Loss of GRB2 associated binding protein 1 in arteriosclerosis obliterans promotes host autophagy / 中华医学杂志(英文版)

BACKGROUND@#Arteriosclerosis obliterans (ASO) is a major cause of adult limb loss worldwide. Autophagy of vascular endothelial cell (VEC) contributes to the ASO progression. However, the molecular mechanism that controls VEC autophagy remains unclear. In this study, we aimed to explore the role of the GRB2 associated binding protein 1 (GAB1) in regulating VEC autophagy.@*METHODS@#In vivo and in vitro studies were applied to determine the loss of adapt protein GAB1 in association with ASO progression. Histological GAB1 expression was measured in sclerotic vascular intima and normal vascular intima. Gain- and loss-of-function of GAB1 were applied in VEC to determine the effect and potential downstream signaling of GAB1.@*RESULTS@#The autophagy repressor p62 was significantly downregulated in ASO intima as compared to that in healthy donor (0.80 vs. 0.20, t = 6.43, P < 0.05). The expression level of GAB1 mRNA (1.00 vs. 0.24, t = 7.41, P < 0.05) and protein (0.72 vs. 0.21, t = 5.97, P < 0.05) was significantly decreased in ASO group as compared with the control group. Loss of GAB1 led to a remarkable decrease in LC3II (1.19 vs. 0.68, t = 5.99, P < 0.05), whereas overexpression of GAB1 significantly led to a decrease in LC3II level (0.41 vs. 0.93, t = 7.12, P < 0.05). Phosphorylation levels of JNK and p38 were significantly associated with gain- and loss-of-function of GAB1 protein.@*CONCLUSION@#Loss of GAB1 promotes VEC autophagy which is associated with ASO. GAB1 and its downstream signaling might be potential therapeutic targets for ASO treatment.

Retrograde tibial approach for endovascular recanalization of popliteal and tibioperoneal artery occlusion / 中华外科杂志

<p><b>OBJECTIVES</b>To describe a procedure of the retrograde approach for endovascular treatment of complex popliteal and/or infrapopliteal occlusions and to determine its safety and efficacy in minimizing failure rates.</p><p><b>METHODS</b>Between January 2010 and March 2012, 28 patients (16 male and 12 female patients) received retrograde tibial approach after failure of antegrade intervention. There were 3 patients with severe claudication (Rutherford category 3) and 25 patients with critical limb ischemia (Rutherford category 4 to 6). From this group, two techniques were employed. Twenty-four patients were treated via a retrograde transpedal access site and 4 patients via a transcollateral loop technique. The clinical and follow-up data of these patients were analyzed retrospectively.</p><p><b>RESULTS</b>The technique success rates were 92.8% (26/28). No major complications and 3 (10.7%) minor sequelaes were documented in this study. Twenty-three patients were followed up for 3 to 29 months, with a mean of (14 ± 9) months. Overall patency was 73.9% (17/23) and 47.8% (11/23) at 6 and 12 months. Overall survival and limb salvage was 95.7% (22/23), ulcer were healed in 9/10 patients.</p><p><b>CONCLUSION</b>The use of retrograde tibial or pedal approach seems feasible and safety in case of failure in antegrade revascularization of popliteal and/or infrapopliteal occlusions.</p>

A single tube modified allele-specific-PCR for rapid detection of erythromycin-resistant Mycoplasma pneumoniae in Beijing / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mycoplasma pneumoniae (M. pneumoniae) is one of the common pathogens causing atypical pneumonia. In recent years, resistance to macrolides has become more common, especially in China. Previous studies have confirmed that the mutation at position 2063 in domain V of the 23S rRNA is the most prevalent, followed by the mutation at position 2064. Reported molecular detection methods for the identification of these mutations include direct sequencing, restriction fragment length polymorphism analysis, real-time polymerase chain reaction (PCR) with high-resolution melt analysis, and nested PCR-linked with capillary electrophoresis, etc. The most commonly used method for monitoring resistance-conferring mutations in M. pneumoniae is direct DNA sequencing of PCR or nested PCR products. However, these methods are time-consuming, labor-intensive or need expensive equipments. Therefore the development of rapid and sensitive methods is very important for monitoring the resistance globally.</p><p><b>METHODS</b>In this study, we reported a fast and cost-effective method for detecting 2063 and/or 2064 macrolide resistant mutations from specimens using a modified allele-specific PCR analysis, and all results were compared with the sequencing data. We also analyzed the clinical courses of these samples to confirm the modified allele-specific PCR results.</p><p><b>RESULTS</b>Among 97 M. pneumoniae specimens, 88 were found to possess mutations by this method, and all modified allele-specific PCR analysis results were consistent with the sequencing data. The data of the clinical courses of these 97 cases showed that they suffered from severe pneumonia. Erythromycin showed better efficacy on cases from which no macrolide resistance mutation was found on their specimens. However, in some cases from which mutations were detected, erythromycin monotherapy had poor efficacy, and on these patients severe symptoms improved only when azithromycin was added to the treatment.</p><p><b>CONCLUSIONS</b>The drug-resistant M. pneumoniae is very common in Beijing, China. Our modified allele-specific PCR analysis can identify erythromycin resistant mutations more rapidly from specimens than any other method currently available. Erythromycin is still effective for treating patients infected with the mutation negative M. pneumoniae, but this treatment fails to work on mutant organisms. This method can facilitate clinicians in selecting appropriate therapy within short timescales.</p>

