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BACKGROUND:We explore the transfection effects of different lentiviruses on bone marrow mesenchymal stem cells (BMSCs) from the same source in order to screen out appropriate cell transfection tools.OBJECTIVE:To explore the difference in the transfection efficiency of three different lentiviruses carrying green fluorescent protein (GFP) to transfect the rabbit BMSCs.METHODS:Bone marrow samples were extracted from the ilium of 2-week-old Japan white rabbits,and then the BMSCs were separated,purified and extended using density gradient centrifugation.Cell surface markers were identified using flow cytometry.pTomo-GFP lentivirus,Ad-GFP lentivirus and lenti-GFP lentivirus were used to transfect the BMSCs,and the transfection efficiency was tested using flow cytometry.The cell viability was detected using cell counting kit-8 before and after transfection.RESULTS AND CONCLUSION:As the positive rates of CD44,CD34 and CD45 were 75.64％,6.14％ and 9.48％,respectively,the cultured cells were identified as BMSCs by the flow cytometry.The expression levels of GFP after transfection by pTomo-GFP,Ad-GFP and ienti-GFP were 2.64％,16.72％ and 45.24％,respectively.The cell growth curve showed that pTomo-GFP lentivirus,Ad-GFP and ienti-GFP lentivirus showed certain effects on cell growth,pTomo-GFP and Ad-GFP had an effect on cell growth at incubation period and logarithmic growth phase at 1-8 days after transfection,which may cause some interference to the viability of early cells,ienti-GFP had little effect on cell growth in the short term,but there was a change in cell growth at day 12 after lenti-GFP transfection,and a long-term effect of lenti-GFP on cell viability was considered.Therefore,lenti-GFP lentivirus could highly transfect the BMSCs and could not impact the normal growth of BMSCs,indicating that lenti-GFP lentivirus is suitable for BMSCs tracing and gene transfection.
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<p><b>OBJECTIVE</b>To evaluate the prevention effect of diabetic retinopathy of Jiangtang Xiaozhi Recipe (, JXR) in streptozotocin (STZ)-induced diabetic rats.</p><p><b>METHODS</b>Sprague-Dawley rats were randomly divided into normal control group and diabetic group. Rats in the diabetic group were induced by intraperitoneal administration of STZ (50 mg/kg), and subdivided into 5 groups. Rats in the diabetic control group were given saline; four treatment groups were given metformin (300 mg/kg), JXR (2, 4 and 8 g/kg) respectively for 8 weeks, while rats in the normal control group were injected with citrate buffer and given the same volume of vehicle. Body weight and food intake were measured every week. The hypoglycaemic effects were determined by testing fasting blood glucose (FBG) every other week, and hemoglobin A1c (HbA1c), insulin, and glucagon at the end of the treatment. The preventive effects of JXR on STZ-induced diabetic rats were determined by histopathological examination with hematoxylin and eosin staining, and periodic acid-schiff staining. The effects were further evaluated by serum superoxide dismutase (SOD) activity and malondialdehyde (MDA).</p><p><b>RESULTS</b>High-dose JXR significantly reduced FBG and HbA1c level at the 8th week of administration (P<0.01, P<0.05). JXR significantly increased insulin level (P<0.05), and decreased glucagon level (P<0.05). JXR showed the antioxidant defense with increased SOD activity and decreased MDA contents in diabetic rats. Histopathological studies revealed that there were no basement membrane thickening and mild destruction in the treated groups. Morphometric measurements of retina microvascular showed that acellular capillary and capillary density decreased in treated rats while pericyte and endothelial cell increasing after the treatment.</p><p><b>CONCLUSION</b>JXR have protective effect of diabetic retinopathy and its mechanism may be associated with the obvious hypoglycemic and antioxidant effect.</p>
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Objective To understand the epidemiological characteristics of physical inactivity among the elderly in Jing'an District during 2002-2011;to quantify the burden of stroke and coronary heart disease attributable to physical inactivity; and to estimate the benefits in disability free life expectancy ( DFLE) from reasonable physical activity. Methods To understand the epidemiological characteristics of physical inactivity among the elderly in Jing'an District, through analyzing DFLE data.To obtain relative risk ( RR) of stoke and coronary heart disease attributable to physical inactivity, via World Health Organi-zation ( WHO) Comparative Risk Assessment ( CRA) project.To calculate population attributable frac-tions ( PAFs) using CRA methodology.To estimate the increase of DFLE attributable to physical activity, based on the results of Jing'an District DFLE research. Results From 2002 to 2011 , the prevalence of physical inactivity and the incidence of stroke and coronary heart disease associated with physical inactivity among the elderly in Jing'an District presented a downward trend, i.e.incidence of stroke at 13.80%(range 9.16%-21.43%) and that of coronary heart disease at 16.83% (11.19%-26.76%).With the elimination of physical inactivity, DFLE is expected to increase by 0.94 (0.11-2.70) years from stroke and 0.65 (0.11-1.91) years from coronary heart disease. Conclusion Increase of physical activity could reduce the incidence of stroke and coronary heart disease among the elderly, increase DFLE , and gain health benefits.
