BACKGROUND:Existing studies have shown that bone marrow mesenchymal stem cel s can significantly improve islet function in diabetic rats to decrease excessively high blood glucose level, which may be related to the enhancement of differentiation ability of autologou pancreatic stem cel s.
OBJECTIVE:To observe the therapeutic efficacy of basic fibroblast growth factor gene eukaryotic expression vector (PEGFP-C3-BFGF) transfection of bone marrow mesenchymal stem cel s in diabetic rats.
METHODS:Recombinant adenovirus (Ad.aFGF) mediated PEGFP-C3-BFGF was transfected into bone marrow mesenchymal stem cel s, and PEGFP-C3-BFGF expression was observed using fluorescence microscopy. Eighty Sprague-Dawley rats were randomly divided into normal control group, diabetes group, transplantation group, gene transfection group, with 20 rats in each group. After modeling, rats in different groups were given portal vein injection of normal saline, PBS, 1 mL of bone marrow mesenchymal stem cel suspension, and 1 mL of PEGFP-C3-BFGF-transfected bone marrow mesenchymal stem cel suspension. RT-PCR method was used to detect mRNA expression of matrix metal oproteinases in pancreatic tissue of rats in each group. Blood glucose levels of rats were detected at 24 hours, 3, 7, 14, 21 days after transplantation. ELISA method was used to detect plasma insulin levels in rats. Pathological changes of the pancreas were observed using hematoxylin-eosin staining.
RESULTS AND CONCLUSION:Under the fluorescence microscope, PEGFP-C3-BFGF transfected into cel s after 48 hours showed significant specific red fluorescence. Two weeks after transplantation, matrix metal oproteinases mRNA expression was significantly increased in the diabetes group compared with the control group (P<0.05), while it was decreased in the transplantation and gene transfection groups compared with the diabetes group (P<0.05). After transplantation, the blood glucose levels in rats were ranked as fol ows:control groupgene transfection group>transplantation group>diabetes group (P<0.05). Pathological findings of the pancreas showed that the transplantation group was superior to the diabetes group, but inferior to the gene transfection group that was similar to the control group. Al these findings indicate that PEGFP-C3-BFGF-transfected bone marrow mesenchymal stem cel transplantation can improve blood glucose levels and stimulate insulin secretion in diabetic rats, which may improve the severity of diabetes mel itus by decreasing the mRNA expression of matrix metal oproteinases.

Objective To investigate pathogens distribution and their drug resistance of hospital acquired pneumonia (HAP) in local elderly paraplegic patients,and to help to gain experience in early using antibiotics.Methods One hundred and thirty six elderly patients diagnosed as HAP from January 2007 to December 2010 in our hospital were selected.Pathogens distribution and their drug resistance were detected.Results One hundred and fifty two pathogens are isolated from the 136 patients,and most of them are gram negative bacteria which accounts for 70.4％.The first three pathogens are Klebsiella pneumonia(24.3％),Escherichia coli (20.4％) and Pseudomonas Aeruginosa (18.4％).Gram-positive cocci accounts for 25.0％ in total pathogens,among them,staphylococcus aureus and streptococcus pneumoniae account for the most,and the number of Fungi is the fewest.Drug resistance rate of gram-negative bacteria is higher than that of Gram-positive bacteria.Gram-negative bacteria has higher resistance to ampicillin,cefoperazone,ciprofloxacin,levofloxacin and eotrimoxazole.Gram-positive bacteria has higher resistance to penicillin,cefazolin and gentamicin.Conclusion To elderly paraplegic HAP patients,the main pathogenic flora is gram-negative bacterium which shows multiple resistances.Being familiar with the features of pathogens and their drug resistance will provide better guidance on early treatment and improve prognosis of elderly paraplegic HAP patients.

