Glucagon-like peptide-1 protects against cardiac microvascular endothelial cells injured by high glucose

OBJECTIVE@#To investigate the protective effect of glucagon-like peptid-1 (GLP-1) against cardiac microvascular endothelial cell (CMECs) injured by high glucose.@*METHODS@#CMECs were isolated and cultured. Superoxide assay kit and dihydroethidine (DHE) staining were used to assess oxidative stress. TUNEL staining and caspase 3 expression were used to assess the apoptosis of CMECs. H89 was used to inhibit cAMP/PKA pathway; fasudil was used to inhibit Rho/ROCK pathway. The protein expressions of Rho, ROCK were examined by Western blot analysis.@*RESULTS@#High glucose increased the production of ROS, the activity of NADPH, the apoptosis rate and the expression level of Rho/ROCK in CMECs, while GLP-1 decreased high glucose-induced ROS production, the NADPH activity and the apoptosis rate and the expression level of Rho/ROCK in CMECs, the difference were statistically significant (P<0.05).@*CONCLUSIONS@#GLP-1 could protect the cardiac microvessels against oxidative stress and apoptosis. The protective effects of GLP-1 are dependent on downstream inhibition of Rho through a cAMP/PKA-dependent manner, resulting in a subsequent decrease in the expression of NADPH oxidase.

Glucagon-like peptide-1 protects against cardiac microvascular endothelial cells injured by high glucose

Objective: To investigate the protective effect of glucagon-like peptid-1 (GLP-1) against cardiac microvascular endothelial cell (CMECs) injured by high glucose. Methods: CMECs were isolated and cultured. Superoxide assay kit and dihydroethidine (DHE) staining were used to assess oxidative stress. TUNEL staining and caspase 3 expression were used to assess the apoptosis of CMECs. H89 was used to inhibit cAMP/PKA pathway; fasudil was used to inhibit Rho/ROCK pathway. The protein expressions of Rho, ROCK were examined by Western blot analysis. Results: High glucose increased the production of ROS, the activity of NADPH, the apoptosis rate and the expression level of Rho/ROCK in CMECs, while GLP-1 decreased high glucose-induced ROS production, the NADPH activity and the apoptosis rate and the expression level of Rho/ROCK in CMECs, the difference were statistically significant (. P<0.05). Conclusions: GLP-1 could protect the cardiac microvessels against oxidative stress and apoptosis. The protective effects of GLP-1 are dependent on downstream inhibition of Rho through a cAMP/PKA-dependent manner, resulting in a subsequent decrease in the expression of NADPH oxidase.

Construction of a lentiviral vector containing human NESG1 gene and its expression in 293FT cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral vector carrying human NESG1-EGFP gene and observe its expression in 293FT cells.</p><p><b>METHODS</b>The CDS region of NESG1 gene was amplified from a plasmid containing the full-length NESG1 sequence and cloned into the lentiviral vector pGC-FU-EGFP by restriction endonuclease AgeI digestion and T(4) DNA ligase ligation. After transformation into competent E. coli cells, the candidate clones were identified by PCR and sequencing. The recombinant plasmid and the two packaging plasmids were co-transfected into human embryonic kidney cell line 293FT cells by lipofectamine 2000 to produce the lentiviral particles, and the viral titer was determined. The 293FT cells were infected by the lentiviral particles obtained and the transfection efficiency was assessed under fluorescent microscope. Western blotting was used to detect the expression of NESG1 protein in the transfected cells.</p><p><b>RESULTS</b>The lentiviral vector pGC-FU-NESG1-EGFP for NESG1 gene was constructed successfully. Strong green fluorescence was observed in 293FT cells under fluorescent microscope after co-transfection of the cells with the 3 plasmids of lentiviral vector. The virus in the supernatant reached a titer of 2×10(7) TU/ml. The transfection efficiency of the collected virus exceeded 90% in 293FT cells with a multiplicity of infection of 1. Western blotting identified the presence of NESG1 expression in the transfected 293FT cells.</p><p><b>CONCLUSION</b>The lentiviral vector for NESG1 has been successfully constructed with a high yield of lentivirus, which facilitate further investigation of the roles of NESG1 gene in the development and progression of nasopharyngeal carcinoma.</p>

