BACKGROUND: Previous studies have demonstrated that the responses of myocardium to short periods of ischemia/reperfusion are associated with changes in the expressions of a variety of genes. However, the characteristics of these changes are unknown.OBJECTIVE: To clone and analyze a stress-inducible gene characteristics in order to study the molecular mechanism of myocardium to short periods of ischemia/reperfusion damage.DESIGN: A controlled animal study.SETTING: Department of Hematology, Xiangya Hospital of Central South University.MATERIALS: Sixteen castrated male German Landrace-type domestic porcines, weighing between 21 and 39 kg, were provided by the laboratory of Department of Experimental of Cardiology at Bad Nauheim in Germany. The porcines were randomly divided into two groups ischemia/reperfusion group (n =14) and control group (n =2). The porcines in the ischemia/reperfusion group were observed at 0, 30 and 90 minutes of reperfusion after reocclusion, and ischemia and non-ischemia myocardial tissues were harvested from each pocrine.METHODS: The experiments were performed in the laboratory of Department of Experimental Cardiology of Max-Planck Institute for Physiological and Clinical Research at Bad Nauheim in Germany and the Department of Hematology, Xiangya Hospital of Central South University from July in 1998 to May in 2007. All the animals were anesthetized and thoracotomized. Following 30 minutes of stabilization, the left anterior descending coronary artery (LAD) was occluded for 10 minutes followed by 30-minute reperfusion and then another 10-minute reocclusion. The porcines were killed immediately at corresponding time points. The sham-operated animals were killed without occlusion. Experimental tissue was removed from the LAD area and control tissue from the region of the left circumflex coronary artery area. Firstly, a porcine heart cDNA library was screened, DNA and deduced amino acid sequences were then analyzed. Meanwhile, short periods of myocardium ischemia/reperfusion was performed by occluding porcine LAD followed by reperfusion as mentioned above, and total RNAs isolated from myocardium or a variety of other tissues were used for Northern blotting. Quantitation of mRNA levels was accomplished by using a PhosphorImager and Image Quant software, the ration of mRNA/18S rRNA was suggested as the level of gene expression.MAIN OUTCOME MEASURES: ① DNA and amino acid sequences analysis of the cloned gene; ② Analysis of the cloned gene expression.RESULTS: All the 16 porcines were involved in the final analysis of results. After screening, a cDNA fragment with 3 461 base pairs was obtained. DNA sequencing and searching revealed that this cDNA shared 86% identity to human structural maintenance of chromosome 6 gene (hSMC6), and 84% identity to Mus musculus SMC6 (mSMC6). Furthermore, a largest polypeptide deduced from this cDNA contained 1007 amino acid residues, and protein homology searching indicated that the predicted polypeptide carried 92% and 89% identity to hSMC6 or mSMC6 protein respectively. Thus, the cloned cDNA was referred to as a porcine homology gene to hSMC6 (pSMC6). The results of Northern blotting showed that the expression of the pSMC6 mRNA in ischemia/reperfusion myocardium was increased, and its mRNA existed in all organs tested. CONCLUSION: A stress-inducible gene of pSMC6 from porcine myocardium has been cloned, which is high homology to hSMC6 gene, and the pSMC6 or along with other molecules is possible involved in the early repair of myocardium to DNA damage caused by ischemia/reperfusion in the porcine heart.

Objective To observe the efifcacy and safety of XELOX plus bevacizumab as a ifrst-ine treatment for Chinese patients with metastatic colorectal cancer. Methods In this study, we retrospectively reviewed cases in which XELOX plus bevacizumab were administered in order to evaluate its efifcacy and safety in clinical practice.In total, 40 patients with mCRC who presented at Fuchu Hospital received XELOX plus bevacizumab as a ifrst?line treatment from September, 2009 to April, 2012. Eligible patients had histologically conifrmed mCRC. XELOX consisted of a 2-h intravenous infusion of oxaliplatin 130 mg/m 2 on day 1 plus oral capecitabine 1,000 mg/m 2 twice daily for two weeks of athree?week cycle. Overall survival (OS) and survival benefit were analyzed when patients continued with XELOX plus bevacizumab beyond disease progression. Results The median progression?free survival (PFS) was 290 days [95%conif-dence interval (CI):222?409 days] and the median OS was 816 days (95%CI:490?842 days). The response rate (RR:complete plus partial response) was 67.5%, and the disease control rate (RR plus stable disease) was 90%. Conclusion XELOX plus bevacizumab may be considered a routine ifrst?line treatment option for patients with mCRC.