Study on small diameter artificial vascular graft with eNOS gene transfected endothelial cells in canine / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the patency efficacy of small diameter artificial vascular graft with eNOS gene transfected endothelial cells in canine.</p><p><b>METHODS</b>By using the two steps high pressure injection, eNOS gene transfected endothelial cells were planted on artificial vascular graft with a diameter of 3 mm. The artificial vascular grafts were transplanted into canine femoral artery, and the patency of the artery was observed through digital subtracted angiography (DSA) and electron telescope 1, 4, 12 and 24 weeks after the operation.</p><p><b>RESULTS</b>The adherence rate of eNOS gene transfected endothelial cells to artificial vascular grafts was up to 90%. For 10 days, the cells extended and formed a continuous intima. One week after the operation, 3/9 grafts were obstructed in the group of simple artificial vascular grafts; 4 weeks after, all of grafts (9/9) were obstructed in the group of simple artificial vascular grafts and almost half grafts (5/10, 4/9) in un-transfected groups were obstructed; 12 weeks after, all of the grafts were obstructed in the group of simple artificial vascular grafts and in un-transfected groups (9/9, 9/10); 24 weeks after, all of the grafts were obstructed in the groups of un-transfected artificial vascular grafts, while nearly all of grafts (8/10) were open in eNOS groups. Electron microscope scanning showed that endothelial cells of artificial vascular grafts in eNOS transfected groups arranged closely and formed a continuous intima; only few red blood cells, leucocytes, platelets deposited at the surface of endothelial cells in the artificial vascular grafts.</p><p><b>CONCLUSIONS</b>The patency efficacy of eNOS gene transfected endothelial cells planting on artificial vascular graft is satisfactory. It could provide an experimental basis for further clinical application of the artificial vascular grafts.</p>

Transfection and Expression of eNOS Gene on Canine Endothelial Cells / 上海第二医科大学学报

Objective To study endothelial nitric oxide synthase (eNOS) gene transfecting endothelial cells (ECs). Methods eNOS gene was transfected into the steady ECs system by cationic liposomal transduction and check the transfection effect by RT-PCR and immunohistochemistry assay. To test the concentrations of NOS, nitric oxide (NO) and von willebrand factor (vWF) in culture media by colorimetry and ELISA, respectively,and transfected EC function was observed. Results The effect of transfection was satisfactory with RT-PCR products electrophoresis, sequence and immunohistochemistry. The concentrations of NOS and NO in transfected EC culture media increased with time (P

Endothelial nitric oxide synthase gene transfected endothelial cells inhibit smooth muscle cell proliferation and platelet aggregation early in vitro / 中华外科杂志

<p><b>OBJECTIVE</b>To test the function of endothelial cells transfected eNOS gene.</p><p><b>METHODS</b>To test the function of eNOS-overexpressing endothelial cells inhibiting smooth muscle cell proliferation and platelet aggregation in vitro by colorimetry, MTT and (3)H-TdR permeate, respectively, in four different endothelial cells groups.</p><p><b>RESULTS</b>The platelet aggregation is significantly decreasing between the groups of EC and eNOS at the different time (P < 0.05), is 42.2 and 32.6 separately at the time of 120 h. The SMC proliferation is significantly decreasing between the groups of EC and eNOS at the different time (P < 0.05). It is 0.28 and 0.22 by MTT test, 4691 and 3995 by (3)H-TdR permeate separately at the time of 120 h.</p><p><b>CONCLUSIONS</b>eNOS-overexpressing endothelial cells inhibited significantly smooth muscle cells proliferation and platelet aggregation in vitro; which shows powerful effect 48 hours post transfection and lasts up to 120 hours at the least; and were held back by L-NAME.</p>