Transcatheter closure of paraprosthetic valve leaks post surgical valve replacements / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility and efficacy of transcatheter closure of paravalvular leak (PVL) with Chinese-made occluder.</p><p><b>METHODS</b>Five PVL patients were involved in this study, 2 out of the 5 patients underwent aortic mechanical valve replacements, 2 underwent mitral bioprosthetic valve replacements, and the remaining 1 underwent double mechanical valve replacement.Left ventricular end diastolic diameter, left atrial diameter and the systolic pulmonary artery pressure were assessed by echocardiography before and post the procedure.</p><p><b>RESULTS</b>Complete occlusion without residual regurgitation was achieved in 2 patients with aortic PVL, for the 3 patients with mitral PVL, there was only tiny or mild mitral paraprosthetic leak remained post closure procedure. Cardiac perforation and pericardium tamponade occurred in 1 patient with aortic PVL during interventional closure and the patient recovered post emergent pericardiocentesis. Transient severe hemolysis and hemoglobinuria occurred in 3 patients with mitral PVL post closure procedure and they recovered after 1 to 3 weeks conservative therapy. During 3 months follow up, left ventricular end diastolic diameter [(52.2 ± 6.8) mm vs. (61.1 ± 7.2) mm, P < 0.05], the systolic pulmonary artery pressure [(40.0 ± 5.4) mm Hg (1 mm Hg = 0.133 kPa) vs. (57.0 ± 3.6) mm Hg, P < 0.05] and left atrial diameter of mitral PVL patient [(49.0 ± 4.3) mm vs. (56.0 ± 6.3) mm, P < 0.05] were significantly reduced compared to before closure procedure.</p><p><b>CONCLUSION</b>Percutaneous or transapical left ventricular access closure of PVL is feasible, effective and relative safe in selected patients.</p>

Efficacy and safety of Firebird sirolimus-eluting stent in treatment of complex coronary lesions in Chinese patients: one-year clinical and eight-month angiographic outcomes from the FIREMAN registry / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Off-label application of drug-eluting stents (DES) during percutaneous coronary intervention (PCI) was not uncommon in daily practice, however DES in treating Chinese patients with complex lesion subset was under-investigated. The primary objective of the FIREMAN registry was to evaluate the long term efficacy and safety of the Firebird sirolimus-eluting stent (SES) in treating patients with complex coronary lesions. Here we report the mid-term of one-year clinical outcomes and eight-month angiographic follow-up results of FIREMAN registry.</p><p><b>METHODS</b>The FIREMAN registry was a prospective multi-center registry, which included 1029 consecutive patients undergoing PCI with Firebird SES implantation between September 2006 and July 2007 in 45 centers in China. The clinical follow-up was designed to be performed at 1, 6, 12, 18, 24, 30 and 36 months post index procedure, and non-mandatory angiographic follow-up at 8 months was planned. One hundred percent site monitoring was conducted.</p><p><b>RESULTS</b>Long lesions (59.2%), multi-vessel disease (50.4%), and small vessel disease (31.6%) were mostly found in angiography. Major adverse cardiac events (MACE) occurred in 51 (5.1%) patients at 1 year clinical follow-up, including cardiac mortality in 6 (0.6%), non-fatal myocardial infarction in 11 (1.1%), and target lesion revascularization in 36 (3.5%) of the patients. Definite and probable stent thrombosis (ST) by Academic Research Consortium (ARC) definition occurred in 12 (1.36%) patients at one-year clinical follow-up. The 8-month binary restenosis rate was 5.7% in-segment and 4.3% in-stent, respectively. Late lumen loss was (0.21 ± 0.40) mm in-segment and (0.23 ± 0.36) mm in-stent, respectively. Furthermore, Cox regression analysis revealed that diabetes, small vessel diameter, and chronic total occlusion were independent predictors of ST.</p><p><b>CONCLUSIONS</b>The results showed that the Firebird SES was effective and safe in treating Chinese patients with complex coronary lesions and occurrence of ST rate at one-year clinical follow-up was acceptable, however further long-term follow-up was still necessary. (NCT00552656)</p>

