AIM:To evaluate the effects of atorvastatin on nitric oxide(NO),endothelin-1(ET-1)and myocardial no-reflow in a rabbit model of acute myocardial infarction and reperfusion(AMI/R). METHODS:Twenty-four rabbits were randomized into 3 groups:8 in AMI/R group,8 in atorvastatin-treated group(5 mg·kg~(-1)·d~(-1))and 8 in sham-operated group. Animals in the former two groups were subjected to 60 min of coronary occlusion followed by 120 min of reperfusion. Data on haemodynamics were collected. NO in blood sample,and in normal,and in infarcted reflow and no-reflow myocardium were evaluated respectively by nitrate reductase method. The levels of ET-1 in blood sample,and in normal,infarcted reflow and no-reflow myocardium were determined by radioimmunoassay. RESULTS:(1)Compared to the baselines,the heart rate(HR),systolic blood pressure(SBP),diastolic blood pressure(DBP),left ventricular systolic pressure(LVSP),maximal rate of increase and decline in left ventricular pressure(±dp/dt_(max))and cardiac output(CO)in AMI/R and atorvastatin-treated groups were significantly declined,whereas left ventricular end-diastolic pressure(LVEDP)was increased after 60 min of coronary occlusion and 120 min of reperfusion(P<0.05 or P<0.01). However,in atorvastatin-treated group,LVSP,LVEDP,±dp/dt_(max) and CO at the time point of 120 min of reperfusion recovered more significantly than those at the time point of 60 min of coronary occlusion(P<0.01),which was more significant than those in AMI/R group(P<0.05 or P<0.01). Compared to AMI/R group,the SBP and DBP were significantly heigher in atorvastatin-treated group(P<0.01).(2)In atorvastatin-treated group,the levels of ET-1 in blood sample were significantly lower than those in AMI/R group(P<0.01),and the levels of NO were significantly higher(P<0.01). Moreover,the levels of NO or ET-1 in infarcted reflow myocardium were significantly lower than that in AMI/R group(P<0.05 or P<0.01).(3)Atorvastatin could ameliorate myocardial function. CONCLUSION:Atorvastatin is effective in increasing NO and reducing ET-1 in blood plasma and local myocardium,and in protection of endothelial cells. Atorvastatin also has a beneficial effect on improving left ventricular function during acute myocardial infarction and reperfusion in rabbits.

Thirty-five patients with perioperative pulmonary embolism during January 2000 and October 2006 were analyzed retrospectively.Disease diagnosis and treatment outcomes were assessed.Of the 35 patients,2 died without any management,and 33 received conventional anti-coagulation and thrombolysis.including 3 percutaneous catheterization.Disease improvement Was seen in 29.Four deaths occurred after no response to the treatment.Thus,early diagnosis and treatment,which miight depend on further understanding of pulmonary embolism,could play an important role in reduced adverse events.Prevention is crucial to avoid perioperative occurrence of pulmonary embolism.

Objective To study the proliferation and migration of vascular smooth muscle cell (VSMC) after transferred angiotensin Ⅱ (AngⅡ) type 2 receptor (AT2R) gene. 　Methods The recombinant adenoviral vector, AdCMV-AT2R, containing rat AT2 receptor gene was constructed by homologous recombination, and transfered to rat VSMC in vitro. The expression of AT2R mR NA was detected by RT-PCR. The rate of expression and the change of cell cycle in VSMC were analysed by flow cytometry. These assays including cell devision index. Incorporation of bromodeoxyuridine(BrdU) and 3-(4,5-dimethyl-thiazol-2-yl)2,5-diphenyltetrazolium bromide( MTT) were used to determine the proliferation of VSMC. The modified Boyden's chamber method was used to test the migration of VSMC. The r e-organization of F-actin in VSMC was analysed by confocal microscopy. 　Results RT-PCR showed that the expression of AT2R mRNA increased substantially in transferred VSMC, and the peak value of expression rate is about 89.51% at 48 hours. When the expression of AT2R is at peak value, the ratio of S, G2 and M periods was reduced from 31.7% to 13.9%(P<0.05). The OD values of MTT and BrdU incorporation were reduced by 61.4% and 51.6% respectively(P<0.0(1). the number of VSMC migration was reduced by 62.2%(P<0.05) and the expression of F-actin was also decreased signific antly. 　Conclusion AdCMV-AT2R induced high level expression of AT2 receptor in cultured rat VSMC and its expression can significantly inhibit the proliferation and migration of rats VSMC in vitro.

