Adolescent ; Cardiomyopathies ; complications ; genetics ; Female ; Heart Ventricles ; abnormalities ; Humans ; Pedigree ; Sick Sinus Syndrome ; complications ; genetics

OBJECTIVETo observe the effect of small interfering RNA (siRNA)-induced MAPK p42 silencing on the survival of HeLa cells.

METHODSTwo siRNAs targeting at the MAPK p42 gene and one random siRNA were synthesized respectively by Silencer siRNA Construction Kit and transfected into HeLa cells by Lipofectamin 2000. The expression of p42(MAPK) in the transfected HeLa cells was analyzed by Western blotting and immunohistochemistry, and the morphology of cells were observed with electron microscope. TUNEL assay and Annexin V/PI staining were employed for detecting the cell apoptosis.

RESULTSThe expression of p42(MAPK) in the HeLa cells was remarkably suppressed after transfection with the two siRNAs, reduced by about 2.5 and 3.2 folds respectively in comparison with the negative control. Chromatin margination in the cell nuclei were observed in the transfected cells, and TUNEL assay and Annexin V/PI staining further confirmed the occurrence of cell apoptosis.

CONCLUSIONIn vitro MAPK p42 siRNA-1 and siRNA-2 transfection can specifically silence the gene expression and induce apoptosis of HeLa cells.

Apoptosis ; physiology ; Gene Silencing ; physiology ; HeLa Cells ; Humans ; Mitogen-Activated Protein Kinase 1 ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo screen for siRNAs that inhibit the expression of p42(MAPK) in HeLa cell line.

METHODSThree p42(MAPK) siRNAs and one random siRNA were synthesized using Silencer siRNA Construction Kit, and labeled with Cy-3 for measurement of transfection effect. SiRNAs were transfected into HeLa cells by Lipofectamin 2000. The expression of p42(MAPK) was analyzed by Western blot. The biological effect of siRNAs on HeLa cell growth was monitored by MTT and flow cytometry.

RESULTSTwo siRNAs (siRNA-2 and siRNA-3) among three tested were identified to be able to downregulate the p42(MAPK) expression. A concurrent growth retardation of HeLa cell line was observed in comparison with the control.

CONCLUSIONInhibition of p42(MAPK) expression with siRNA technique can inhibit the proliferation of HeLa cells.

Cell Cycle ; Cell Proliferation ; Cell Survival ; Flow Cytometry ; HeLa Cells ; Humans ; Mitogen-Activated Protein Kinase 1 ; biosynthesis ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

This study was aimed to explore the effect of BCL11A gene on transcription of γ-globin gene in K562 cells. B-cell lymphoma/leukemia 11A (BCL11A) gene was silenced by small interfering RNA (siRNA) expression vectors in K562 cells (human erythroblastic leukemia cell line). Gamma-globin mRNA level in K562 cells was determined by RT-PCR. Association between the BCL11A gene and γ-globin gene transcription was explored by comparison of mRNA levels. The results indicated that the silence rate of the BCL11A gene in K562 cells by 4 siRNA expression vectors was 49.7%, 55.4%, 78.2%, and 84.1%, respectively. The siRNA expression vector with 84.1% silence rate was transfected into K562 cells, transcription level of γ-globin mRNA in K562 cells transfected with siRNA expression vector increased 2.4 times as compared with control K562 cells. It is concluded that level of γ-globin mRNA increases when the BCL11A gene is silenced. It indicates that the BCL11A gene may be a negative regulator for γ-globin gene expression.
Carrier Proteins ; genetics ; Gene Expression Regulation, Leukemic ; Genes, Regulator ; Genetic Vectors ; Humans ; K562 Cells ; Nuclear Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transcription, Genetic ; Transfection ; gamma-Globins ; genetics

