)infection and explore the relationship between TLR4 and chronic HBV infec- tion.Methods The expression level and transcription level of TLR4 were determined by flow cytometre and reverse transcriptase polymerase chain reaction respectively in PBMC from 37 chronic hepatitis patients,28 liver cirrhosis patients,31 severe hepatitis patients and 27 healthy controls. Meanwhile,liver function,as well as blood routine test,prothrombin test activity(PTA)and HBV DNA was measured.Results The expression level and transcription level of TLR4 in patients were higher than those in healthy controls(P

Genomic DNA of two fungi Thamnidium elegans and Umbelopsis isabellina were extracted with an amended Cetyltrimethyl Ammonium Bromide (CTAB) method. This modified method uses repeated freezing in liquid nitrogen and thawing with combination of shocking with glass beads to replace of the tra- ditional method. Quality and concentration of DNA extracted by the modified methodwere tested. Compared with the traditional method, higher yield and purity of genomic DNA were obtained with less amount ofmy- celium. The result indicted that this is a simple and highly efficient method, which is suitable to treat many samples at one time and for basic molecular experiments, such as restriction endonuclease reaction and PCR.

Adult ; Hepatitis C, Chronic ; complications ; Humans ; Interferons ; adverse effects ; Male ; Thyroid Diseases ; chemically induced

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.

Objective To compare the difference of the protein about the patient of hepatitis B who received adefovir dipivoxil(ADV)therapy,and seek the useful biomarker of effective therapy.Methods We used the two-dimensional gel electrophoresis technology to examine HBV infected serum samples aiming at searching protein's alteration after ADV therapy.Results After 1 year's treatment,haptoglobin, haptoglobin 2-alpha raised and alpha-l-antitrypsin precursor,Factor B,Chain B,transthyretin,glutathione peroxidase,alpha-2-HS-glycoprotein,retina]binding protein,retinol-binding protein precursor, apolipoprotein,apolipoprotein A-I precursor fell in viral response patients.Transthyretin raised and leucine- rich alpha-2-glyeoprotein,haptoglobin,alpha-2-actin,apolipoprotein A-I precursor fell in none viral response patients.To compare two groups:apolipoprotein A-I have the same change and haptoglobin, transthyretin have the opposite change.Conclusion Proteomics study can find the alteration of protein during the ADV treatment,and is helpful to searching the predictable biomarker to ADV.

AIM:To investigate the effect of over-expression of peroxisome proliferator-activated receptor γ co-activator-1α (PGC-1α) on mitochondrial morphology and cell apoptosis in the cortical neurons with oxygen glucose depriva-tion/reoxygenation(OGD/R). METHODS:The whole gene sequence of PGC-1α was obtained from the cerebral cortex of C57BL/6 mice by RT-PCR and cloned into the eukaryotic expression vector pEGFP-N1. The pEGFP-N1-PGC-1α was iden-tified by PCR,and transfected into cortical neurons. The level of PGC-1α expression was identified by Western blot. The cortical neurons transfected with pEGFP-N1 and pEGFP-N1-PGC-1α vectors were treated with OGD/R. The mitochondrial mass,reactive oxygen species (ROS) and ATP production,cell apoptosis and changes of cleaved caspase-3 were detected by MitoTracker Red staining,flow cytometry,ATP metabolic assay kit and TUNEL. RESULTS:Over-expression of PGC-1α inhibited the decrease in mitochondrial biogenesis capacity and the ROS formation of OGD/R neurons(P<0.05),en-hanced the ability of ATP synthesis (P<0.01),inhibited neuronal apoptosis (P<0.01) and decreased the activation of caspase-3 (P<0.01). CONCLUSION:PGC-1α over-expression inhibits neuronal apoptosis with OGD/R treatment by promoting mitochondrial biogenesis,inhibiting the production of ROS and maintaining mitochondrial function. PGC-1α may be used as a target for the development of cerebral ischemia/reperfusion injury drugs.

OBJECTIVETo investigate the inhibition of COX-2 gene expression and its effects on malignant proliferation of human lung adenocarcinoma A549 cells after interfering at different target sites in vitro.

METHODSThe 3rd, 7th and 10th exon of COX-2 were selected as the targets and three COX-2 siRNA expression vectors with human U6 promoter were constructed. Three siRNA expression vectors and two vacant vectors were transfected into A549 cells expressing COX-2 with lipofectamine, respectively. The transfected cell strains were constructed and the change of COX-2 expression levels was examined by Western blot and RT-PCR. The effects on the proliferation of A549 cells after interfering at different target sites were studied by cell growth curve and colony formation assay in vitro.

RESULTSThe three siRNAs and U6 promoter were validated by PCR, restriction endonuclease digestion, DNA sequencing and BLAST alignment, and cloned into the pEGFP vector. The cell strains transfected were named as A549-3, A549-7, A549-10, A549-p and A549-pU6, respectively. A549-p cells showed expression of GFP and A549-3, A549-7, A549-10, A549-p and A549-pU6 cells did not show at 24, 48 and 72 hours after transfection. The results of RT-PCR and Western blot showed an inhibition of COX-2 expression after interfering at three target sites (3rd, 7th and 10th exons). In contrast to A549 cells, the levels of COX-2 mRNA of A549-3, A549-7 and A549-10 cells were reduced by 10.6%, 33.4% and 61.2%, respectively. The levels of COX-2 protein of A549-3, A549-7 and A549-10 cells were reduced by 26.7%, 44.7% and 56.2%, respectively. The results of cell growth curve and colony formation assay showed a slowing down of the growth of A549-10 cells and reduction of their colony formation rate. The other two targets had no apparent effect on the growth of A549 cells.

CONCLUSIONThere is a significant inhibiting effect of RNA interference on the malignant proliferation of A549 cells in vitro, and the most striking effect can be seen when the 10th exon of COX-2 is taken as the interference target.

Adenocarcinoma ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Cyclooxygenase 2 ; genetics ; metabolism ; physiology ; Exons ; Genetic Vectors ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Promoter Regions, Genetic ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo investigate the vertical bone height and the bone density of the palate for implants placement using cone beam CT(CBCT) and to provide references to the safe and stable placement of palatal implants.

METHODSThree-dimensional reformatting images were reconstructed with the selected CBCT scanning data of 34 patients aged 18 to 35 yeras, by means of EZ implant software. The vertical bone height was measured at 20 interesting sites of palate. Bone density was measured at 10 sites that could support 3.0 mm long implants. The data of the vertical bone height and bone density were analyzed by K-means cluster analysis.

RESULTSAccording to the cluster analysis results, the 10 sites were classified into 3 clusters. There were statistical differences among these three clusters in bone height and bone density (P < 0.05). The LSD result showed that the greatest mean value of vertical bone height was obtained in cluster 2, followed by cluster 1 and cluster 3; the highest bone density was founded in cluster 3, followed by cluster 1 and cluster 2.

CONCLUSIONSEvaluation of the sites for palatal implant placement with cone beam CT would be helpful in safe and stable implantation.

Adolescent ; Adult ; Bone Density ; Cone-Beam Computed Tomography ; Dental Implantation, Endosseous ; Female ; Humans ; Male ; Palate ; diagnostic imaging ; Software ; Young Adult