The objective of this study is to combine troponin and indicators of cardiac acoustics for synthetically evaluating cardiac fatigue of rabbits, analyzing exercise-induced cardiac fatigue (EICF) and exercise-induced cardiac damage (EICD). New Zealand white rabbits were used to conduct a multi-step swimming experiments with load, reaching an exhaustive state for evaluating if the amplitude ratio of the first to second heart sound (S1/S2) and heart rate (HR) during the exhaustive exercise would decrease or not and if they would be recovered 24-48 h after exhaustive exercise. The experimental end point was to complete 3 times of exhaustions or death from exhaustion. Circulating troponin I (cTnI) were detected from all of the experimental rabbits at rest [(0. 02±0. 01) ng/mL], which, in general, indicated that there existed a physiological release of troponin. After the first exhaustive swim, cTnI of the rabbits increased. However, with 24-hour rest, S1/S2, HR, and cTnI of the tested rabbits all returned toward baseline levels, which meant that the experimental rabbits experienced a cardiac fatigue process. After repeated exhaustion, overloading phenomena were observed, which led to death in 3 out of 11 rabbits, indicating their cardiac damage; the troponin elevation under this condition could be interpreted by pathological release. Evaluation of myocardial damage can not be based on the troponin levels alone, but can only be based on a comprehensive analysis.
Animals ; Fatigue ; Heart ; physiopathology ; Heart Rate ; Myocardium ; pathology ; Rabbits ; Swimming ; Troponin I ; blood

Objective To constructed the cell model transferred angiotensin Ⅱ (AngⅡ ) type 2 receptor (AT2R) in vascular smooth muscle cells(VSMC) Method The VSMCs, isolated from the aorta of rat, were cultured by routine method. Recombinant adenoviral vector, AdCMV-AT2R, containing rat AT2 receptor gene was constructed by homologous recombination, and then it was used to transfer AT2 receptor gene to VSMC in vitro. The rate of AT2R expression in VSMC was analysed by flow cytometry. The expression of mRNA, protein were detected by RT-PCR, Western blot and immunohistochemistry respectively. The angiotensin Ⅱ (AngⅡ) type 1 receptor (A T1R) was determined as well. Result The expression rate of AT2R in VSMC was increased signific antly after transferred by AdCMV-AT2R with time, and the peak value detected by flow cytometry was about 89.51% at 48 hours. RT-PCR, Western blot and immunohistochemistry showed that the expression of AT2R mRNA and protein were increased obviously in transferred VSMC. There were no significantly change of AT1R expression during AT2R expression. Conclusion Our study indicates that AdCMV-AT2R did generate high level expression of AT2 receptor and its expression did not affect AT1R exp ression in cultured VSMC. The VSMCs transferred AT2R gene may be used as a good model to study the effect of AT2R on their biological action such as proliferation, migration and apoptosis.

Objective To analyze the direct and indirect effects of influencing factors on suicide ideation among college students based on the structural equation model. Methods 1505 college students were investigated with ASLEC, Simplified Coping Style Questionnaire, SSRS, QSA and BDI. Results Incidence rate of college students' suicidal ideations during the past year was 6.67%. The goodness of fit for the structural equation model was satisfactory and 3 major indices( x2/df = 2.23, GFI = 0. 982, RMSEA = 0. 029 ) had met corresponding requirements. The depression was directly influencing factor on suicide ideation, while four factors including passive coping style, social support, positive coping style and suicide attitude, had indirect impacts. Negative life event not only directly affected suicide ideation, but also had indirect effects. According to the percentages of their contribution, the risk factors were ranked as follows:depression (41.08%), negativity life event (35.35%) and passive coping style (6.05%). Similarly, the top protective factor was: social support ( 11.94% ), followed by positive coping style (4.94%) and suicide attitude (0.63%). Conclusion Depression is an important risk factor of suicide,and has a direct impact. So, not only strengthen the mental health of college students, but also train students to face up the difficulties with a positive style, and make the college students get social supports sufficiently.

In producing transgenic livestock, selectable marker genes (SMGs) are usually used to screen transgenic cells from numerous normal cells. That results in SMGs integrating into the genome and transmitting to offspring. In fact, SMGs could dramatically affect gene regulation at integration sites and also make the safety evaluation of transgenic animals complicated. In order to determine the deletion time and methods in the process of producing transgenic goats, the feasibility of deleting SMGs was explored by Cre/LoxP before or after somatic cell cloning. In addition, we compared the efficiency of protein transduction with plasmids co-transduction. We could delete 43.9% SMGs after screening out the transgenic cell clones, but these cells could not be applied to somatic cells cloning because of serious aging after two gene modifications. The SMG-free cells suitable for nuclear transfer were accessible by using the cells of transgenic goats, but this approach was more time consuming. Finally, we found that the Cre plasmid could delete SMGs with an efficiency of 7.81%, but about 30% in SMG-free cells had sequences of Cre plasmid. Compared with Cre plasmid, the integration of new exogenous gene could be avoided by TAT-CRE protein transduction, and the deletion rate of TAT-CRE transduction was between 43.9 and 72.8%. Therefore, TAT-Cre transduction could be an effective method for deleting selectable marker genes.
Animals ; Animals, Genetically Modified ; genetics ; Cloning, Organism ; veterinary ; Gene Knockout Techniques ; Gene Targeting ; methods ; Genes, Reporter ; Genetic Engineering ; Genetic Vectors ; genetics ; Goats ; genetics ; Integrases ; chemistry ; metabolism ; Recombination, Genetic ; Transgenes ; genetics

BACKGROUNDPathology is the "gold standard" of the diagnosis of lung cancer, but at present there are few articles about evaluating the value of cytopathological check. The aim of this study is to evaluate the value of cytopathological check in the diagnosis of primary bronchogenic carcinoma of the lung.

