Objective To study the expression changes of matrix metalloproteinases (MMPs), protein kinase C (PKC) and nuclear factor-?B (NF-?B ) on the injured rabbit vascular smooth muscular cells (VSMC) transfected with tissue inhibitor of metalloproteinase-2 (TIMP-2) vector. Methods Lipofectin method was used to transfect TIMP-2 vector into VSMC, Western blot analysis to detect TIMP-2 peptides and zymography assay to determine MMPs. The activities of MMPs and NF-?B were detected by electrophoretic mobility shift assay (EMSA). The mRNA and protein expressions of PKC? were determined by RT-PCR and immunocytochemistry. Results The injured VSMC showed increased enzyme activity of MMP-2. There was very lower level expressions of PKC? and NF-?B in the normal VSMC but high in the injured VSMC. However, in injured VSMC transfected with TIMP-2 vector, the activity of MMP2/9 was suppressed and the expressions of PKC? and NF-?B decreased (P

Objective: To evaluate value of application of intravascular ultrasound imaging (IVUS) in diagnosing borderline lesions in left anterior descending artery (LAD) and left main coronary artery (LM). Methods: According to results of coronary angiography (CAG) in 60 cases with coronary borderline lesions, including 20 cases in LM, 20 cases in proximal segment of LAD and 20 cases in middle segment of LAD, the diagnostic value of IVUS in coronary borderline lesion was evaluated. Results: Compared with CAG, mean diameter stenosis rate of each coronary artery [LM: (65.3l±7.81) % vs. (75.28±8.89) %，proximal segment of LAD: (66.67±8.79) % vs. (78.89±7.88) %，middle segment of LAD: (71.55±6.83) % vs. (75.3l±7.81) %, P<0.01 all] significantly increased in IVUS. The differences of detection rate of plaque calcification and plaque rupture were no significant between CAG and IVUS（＞0.05）. Conclusion: Different degrees of underestimation of coronary artery stenosis exist in CAG, especially in proximal segment of LAD. IVUS can be an effective complement to CAG.

Objective To investigate the effects of electrical stimulation on the expression of vascular endothelial growth factor(VEGF)mRNA and receptor FLK 1/KDR in a ischemic model of rat hindlimb. Methods The model of hindlimb ischemia on the right side was established by ligation of the superficial femoral artery in 10 rats. The rats were then randomized into an experimental group and a control group. The rats in the experimental group were intervened with electrical stimulation ( 25 Hz, 0.1 V) on the tibialis anterior (TA) of the right side, while those in the control group were not. RT PCR and immunohistological methods were used to detect the expressions of VEGF mRNA and protein in TA muscles. FLK 1/KDR was detected by means of Western blot and immunofluorescence. Results After 7 days of continuous stimulation, there was a significant increase in blood flow within the muscle. VEGF mRNA and VEGF protein had 4 fold and 2 fold increases, respectively, in the stimulated TA muscles as compared to the control(2.58 vs 0.93, 0.48 vs 0.24, P

Juvenile idiopathic arthritis(JIA)is the most common rheumatology disease in childhood period with poor prognosis.The biological agents are newly developed drugs with features of clear therapeutic targets and fast effects.But its use in JIA is still limited,so this article focuses on the clinical use experience,timming and sideffects of the biological agents on JIA.

Objective To study the effect of Fas gene transfection on rectal carcinoma cells in vitro. Methods By using RT-PCR technique, a full length of Fas gene 1007 bp was cloned from actived peripheral mononuclear cells of healthy donors. The fragment was ligated with the pGEM-T Easy and sequenced. The constructed vector was transfected into 8348 cells with lipofectin, the change in expression of Fas gene was determined by RT-PCR. The apoptosis and proliferation of rectal carcinoma cells pre- and posttransfection induced by cisplatin were analysed by ladder and MTT methods. Results Transfection of Fas gene significantly upregulates the expression of Fas in human rectal carcinoma 8348 cells. With the concentration of cisplatin at the level of 1, 5, 10, 20 and 40 mg/L , respectively, the suppression rates of Fas transfection group and control group were 47.2%51.8%57.2%65.4%71.0% and 29.6%33.0%37.8%41.4%47.0% respectively(t=15.33, P