Construction of a lentivirus interfering vector targeting Cyc1 and its interfering efficiency in nasopharyngeal carcinoma cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the expression of Cyc1 in nasopharyngeal carcinoma (NPC) and evaluate the interfering efficiency of a lentivirus interfering vector targeting Cyc1 in NPC cells.</p><p><b>METHODS</b>Microarray technique was used to examine the expression of Cyc1 in NPC tissues. Real-time PCR was utilized to confirm the high expression of Cyc1 in NPC tissues and NPC cell lines. The recombinant Cyc1 shRNA-expressing plasmid (pLentiU6/Cyc1-shRNA) was stably transfected into NPC cells, and the interfering efficiency against Cyc1 was evaluated by quantitative RT-PCR.</p><p><b>RESULTS</b>The result of microarray showed that Cyc1 was highly expressed in NPC tissues compared to noncancerous nasopharyngeal tissues, as confirmed by Real-time PCR. All of the 8 NPC cells showed a high expression of Cyc1, among which 5-8F cells showed the highest expression. Sequence analysis indicated that the recombinant plasmid pLentiU6/Cyc1-shRNA was successfully constructed and could significantly and stably suppress the expression of Cyc1 in NPC cells.</p><p><b>CONCLUSION</b>Cyc1 is highly expressed in NPC cells. The lentivirus vector constructed can markedly inhibit the expression of Cyc1 in NPC cells, which provides assistance in the investigation of the function and molecular mechanism of Cyc1 in NPC.</p>

Inhibition of the growth of hepatocarcinoma xenograft in Balb/c mice induced by dendritic cells immunized with AFP cDNA fragement / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the antitumor immune response induced by dendritic cells vaccine coding AFPcDNA fragment with signal peptide (AFP(1)) and without signal peptide (AFP(2)), and to determine the inhibiting effect of the vaccine on the growth of hepatocarcinoma xenograft in Balb/c mice.</p><p><b>METHODS</b>pcDNA3.1/AFP(1) and pcDNA3.1/AFP(2) were transfected into dendritic cells (DCs) by calcium phosphate nanoparticles and became DCs vaccine. Mouse spleen lymphocytes were stimulated by AFP(1)/DC and AFP(2)/DC. A Balb/c mouse model bearing mouse HCC xenograft was established on the day 14 after transplantation. Forty mice were divided equally into AFP(2)/DC group, AFP(1)/DC group and plasmid control group. The treated mice received DCs vaccine and the same amount of control plasmid.</p><p><b>RESULTS</b>AFP(2)/DC stimulated T lymphocytel proliferation in vitro and improved CTL activity. The effects were better than AFP(1)/DC. The tumor-bearing mice injected intralesionally with AFP(1)/DC and AFP(2)/DC at a dose of 0.5 ml per mouse showed inhibition of tumor growth and prolongation of survival time. The tumor inhibition rate of the AFP(2)/DC group was 79.2% and the AFP(1)/DC group was 39.7% at 2 weeks after treatment. The tumor volume of AFP(2)/DC group was (726.7 +/- 298.2) mm(3), significantly smaller than the (1486.2 +/- 457.2) mm(3) of the AFP(1)/DC group and (2137.2 +/- 547.2) mm(3) of the plasmid control group (P < 0.05). The mean survival time of mice in the AFP(2)/DC group [(58.5 +/- 4.2) d] and AFP(1)/DC group [(45.2 +/- 4.8) d] were significantly longer than that of plasmid control group [(30.6 +/- 6.2) d, P < 0.05]. Bax-positive cell percentage was increased in the xenografts of AFP(2)/DC-treatment group compare with that of plasmid control group.</p><p><b>CONCLUSION</b>AFP(2)/DC and AFP(1)/DC vaccines show evident inhibiting effect on the growth of H22 xenograft in Balb/c mice through inducing efficient and specific immune response against the hepatocarcinoma cells.</p>