To investigate the possible mechanism of apoptosis induced by indole-3-acetic acid (IAA) combined with horseradish peroxidase in leukemia cell line K562, cell proliferation and apoptosis of K562 cell were examined by MTT assay and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), respectively; the activity of superoxide dismutase (SOD) and the quantitative change of MDA were measured by biochemical method; changes of free radical were determined by 2, 7-dichlorofluorescin diacetate (DCFH-DA) probe with confocal microscopy. The results showed that of MTT assay and TUNEL indicated that IAA/HRP could significantly inhibit cell proliferation (P < 0.05) and induce apoptosis of K562 cell (P < 0.01), at the same time a positive correlation was found between apoptosis rate and IAA concentration (r = 0.971, P < 0.01). The activity of SOD and the quantitative of MDA increased, accompanied with a rise in IAA concentration. Results detected by DCFH-DA probe indicated that the fluorescence intensity of intracellular free radical increased, as compared with control, and a positive correlation was found. It is concluded that IAA/HRP can inhibit proliferation of K562 cells and induce K562 cell apoptosis, its mechanism may be related with the increase of intracellular free radical due to the effects of IAA/HRP.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Drug Synergism ; Horseradish Peroxidase ; pharmacology ; Humans ; In Situ Nick-End Labeling ; Indoleacetic Acids ; pharmacology ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Microscopy, Confocal ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

Objective To provide the theoretical basis for the application of cortical somatosensory evoked potential (CSEP) in monitoring the function of the spinal cord to prevent postoperative neurological dysfunction. Methods Thirty-three New Zealand rabbits were randomly divided into 6 groups: 8 were chosen as control group to eliminate the influence of anesthesia and surgery on the evoked potential; the other 25 were assigned to 5 sub-experimental groups (n=5) according to the artery number being ligatured in the left renal arteries and the spinal arteries. Baseline evoked potential in each group was noted immediately after anesthesia; the CSEP were recorded at different time points (before vascular ligation, 30 min and 2 d after vascular ligation). Motor functions were assessed after narcotic conscious and 2 d after vascular ligation. The specimens were taken for HE staining. Results The latency was not sensitive to spinal cord ischemia and no significant difference of that was found between the experimental groups and the control group (P>0.05); except that, the changes of theamplitudes were very complex and the specificity of motor function was decreased. The amplitude reduced and then gradually restored in the 2, 3 and 4 levels of ligation. The changes of amplitude could indicate the degree of pathological damage in the spinal cord and its motor function. Conclusion Complex amplitude of somatosensory evoked potential can be found in the acute phase of ischemia in the spinal cord. Specificity of motor function is poor resulting from its signal averaging process. Motor evoked potential monitoring in the operation should also be added in the detection of the spinal cord.

OBJECTIVETo explore and evaluate the combined conservative managements in the treatment of cervical chylous leakage.

METHODSThirty nine cases of cervical chylous leakage from June 1992 to June 2008 were retrospectively analyzed in this hospital. All of the 39 cases were cured by treating with conservative individualized therapy, including the applying of diet with high calorie, high protein and low fat and fatty food should only contains medium-chain triglycerides, total parenteral nutrition, keep the balance of hydrogen and electrolyte and correct hypoproteinemia, local pressure dressing, high persistent vacuum drainage (-50 approximately -80 kPa) and/or somatostatin analogue.

RESULTSAll the cases of chylous leakage happened 2nd to 5th days after the operation. Among the 39 cases, 7 were high flow (drainage>or=500 ml/d) chylous leakage, the amount of drainage reached as high as 1440 ml per day. The time of chylous leakage closure was 3 approximately 12 days, and the mean time was 7 days. No one experienced re-operation, wound hydrops or wound infection.

CONCLUSIONSThe conservative individualized therapy may play a key role in the treatment of cervical chylous leakage.