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<p><b>OBJECTIVE</b>To report the operative techniques and clinical results of modified superficial peroneal neurocutaneous propeller adipofascial-cutaneous flap for reconstruction of donor site defects at foot dorsum.</p><p><b>METHODS</b>A propeller adipofascial flap with a skin pedicle (4-6 cm in width) based on the lateral superamalleolar perforating artery which vascularized the flap through the nutrient vessel chain of the superficial peroneal nerve was designed to repair defects after harvesting of foot pedicled dorsal flap. The defects at donor site of the leg was closed directly and split-thickness skin grafting was performed on the adipofascial surface of the flap primarily or secondarily.</p><p><b>RESULTS</b>From May 2007 to Oct. 2011, 7 cases were treated. All flaps were transplanted successfully with satisfactory cosmetic and functional results. The flaps size ranged from 19 cm x 8 cm to 30 cm x 11 cm.</p><p><b>CONCLUSIONS</b>The flap has reliable blood supply with a relatively large vascularized area, long rotation are and minimum donor-site cosmetic morbidity. It' s a simple and safe procedure which is suitable for covering donor sites defects after harvesting foot pedicled dorsal flap.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Foot Injuries , General Surgery , Peroneal Nerve , Transplantation , Plastic Surgery Procedures , Methods , Skin Transplantation , Methods , Surgical FlapsABSTRACT
<p><b>OBJECTIVE</b>To report the operative techniques and clinical results of specially designed sural neurocutaneous vascular flap pedicled on a dominant perforator (the diameter > or = 0.8 mm) of the peroneal artery for coverage of soft tissue defects overlying the Achilles tendon.</p><p><b>METHODS</b>An approximately rectangular sural neurocutaneous vascular flap pedicled on the lowest dominant perforator arising from the peroneal artery was designed and harvested to repair defects over the Achilles tendon. The pedicle was located at a certain part of the flap, which divided the flap into the distal and the proximal parts. After the tendon was repaired, the flap was rotated 180 degrees based on the perforator and the position of the distal and proximal parts of the flap was changed to cover the defects and part of the donor site respectively. In most cases, skin graft was not needed.</p><p><b>RESULTS</b>The modified flaps were applied in 15 cases. All flaps (ranged from 13 cm x 15 cm - 18 cm x 9 cm ) were transplanted successfully without necrosis, and no vascular problems occurred. Following up for 10-17 months showed both satisfactory functional and cosmetic results.</p><p><b>CONCLUSIONS</b>The modified flap has reliable blood supply and the special design provides nearly normal outline of the ankle which favorites shoe wearing. It' s an excellent option for covering defects overlying the Achilles tendon.</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Achilles Tendon , Wounds and Injuries , General Surgery , Arteries , Transplantation , Skin Transplantation , Methods , Sural Nerve , Surgical Flaps , Wound HealingABSTRACT
Objective To evaluate the preventive and therapeutic effect of oral solution of Niao Du Kang (尿毒康) on acute renal tubular necrosis (ATN) in Sprague-Dawley (SD) rats,and preliminarily approach its mechanisms.Methods The oral solution was composed of traditional Chinese medicinal herbs of Radix et Rhizoma Rhei (大黄),Radix Sanguisorbae Offieinalis (地榆),Radix Salviae Miltiorrhizae (丹参), Radix Astragali seu Hedysari (黄芪),Flos Carthami Tinctorii (红花) and so on.Sixty-four SD male rats were randomly divided into 4 groups:normal control group,ATN model group,verapamil treatment group and traditional Chinese medicine,oral solution of Niao Du Kang treatment group,every group having 16 rats.. The