Catheter Ablation ; methods ; Humans ; Hypertension ; surgery ; Renal Artery ; Sympathectomy ; methods

Experiment teaching is a most important item of college teaching, and plays a vital and unexchangeable role to train students for their ability to practice and to innovate. To construct a feasible and scientific examination system of experiment teaching, will significantly help deepen the reformation of experiment teaching and improve the teaching quality. So according to the request for cultivating qualified ap- plication person, we preliminarily constitute the valuation system throughout the whole course and in the final, of theoretic examination and practice examination, and combined with students’ self-valuation and valuation from teachers. In fact, the system works well with a perfect effect.

Aged ; Female ; Heart Atria ; pathology ; Humans ; Middle Aged

OBJECTIVETo observe the changes in matrix metalloproteinase-2,7 (MMP-2,7) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in deep partial thickness burn during the process of wound healing, and the effects of bFGF on wound healing.

METHODSThe rats inflicted by 30% TBSA deep partial thickness burn were randomly divided into simple scald and bFGF treatment groups. Biopsies from wound skin were harvested at 3 and 6PBHs and 1, 3, 7, 14 PBDs for the detection of the epithelialization rate and collagen content. The above indices were also detected in the skin of another 6 normal rats as normal control.

RESULTS(1) The epithelialization rate in bFGF treatment group was higher than that in simple scald group during 3PBH to 14 PBD. (2) The collagen contents in both bFGF treatment group and simple scald group were continually decreased during 3 PBH to 3 PBD, and increased from 7 to 14 PBD, but still lower than that in normal control (P < 0.05). (3) The expression of MMP-2,7 and TIMP-2 in simple scald group enhanced from 1 to 14 PBD, and peaked on 7 PBD. (4) The expression of MMP-2,7 in bFGF treatment group was similar to that in simple scald group from 3 to 6 PBH, while the expressions of MMP-2,7 and TIMP-2 was higher than those in simple scald group from 1 to 14 PBD.

CONCLUSIONThe collagen deposition would be affected by the activities of extracellular matrix in scald wound in rats. Changes in MMP-2,7 and TIMP-2 expressions were an important process of wound repair, which was closely related to the acceleration of wound healing by the application of bFGF.

Animals ; Burns ; metabolism ; Collagen ; analysis ; Epithelium ; physiology ; Fibroblast Growth Factor 2 ; pharmacology ; Male ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 7 ; analysis ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; analysis ; Wound Healing

OBJECTIVETo determine the correlation between methylation of p16 gene in promoter region and the carcinogenesis and progression of squamous cell carcinoma (SCC) of buccal mucosa.

METHODSMethylation of pl6 gene in SCC and leukoplakia of buccal mucosa was investigated by MSP and pl6 protein was analyzed by Western blot.

RESULTSThe methylation of p16 gene was found in 15 of 30 cases SCC and 1 of 10 cases of leukoplakia of buccal mucosa (P < 0.05). Methylation of p16 gene in SCC of buccal mucosa was not related with age, sex, cell differentiation and clinical stage. But methylation of p16 in the cases with lymph node-metastasis was higher than that in the cases without lymph node-metastasis protein (P < 0.05). Meanwhile Methylation of p16 gene was positively correlated with no-expression of p16 protein (P < 0.01).

CONCLUSIONSThe methylation of p16 gene leaded to the inactivation of p16 gene and was related with the carcinogenesis and progress of SCC of buccal mucosa.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cheek ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA Methylation ; Genes, p16 ; Humans ; Leukoplakia, Oral ; embryology ; genetics ; pathology ; Mouth Neoplasms ; genetics ; metabolism ; pathology ; Promoter Regions, Genetic

OBJECTIVETo examine the effects of H-ras gene silence on cell cycle, proliferation and apoptosis of salivary adenoid cystic carcinoma -M (SACC-M) cell lines.

METHODSThe plasmid H-ras-shRNA, containing the sequence of shRNA targeting H-ras, and HK-shRNA (without interfering effect) were constructed and transfected into SACC-M cells. The cell line with shRNA plasmid stable expression was isolated by G418. The expression levels of H-ras were detected by RT-PCR and protein immunofluorescent assay; cell cycle and cell apoptosis were analyzed by flow cytometry (FCM). The proliferation of cell was also determined by subcutaneous tumor formation in nude mice.

