OBJECTIVESTo evaluate the effects of cryopreservation on human sperm chromosome.

METHODSperm chromosome were acquired using in vitro fertilization of zona-free hamster oocytes and human sperm. The frequency of sperm chromosome anomalies and sex chromosomes ratio before and after freezing and with different freezing methods were compared.

RESULTSThere were no significant differences of frequency of sperm chromosome anomalies among fresh, fast frozen and slow frozen sperm (9.40%, 7.48% and 8.74%) (P > 0.75) or ratios of sex chromosomes (P > 0.90).

CONCLUSIONSThese studies indicate that cryopreservation does not exert effects on human sperm chromosome.

Chromosomes ; Cryopreservation ; Humans ; Male ; Spermatozoa ; physiology

OBJECTIVETo explore the significance of C4d deposition in follicular lymphoma (FL).

METHODSThe deposition of C4d was detected in samples from 133 cases of lymphoma by immunohistochemistry and FL was studied by the double stainings of CD35/C4d, CD21/C4d and Bcl-2/C4d,respectively.

RESULTSAmong the 26 FL tissues, irregular C4d deposition was seen in 19 tumor tissues. Double staining for CD35, CD21 or Bcl-2 showed the C4d deposition was around the follicular dendritic cells (FDC). There was no significant difference between the positive rate of C4d and the degree of lymphoma. No deposition was found in the diffuse areas of FL and other type lymphomas.

CONCLUSIONC4d deposition around the follicular dendritic cell in the neoplastic follicles is a specific indicator of follicular lymphoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Complement C4b ; immunology ; Female ; Humans ; Immunohistochemistry ; Lymphoma, Follicular ; immunology ; pathology ; Male ; Middle Aged ; Peptide Fragments ; immunology

OBJECTIVETo evaluate the effect of adenosine on endothelin-1 (ET-1) after acute myocardial infarction (AMI) and reperfusion and explore the possible mechanism of no-reflow.

METHODSTwenty-four mini-swine were randomized into three study groups: control group (n=8), adenosine treated group (n=8), and sham-operated group (n=8). The mini-swine in the groups were subjected to 3 hours of coronary occlusion, followed by 60 minutes of reperfusion except those in the sham-operated group. The levels of ET-1 in blood sample, normal, infracted reflow and no-reflow myocardium were evaluated by radioimmuno-assay (RIA). The gene expressions of ET-1 in normal, infracted reflow and no-reflow myocardium were quantified by reverse transcription-polymerase chain reaction.

RESULTSIn both control group and adenosine group, compared with that at the baseline, ET-1 in blood sample significantly increased at 5 minutes and 180 minutes of left anterior descending coronary artery occlusion, as well as 5 and 60 minutes of reperfusion (all P < 0.01). In adenosine group, the levels of ET-1 were significantly lower than those in the control group (P < 0.05, P < 0.01). In both control group and adenosine group, compared with that in normal myocardium, ET-1 levels in both infarcted reflow and no-reflow myocardium significantly increased (both P < 0.01), with the level of ET-1 in no-reflow myocardium significantly higher than that in infarcted reflow myocardium (P < 0.01). In adenosine group, the level of ET-1 in infarcted reflow myocardium was significantly lower than that in the control group (P < 0.01). In both control and adenosine groups, compared with that in normal myocardium, the gene expression of ET-1 in infarcted reflow myocardium was significantly up-regulated (P < 0.01), while that of ET-1 in. no-reflow myocardium significantly down-regulated (P < 0.01). In adenosine group, the level of ET-1 in infarcted reflow myocardium was significantly lower than that in the control group (P < 0.01).

CONCLUSIONThe endothelium injury may be one of the important mechanisms for no-reflow phenomenon. Adenosine cay prevent endothelium from injury to reduce no-reflow.