ATN model of SD rats was induced by intramuscular injection of 50% glycerin mixed with isotonic saline solution at the back of bilateral lower extremities.After the model was established in each group ,the levels of blood urea nitrogen (BUN),levels of blood serum creatinine (SCr),renal failure index (RFI) and renal pathological changes were detected at different time points after modeling for 12 and 24 hours,and the therapeutic effect of oral solution of Niao Du Kang was observed.Results After the model establishment, Niao Du Kang oral solution could lower the elevation of blood BUN,SCr,RFI in the model,and compared with the normal control group,there were statistical significances (all P
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<p><b>OBJECTIVE</b>The aim of this study is to identify short-term result of cell transplantation in idiopathic dilated cardiomyopathy (IDC) patients who were treated by intracoronary transplantation of autologous mononuclear bone marrow cells (BMCs) in addition to standard therapy.</p><p><b>METHODS</b>Based on given standard therapy, eighteen patients with idiopathic dilated cardiomyopathy were enrolled and divided into transplantation group and control group. The clinical characteristics of two groups were comparable. Among these patients, 10 patients were performed percutaneous coronary autologous BMCs transplantation. Blood routine test, hepatic function, renal function, glucose, triglyceride (TG), cholesterol, low density cholesterol (LDL), high density cholesterol (HDL), uric acid (UA) and high sensitive C-reactive protein (hsCRP) were measured at the time point of pre-operation and some time after transplantation. All patients were monitored under ultrasonic cardiography, Holter, six-minute-walk test and magnetic resonance imaging over a period of at least 6 months. Annual hospital days were recorded during two-year follow-up.</p><p><b>RESULTS</b>Blood routine test, hepatic function, renal function, glucose, TG, cholesterol, LDL, HDL, UA and hsCRP had no significant differences among 48 hours, 3 months and 6 months after transplantation compared with control and pre-transplantation (P > 0.05). Six-minute-walk distance elevated significantly six months after BMCs transplantation compared with control and pre-transplantation [(494.3 +/- 62.8) m vs (307.2 +/- 75.0) m, (321.5 +/- 63.7) m, P < 0.05]. Left ventricular ejection fraction (LVEF) and the sizes of LVEDd had no significant changes compared with that of control and pre-transplantation (P > 0.05). Myocardium lesion area measured by (MRI) seemed decrease in transplantation group compared with that of control and pre-operation [(4.96 +/- 0.47) cm(2) vs (5.12 +/- 0.54) cm(2), (5.02 +/- 0.39) cm(2), P > 0.05], but there was no significance. None of proarrhythmias and side effects had been observed around transplantation and 2 years follow-up. There was no significant difference in survival between two groups in 2 years follow-up. Interestingly, annual hospital day in BMCs transplantation patients was significantly shorter than that in control group [(30.2 +/- 11.2) d vs (43.6 +/- 9.8) d, P < 0.05].</p><p><b>CONCLUSIONS</b>Autologous bone marrow mononuclear cells transplantation can prolong six-minute-walk, decrease re-hospitalization rate, elevate exercise ability and help to improve cardiac function in patients with IDC. In addition, it was demonstrated that cell transplantation is safe.</p>
Subject(s)