METHODSA total of 552 samples' cytopathological results of 248 patients from January 2003 to May 2005 in this hospital were retrospectively analyzed. The samples included pleural fluids (110 for 68 patients), materials from lung puncture (33 for 31 patients), smears of transcatheter bronchial brushing (152 for 138 patients), bronchoalveolar lavage fluids (30 for 26 patients) and sputa (227 for 118 patients).

RESULTSThe positive results of different specimens were as follow: the pleural fluids was 69.12%, the materials from lung puncture 67.74%, the smears of transcatheter bronchial brushing 65.22%, the bronchoalveolar lavage fluids 23.08% and sputa 21.19% respectively. The positive rates of the pleural fluids, the materials from lung puncture and the smears of transcatheter bronchial brushing were higher than that of the bronchoalveolar lavage fluids and sputa (P < 0.01). There was no significant difference among the positive rates of the pleural fluids, the materials from lung puncture and the smears of transcatheter bronchial brushing, and between the positive rates of the bronchoalveolar lavage fluids and sputa (P > 0.05).

CONCLUSIONSThe cytopathological examination is helpful for the diagnosis of primary bronchogenic carcinoma of the lung, and is one of the important ways to the diagnosis of primary bronchogenic carcinoma of the lung.

In the application of therapeutic antibodies, large molecular weight of antibodies is always a problem that prevents them from penetrating into tissues or binding to antigenic determinants. To overcome this problem, we investigated the function of the heavy chain variable domain of a monoclonal anti-VEGF human IgM antibody derived from the Five-Feature Translocus Mice. We cloned the cDNA of the heavy chain variable domain, which was then inserted into pET28a vector and expressed in Escherichia coli. After purification and renaturation of the denatured recombinant protein, we obtained a 16 kDa antibody fragment, which is named as rhVVH. By immunoassaying its VEGF-binding capability in vitro, we proved that rhVVH retains this activity as the complete IgM. Importantly, rhVVH is shown to inhibit the HUVEC cell proliferation in a concentration-dependent manner. Our results indicate that the single heavy chain variable domain might inherit part of the biological function of the complete IgM antibody, which provided a valuable potential in further research on antibody miniaturisation.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; Mice ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; Single-Chain Antibodies ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo determine the biotic effects of angiotensin II (Ang II) on the migration of rat smooth muscle cells (VSMCs) and investigate the mechanisms involved in the development of vascular injury.

METHODSVSMCs isolated from aortic media of Wistar rats and cultured by the modified explant method were adopted. In the presence and absence of Ang II, the expression of Ang II receptor (ATR) and reorganization of the actin cytoskeleton and focal adhesion of VSMCs were studied by an immunocytochemistry technique and fluorocytochemistry technique. Migration assays were performed with a modified Boyden's chamber. The effects of AT(1)R antagonist (CV-11974), AT(2)R antagonist (PD123319) on the aforementioned target were studied.

RESULTSVSMCs migration was stimulated by adding Ang II. The dynamic reorganization of actin cytoskeleton and focal adhesions may be an important mechanism by which Ang II facilitates VSMCs motility. The expression of AT(1)R in VSMCs could be upregulated initially after treatment with Ang II, then decreased gradually. The expression of AT(1)R was downregulated by AT(1)R antagonists. The effect of Ang II on VSMCs migration was mediated by AT(1)R, while AT(2)R had no significant effect.

CONCLUSIONSThe dynamic reorganization of focal adhesions and the actin cytoskeleton is required for Ang II-induced VSMCs migration. This effect is mediated by AT(1)R.

Actins ; drug effects ; metabolism ; Angiotensin II ; pharmacology ; Angiotensin Receptor Antagonists ; Animals ; Benzimidazoles ; pharmacology ; Cell Movement ; drug effects ; Cells, Cultured ; Cytoskeleton ; drug effects ; Dose-Response Relationship, Drug ; Female ; Focal Adhesions ; drug effects ; metabolism ; Imidazoles ; pharmacology ; Immunohistochemistry ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Pyridines ; pharmacology ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; Receptors, Angiotensin ; metabolism ; Tetrazoles ; pharmacology