Thorombolytic therapy with rescue percutaneous coronary intervention versus primary percutaneous coronary intervention in patients with acute myocardial infarction: a multicenter randomized clinical trial / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Although thrombolytic therapy with rescue percutaneous coronary intervention (PCI) is a common treatment strategy for ST-segment elevation acute myocardial infarction (STEMI), scant data are available on its efficacy relative to primary PCI, and comparison was therefore the aim of this study.</p><p><b>METHODS</b>This multicenter, open-label, randomized, parallel trial was conducted in 12 hospitals on patients (age < or = 70 years) with STEMI who presented within 12 hours of symptom onset (mean interval > 3 hours). Patients were randomized to three groups: primary PCI group (n = 101); recombinant staphylokinase (r-Sak) group (n = 104); and recombinant tissue-type plasminogen activator (rt-PA) group (n = 106). For all patients allocated to the thrombolytic therapy arm, coronary angiography was performed at 90 minutes after drug therapy to confirm infarct-related artery (IRA) patency; rescue PCI was performed in cases with TIMI flow grade < or = 2. Bare-metal stent implantation was planned for all patients.</p><p><b>RESULTS</b>After randomization it required an average of 113.4 minutes to start thrombolytic therapy (door-to-needle time) and 141.2 minutes to perform first balloon inflation in the IRA (door to balloon time). Rates of IRA patency (TIMI flow grade 2 or 3) and TIMI flow grade 3 were significantly lower in the thrombolysis group at 90 minutes after drug therapy than in the primary PCI group at the end of the procedure (70.5% vs. 98.0%, P < 0.0001, and 53.0% vs. 85.9%, P < 0.0001, respectively). Rescue PCI with stenting was performed in 117 patients (55.7%) in the thrombolytic therapy arm. Rates of patency and TIMI flow grade 3 were still significantly lower in the rescue PCI than in the primary PCI group (88.9% vs. 97.9%, P = 0.0222, and 68.4% vs. 85.0%, P = 0.0190, respectively). At 30 days post-therapy, mortality rate was significantly higher in the thrombolysis combined with rescue PCI group than in primary PCI group (7.1% vs. 0, P = 0.0034). Rates of death/MI and bleeding complications were significantly higher in the thrombolysis with rescue PCI group than in the primary PCI group (10.0% vs. 1.0%, P = 0.0380, and 28.10% vs. 8.91%, P = 0.0001, respectively).</p><p><b>CONCLUSIONS</b>Thrombolytic therapy with rescue PCI was associated with significantly lower rates of coronary patency and TIMI flow grade 3, but with significantly higher rates of mortality, death/MI and hemorrhagic complications at 30 days, as compared with primary PCI in this group of Chinese STEMI patients with late presentation and delayed treatments.</p>

Effects of Apelin-13 on rat myocardial ischemia-reperfusion injury in vivo and its signal transduction pathway / 上海第二医科大学学报

Objective To investigate the role of apelin-13, a vasoactive peptide, in rat myocardial ischemia-reperfusion injury in vivo and explore its signal transduction pathway. Methods Rats were randomly divided into control group (n=10) and Apelin-13 group (n=15), and in vivo models of rat myocardial ischemia-reperfusion injury were established. Normal saline (control group) or Apelin-13 (Apelin-13 group) was administered intravenously 5 min before reperfusion. TTC and Evan's blue staining were used to determine the infarction size (IS) and area at risk (AAR), apoptotic cells were quantified by TUNEL method, and the expression of ERK1/2 was determined by Western blotting. Results IS/AAR and apoptosis index of Apelin-13 group were significantly lower than those in control group [(38.33±12.95) % vs (52.61±11.00)% and (0.21±0.02) vs (0.31±0.05)](P <0.05). The expression of p-ERK1/2 in Apelin-13 group was significantly increased than that in control group [(1.15±0.16) vs (0.63±0.07)](P < 0.05). Conclusion Apelin-13 may protect rat hearts from in vivo ischemia-reperfusion injury, reduce infarction size and attenuate myocardial apoptosis, which may be mediated by the activation of ERK1/2 MAPK signal transduction pathway.

