Objective To evaluate the effects of sevoflurane postconditioning on mitophagy during myocardial ischemia-reperfusion (I/R) in rats.Methods Forty-two pathogen-free adult male SpragueDawley rats, weighing 250-300 g, were randomly divided into 3 groups (n=14 each) using a random number table: sham operation group (group S) , I/R group and sevoflurane postconditioning group (group SP).Myocardial I/R was induced by 30 min ligation of the left anterior descending branch of the coronary artery followed by 2 h of reperfusion.In group SP, 2.4％ sevoflurane was inhaled for 15 min starting from the onset of reperfusion, while 33％ oxygen was inhaled in group I/R.The rats were sacrificed at the end of reperfusion, and the hearts were removed for measurement of myocardial infarct size (by 1％ 2, 3, 5 triphenyltetrazolium chloride) , expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1, p62 and Parkin (by Western blot) ,and mitochondrial menbrane potential (by using JC-1 probe) , and for examination of the uhrastructure of cardiomyocytes (with transmission electron microscope).Results Compared with group S, the myocardial infarct size was significantly increased, mitochondrial membrane potential was decreased, the expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1 and Parkin was up-regulated, and the expression of p62 was down-regulated in group I/R.Compared with group I/R, the myocardial infarct size was significantly decreased, the mitochondrial membrane potential was increased, and the expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1, p62 and Parkin was down-regulated in group SP.Conclusion Sevoflurane postconditioning can mitigate I/R injury in rats, and inhibition of excessive activation of mitophagy may be involved in the nechanism.

BACKGROUND:Drug therapy can partly reduce and delay the progress of Alzheimer’s disease, but only 30%with the single drug treatment obtain clinical cure. OBJECTIVE:To study the therapeutic effect of bone marrow mesenchymal stem cel s transplantation for rats with Alzheimer’s disease. METHODS:Amyloidβ-protein was injected into the hippocampus of Sprague-Dawley rats to construct the model of Alzheimer’s disease. And bone marrow stromal stem cel s were transplanted into the hippocampus of the rat models. RESULTS AND CONCLUSION:At 2 weeks after modeling, compared with the control group, the escape latency in the model and experimental groups was significantly longer (P<0.05), which indicating that Alzheimer’s disease models were successful y established. At 4 weeks after cel transplantation, compared with the model group, the average escape latency in the experimental group was significantly decreased, but retention time on the platform quadrant was significantly prolonged (P<0.05). Besides, at 4 weeks after cel transplantation, expression of choline acetyltransferase in the experimental group was significantly higher than that in the model group (P<0.05). In conclusion, bone marrow mesenchymal stem cel s cannot only differentiate and survive in the hippocampus of rats with Alzheimer’s disease, but also improve the learning and memory ability.

Objective To investigate whether and what staining techniques are applied to the ultrathin sheet plastination slice and whether the stained specimen is of autofluorescences .Methods A cadaveric hand block was plastinated and then sectioned as a series of 300-400μm thick transverse sections .A total of 56 slices in total .Alternative sections were stained with hematoxylin -eosin staining ( HE) , Verhoeff -Van Gieson staining ( VVG) or methylene blue and azureⅡstaining(MA).The stained slices were examined under a light microscope and a confocal microscope .Results The plastinated slices were stained with the three staining methods .HE staining revealed the muscle and connective tissues were red or violet , bone was violet or blue;VVG staining showed the elastic fibers was black , the collagen was red , and other tissues were yellow .MA staining showed the tendon was violet , the bone was pink , cartilage was violet , and other tissues were purple.However, the intracellular structures appeared not very well stained .The collagen, elastin and muscular structures in the stained slices were observed under a confocal microscope .Conclusion The commonly used histology staining methods can be used to stain the ultrathin sheet plastination slices .The staining provides a better observation of various tissues in the slice than the unstained slice .After staining, those autofluorescent structures in the plastinated slice are detectable under a confocal microscope .

Adult ; Enzyme-Linked Immunosorbent Assay ; Female ; HSP70 Heat-Shock Proteins ; blood ; Humans ; Lead ; toxicity ; Male ; Occupational Exposure

Objective To evaluate the forecasting capability of serum N-terminal pro-brain natriuretic peptide ( NT-proBNP) level for Left ventricular ( LV) remodeling of patients with acute myocardial infarction (AMI) through observation of the relationship between serum NT-proBNP on the 3rd day and LV remodeling following onset of AMI. Methods Electrochemilumin-escence was adopted to determine the serum NT-proBNP level on the 3rd day after AMI attack for 106 cases of patients with anterior, anteroseptal and anterolateral AMI, with echocardiography performed to detect LV end-diastolic diameter (LVDd) and leftventricular ejection fraction (LVEF) respectively on the 3rd day and 3 months after the attack. Results The serum NT-proBNP level of AMI patients on the 3rd day averaged 1039. 28 (241. 50-1 184. 25) ng/L For AMI patients on the 3rd day and 3 months after, LVDd rose to (53?7) mm (P 0.05) from (53?8) %. NT-proBNP concentration on the 3rd day after AMI onset was positively correlated with△LVDd significantly, r =0. 403 (P 5mm during the 3 months and LVEF at the 3rd month≤40 % , AUC was 0. 893. Conclusion The test results showed serum NT-proBNP level for AMI patients on the 3rd day may be used as one of forecasting indexes of LV remodeling for AMI of advanced stage.