Humans , Bone Marrow Transplantation , Cardiomyopathy, Dilated , General Surgery , Therapeutics , Transplantation, Autologous , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To investigate the safety of autologous bone marrow mononuclear cell (BM-MNCs) transplantation by intracoronary infusion in patients with acute myocardial infarction (AMI).</p><p><b>METHODS</b>One hundred and eighty-four patients with AMI treated with percutaneous coronary intervention (PCI) were randomized in a 1:1 way to either intracoronary transplantation of autologous BM-MNCs (n = 92) right after PCI or to sodium chloride concluding heparin (controlled, n = 92) via a micro infusion catheter. In the process of the intracoronary infusion of BM-MNCs, the complications should be recorded, which were aberration reflect (including of pale, syncope, nausea, hypotension and shock), deterioration of angina or heart failure, arrhythmias (including of bradycardia, sinus arrest or atrial ventricular block or ventricular fibrillation), embolism etc. Body temperature, blood pressure and heart rates should be monitored during the first week after transplantation. Holter, coronary angiography and ultrasonic cardiography were performed at the designed time points. Main heart accidents, restenosis and tumor were recorded during 2-years follow up.</p><p><b>RESULTS</b>During the period of bone marrow puncture and intracoronary infusion of BM-MNCs, few patients occurred pale, dizziness, bradycardia and hypotension, which were transient and due to vagus reflect. No stem cell-related arrhythmias, deterioration of angina were noted. In BM-MNCs group one patient developed in-stent reocclusion in one week after transplantation, five developed in-stent restenosis during further follow-up 30 months, which were similar with control group. There were no deaths, major adverse cardiac events, tumor and other late adverse events during follow-up period in both groups.</p><p><b>CONCLUSION</b>Intracoronary transplantation of autologous BM-MNCs in the acute phase after AMI is feasible and seems safe in the 30 months of follow-up.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Bone Marrow Transplantation , Methods , Coronary Vessels , Follow-Up Studies , Mesenchymal Stem Cell Transplantation , Methods , Myocardial Infarction , General Surgery , Transplantation, AutologousABSTRACT
<p><b>OBJECTIVE</b>To evaluate the value of quantitative real-time myocardial contrast echocardiography (RT-MCE) combined with low dose dobutamine stress test in the detection of coronary artery disease (CAD), and to assess the contribution of collateral blood flow (CBF) to myocardial perfusion.</p><p><b>METHODS</b>Twenty-six hospitalized patients referred for coronary angiography and subsequent revascularization underwent routine echocardiography, RT-MCE at baseline and after low dose dobutamine administration. The images of RT-MCE were analyzed quantitatively from microbubble replenishment curves for myocardial perfusion and its reserve by using the QLab software.</p><p><b>RESULTS</b>At baseline, both beta and A x beta (but not A) were decreased with the increase of severity of coronary stenosis (P < 0.01). Under dobutamine stress, A, beta and A x beta values were decreased with the increase of severity of coronary stenosis (P < 0.01), Graded decreasing in the reserves of A, beta and A x beta were observed with increasing coronary stenosis severity (P < 0.01). Furthermore, significant differences in beta, A x beta, and WMS were observed between segments with CBF and those without.</p><p><b>CONCLUSION</b>Quantitative RT-MCE in conjunction with dobutamine stress can be used as a sensitive measure to identify and stratify CAD as well as to assess the contribution of collateral blood flow.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Collateral Circulation , Contrast Media , Coronary Circulation , Coronary Disease , Diagnostic Imaging , Echocardiography, Stress , MethodsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the role of contrast-enhancement magnetic resonance imaging (CeMRI) in patients with myocardial infarction (MI).