Dual-phase contrast-enhancement multislice computed tomography imaging in assessment of acute myocardial infarction-reperfusion injury / 上海第二医科大学学报

Objective To explore the value of dual-phase contrast-enhancement multislice computed tomography (MSCT) in the assessment of acute myocardial infarction volume and perfusion in porcine models. Methods The distal left anterior descending coronary arteries of 5 pigs were balloon-occluded for 90 min and followed by reperfusion. MSCT was performed 1 min (early phase) and 5 min (delayed phase) after administration bolus of 100 mL of iodinated contrast material 30 min after reperfusion. On the same day, hearts were excised, sectioned in 8 mm short-axis slices, and stained with TTC. Infarction volume was defined as the sum of the hyper-enhanced area and surrounding hypo-enhanced area in all slices on delay enhanced phase of MSCT and the TTC-negative area on TTC staining slices. Infarction volume was expressed as percentage of total slice volume. Results Acute infarction detected by MSCT was characterized by early myocardial perfasion defects in the early phase of the contrast bolus (early defects) with surrounding residual defects and late enhancement observed in the late phase. Mean CT attenuation value of early defects was significantly different from CT attenuation value of remote myocardium [(213±55)HU vs (304±30)HU](P < 0.05), CT attenuation values of residual defects and late enhancement were also significantly different from those of remote myocardium [(360±75) HU vs (90±37) HU and (152±23) HU vs (190±37) HU, repectively](P < 0.01, P < 0.05). The mean infarction volume was (8.9± 1.0)% on MSCT and (9.2±1.4)% on TTC pathology images. The infarction volume assessed by MSCT compared well with TTC staining slices. Conclusion Acute reperfused myocardial infarction zone has specific enhancement pattens different to remote normal zone on dual phase MDCT, which is in good agreement with in vivo Trc pathology in the assessment of acute reperfused myocardial infarction shortly offer reperfusion.

Preparation and application of Epstein-Barr virus genome-wide genechip / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare an Epstein-Barr virus (EBV) microarray using known and predicted EBV-coded genes as the cDNA probes to detect the EBV gene expression in nasopharyngeal carcinoma (NPC) tissues.</p><p><b>METHODS</b>The EBV gene probes were amplified by PCR using a pair of primers designed in both sides of the multiple clone site (MCS) of the T/A vector. After purification of the PCR products, 85 EBV genes and 8000 human genes were printed onto the same slide as the detection chip consisting of both EBV and human genes. This genechip was used to detect the differential gene expression in NPC and non-cancerous nasopharynx (NP) tissues.</p><p><b>RESULTS</b>Detection of the human gene expression profile using the prepared genechip resulted in the identification of numerous human genes in the tissue specimens. Some EBV genes were also detected in the tissues using the genechip, but the signals of the genes appeared rather weak without distinctly visible fluorescence, and were not comparable to the strong signal intensities of the human genes.</p><p><b>CONCLUSION</b>The EBV microarray, though constructed successfully, can not meet the needs for clinical application due to the limited detection sensitivity and the relative small quantity of EBV gene expression in NPC samples. Further improvements of the research methods are warranted.</p>

Construction and identification of a recombinant lentivirus harboring small interfering RNA targeting mouse CD99 antigen-like 2 gene / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant lentivirus harboring RNA interference sequence targeting mouse CD99 antigen-like 2 (mCD99L2) gene and observe its infection efficiency of 293FT cells.</p><p><b>METHODS</b>Four pairs of small interfering RNAs (siRNAs) targeting mCD99L2 cDNA were designed, synthesized and linked to the lentivirus vector SD1259 to construct the lentivirus shuttle plasmids. After sequencing, the 4 lentivirus shuttle plasmids were transfected into 293FT cells in the presence of packaging plasmids. Forty-eight hours later, the supernatant was collected and the titer and infection efficiency of the recombinant lentivirus were determined according to the expression of the reporter gene enhanced green fluorescent protein (EGFP) under fluorescent microscope.</p><p><b>RESULTS</b>DNA sequencing demonstrated that mCD99L2 siRNAs were successfully cloned to the lentiviral vector SD1259. The titer of concentrated virus was 1x10(7)/ml in the supernatant of the infected cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus containing siRNA targeting mCD99L2 gene has been successfully constructed, which provide the basis for future establishment of visualized cell model and animal model of Hodgkin's lymphoma.</p>