Objective To detect the expression of integrin ?5?1 in restenosed human vein grafts.Methods The expression of integrin ?5?1 and ?-smooth muscle actin(?-SMA) in 30 resected restenosed human vein grafts(from Kerfuff clinical hospital of Germany) was detected by confocal immunofluorescence with specific antibodies against ?5?1 and ?-SMA.Images were processed with Silicon Graphics Octane.Results In normal veins,integrin ?5?1 expression was very weak in media smooth muscle cells and in endothelium,and ?-SMA expression was present in media smooth muscle cells.In the restenosed vein grafts,integrin ?5?1 was strongly stained in the media smooth muscle cells and intimal endothelial cells,and moderately in the intimal smooth muscle cells,?-SMA was present in media smooth muscle cells and in the intimal smooth muscle cells.Conclusion Our research reveals that integrin ?5?1 is significantly upregulated in the media smooth muscle cells and intimal endothelial cells in the restenosed human vein grafts,suggesting the participation of integrin ?5?1 in the restenosis formation of human vein grafts.

By means of cell hybridization in situ with digoxigenin labeled probe, we detected the expression of C-myc protooncogene in peripheral blood mononuclear cells from 31 SLE patients and 10 healthy persons. The results showed that the level of C-myc mRNA in SLE patients was significantly higher than that in normal controls, and the increased level correlated with disease activity. There was also a positive correlation between C-myc mRNA and serum levels of ANA, anti-dsDNA, IgG, C3. Our study indicates that C-myc gene is important for the pathogenesis of SLE.

The distribution of TrkA and the postnatal development(PD) of TrkA and ChAT-immunoreactive(-IR) neurons andthe relation between them in the basal nucleus of Meynert of rats were studied with immunohistochemical method. The number,mean profile areas and grey degree of TrkA-IR and ChAT-IR neurons were examined with image analyser. The data revealed thatTrkA-IR neurons were localized in the basal forebrain of rats. TrkA immunostaining was present at PDI, but ChAT was not.ChAT immunostaining was present at PD5. Most densely stained TrkA and ChAT neuronal bodies and fibers were present atPD20, the mean grey degrees of TrkA-IR and ChAT-IR neuronal profiles reached its peak. Both TrkA and ChAT neurons beganto cline at PD30 and maintained a relatively higher level in the adult. However, during aging both TrkA and ChAT-IR neuronsatrophy and became smaller than that in the adult. The number of TrkA-IR and ChAT-IR neurons were decreased by 41.38% and 51.61%; the mean profile areas decreased by 15.7% and 12.8%; and the mean grey degrees by 29.9% and 9.9%, respec-tively. The mean profile areas of TrkA-IR and ChAT-IR neurons from PD5 to aged rats were positively correlated. The resultsindicated that the expression of TrkA was earlier than ChAT. The expression of TrkA and ChAT followed a very similar tempo-ral pattern in the basal nucleus of Meynert from PD5 to aged rats, suggesting that TrkA might participate the regulation ofChAT-IR neuronal development, differentiation, maturation, and ageing. The down-regulation of TrkA and ChAT of aged ratsis associated with neuronal atrophy and loss and may contribute to the pronounced vulnerability of these neurons to degenerationin aging animals and Alzheimer's disease.