Objective To evaluate the myocardial protective effects of pinacidil combined with pyrrolidine dithiocarbamate (PDTC) against ischemia-reperfusion (I/R) injury to the isolated rabbit hearts and investigate its mechanisms. Methods One-hundred and twelve Japanese long-ear white rabbits of both sexes weighing 1.8-8.2 kg were killed by a knock to the head after heparinization. Their hearts were immediately removed and mounted on Langendorff apparatus and perfused with oxygenated K-H solution at 371 . Of the 112 isolated hearts 96 were randomized into 6 groups with 16 hearts in each group of which 8 hearts underwent 60 min reperfusion and another 8 hearts 120min reperfusion after 40min global myocardial ischemia: the hearts were perfused with K-H solution in group Ⅰ(K); with St Thomas Ⅱ solution in group Ⅱ(S); with pinacidil in group Ⅲ(P); with PDTC + K-H solution in group Ⅳ(PK); with PDTC + St Thomas Ⅱ solution in group Ⅴ(PS) and with PDTC + pinacidil in group Ⅵ(PP) . The rest of the 112 hearts (16 hearts) were perfused with K-H solution for 10 min. Then myocardial tissue was obtained for immuno-histochemical examination (SABC staining) used as normal control value.(1) Time of resumption of heart beat (from the beginning of reperfusion to the resumption of heart beat) was recorded; (2) left ventricular systolic and end-diastolic pressure (LVSP, LVEDP) and + dp/dtmax were monitored; (3) effluent from coronary sinus was collected at 60 min of reperfusion for determination of TNF-? concentration and (4) myocardial tissue was obtained at the end of reperfusion for determination of expression of NF-?B p65 and ICAM-1 in myocardium. Results (1) The heart beat resumption time was significantly shorter in group PP, PS and P than in the other 3 groups (P

Objective To evaluate the role of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) pathway in cardio-protection by ischemic or pinacidil postconditioning ( IP,PP) against ischemia-reperfusion (I/R) injury in isolated rat hearts.Methods Fifty-six male SD rats of both sexes weighing 200-250 g were anesthetized with intraperitoneal amobarbital sodium.The isolated rat hearts were perfused in a Langendorff apparatus with Krebs-Hensleit buffer (K-H).Fifty-six isolated rat hearts with I/R injury were randomly divided into 7 groups ( n =8 each):normal control group (group C) ; group I/R; group IP and group PP1-4 postconditioning with 4 different concentrations of pinacidil.After 20 min of equilibration,the perfusion was suspended for 40 min (global ischemia) followed by 60 min of reperfusion in group I/R.In group IP after 40 min of global ischemia,the isolated hearts underwent 6 cycles of 10 s reperfusion and 10 s ischemia followed by 58 min of reperfusion.In group PP1-4 at the end of 40 min of global ischemia,the isolated hearts were perfused with K-H containing pinacidil 5,10,30 and 50μmol/L for 5 min respectively followed by 55 min reperfusion with regular K-H.Left ventricular developed pressure (LVDP) and LVEDP were measured immediately before global ischemia and at the end of 60 min reperfusion.Myocardial specimens were obtained at the end of reperfusion for detection of Nrf2,quinopeoxidoreductase (NQO1),HO-1 and SOD1 mRNA (by RT-PCR) and protein (by Western blot) expression.Results I/R significantly up-regulated Nrf2,NQO1,HO-1 and SODI mRNA and protein expression,decreased LVDP and increased LVEDP in group I/R as compared with group C.IP and 30,50 μmol/L pinacidil postconditioning further significantly increased Nrf2,NQO1,HO-1 and SOD1 mRNA and protein expression and IP,5,10,30,50 μmol/L pinacidil postconditioning significantly increased LVDP and decreased LVEDP as compared with group I/R.Conclusion Ischemic or pinacidil postconditioning can attenuate I/R injury by activating Nrf2-ARE pathway in isolated rat hearts.

With the extensive application of informationalized management systems for barcode specimens , the degree of informatization is becoming higher and higher in medical laboratory.Informationalized management combines modern information technology and advanced management concepts , transforms or reengineers the laboratory operation and business process.Test specimen turnaround time ( TAT) is an important factor affecting the quality of the inspection.By analyzing the test process of each time node , establish the suitable specimens monitoring program for clinical requirements and real -time monitor the key nodes in test processes ,which will effectively shorten TAT , improve reporting timeliness rate and avoid clinical complaints.