Adenosine ; pharmacology ; therapeutic use ; Animals ; Disease Models, Animal ; Endothelin-1 ; genetics ; metabolism ; Female ; Male ; Myocardial Infarction ; drug therapy ; physiopathology ; Myocardial Reperfusion ; Swine ; Swine, Miniature

OBJECTIVETo evaluate the effects of ischemic preconditioning (IPC) on myocardial no-reflow in a mini-swine model of acute myocardial infarction (AMI) and reperfusion.

METHODSTwenty-four mini-swines were randomized into 3 study groups: 8 in control, 8 in IPC and 8 in sham-operated. Animals in the former two groups were subjected to 3 hours of coronary occlusion followed by 1 hour of reperfusion. Data on hemodynamics and coronary blood flow volume (CBV) were collected, and the area of no-reflow (ANR) was evaluated with both myocardial contrast echocardiography (MCE) in vivo and pathological means. Necrosis area (NA) was measured with triphenyltetrazolium chloride (TTC) staining.

RESULTSIn control group, left ventricular systolic pressure (LVSP), the maximum change rate of left ventricular pressure rise and fall (+/-dp/dtmax) and cardiac output (CO) significantly declined (P < 0.05, P < 0.01), while left ventricular end-diastolic pressure (LVEDP) significantly increased at the end of 3 hours of left anterior descending coronary artery occlusion (both P < 0.01), with +/-dp/dtmax further significantly declined (both P <0.05) at 1 hour of reperfusion. In IPC group, LVSP, +/-dp/dtmax, CO and LVEDP significantly recovered at 1 hour of reperfusion, compared with those in control group. In IPC group, the coronary ligation area was similar on both MCE in vivo and pathological evaluation (P > 0.05), and ANR was both also similarly as high as (16.4 +/- 2.24) % and (17.5 +/- 2.87) %, respectively, with final necrosis area (NA) reaching (78.4 +/- 3.62) %. In IPC group, ANR and final NA were significantly lower than those in control group (P < 0.05, P < 0.01). In the control group, coronary blood flow volumn immediately after release of 3 hours occlusion and at 1 hour of reperfusion were significantly lower than the baseline (both P < 0.01). In IPC group, coronary blood flow volumn were significantly higher than those in the control group (both P < 0.01).

CONCLUSIONIPC is effective to prevent myocardial no-reflow, improve left ventricular function and decrease infarct area.

Animals ; Blood Flow Velocity ; Coronary Circulation ; physiology ; Echocardiography ; Hemodynamics ; Ischemic Preconditioning ; Myocardial Infarction ; diagnostic imaging ; physiopathology ; Myocardial Reperfusion ; methods ; Myocardial Reperfusion Injury ; diagnostic imaging ; physiopathology ; prevention & control ; Random Allocation ; Swine ; Swine, Miniature

OBJECTIVETo study the growth and differentiation of adult canine autologous skeletal myoblasts after being transplanted into acute myocardial infarction (AMI) region by intramyocardium injection (IMI) and intracoronary infusion (ICI).

METHODSAutologous skeletal myoblasts were procured by a modified method. AMI model of adult canine was obtained through left anterior descending branch ligation and was divided into 4 groups (n = 5 for each group). Autologous skeletal myoblasts (1.0 - 1.4 x 10(8) cells) were injected locally into AMI region or infused into infarction-related coronary artery. Specimens were harvested 4 weeks after cellular transplantation for histological study including HE, PTH, immunochemical stain and transmission electronmicroscope.

RESULTSIn both two transplantation groups, newborn muscle-derived cells, striated muscle tissue and mature skeletal myofibril were demonstrated existing in MI region by electronmicroscope, PTH stain or anti-skeletal myosin heavy chain (slow) immunochemical stain, respectively. Newborn striated muscle tissues arranged in order of consistency with host myocardial fibers in two treatment groups. Newborn striated muscle tissue was more inclined to gather in MI region in the local injection group but distracted from each other in the intracoronary infusion group.

CONCLUSIONAutologous skeletal myoblasts appears to live and differentiate into mature striated muscle tissue after transplanting into AMI region by IMI or ICI routes.