Effect of coronary bifurcation angle on clinical outcomes in Chinese patients treated with crush stenting: a subgroup analysis from DKCRUSH-1 bifurcation study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Bifurcation angles may have an impact on the clinical outcomes of crush stenting. We sought to compare high (> or = 60 degrees ) with low (< 60 degrees ) bifurcation angle in patients who underwent either classical or double kissing (DK) crush stenting for bifurcation lesions from the DKCRUSH-1 data base.</p><p><b>METHODS</b>There were 212 patients with 220 lesions, some with low-angle (n = 138) and some with high-angle (n = 74). Angiography was indexed at 8-month after procedure. Primary endpoint was the occurrence of major adverse cardiac events (MACEs), defined as cardiac death, myocardial infarction and target lesion revascularization (TLR). Secondary endpoint included late lumen loss, the rate of restenosis, and final kissing balloon inflation (FKBI).</p><p><b>RESULTS</b>At 8 months, clinical follow-up was 100%; angiographic follow-up was 75% in the low-angle group and 83.3% in the high-angle group. There were no significant differences in the FKBI between the high-angle group (91.43%) and the low-angle group (82.39%). In the high angle group, there was a significant difference in contrast volume used (P = 0.005) but no significant difference in acute gain, minimum lumen diameter (MLD), late loss and diameter stenosis in the pre-bifurcation segment, post-bifurcation segment or side branch. When lesions were assigned into with-(n = 133) and without-FKBI (n = 42), significant side-branch late loss was seen in the group without-FKBI ((0.65 +/- 0.49) mm vs (0.47 +/- 0.62) mm, P = 0.02), with a resultant greater restenosis rate (37.68% vs 18.32%, P = 0.001). No difference was detected in the MACE free survival rate between the high and low angle groups (82.39% vs 82.36%, P = 0.84). The rate of stent thrombosis tended to be higher in the lower-angle group although there was no significant difference (P = 0.38). The TLR free survival rate was 87.2% in the with-FKBI group vs 73.5% in the without-FKBI group (P = 0.001). Cox regression analysis showed that the independent predictors for target vessel revascularization were the side branch stent MLD post stenting (hazard ratios (HR) 1.028, 95% CI 2.357 - 16.233, P = 0.002), lack of FKBI (HR 4.910, 95% CI 4.706 - 8.459, P = 0.001) and unsatisfactory kissing (HR 3.120, 95% CI 2.975 - 5.431, P = 0.001).</p><p><b>CONCLUSIONS</b>Bifurcation angles do not influence the clinical outcome of crush stenting. Successful final kissing balloon inflation, regardless of bifurcation angles, can predict TLR.</p>

Construction and identification of a recombinant lentivirus vector of latent membrane protein 1 / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant lentivirus vector of latent membrane protein 1 (LMP1) and detect the expression of LMP1 in vitro.</p><p><b>METHODS</b>The LMP1 fragment including all the exons was amplified by PCR and inserted to the downstream of CMV promoter in the lentivirus vector pCDF. The three plasmids (packaging plasmid pFIV-34N, envelope plasmid pVSV-G and target plasmid pCDF-LMP1) were packaged into 293FT cells via liposome. The virus supernatant was harvested, concentrated and titrated. Mouse B lymphoma cell line A20 was transfected with the recombinant lentivirus vector of LMP1, and the expression of LMP1 in A20 cells was detected by RT-PCR and Western blotting.</p><p><b>RESULTS</b>DNA sequencing confirmed that the sequence of PCR-amplified LMP1 was consistent with the GenBank data. The LMP1 gene fragment was cloned into pCDF in the right direction, and the open reading frame of LMP1 was maintained. The 3 plasmids were effectively transferred into 293FT cells, which emitted green fluorescence in the cytoplasm and on the cell membrane under fluorescence microscope. The titer of the lentivirus vector reached 10(7) Tu/ml with a transfection efficiency 90% in A20 cells. LMP1 expression was detected by RT-PCR and Western blotting in transfected A20 cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus vector of LMP1 constructed can be effectively transfected into A20 cells, which provides a basis for exploring the role of LMP1 in the pathogenesis of lymphoma.</p>