Objective To investigate the clinical significance and relationship of preS1, HBV DNA and HBV-M. Methods PreS1 and HBV-M was detected by ELISA method,and HBV DNA was detected by PCR. Then the results were analyzed. Results In HBV patients,the positive rates of preS1 and HBV DNA had no statistically significant ,they had fine dependability. The detection rate of preS1 in HBeAg(+) group(80.3%) and HBeAg(+)group( 56.3% ) had statistically significant. In some patients,though HBeAg had become negative, HBV still replicated. In HBV DNA replicated patients(≥103 copies/ml) ,the detection rate of HBeAg and preS1 were 51.5% and 70.9% ,they had statistically significant. Conclusion HBV DNA and PreS1 had fine dependability,preSl could reflect the replication of HBV sensitively than HBeAg,it could be used as a reliable new marker of HBV replication in vivo.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Biopsy ; CD56 Antigen ; metabolism ; DNA-Binding Proteins ; metabolism ; Gene Deletion ; Humans ; Keratin-7 ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Small Cell Lung Carcinoma ; genetics ; metabolism ; pathology ; Synaptophysin ; metabolism ; Transcription Factors

Objective To investigate the relationship between the mechanism of ischemic postconditioning-induced activation of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway during myocardial ischemia-reperfusion (I/R) and reactive oxygen species (ROS).Methods Healthy male Sprague-Dawley rats, aged 16-20 weeks, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1％ pentobarbital sodium 40 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution.Thirty-two isolated rat hearts were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C) , group I/R,ischemic postconditioning group (group IPO) , and N-(2-mercaptopropionyl)-glycine (a ROS scavenger) + IPO group (group M + IPO).After 20 min of equilibration, group C was continuously perfused with K-H solution for 100 min, and the isolated hearts received the drugs via the perfusion system in the other groups.Group I/R was perfused with cardioplegic solution 4 ℃ St.Thomas, and then was subjected to 40 min of ischemia at 32 ℃ followed by 60 min of reperfusion.In group IPO, ischemic postconditioning was induced by 6 cycles of 10 s reperfusion followed by 10 s limb ischemia starting from the onset of reperfusion, and the hearts were then perfused for 58 min.In group M + IPO, the hearts were perfused with K-H solution containing N-(2-mercaptopropionyl)-glycine 2 m mol/L for 3 min starting from the onset of reperfusion,underwent 2 min of ischemic postconditioning, and then was perfused for 55 min.Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP),and positive maximal pressure of left ventricular increase (+dp/dtmax) were recorded at the end of equilibration and of reperfusion.At 5 min of reperfusion and the end of reperfusion, myocardial specimens were obtained from the left ventricle for determination of ROS content by enzyme-linked immunosorbent assay.At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase-1 (HO-1) , quinone oxidoreductase 1 (NQO1), and superoxide dismutase 1 (SOD1) mRNA and protein expression (by using Western blot and real-time polymerase chain reaction).The damage to myocardial mitochondria was assessed using Flameng scoring.Results Compared with group C, HR, +dp/dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I/R and M+IPO groups, HR and LVDP were decreased, LVEDP was increased, and no significant changes were found in +dp/dtmax at the end of reperfusion in IPO group, Flameng score was increased in I/R, IPO and M+IPO groups , the ROS content was increased at the end of reperfusion in I/R, IPO and M+IPO groups, and Nrf2, HO-1,NQO1 and SOD1 mRNA and protein expression was down-regulated at the end of reperfusion in I/R, IPO and M+IPO groups.Compared with group I/R, HR, +dp/dtmax and LVDP were significantly increased, and LVEDP and ROS content were decreased at the end of reperfusion, Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was up-regulated at the end of reperfusion in IPO and M+IPO groups, Flameng score was decreased in IPO group, there was no significant change in Flameng score in M+IPO group.Compared with group IPO, HR, +dp/dtmax and LVDP were significantly decreased, LVEDP and ROS content were increased at the end of reperfusion, Flameng score was increased, and Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was down-regulated in M+IPO group.Conclusion Ischemic postconditioning can regulate ROS level and activate Nrf2-ARE signaling pathway, thus attenuating myocardial I/R injury in rats.