Animals ; Cell Transplantation ; methods ; Cells, Cultured ; Dogs ; Female ; Male ; Myoblasts, Skeletal ; cytology ; transplantation ; Myocardial Infarction ; surgery ; Transplantation, Autologous

OBJECTIVETo investigate the therapeutic effectiveness of intracoronary transplantation of autologous bone marrow mononuclear cells (BM-MNC) on myocardial ischemia reperfusion injury in mini-swine model.

METHODSMyocardial ischemia reperfusion injury model was established by ligating in 16 mini-swines, which were further randomized into two groups: (3.54 +/- 0.90) x 10(8) BM-MNC was intracoronarily transplanted in BM-MNC group (n = 9), and phosphate buffer saline was intracoronarily applied in the control group (n = 7). Ultrasonic cardiograhpy, hemodynamics, neovascular density, and myocardium infarction size were evaluated before and 4 weeks after transplantation.

RESULTSIn BM-MNC group, left ventricular ejection fraction (LVEF), intra-ventricular septa, lateral wall and anterior wall, cardiac output (CO) and + dp/dt(max) had no significant differences before and 4 weeks after transplantation (P > 0.05). In the control group, LVEF, intraventricular septa, lateral wall and anterior wall, CO, and + dp/dt(max) significantly decreased 4 weeks after transplantation (P < 0.05). Left ventricular end-diastolic pressure and- dp/dt(max) had no significant differences before and after cell transplantation. Capillary density was significantly larger in the BM-MNC group than in the control group [(13.39 +/- 6.96) /HP vs. (3.50 +/- 1.90) /HP]. The percentage and size of myocardial infarction was significantly lower in the BM-MNC group than in the control group.

CONCLUSIONTransplantation of BM-MNC into the myocardial ischemic reperfusion-injury area can increase capillary density and decrease infarction area, and thus remarkably improve cardiac systolic function.

Animals ; Bone Marrow Transplantation ; Coronary Vessels ; Myocardial Reperfusion Injury ; pathology ; physiopathology ; therapy ; Myocardium ; pathology ; Random Allocation ; Swine ; Swine, Miniature

OBJECTIVETo compare the effects of intracoronary transplantation of autologous bone marrow mononuclear cells (BM-MNC) or peripheral endothelial progenitor cells (EPC) in mini-swine model of myocardial ischemia-reperfusion.

METHODSThe Mini-swine acute myocardial infarction and reperfusion model was created with 90 min occlusion of the left anterior descending coronary artery followed by reperfusion and the animals were then divided into BM-MNC group (3.54 x 10(8) +/- 0.90 x 10(8), n = 9), EPC group (1.16 x 10(7) +/- 1.07 x 10(7), n = 7) and control group (saline, n = 7). Echocardiography, hemodynamic measurements and myocardium infarction size were evaluated before and 4 weeks after intracoronary cell transplantations.

RESULTSThe net decrease from baseline to 4 weeks after transplantation of left ventricular ejection fraction (LVEF), left ventricular end systolic pressure, cardiac output and +dp/dt(max) were significantly attenuated post BM-MNC and EPC therapy compared to control group (all P < 0.05) and were similar between BM-MNC and EPC groups. Transplantation of BM-MNC and EPC also significantly decreased myocardial infarction size compared to control group.

CONCLUSIONAutologous intracoronary transplantation of BM-MNC or EPC in this model equally improved cardiac systolic function and reduced infarction area.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Coronary Circulation ; Disease Models, Animal ; Endothelial Cells ; cytology ; Female ; Male ; Myocardial Reperfusion Injury ; therapy ; Stem Cells ; cytology ; Swine ; Swine, Miniature ; Transplantation, Autologous

OBJECTIVETo investigate the differentiation status of autologous bone marrow mononuclear cells (BM-MNC) and peripheral endothelial progenitor cells (EPC) transplanted into myocardial ischemia reperfusion injury region in swine.