Adolescent ; Adult ; Aged ; Chylous Ascites ; etiology ; therapy ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Parenteral Nutrition, Total ; Postoperative Complications ; therapy ; Retrospective Studies ; Young Adult

OBJECTIVETo obtain allele and genotype frequencies and related forensic data of CF1PO, TPOX and TH01 loci in Chinese Xinjiang Sibo population.

METHODSGenomic DNA from peripheral blood mononuclear cells of normal Chinese Xinjing Sibo population was used as template, and CSF1PO, TPOX and TH01 fragments were amplified by PCR. The PCR products were analyzed by 4% denaturing PAGE and detected using silver stain detection.

RESULTSNine alleles were found at CSF1PO locus, eight alleles at TPOX locus and eight alleles at TH01 locus in Chinese Sibo population. All the 3 loci complied with Hardy-Weinberg equilibrium. The heterozygosities were 0.9426, 0.8361 and 0.8853, and the polymorphism information contents were 0.8298, 0.7213 and 0.7626 for CSF1PO, TPOX and TH01, respectively.

CONCLUSIONThe data on the alleles frequency of these 3 STR loci might be used for individual identification and paternity identification and for genetic researches in Chinese Sibo population.

Alleles ; China ; DNA ; genetics ; Gene Frequency ; Genetic Markers ; genetics ; Genotype ; Humans ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics

OBJECTIVETo compare the efficacy of three-dimensional (3D) and two-dimensional (2D) quantitative coronary X-ray angiography (QCA) and visual estimation in the assessment of target vessels.

METHODSThe radiographic data of 60 patients (65 vessel segments) receiving coronary angiography and interventional stent placement were retrospectively analyzed. The area stenosis, diameter stenosis, lesion length, and reference diameter assessed by Medis 3D QCA, Siemens 2D QCA and visual estimation were compared.

RESULTSThree-dimensional reconstruction was successfully performed for 65 vessel segments, and 3 target vessel were excluded due to the lack of a second angiographic view for 3D reconstruction. There were significant differences in the assessments of the area stenosis [(73.87 ∓ 8.98)% vs (79.10 ∓ 8.06)% vs (83.53 ∓ 8.19)%, P<0.001], lesion length (28.95 ∓ 17.31 mm vs 26.20 ∓ 16.04 mm vs 27.21 ∓ 16.58 mm, P<0.001), reference diameter (28.95 ∓ 17.31 mm vs 26.2 ∓ 16.04 mm vs 27.21∓16.58 mm, P<0.001) by 3D QCA, 2D QCA and visual estimation; the diameter stenosis assessed by 3D [(54.21 ∓ 9.48)%] and 2D QCA [(57.84 ∓ 10.17)%] also differed significantly (P=0.016).

CONCLUSION3D QCA allows successful three-dimensional reconstruction of the target vessel and restores the actual dimensions of the vessel for a more accurate assessment of coronary artery disease than 2D QCA and visual estimation.

Aged ; Coronary Angiography ; methods ; Coronary Disease ; diagnostic imaging ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo investigate the roles of STAT1 and STAT2 in growth inhibition induced by phosphatidylethanolamine (PE) in human hepatoma HepG2 cells.

METHODSThe growth of HepG2 cells exposed to 0.125, 0.25, 0.5 and 1.0 mmol/L PE was assessed by MTT assay, and the expressions of STAT1 and STAT2 were analyzed using immunocytochemical assay.

RESULTSPE inhibited the growth of HepG2 cells in a dose-dependent manner and increased the expression of STAT1 and STAT2 in comparison with those in the control group. AG490, an inhibitor of JAKs, partially reversed PE-induced growth inhibition of HepG2 cells.

CONCLUSIONSTAT1 and STAT2 are involved in the growth inhibition of human hepatoma HepG2 cells induced by PE.

Apoptosis ; drug effects ; Cell Proliferation ; Hep G2 Cells ; Humans ; Phosphatidylethanolamines ; pharmacology ; STAT1 Transcription Factor ; metabolism ; STAT2 Transcription Factor ; metabolism