Establishment of a stable nasopharyngeal carcinoma cell line with lentivirus-mediated RNA interference for EIF4G1 gene silencing / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a nasopharyngeal carcinoma (NPC) cell line with stable EIF4G1 gene silencing induced by small interfering RNA (siRNA).</p><p><b>METHODS</b>The EIF4G1 mRNA levels in 8 NPC cell lines including 5-8F, 6-10B, C666-1, CNE1, CNE2, HNE1, HONE1, and SUNE1 were detected by fluorescence quantitative RT-PCR (QRT-PCR). The recombinant lentivirus shRNA expression plasmid targeting EIF4G1 gene was packaged into mature lentivirus by 293FT cells and used to infect 5-8F cells. After blasticidin selection of NPC cells with constant expression of the EIF4G1-siRNA, the efficiency of EIF4G1 mRNA expression interference was determined using QRT-PCR.</p><p><b>RESULTS</b>The 8 NPC cell lines showed differential expression of EIF4G1 mRNA, among which 5-8F cells had the highest EIF4G1 expression. The recombinant lentivirus plasmid pLenti6/BLOCK-iT-DEST/EIF4G1-shRNA was successfully constructed and verified by PCR and sequencing. The EIF4G1 mRNA level of 5-8F cells infected with shRNA-EIF4G1 lentivirus was significantly reduced as compared with the negative control and the blank control cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus vector pLenti6/BLOCK- iT-DEST/EIF4G1-shRNA we constructed results in marked downregulation of EIF4G1 mRNA expression and constant expression of EIF4G1-siRNA after infection of 5-8F cells.</p>

Crush stenting in treating coronary bifurcate lesions: paclitaxel eluting stents versus sirolimus eluting stents / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Because no data regarding the comparison of crush stenting with paclitaxel (PES) or sirolimus eluting stents (SES) for coronary bifurcate lesions have been reported, we compared the clinical outcomes of these two types of stents.</p><p><b>METHODS</b>Two hundred and thirty patients with 242 bifurcate lesions were enrolled in a prospective, nonrandomized trial. Primary endpoints included myocardial infarction, cardiac death and target vessel revascularization at 8 months.</p><p><b>RESULTS</b>All patients were followed up clinically and 82% angiographically at 8 months. Final kissing balloon inflation was performed in 72% in the PES and 75% in the SES groups (P>0.05). Compared to the SES group, PES group had a higher late loss and incidence of restenosis (P=0.04) in the prebifurcation vessel segment. The postbifurcation vessel segment in the PES group had a greater late loss ((0.7+/-0.6) mm vs (0.3+/-0.4) mm, P<0.001) and higher restenosis in the side branch (25.5% vs 15.6%, P=0.04) when compared to the SES group. There was significant difference of insegment restenosis in the entire main vessel between PES and SES groups (P=0.004). Target lesion revascularization was more frequently seen in the PES group as compared to the SES group (P=0.01). There was significant difference in the accumulative MACE between these two groups (P=0.01). The survival rate free from target lesion revascularization was significantly higher in the SES group when compared to the PES group (P<0.001).</p><p><b>CONCLUSION</b>SES is superior to PES in reducing restenosis and target lesion revascularization by 8-month follow-up after crush stenting for bifurcate lesions.</p>

AF172993 sequence of Plunc in GenBank database is not the complete CDS / 南方医科大学学报

<p><b>OBJECTIVE</b>To determine AF172993 sequence is either the complete CDS or a transcript variant.</p><p><b>METHODS</b>RT-PCR was used to amplify the CDS sequence of Plunc, which was subsequently cloned into the pEGFP-N1 eukaryotic expression vector. After bi-directional sequence analysis, the sequence obtained was blasted against the AF172993 sequence, nr database and human genome database.</p><p><b>RESULTS</b>In CDS of the new cloned sequence, the 658 base A in the AF172993 sequence was replaced by C, and the corresponding genetic code was also converted from AAG to CAG, leading to the alteration of the amino acid Gln to Lys. In addition, the base C at the 658 position of the CDS showed perfect match with the base C at 2094188 position in human chromosome 20.</p><p><b>CONCLUSION</b>The base A at the 658 position of AF172993 sequence of Plunc is a mutation site, which alters the coding of the amino acid. AF172993 sequence is actually a transcript variant of Plunc, and the annotation to AF172993 in GenBank database is not correct and need to be revised.</p>