Objective To evaluate the role of nitric oxide in sevoflurane postconditioning-induced mitigation of autophagy and cell apoptosis during ischemia/reperfusion (I/R) in isolated rat hearts.Methods The hearts of male Sprague-Dawley rats,aged 2-3 months,weighing 250-300 g,were excised and retrogradely perfused in a Langendorff apparatus.One hundred and eight isolated rat hearts,which were successfully perfused in a Langendorff apparatus,were equally and randomly divided into 6 groups:control group (C group),sevoflurane group (S group),I/R group,sevoflurane postconditioning group (SSP group),sevoflurane postconditioning + L-NAME (non-selective nitric oxide synthase (NOS) inhibitor group (SSP + L group),and L-NAME group (L group).The hearts were perfused with K-H solution for 150 min in C group.The hearts were continuously perfused for 180 min and perfused with K-H solution containing 3％ sevoflurane for 15 min starting from 60 min of perfusion in S group.After being perfused with K-H solution for 30 min,the hearts were subjected to occlusion for 30 min followed by reperfusion for 120 min in the other groups except C and S groups.After onset of reperfusion,the hearts were perfused with K-H solution containing 3％ sevoflurane for 15 min in SSP group,the hearts were perfused with K-H solution containing 3％ sevoflurane and L-NAME 100 μmol/L for 15 and 60 min,respectively,in SSP + L group,and the hearts were perfused with K-H solution containing L-NAME 100μmol/L for 60 min in L group.Inn ediately before ischemia,and at 30,60,90 and 120 min of reperfusion,each parameter of cardiac function was recorded.At the end of reperfusion,myocardial specimens were obtained at the end of reperfusion for measurement of the infarct size,NOS activity,NO content,and expression of Bcl-2,Beclin 1 and caspase-3,for observation of formation of autophagosomes,and for examination of the pathological changes.Results Compared with C group,LVSP,+ dp/dtmax,-dp/dtmax,NOS activity and NO content were significantly decreased,and LVEDP was increased in I/R and SSP groups.Compared with I/R group,LVSP,+ dp/dtmax,-dp/dtmax,NOS activity and NO content were significantly increased,LVEDP was decreased,Bcl-2 expression was down-regulated,and the expression of Beclin 1 and caspase-3 was up-regulated in SSP group,and no significant changes were found in each index in SSP+ L and L groups.Compared with SSP group,LVSP,+ dp/dtmax,-dp/dtmax,NOS activity and NO content were significantly decreased,LVEDP was increased,Bcl-2 expression was down-regulated,and the expression of Beclin 1 and caspase-3 was up-regulated in SSP + L group.Conclusion The mechanism by which sevoflurane postconditioning reduces I/R injury may be related to promoted NO product and inhibited autophagy and cell apoptosis in isolated rat hearts.

Objective To evaluate the effects of sevoflurane postconditioning on the expression of 70 kD aribosomalprotein S6 kinase (p70S6K) during ischemia-reperfusion (I/R) in isolated rat hearts.Methods Pathogen-free male Sprague-Dawley rats,aged 3 months,weighing 270-350 g,were used in the study.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95 ％ O2-5 ％ CO2.Ninety isolated rat hearts with I/R injury were randomly divided into 3 groups (n =30 each):sham operation group (group S),I/R group (group I/R),and sevoflurane postconditioning group (group SP).The hearts were subjected to ischemia for 30 min followed by 2 h reperfusion.In group SP,the hearts were perfused with K-H solution saturated with 3.0％ sevoflurane for 15 min starting from the end of ischemia until 15 min of reperfusion,and then with plain K-H solution for 105 min.At 2 h of reperfusion,myocardial infarct size was measured,the percentage of myocardial infarct size was calculated,and the phosphorylated p70S6K (p-p70S6K)/total p70S6K (tp70S6K) ratio,and cytoplasm,cytochrome C,and caspase-8 expression was measured.Results Compared with group S,the percentage of myocardial infarct size and p-p70S6K/t-p70S6K ratio were significantly increased,the expression of cytochrome C,and caspase-8 was up-regulated,and the expression of cytochrome C was downregulated in I/R group.Compared with I/R group,the percentage of myocardial infarct size was significantly decreased,the ratio of p-p70S6K/t-p70S6K was increased and cytochrome C expression was up-regulated,and the expression of cytochrome C and caspase-8 was down-regulated in SP group.Conclusion Sevoflurane postconditioning can mitigate I/R injury to isolated rat hearts,and up-regulation of p-p70S6K expression,inhibition of transfer of cytochrome C from mitochondria to cytoplasm,and reduced cell apoptosis are involved in the mechanism.