RESULTSAfter transfection of H-ras-shRNA plasmid, the mRNA expression of H-ras in SACC-M cells was down-regulated by 61.80% and protein expression of H-ras was inhibited by 62.76%; the cell proliferation was inhibited obviously; the G0G1 phase cells were increased. The cell apoptosis rate of H-ras-shRNA group was significantly higher than that of HK-shRNA group (P <0.05). The volume of subcutaneous tumor in nude mice was significantly smaller in Hras-shRNA group than in control group.

CONCLUSIONSThe recombinant plasmid HRAS-shRNA could efficiently down-regulate the expression of H-ras gene and protein, induce apoptosis of SACC-M cells and simultaneously inhibit proliferation of these cells in vitro and in vivo.

Animals ; Apoptosis ; Carcinoma, Adenoid Cystic ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Gene Silencing ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins p21(ras) ; genetics ; RNA, Small Interfering ; genetics ; Salivary Gland Neoplasms ; genetics ; pathology ; Transfection

OBJECTIVETo evaluate the expression of dental matrix protein-l (DMP1) in porcine oral mucosa fibroblasts (POMF) transfected by DMP1 and the influences of the transfection.

METHODSThe full length of porcine DMP1 cDNA was linked into an eukaryotic expression vector pEGFP-C1. POMF and mesenchymal stem cells (MSC) were transfected with the pEGFP-DMP1. The expression of DMP1, dental sialoprotein (DSP), amelin and ameloblastin (Ambn) gene of transfected POMF and MSC were detected by RT-PCR. The expression of DMP1 and DSP protein was examined by immunocytochemical staining. The formation ratio of mineralized nodules of transfected cells was compared with un-transfected ones after mineralized induction. The formation of mineralized nodules of three-dimensional pellet transfected cells was compared with un-transfected ones after hematoxylin and eosin staining.

RESULTSThe constructed pEGFP-DMP1 could produce 4.7 kb and 1.5 kb fragments. DMP1 gene, DSP gene and Ambn gene were expressed by POMF after transfection. Immunohistochemical staining and the quantitative analysis of protein showed that DMP1 and DSP protein was positive in transfected POMF and MSC. The formation ratio of mineralized nodules of transfected POMF and MSC was higher than that of un-transfected ones (P < 0.05).

CONCLUSIONSThe expression of porcine DMP1 in POME after gene transfection can induce the expression of tooth-development-associated gene Ambn and DSP and enhance the formation of mineralized nodules.

Animals ; Calcification, Physiologic ; Cell Differentiation ; Cells, Cultured ; Extracellular Matrix Proteins ; genetics ; metabolism ; Fibroblasts ; cytology ; metabolism ; Genetic Vectors ; Mouth Mucosa ; cytology ; Phosphoproteins ; genetics ; metabolism ; Swine ; Transfection

OBJECTIVETo study the feature of apoptosis of salivary adenoid cystic carcinoma (SACC) induced by recombined human tumor necrosis factor-alpha (rhTNF-alpha) in nude mice, and to evaluate the related genes expression of apoptosis.

METHODSTwelve SPF grade 4 approximately 5 weeks old female Balb/c nude mice were selected in this study. SACC-83 cells were collected to 6 x 10(7) per milliliter and injected subcutaneously. Group A and B were experimental group which was given 100 x 10(4) IU/kg TNF-alpha or 10 x 10(4) IU/kg TNF-alpha respectively. Group C was only given normal saline and used as normal control. The investigations were adopted by using both light and transmission electron microscope (LM and TEM), flow cytometer and In Situ Cell Death Detection Kit. The evaluations of bax and bcl-2 expression were utilized by immunohistochemistry.

RESULTSThe percentage of apoptosis of transplanted tumors was much higher than that of the control (P<0.01). Apoptotic cells were calcified and grit bodies were formed. Apoptotic cells expressed and contained significantly higher proportions of both bax and bcl-2 proteins (P<0.05).

CONCLUSIONSIt is suggested that calcification may be the obvious feature and the last outcome of the apoptosis of SACC transplanted tumors. Apoptosis induced by TNF-alpha can increase the expressions of bax and bcl-2.