METHODSBM-MNC marked with PKH26 (n = 9), EPC marked with CM-DiI (n = 7), phosphate buffer saline (control, n = 7) were transplanted into myocardial ischemia reperfusion injury region of swine by intracoronary artery injection. Specimens were harvested 4 weeks after injection for histological analysis (HE, immunochemical stain for vWF, alpha-sarcomeric-actin and fibronectin antibody). Cell differentiation was observed under transmission electronmicroscope.

RESULTSThe number of small blood vessels was similar between BM-MNC group and EPC group (13.39 +/- 6.96/HP vs.12.39 +/- 4.72/HP, P < 0.05), but was significantly higher than that of control group (P < 0.05). Responsive intensity of immunochemical stain for fibronectin antibody was significantly lower in BM-MNC and EPC groups than that in control group. Responsive intensity of immunochemical stain for alpha-sarcomeric-actin antibody was similar among the three groups. Cluster cells were observed in one swine from BM-MNC group which might relate to the proliferation of stem cells in situ. Immature endothelial cells and myocytes were also detected by transmission electronmicroscope in BM-MNC and EPC group.

CONCLUSIONBM-MNC and EPC transplanted into myocardial ischemia reperfusion injury region in swine stimulated the formation of blood vessels and inhibited fibrogenesis.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Survival ; Cells, Cultured ; Disease Models, Animal ; Endothelial Cells ; cytology ; transplantation ; Mesenchymal Stem Cell Transplantation ; Monocytes ; transplantation ; Myocardial Reperfusion Injury ; blood ; Stem Cells ; cytology ; Swine ; Swine, Miniature ; Transplantation, Autologous

OBJECTIVETo evaluate the effects of fosinopril on myocardial no-reflow in a mini-swine model of acute myocardial infarction and reperfusion.

METHODSTwenty-four mini-swines were randomized into 3 study groups: 8 in control group, 8 in fosinopril-treated group (1 mg.kg(-1).d(-1)) and 8 in sham-operated group. Animals in the former two groups were subjected to 3 hours of coronary occlusion followed by 60 minutes of reperfusion. Data on haemodynamics and coronary blood flow volume (CBV) were collected, and the area of no-reflow was evaluated with both myocardial contrast echocardiography (MCE) in vivo and pathological means. Necrosis area was measured with triphenyltetrazolium chloride (TTC) staining.

RESULTS(1) In the control group, systolic and diastolic blood pressure (SBP and DBP), left ventricular systolic pressure (LVSP), maximal rate of increase and decrease in left ventricular pressure (+/- dp/dt(max)) and cardiac output (CO) significantly declined (P < 0.05-0.01), while left ventricular end-diastolic pressure (LVEDP) and pulmonary capillary wedge pressure (PCWP) significantly increased at the end of 3 hours occlusion of left anterior descending artery (both P < 0.01). Compared with those at the end of 3 hours of occlusion, +/- dp/dt(max) further significantly declined (P < 0.05) at 60 minutes of reperfusion. In the fosinopril group, the changes of SBP and DBP, LVSP, +/- dp/dt(max), CO, LVEDP and PCWP were similar as those in the control group after 3 hours of acute myocardial infarction. In contrast, LVSP, +/- dp/dt(max), CO, LVEDP and PCWP recovered significantly at 60 minutes of reperfusion. (2) In the control group, the coronary ligation area was similar on both MCE in vivo and pathological evaluation, and the area of no-reflow was similarly as high as 78.5% and 82.3%, respectively, with final necrosis area reaching 99% of ligation area. In the fosinopril group, there was no significant difference in ligation area on both MCE and pathological evaluations between the fosinopril and control groups, although the area of no-reflow on both methods was significantly decreased to 24.5% and 25.2%, respectively, (P < 0.01) with final necrosis area of pathological evaluation being also significantly decreased to 88.9% of LA (P < 0.05). (3) In the control group, CBV was significantly declined to 45.8% and 50.6% from at baseline, immediately after release of occlusion (3 hours) and at 60 minutes of reperfusion (P < 0.01). In the fosinopril group, CBV was also significantly declined immediately after release of occlusion (3 hours), and at 60 minutes of reperfusion (P < 0.05), but significantly increased to 69.1% and 72.1% from at baseline, that were significantly greater than those in the control group (both P < 0.01).