Simvastatin downregulates CD40L induced vascular cell adhesion molecule-1 expression and adhesive function in human umbilical vein endothelial cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of simvastatin on vascular cell adhesion molecule-1 (VCAM-1) expression and adhesive function in ECV-304 cells treated with CD40L.</p><p><b>METHODS</b>Human umbilical vein endothelial cell (HUVEC) was cultured and treated with various concentrations CD40L alone or in combination with various concentrations simvastatin in the absence or presence of mevalonic acid (400 micromol/L). RT-PCR and FCM analysis were used to determine VCAM-1 expression and lymphocytes adhesion to endothelial cells.</p><p><b>RESULTS</b>Simvastatin (0 - 10 micromol/L) decreased in a concentration-dependent manner the expression of VCAM-1 induced by CD40L and this effect could be blocked by cotreatment with mevalonic acid. Moreover, Simvastatin also significantly decreased adhesion capacity of lymphocytes to endothelial cells induced by CD40L.</p><p><b>CONCLUSION</b>Simvastatin downregulates VCAM-1 expression and adhesive capacity of lymphocytes to endothelial cells induced by CD40L.</p>

Differentially expressed transcription factor-related genes in nasopharyngeal carcinoma and nasopharyngeal tissues / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the gene expression profiles of transcription factor-related genes in nasopharyngeal carcinoma (NPC) tissues and normal nasopharyngeal tissues using a cDNA microarray membrane for exploring the regulatory mechanism of differential gene express in NPC tissues.</p><p><b>METHODS</b>The total RNAs from 24 NPC tissues and 24 pooled normal nasopharyngeal tissues were reverse transcribed and labeled with alpha-(32)P-dCTP. The resultant cDNAs were hybridized to GF211 microarray, and the signals were analyzed by Pathway 4.0 software. RT-PCR was carried out to confirm the results.</p><p><b>RESULTS</b>Among the 1625 differentially expressed genes detected in NPC and nasopharyngeal tissues, 35 transcription factor-related genes were identified with either up- or down-regulation.</p><p><b>CONCLUSION</b>These differentially expressed transcription factor-related genes in NPC tissues might play a role in the regulation of NPC-related gene expression.</p>

QT hysteresis in long-QT syndrome children with exercise testing / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Congenital long QT syndrome (LQTS) is an inherited ion channel disorder resulting in abnormal cardiac repolarization that can cause syncope and sudden death associated with a prolonged rate-corrected QT interval and polymorphic ventricular tachycardia. Several studies in adults showed that LQTS patients have altered QT adaptation to heart rate changes compared with normal subjects which forming a "hysteresis loop" in the QT-circle length plot. This study was to observe the QT interval changing during exercise testing in children long QT syndrome (LQTS) patients, explore the new diagnosis methods of LQTS.</p><p><b>METHODS</b>The subjects were divided into 3 groups according to 1993 LQTS diagnostic criteria. Group 1: LQTS group (n = 17) who scored > or = 4 points indicating definite LQTS. Group 2: Middle group (n = 16), patients who have prolonged QT interval but scored 1.5 to 3.5. Group 3: Normal control group (n = 18). The average age of all study population is (12.3 +/- 5.8) years. No case had beta-adrenergic antagonists administration before exercise testing. All subjects were underwent tread mill exercise testing and electrocardiograph in whole exercise testing and recovery were recorded. QT and heart rate changing during whole exercise testing period were recorded. DeltaQT, the QT interval at 1, 2, 4, 6 minutes into recovery subtract from the QT interval at a similar heart rate during exercise, were calculated.</p><p><b>RESULTS</b>In all three groups, QT intervals were shortening with the increasing of heart rate, but QTc had no significant change. DeltaQT at 1 minute ((45 +/- 11) ms), 2 minutes ((37 +/- 15) ms), 4 minutes ((23 +/- 12) ms) into recovery in LQTS group were significantly greater than that of the other two groups (P < 0.05, P < 0.01, P < 0.01, respectively). There was no DeltaQT significant difference between middle group and normal control group at recovery time. During the recovery phase in LQTS group, the QT interval remained shortened despite a decelerating heart rate, forming a hysteresis "loop" in the curve relating the QT interval to the cycle length.</p><p><b>CONCLUSIONS</b>In children LQTS patients, there is significant QT hysteresis loop in the relation of QT interval with heart rate during recovery of exercise testing, which could be useful to the early diagnosis for LQTS.</p>