</p><p><b>METHODS</b>There were twenty-three patients enrolled in this study. After dynamic observation, there were 20 patients who were diagnosed as MI. All those patients underwent coronary artery angiography and CeMRI. MRI was performed with a 1.5-T magnet (AVANTO, SIMENS). After tagged images were acquired, the patients received an intravenous bolus of 0.1 mmol/kg Gd-DTPA at a rate of 5 ml/s. A first-pass perfusion scan was acquired simultaneously with a bolus injection. A second bolus of 0.3 mmol/kg Gd-DTPA was given following the first-pass images. Delayed images were acquired 5 minutes after the second bolus by using an inversion-recovery prepared gated fast-gradient echo-pulse sequence.</p><p><b>RESULTS</b>Hypoenhancement was seen in 20 patients at the first-pass perfusion at the myocardial infarction site, while hyperenhancement was seen at delayed CeMRI. Myocardial infarction area in delayed CeMRI was 16.58% +/- 9.73%, which was correlated positively with peak CK and cTnT (r = 0.821, P < 0.01 and r = 0.565, P < 0.05), respectively. The ejection fraction (EF) detected by MRI was 0.46 +/- 0.13, while the left ventricular EF (LVEF) detected by left ventriculography was 0.49 +/- 0.16. There was no difference between two parameters.</p><p><b>CONCLUSIONS</b>CeMRI may play an important role in the diagnosis and prognosis of patients with MI.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Contrast Media , Gadolinium DTPA , Image Enhancement , Methods , Magnetic Resonance Imaging , Methods , Myocardial Infarction , DiagnosisABSTRACT
<p><b>BACKGROUND</b>Emerging evidence suggests that stem cells can be used to improve cardiac function in patients after acute myocardial infarction. In this randomized trial, we aimed to use Doppler tissue tracking and strain imaging to assess left ventricular segmental function after intracoronary transfer of autologous bone-marrow stem cells (BMCs) for 6 months' follow up.</p><p><b>METHODS</b>Twenty patients with acute myocardial infarction and anterior descending coronary artery occlusion proven by angiography were [corrected] randomized into intracoronary injection of bone-marrow cell (treated, n = 9) or diluted serum (control, n = 11) groups. GE vivid 7 and Q-analyze software were used to perform echocardiogram in both groups 1 week, 3 months and 6 months after treatment. Three apical views of tissue Doppler imaging were acquired to measure peak systolic displacement (Ds) and peak systolic strain (epsilonpeak) from 12 segments of LV walls. Left ventricular ejection fraction (LVEF), end-diastolic volume (EDV) and end-systolic volume (ESV) were obtained by Simposon's biplane method.</p><p><b>RESULTS</b>(1) 3 months later, Ds and epsilonpeak over the infract-related region clearly increased in the BMCs group [Ds: (4.49 +/- 2.71) mm vs (7.56 +/- 2.95) mm, P < 0.01; epsilonpeak: (-13.40 +/- 6.00)% vs (-17.06 +/- 6.05)%, P < 0.01], but not in the control group [Ds: (4.74 +/- 2.67) mm vs (5.01 +/- 3.23) mm, P > 0.05; epsilonpeak: (-13.84 +/- 6.05)% vs (-15.04 +/- 6.75)%, P > 0.05]. At the same time, Ds over the normal region also increased, but the Ds enhancement was markedly higher in the BMCs group than that in the control group [(3.21 +/- 3.17) mm vs (0.76 +/- 1.94) mm, P < 0.01]. Parameters remained steady from the 3rd to 6th month in either group (P > 0.05). (2) LVEF in treated and control groups were almost the same at baseline (1st week after PCI) [(53.37 +/- 8.92)% vs (53.51 +/- 5.84)%, P > 0.05]. But 6 months later, LVEF in the BMCs group were clearly higher than that in the control group [(59.33 +/- 12.91)% vs (50.30 +/- 8.30)%, P < 0.05]. (3) There were no evident difference in EDV or ESV between two groups at baseline [EDV: (113.74 +/- 23.24) ml vs (129.94 +/- 32.72) ml, P > 0.05; ESV: (57.12 +/- 18.66) ml vs (62.09 +/- 17.68) ml, P > 0.05]. Three months later, EDV and ESV in the control group were markedly greater than those in the BMCs group [EDV: (154.89 +/- 46.34) ml vs (104.85 +/- 33.21) ml, P < 0.05; ESV: (82.91 +/- 35.79) ml vs (49.54 +/- 23.32) ml, P < 0.05]. But EDV and ESV did not change much from 3rd to 6th month in either group (P > 0.05).