CONCLUSIONFosinopril is effective in preventing myocardial no-reflow, improving left ventricular function, and reducing infarct area during acute myocardial infarction and reperfusion in mini-swine.

Animals ; Blood Flow Velocity ; drug effects ; Disease Models, Animal ; Female ; Fosinopril ; pharmacology ; therapeutic use ; Male ; Myocardial Infarction ; drug therapy ; Myocardial Reperfusion ; methods ; Myocardial Reperfusion Injury ; prevention & control ; Swine ; Swine, Miniature

OBJECTIVETo evaluate the beneficial effects of adenosine on myocardial no-reflow in a mini-swine model of acute myocardial infarction (AMI) and reperfusion.

METHODSTwenty-four animals were randomly assigned to 3 groups: 8 in controls, 8 in adenosine-treated and 8 in sham-operated. The groups were subjected to 3 hours of coronary occlusion followed by 60 minutes of reperfusion except the sham-operated group. Data on hemodynamics and coronary blood flow volume (CBV) were collected. The area of no-reflow was evaluated by both myocardial contrast echocardiography (MCE) in vivo and histopathological means and necrosis area was measured with triphenyltetrazolium chloride staining.

RESULTS(1) In control group, systolic and diastolic blood pressure (SBP and DBP), left ventricular systolic pressure, maximal rate of increase and decline in left ventricular pressure (+/- dp/dtmax) and cardiac output significantly declined (P < 0.05-0.01), while left ventricular end-diastolic pressure (LVEDP) and pulmonary capillary wedge pressure (PCWP) significantly increased at the end of 3 hours of LAD occlusion (both P < 0.01), with +/- dp/dtmax further significantly declined (both P < 0.05) at 60 minutes of reperfusion. In adenosine treated group, the changes of SBP and DBP, left ventricular systolic pressure, +/- dp/dtmax, cardiac output, LVEDP and PCWP were the same as those in the control group after AMI and reperfusion, while left ventricular systolic pressure, +/- dp/dtmax, cardiac output, LVEDP and PCWP recovered significantly at 60 minutes of reperfusion compared with those at 6 hours AMI. (2) In control group, the coronary ligation areas (LA) were similar (P > 0.05) detected by MCE in vivo and histopathological evaluation, and the areas of no-reflow were both as high as 67.5% and 69.3%, respectively. The final necrosis area reached 99% of LA. Compared with those in the control group, there was no significant difference in LA on both MCE and histopathological evaluation in the adenosine-treated group, though the areas of no-reflow on both methods were significantly decreased to 21% and 22% (both P < 0.01) and final necrosis area was also significantly decreased to 75% of LA (P < 0.05). (3) In the control group, CBV were significantly declined to 45.8% and 50.6% of the baseline at immediately after release of 3 hours occlusion and at 60 minutes of reperfusion, respectively (both P < 0.01). In the adenosine-treated group, CBV were also significantly declined at immediately after release of 3 hours occlusion, and at 60 minutes of reperfusion (both P < 0.05), though significantly increased to 79.5% and 79.9% of the baseline which were both significantly higher than those in the control group.

CONCLUSIONAdenosine has an effective role in preventing myocardial no-reflow, improving left ventricular function and reducing infarct area during AMI and reperfusion in mini-swine.

Adenosine ; pharmacology ; therapeutic use ; Animals ; Cardiac Output ; drug effects ; Coronary Circulation ; drug effects ; Disease Models, Animal ; Female ; Male ; Myocardial Infarction ; drug therapy ; physiopathology ; Myocardial Reperfusion Injury ; prevention & control ; Pulmonary Wedge Pressure ; drug effects ; Swine ; Swine, Miniature