AIM: To evaluate the expression of iNOS mRNA in different invasion ability colon carcinoma cell strains. METHODS: MTT was used to detect the growth and reproduction of colon cancer cell strain CW-2 and LS174T. RT-PCR and Northern blot were used to detect expression of iNOS mRNA in colon cancer. RESULTS: MTT growth curve displayed that colon cancer cell strain LS174T grew and reproduced faster than cell strain CW-2. RT-PCR showed that iNOS mRNA expressed strongly in CW-2 cell strain, while iNOS mRNA expressed weakly in LS174T cell strain. Northern blot detected that iNOS mRNA expressed obviously in CW-2 cell strain, but cell strain LS174T have no obvious iNOS mRNA expression. All-trans retinoic acid (ATRA) had no obvious effect on iNOS mRNA expression in CW-2 cell strain of colon cancer. CONCLUSIONS: ATRA has no obvious effect on iNOS mRNA expression. iNOS has a dual effect on tumor growth. In low-metastatic colon carcinoma CW-2, iNOS may exert a anti-tumor influence by cytotoxicity or inducing cell apoptosis. In high-metastatic colon cancer LS174T, iNOS produced low concentration of NO, which may be an important signal-transduction molecule for increasing blood supply and angiogenesis, which improve the growth, invasion and metastasis of tumor.

Cholangiopancreatography, Endoscopic Retrograde ; Duodenum ; surgery ; Humans ; Retrospective Studies

NS2 is a nonstructural protein of Periplaneta fuliginosa densovirus(PfDNV) with a molecular mass of 30 kD,whose function is not yet clearly understood. In order to study the expression,subcellular distribution and the function of NS2 protein,the coding region of NS2 was amplified from the hindgut tissue of cockroaches infected with PfDNV by RT-PCR and then the recombinant prokaryotic expression vector pET28a-NS2 was constructed. The recombinant plasmid was transformed into E. coli BL21(DE3) to express the 6?His fusion protein in the bacteria. After purification,the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody. The specificity of the anti-NS2 antibody was successfullyproved by western blotting on the eukaryotic expressed products of NS2 protein.Meanwhile,the full sequence of ns2 gene was also cloned into the eukaryotic expression vector pAC. The recombinant plasmid pAC-NS2 was then transfected into Schneider line 2(S2) cells to express NS2 protein in the insect cells. The subcellular localization of NS2 in the insect cells was then investigated by indirect immunofluorescence technique using the anti-NS2 polyclonal antiserum. The confocal laser scanning microscope observation showed that NS2 protein was located primarily in the cytoplasm with some punctate nuclear staining.

Objective To investigate the prottective effect of ulinstatin on lung of rats with sepsis and its mechanism of ameliorating cell apoptosis. Method A total of 40 female SD rats were randomly (random number)divided into the control group and the therapy group (ulinastatin 300 000 u/kg). The rat models of sepsis were produced by the classical method of cecal ligature and puncture (CLP), and the designed doses ulinastatin were given intra-peritoneally to the rats of the ulinastatin group and the same amounts of PBS (phosphate buffered solution) instead of ulinastatin were administered intra-peritoneally to the rats of the control group when the sepsis symptoms appeared usually in 3 hours after modeling. In 12 hours after treatment, lung tissues of rats in two groups were taken for observation under the transmission electron microscopy and detecting the levels of Bcl-2 and Bax protein in lung tissues by using immunohistochemical technique. The levels of the integrated optical density(IOD)of Bcl-2 and Bax protein were detected by using Image-pro plus software and the ratio of Bcl-2/Bax was calculated. Results Transmission electron microscope showed that lung tissue in control group had inflmmatory changes such as severe congestion and consolidation, and those changes in ulinastatin treatment group (300 000 u/kg)were significantly slighter. There were significant differences in the levels of antiapoptotic protein Bcl-2 in lung tissue of rats between two groups(P＜0.01), and the level of protein Bcl-2 in ulinas tatin group were higher than those in control group. The level of pro-apoptotic protein Bax in control group were higher than that in ulinastatin group (P＜0.01). The ratio of Bcl-2/Bax in ulinas tatin group was higher than that in control group (P ＜ 0.05).Conclusions Ulinastatin has protective effect on lung tissue in septic rats, and it may inhibit the inflammatory response and in the same time plays a role in inhibiting cell apoptosis.