Background and purpose:Stromal tumor is one of the common gastrointestinal mesenchymal tumors. There is certain understanding about the typical cases. However, the diagnosis of those occurring in rare location or with rare imaging findings is often difficult. This research summarized this kind of cases,in order to increase the radiological knowledge of the disease.Methods:This study retrospectively analyzed clinical, radiological and pathological data from 550 patients who had stromal tumor conifrmed by pathology in our hospital. Those with incomplete data were eliminated. Forty-nine patients were selected for further study according to the typical imaging findings.Results:Among these 49 patients, 9 were pathologically confirmed to have extra-gastrointestinal stromal tumor, while 40 patients had gastrointestinal stromal tumor. Among the patients with gastrointestinal stromal tumor, 22 were found in rare locations, 12 in retroperitoneal space, 3 in omentum majus and mesenterium, 5 in esophagus, and 2 in prostate. Obvious cystic degeneration was found in 16 patients. Bulky calciifcation, such as lfake or annulus, was found in 7 patients. The analysis result of risk-stratiifcation showed 19 patients were conifrmed as high-grade among the patients with tumors found in rare locations, 15 as high grade among those with obvious cystic degeneration, and 7 as high-grade among those with extra-gastrointestinal stromal tumor.Conclusion:Rare stromal tumor often occurs in the locations, such as retroperitoneal space, omentum majus and mesenterium. Obvious cystic degeneration and bulky calciifcation can be seen. The risk-stratiifcation of these patients often showed high-grade. Comprehensively analyzing its clinical features and imaging ifndings can help improve the diagnostic accuracy.

Objective To investigate the postnatal developmental rule of TrkA and ChAT\|immunoreactive(ChAT\|ir) neurons and the relationship between TrkA and ChAT\|ir neurons in the horizontal limb of diagonal band(HDB) of rats. Methods Immunohistochemistry technique combined with image analyser were used. Results TrkA and ChAT\|ir neurons localized in the neurons of basal forebrain of rats. TrkA immunostaining was present at postnatal day 1(PD1), but ChAT immunostaining was present at PD5 Most densely stained TrkA and ChAT neuronal bodies and fibers were present at PD20, while the mean grey degrees of TrkA and ChAT\|ir neurons reached to the peak. Both TrkA and ChAT began to decline at PD30 and maintain a relatively higher level in the adult. However, during aging both TrkA and ChAT\|ir neurons atrophied and became smaller than that of adult. The number of TrkA and ChAT\|ir neurons decreased 39 8% and 33 3%;the mean areas 15 7% and 12 8%; the mean grey degree values were 29 9% and 9 9%, respectively. The mean areas, grey degrees and numbers of TrkA and ChAT\|ir neurons from PD5 to aged rats had positive correlation. Conclusion The results indicate that the expression of TrkA was earlier than ChAT. The expression of TrkA and ChAT followed a very similar temporal pattern in HDB from PD5 to aged rats, suggesting that TrkA may participate in the regulation of ChAT\|ir neuronal development, differentiation, maturation and aging. The down\|regulation of TrkA and ChAT of aged rats is associated with neuronal atrophy and loss and may contribute to the pronounced vulnerability of these neurons to degeneration in aging animals and Alzheimer disease.\;