Animals ; Apoptosis ; Carcinoma, Adenoid Cystic ; chemistry ; pathology ; therapy ; Female ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Salivary Gland Neoplasms ; chemistry ; pathology ; therapy ; Tumor Necrosis Factor-alpha ; pharmacology ; bcl-2-Associated X Protein

OBJECTIVESerum pregnancy-associated plasma protein A (PAPP-A) is increased in acute coronary syndrome patients and related to prognosis. We investigated the effects of C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-alpha) on PAPP-A mRNA expression in monocytes.

METHODSMonocytes were isolated by Ficoll density gradient centrifugation from blood of healthy volunteers. The PAPP-A expressions at mRNA level post CRP or rhTNF-alpha stimulation were measured by RT-PCR.

RESULTSPAPP-A mRNA expression in peripheral blood monocytes increased 2 hours (0.2128 +/- 0.0136) and peaked 24 hours (0.6837 +/- 0.1360) after CRP (20 mg/L) stimulation compared with control group (0.1842 +/- 0.0101). PAPP-A mRNA expression increased rapidly, peaked 2 hours (1.2546 +/- 0.0866) and remained elevated up to 24 hours (0.8203 +/- 0.0413) after rhTNF-alpha (100 ng/ml) stimulation. The effects of CRP and TNF-alpha were dose-dependent. PAPP-A mRNA expression of monocytes were 0.2544 +/- 0.0611, 0.4177 +/- 0.1200, 0.5828 +/- 0.0152, 0.6837 +/- 0.1360 after stimulated with CRP (1, 5, 10, 20 mg/L), and 0.2424 +/- 0.1378, 0.3335 +/- 0.0196, 0.5742 +/- 0.0131, 0.6913 +/- 0.0219 and 0.8203 +/- 0.0413 after stimulated with rhTNF-alpha (5, 10, 25, 50 and 100 ng/ml). Actinomycin D, the DNA-directed RNA polymerase inhibitor, completely blocked CRP and TNF-alpha induced PAPP-A expression.

CONCLUSIONSPAPP-A mRNA expression could be stimulated by CRP and TNF-alpha in human peripheral blood monocytes which might be responsible for the increased serum PAPP-A level in patients with acute coronary syndromes.

C-Reactive Protein ; adverse effects ; pharmacology ; Cells, Cultured ; Humans ; Monocytes ; drug effects ; metabolism ; Pregnancy-Associated Plasma Protein-A ; metabolism ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVESProgrammed death-1 (PD-1) up-regulation impairs virus-specific CD8+ T-cell responses during chronic viral infection. Whether PD-1 expression influences the virus-specific CD8+ T cells in humans with acute viral infection remains largely undefined. This study aims to characterize the PD-1 expression during acute hepatitis B (AHB), and further addresses the association between the PD-1 dynamics and memory T-cell formation during acute HBV infection.

METHODSPeripheral HBV-specific CD8+ T cells from 11 HLA-A2-positive AHB patients were longitudinally quantitatively analyzed, and PD-1, memory markers CCR7, CD45RA and CD127 and activation marker CD38 on HBV-specific CD8+ T cells were measured using flow cytometric assay. Serum ALT, HBsAg, HBsAb and HBV-DNA levels were evaluated for each subject.

RESULTSAll 11 AHB patients examined had multiple pentamer-positive CD8+ T-cell responses in their early phase of HBV infection. Specifically, their PD-1 on pentamer-positive CD8+ T-cells was significantly up-regulated at the onset of their disease. Following their disease resolution, the dynamic decrease in PD-1 expression was found to correlate with the phenotypic development of memory CD8+ T cells, indicated by the increases in CCR7, CD45RA and CD127 and decrease in CD38.

CONCLUSIONPD-1-mediated negative signaling may be closely associated with memory T-cell formation during acute self-limited hepatitis B.

Acute Disease ; Adult ; Antigens, CD ; metabolism ; Apoptosis Regulatory Proteins ; metabolism ; Female ; Hepatitis B ; immunology ; metabolism ; Humans ; Immunologic Memory ; Male ; Middle Aged ; Programmed Cell Death 1 Receptor ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; Young Adult