ABSTRACT
<p><b>BACKGROUND</b>The calcium clearance and reactive oxygen species (ROS) generations in the coronary artery smooth muscle cells in chronic heart failure (HF) have not been fully investigated. Therefore, we attempted to understand the gene expressions underlying the mishandling of calcium clearance and the accumulations of ROS.</p><p><b>METHODS</b>We initially established an animal model of chronic HF by making the left anterior descending coronary artery ligation (CAL) in rats, and then isolated the coronary artery vascular smooth muscle cells from the ischemic and the nonischemic parts of the coronary artery vessels in 12 weeks after CAL operation. The intracellular calcium concentration and ROS level were measured using flow cytometry, and the gene expressions of sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2a), encoding sarcoplasmic reticulum Ca2+-ATPase 2a, encoding sodium-calcium exchanger (NCX), and p47phox encoding a subunit of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase were examined using real-time quantitative reverse transcription polymerase chain reaction and Western blotting, respectively.</p><p><b>RESULTS</b>We found that the calcium accumulation and ROS generation in the coronary artery smooth muscle cells isolated from either the ischemic or the nonischemic part of the CAL coronary artery vessel were significantly increased irrespective of blood supply (all P < 0.01). Moreover, these were accompanied by the increased expressions of NCX and p47phox, the decreased expression of SERCA2a, and the increased amount of phosphorylated forms of p47phox in NADPH oxidase (all P < 0.05).</p><p><b>CONCLUSIONS</b>Our results demonstrated that the disordered calcium clearance and the increased ROS generation occurred in the coronary artery smooth muscle cells in rats with chronic HF produced by ligation of the left anterior descending coronary artery (CAL), and which was found to be disassociated from blood supply, and the increased generation of ROS in the cells was found to make concomitancy to the increased activity of NADPH oxidase in cytoplasm.</p>
Subject(s)
Animals , Male , Rats , Calcium , Metabolism , Cells, Cultured , Coronary Vessels , Metabolism , Disease Models, Animal , Flow Cytometry , Heart Failure , Metabolism , Myocytes, Smooth Muscle , Metabolism , Random Allocation , Rats, Sprague-Dawley , Reactive Oxygen Species , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To establish a method for isolation, cryopreservation and recovery of the highly viable human peripheral blood monomuclear cells (PBMNCs) so as to achieve the long-term preservation of PBMNCs.</p><p><b>METHODS</b>A total of 80-100 ml peripheral blood were collected from the healthy volumteers aged over 50 years old. The PBMNCs were isolated by the Ficoll density gradient technique and cryopreserved gradually by program control method in liquid nitrogen freezer of -196 °C. The serum-free medium and autoloqous plasma medium were test for preservation of PBMNCs. The cell viability was assessed at time point of 1, 2, 4, 8, 12 and 24 months after thawing. Finally, the proliferation ability, purity and cytotoxicity were compared between the autologous immune lymphocytes (AIL) induced from cryopreserved PBMNCs and AIL as control from fresh PBMNCs.</p><p><b>RESULTS</b>After separating, the cell viability was 99.6%±0.4%, and the recovery rate of lymphocytes was 58.4%±6.52%. The cell recovery rate of lymphocyte was 89.7%±3.82% at 24 months. The quality assurance program was reliable within 2 years of running. The AIL cells induced with cryopreserved PBMNCs were not significantly different from those induced from fresh PBMNCs in terms of proliferative action, purity and cytotoxicity(CD3(+)CD8(+) ≥45%,CD3(+)CD56(+) NKT≥10%,CD4(+)CD25(+) NKT≤10%).</p><p><b>CONCLUSION</b>Manual separation of lymphocytes in vitro can get enough high-quality PBMNCs. The long-term cryopreserved PBMNC still maintain their high viability. The reinfusion of the clinical autologous immune cells would be advantageous for early tumor immunotherapy. Human AIL induced from cryopreserved PBMNC maintain their anti-tumor ability. These findings have the important implications for the application of these cells to adoptive cellular therapy.</p>
Subject(s)
Humans , Cell Survival , Cryopreservation , Immunocompetence , Leukocytes, MononuclearABSTRACT
? AIM: To investigate the efficacy and safety of phacoemulsification combined with intraocular lens implantation in the treatment of shallow anterior chamber with cataract.?METHODS: Retrospective case series. From February 2014 to July 2015 in our hospital,65 eyes in 65 patients with cataract were enrolled and divided into mild and high risk of shallow anterior chamber group. Best-corrected visual acuity ( BCVA ) , intraocular pressure ( IOP ) , central anterior chamber dept ( CACD ) , angle opening distance ( AOD ) , complications pre- and post treatment, were observed and analyzed as outcome measures.?RESULTS: In this study, the mild shallow anterior chamber group included 34 eyes; postoperative BCVA were improved in 29 eyes, with 4 eyes remaining stable and decreased in 1 eye; BCVA was improved in 16 eyes, with 10 eyes remaining stable and decreased in 5 eyes in high risk of shallow anterior chamber group postoperatively. BCVA had a better prognosis in the mild shallow anterior chamber group than another group ( t=-2. 956, P<0. 05). Meanwhile, IOP decreased by 5. 71± 2. 07mmHg and CACD increased by 1. 37 ± 0. 38mm in the mild shallow anterior chamber group, by 9. 77±4. 04mmHg and 1. 67±0. 43mm respectively in high risk group, and the difference has statistical significance ( t=-5. 02,-3. 04; P<0. 05). The mean preoperative nasal AOD500 was 200. 57± 33. 74μm, and they were 346. 62 ± 101. 37μm and 410. 75 ± 137. 48μm and 398. 69 ± 122. 28μm respectively at postoperative 1d, 1 and 3mo, and all nAOD500 comparing with preoperative were increased obviously, and the difference has statistical significance (F=203. 75, P<0. 01). And AOD500 at temporal, superior and inferior presented similar trends. Complications were corneal edema ( 5 eyes ) , transient intraocular hypertension ( 2 eyes ) , posterior capsular opacification ( 4 eyes ) , and posterior capsular rupture (1 eye).?CONCLUSION:Micro incision cataract surgery is useful, effective and safe in patients with cataract and shallow anterior chamber which can stabilize or improve BCVA, reduce IOP, deepen CACD and open the anterior chamber angle.