Objective To establish a new rat model of biliary atresia by pure ethanol injection into the common bile duct.Methods A catheter was inserted and fixed in the common bile duct in male SD rats .Saline (8 rats) or pure ethanol (16 rats) was injected through the catheter ,respectively, and the biochemical and pathological changes in the rats were examined .Results SD rats in the experimental group were divided into a persistent injury and a restoration of liver dysfunction groups according to pathological and biochemical detection .In the persistent injury group , biochemical impair-ments were significantly higher at 8 weeks after ethanol injection than those in the control group and restoration group .Dis-tinct pathological changes in the liver were observed using HE , SMA, and Masson staining .Conclusions It is a reliable animal model of biliary atresia induced by injection of pure ethanol into the common bile duct in the rat .It will provide a useful tool in future studies of biliary atresia .

Objective To discuss the experience in the diagnosis, treatment, complications and follow up of carotid body tumor. Methods All the 25 cases were diagnosised by DSA and CTA. The tumor was resected under carotid adventitial plane in 18 cases, with external carotid artery resection in 4cases, and in 3 cases, internal carotid artery (ICA) and external carotid artery (ECA) were resected simultaneously in which internal carotid artery was reconstructed in two cases including using self vein bypass in one and anastomosis between ICA and ECA in the other. ICA was ligated in the third case. Results No cases died perioperatively. ALL CBTs were treated successfully. Horner syndrome and trachyphonia were relieved after operation. Postoperative trachyphonia, bucking and lingual paralysis developed in 3 cases, and in one case with vagus resection caused dyspnea tracheotomy was performed. The rate of nerves injuries was 12% but no semiplegia and aphasia occurred. Follow up period was from 4 to 90 months (average 44 ±6 months) for 21 cases. The trachyphonia and bucking were improved during follow up but the lingual paralysis persists, and tumor recurred in two cases with one dying. Conclusions CBT treatment should include active surgery, sufficient preoperative preparation and avoiding the postoperative nervous complications.

BACKGROUND: At present, the commercial artificial small vascular grafts (diameter < 6 mm) are still unsatisfactory, due to poor blocompatibility and low long-term patency rate. Therefore, finding a vascular substitute with normal biological function and studying construction and function of tissue-engineered blood vessel have become hot topics recently.OBJECTIVE: To construct a novel tissue-engineered blood vessel by rabbit bone marrow mesenchymal stem cells (MSCs) and vascular acellular matrix, and to investigate the biocompatibility and patency rate of tissue-engineered blood vessels.DESIGN, TIME AND SETTING: An in vitro randomized controlled study at level of cytology and histopathology was performed at the Laboratory of Affiliated Drum Tower Hospital of Nanjing University Medical School from January 2006 to June 2008.MATERIALS: The decellularized vascular acellular matrix was obtained by a detergent-enzymatic procedure. MSCs from rabbits were isolated using density gradient centrifugation method and cultured in culture flasks coated with fibronectin. Subsequently, the expanded MSCs were seeded on the decellularized scaffolds, and then co-cultured in the self-made bioreactor to construct the tissue-engineered blood vessels.METHODS: Sixty rabbits were randomly divided into three groups. A1.0-cm abdominal aorta was sheared, and a tissue-engineered blood vessel was transplanted on the abdominal aorta using 8/0 polypropylene thread. Tissue-engineered blood vessel group: Tissue-engineered blood vessel was considered as the transplanted vessel; vascular acellular matrix group:Xenoma artery treated by vascular acellular matrix was considered as the transplanted vessel; xenoma artery group: Fresh xenoma artery was considered as the transplanted vessel.MAIN OUTCOME MEASURES: Immunocytochemical staining was used to identify the cultured MSCs. After 3 months of transplantation, the grafts were retrieved for digital subtraction angiography, pathological test and scanning electron microscope examination.RESULTS: Rabbits MSCs presented a whirlpool-like appearance at 8 days after culture. The immunocytochemistry results were consistent with the phenotype of MSCs. After high proliferation, MSCs were seeded onto the vascular acellular matrix for 12 days,and seed cells attached to well in the lumen of blood vessels. Three months after implantation, the patency rate was 90% of tissue-engineered blood vessel group and 80% of vascular acellular matrix group, which was superior to xenoma artery group (25%). At three months after transplantation, HE staining and scanning electron microscope demonstrated that internal, middle,and external membrane were clearly observed in the tissue-engineered blood vessel group, and the membrane morphology was similar to normal artery. The endothelial cells were covered completely. However, the endothelial cells were not covered completely in the vascular acellular matrix group, while mural thrombosis, mild proliferation of intima, and inflammatory cell infiltration were observed. The intima was thick and necrotic in the xenoma artery group, while lumens were stenotic and accompanied with a certain degree of thrombus organization.CONCLUSION: This study provides a new strategy to develop a tissue-engineered blood vessel with excellent biocompatibility and high patency rate constructed by rabbit MSCs and vascular acellular matrix.