</p><p><b>CONCLUSIONS</b>Emergency transplantation of autologous BMCs in patients with acute myocardial infarction helps to improve global and regional contractility and attenuate post-infarction left ventricular remodeling. Tissue tracking and strain imaging provide quick, simple and noninvasive methods for quantifying left ventricular segmental function in humans.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Double-Blind Method , Hematopoietic Stem Cell Transplantation , Myocardial Infarction , Therapeutics , Transplantation, Autologous , Ventricular Function, Left , Ventricular RemodelingABSTRACT
<p><b>BACKGROUND</b>Quantitatively assessing myocardial perfusion and its reserve is of great importance for the diagnosis and stratification of patients with coronary artery disease (CAD), and represents an important goal of myocardial contrast echocardiography. In this study we sought to test the usefulness of low dose dobutamine stress real-time myocardial contrast echocardiography (RT-MCE) in the assessment of CAD, and to explore the relationship between perfusion reserve and contractile reserve.</p><p><b>METHODS</b>Twenty-six patients with suspected or clinical diagnosed CAD were enrolled and underwent RT-MCE at baseline and under low dose dobutamine stress, and subsequent coronary angiography. RT-MCE images were analyzed quantitatively from microbubble replenishment curves for myocardial perfusion and its reserve.</p><p><b>RESULTS</b>At baseline, significant differences in beta (0.28 +/- 0.12, 0.25 +/- 0.09, 0.22 +/- 0.06, 0.20 +/- 0.07 respectively, P < 0.01) and A x beta (1.37 +/- 0.46, 1.28 +/- 0.47, 1.13 +/- 0.37, 0.91 +/- 0.32, respectively, P < 0.01) were observed among four segment groups with graded coronary artery stenosis severity (normal; 30% - 69% stenosis; 70% - 90% stenosis; and beyond 90% stenosis), but not observed in parameter A. When under stress, significant differences in A (5.73 +/- 1.28, 5.63 +/- 1.01, 4.96 +/- 0.81, 4.57 +/- 0.62, respectively, P < 0.01), beta (0.67 +/- 0.17, 0.55 +/- 0.19, 0.32 +/- 0.13, 0.25 +/- 0.08, respectively, P < 0.01) and A x beta (3.81 +/- 1.20, 3.11 +/- 1.17, 1.59 +/- 0.82, 1.12 +/- 0.37, respectively, P < 0.01) were observed among the formerly mentioned groups. Graded decreases in A reserve (1.20 +/- 0.53, 1.11 +/- 0.16, 0.98 +/- 0.12, 0.99 +/- 0.13, respectively, P < 0.01), beta reserve (2.65 +/- 1.07, 2.32 +/- 0.82, 1.44 +/- 0.40, 1.29 +/- 0.34, respectively, P < 0.01) and A x beta reserve (3.05 +/- 1.63, 2.59 +/- 1.01, 1.42 +/- 0.44, 1.27 +/- 0.34, respectively, P < 0.01) could also be observed with increasing coronary stenosis severity. In five segments groups scored by WMS (1 - 5), concordance between contractile function and myocardial perfusion could be found both at rest (beta: 0.28 +/- 0.11, 0.22 +/- 0.08, 0.21 +/- 0.05, 0.17 +/- 0.05, 0.19 +/- 0.06, respectively, P < 0.01; A x beta: 1.29 +/- 0.48, 0.98 +/- 0.45, 0.94 +/- 0.29, 0.76 +/- 0.30, 0.92 +/- 0.32, respectively, P < 0.01) and under stress (beta: 0.59 +/- 0.20, 0.35 +/- 0.15, 0.27 +/- 0.08, 0.17 +/- 0.05, 0.20 +/- 0.05, respectively, P < 0.01; A x beta: 3.07 +/- 1.38, 1.62 +/- 0.82, 1.28 +/- 0.40, 0.78 +/- 0.24, 0.93 +/- 0.22, respectively, P < 0.01). This concordance is also valid in terms of the reserves, and the MCE parameters in segments with ameliorated contractile function are significantly higher than in those without.</p><p><b>CONCLUSIONS</b>Quantitative RT-MCE in conjunction with dobutamine stress shows promise in identifying and stratifying CAD and in exploring the perfusion-contractile correlation.</p>