BACKGROUND:Previous research has proved good therapeutic effect on the acute hepatic failure by the treatment of polylactic acid-O-carboxymethylated chitosan(PLA-O-CMC) nanoparticles-cultured porcine hepatocytes embedded in type I collagen.OBJECTIVE:To investigate the effect of PLA-O-CMC nanoparticles on immunological rejection in intraperitoneal porcine hepatocyte xenotransplantation for treatment of acute liver failure rats.DESIGN,TIME AND SETTING:Randomized control animal experiments were carried out at the Institute of Hepatobiliary Surgery and Laboratory of Nerve Regeneration in Nantong University from May 2005 to May 2006.MATERIALS:In situ recirculating collagenase perfusion method was used to isolate porcine hepatocytes.64 SD rats were intraperitoneally injected with D-galactosamine to induce acute hepatic failure models.METHODS:Totally 64 rats were divided into four groups:Model group received no intervention;Nano-collagen hepatocytes group was transplanted with PLA-O-CMC nanoparticles-cultured porcine hepatocytes embedded in collagen gel,which was then wrapped up using greater omentum;Porcine hepatocytes embedded in collagen gel(collagen hepatocytes group) and free porcine hepatocytes suspension(pure hepatocytes group) were transplanted into peritoneal cavity of SD rats,respectively.At 48 hours following the modeling,the porcine hepatocytes transplantations were performed and the number of transplanted cells was 5.0?107 hepatocytes in all groups.MAIN OUTCOME MEASURES:The serum interleukin-2,interferon-?,IgG and IgM levels of rats were determined at days 1,2,3,5 and 7.The pathological changes of transplants were observed under microscope at days 1,3 and 7.RESULTS:There were no significant differences in serum interleukin-2 and interferon-? levels of all groups during 7 days post-transplantation.Serum IgG levels increased with a peak at day 3 after hepatocytes transplantation,and then began to decline.Serum IgG levels in nano-collagen hepatocytes group were lower than those in collagen hepatocytes group and pure hepatocytes group at days 2 and 3(P

Objective To investigate the changes of expression of apoptosis-related proteins Bcl-2,Bax and Fas of the transplantated hepatocytes after intraperitoneal transplantation in rats using porcine hepatocytes cultured with polylactic acid-O-carboxymethylated chitosan nanoparticles.Methods Recirculating collagenase perfusion method was used to isolate porcine hepatocytes.At 48h after D-galactosamine intraperitoneal injection,5 mL polylactic acid-O-carboxymethylated chitosan nanoparticles-cultured porcine hepatocytes embedded in collagen gel(group A) and 5 mL porcine hepatocytes embedded in collagen gel(group B) were transplanted into peritoneal cavity of SD rats,respectively.The amount of transplanted porcine hepatocytes was 5.0?107 hepatocytes in two groups.The expression of Bcl-2,Bax and Fas in transplanted hepatocytes three days after transplantation was detected with immunohistochemistry.Results At 1,2 and 3d after transplantation,the rates of Bcl-2 positive cells in group A was significant higher than those in group B,whereas the rates of Bax and Fas positive cells in group A were significant lower than those in group B(P

Objective To investigate the nutrition statuses of thiamine and riboflavin and their relationship with the antioxidant capacity and blood glucose and lipids levels in the middle-aged and elderly. Methods Totally 314 subjects aged above 40 years old from communities in Shanghai were enrolled. The 3-day survey of 24-hour's diet recall was made to evaluate the daily nutrient intakes using a nutrition assessment software provided by CDC China. Thiamine and riboflavin levels were detected by urine load test. Fasting blood glucose ( FBG), total choles-terol (TC), triglyceride (TG), superoxide dismutase (SOD) activity and malondialdehyde (MDA) were deter-mined by commercial kits. Results The average daily intakes of thiamine and riboflavin were (0. 82 + 0. 36) and (0. 91 +0.48) rag, respectively. The middle-aged and elderly whose intakes of thiamine and riboflavin reached recommended nutrient intake (RNI) level accounted for 11.8% and 17.2%, while those lower than 60% RNI ac-counted for 51.3% and 49. 4%, respectively. Urine load test showed the deficiency rates of thiamine and ribofla-vin were 65.0% and 58.6%, respectively. Compared with the normal group, plasma MDA, FBG, TC, and TG levels progressively increased and SOD activity decreased in insufficient and deficient groups; the plasma MDA, FBG, and TC levels were significantly different between normal group and thiamine deficient group ( P < 0. 05 ).The FBG and TC levels were negatively correlated with the status of thiamine ( r = - 0. 246, r = - 0. 154, P <0. 05). The MDA and TG levels were negatively correlated with the status of riboflavin ( r = - 0. 136, r =-0. 297, P <0. 05). Conclusions The nutrition statuses of thiamine and riboflavin are poor in the middle-aged and elderly in Shanghai. The nutritional statuses of thiamine and riboflavin are positively correlated with the antioxi- dant capacities and are negatively correlated with the level of FBG, TC, and TG.