Objective To observe the activation mechanism of Nrf2-ARE pathway in protective effect of ischemia and pinacidil postconditioning on isolated rat hearts.Methods The hearts of adult male Sprague Dawley rats were established ischemia-reperfusion injury model,and devided into six groups(n =8,each group),i.e.Normal group(Group N),ischemiareperfusion group (Group Con,I/R),ischemic postconditioning group (Group IPO),pinacidil postconditioning group (Group P50),N-(2-mercaptopropionyl)-glycine(MPG,2mmol/L) + IPO group(Group M + IPO),MPG + P50 group(Group M + P50).Rat hearts were perfused with Krebs-Henseleit(K-H) buffer for 20 minutes for equilibration.Subsequently,Group N was perfused with K-H buffer for 100 minutes after equilibration,Group Con was perfused with 4℃ ST.Thomas solution to stop the heart beating after equilibration,then the hearts were underwent 40 minutes global ischemia under 32℃,and followed by the K-H solution for 60 minutes.Group IPO after global ischemia period,the hearts were subjected to six 10-seconds cycles of ischemia/reperfusion at the beginning of reperfusion,then were reperfused for 58 minutes.Group P50 after global ischemia,rat hearts were perfused with K-H buffer containing pinacidil(50.μmol/L) for 2 minutes before reperfusion.Group M + IPO after global ischemia,the hearts were subjected to perfuse with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then underwent six 10-seconds cycles of ischemia/reperfusion before reperfusion.Group M + P50 after global ischemia,the hearts were perfused with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then subjected to perfuse with K-H buffer containing pinacidil(50 μmol/L) for 2 minutes before reperfusion.Cardiac function indexes(such as HR,LVDP,LVEDP,and the Max dp/dt) at the end point of equilibration and repeffusion were observed and recorded.The ultrastructure of myocardial tissue was observed by electron microscopy and the mitochondrial Flameng score was calculated.RT-PCR and western-blot were applied to detect the gene transcription and protein expression of HO-1,NQO1,SOD1,and Nrf2 in left ventricular myocardial tissue after reperfusion.Results The HR,LVDP and + dp/dtmax at the end of reperfusion:the cardiac function indexes are lower among each group compared with group N,group 1PO and group P50 are better than group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group group P50,but group M + IPO is obviously decreased(P ＜ 0.05).Compared with group P50,group M + P50 index is decreased significantly(P ＜ 0.05).The LVEDP at the end of reperfusion is lower than that among each group as compared with group Con,which is significantly increased in group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group P50,but group M + IPO is significantly increased(P ＜ 0.05).Compared with group P50,the group M + P50 is obviously decreased(P ＜ 0.05).The ultrastructure of myocardial tissue in group N is mostly normal,group Con presence serious damage.The ultrastructure damage of myocardial tissue is improved in group IPO and group P50 as compared with that in group Con,while group M + IPO is more serious than group IPO,group M + P50 is more serious group P50.The mitochondrial Flameng score is higher among each group as compared with group N (P ＜ 0.05),the score is lower in group IPO and group P50 as compared with group Con and corresponding nonblocking group (M + IPO,M + P50,P ＜0.05).The mRNA and the protein expressions of HO-1,NQO1,SOD1 and Nrf2 among each group are lower as compared with group N(P ＜0.05).Compared with those in group Con,the mRNA and the protein expressions in group IPO and group P50 are obviously increased(P ＜ 0.05),group IPO and group P50 are higher than those in group adding active oxygen scavenger(MPG) (P ＜ 0.05).Conclusion Ischemic postconditioning and pinacidil postconditioning have protective effect of myocardial tissue from ischemia reperfusion injury,while improve the cardiac function index.The cardiac protective effect of Ischemic and Pinacidil postconditioning methods may be involved the ROS in early reperfusion,which activate the Nrf2-ARE pathway,and up-regulate the expression downstream antioxidant protein and phase Ⅱ detoxifying enzyme,ultimately improve the cardiac function index during the reperfusion period.

Objective To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway in mitigation of anoxia/reoxygenation (A/R)-induced damage to rat cardiomyocytes by emulsified isoflurane postconditioning.Methods Primarily cultured cardiomyocytes obtained from Sprague-Dawley rats,aged 16-20 weeks,were seeded into the laminin pre-treated 6-well plates at a density of 104/cm2 and cultured for 20 h.The cells were randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),group A/R,and emulsified isoflurane postconditioning group (group EI).The cells were cultured for 110 min without any treatment in group C.The cells were exposed to 45 min anoxia followed by 60 min reoxygenation in A/R and EI groups.After 45 min of hypoxia,the cells were cultured with emulsified isoflurane 1.68 mmol/L for 5 min before onset of reoxygenation in group EI.At the end of reoxygenation,transmission electron microscope was used to observe the ultrastructure of myocardial cells,and mitochondrial function was scored.The mRNA and protein expression of Nrf2,heme oxygenase-1 (HO-1),superoxide dismutase 1 (SOD1),and quinone oxidoreductase 1 (NQO1) was detected by real-time PCR and Western blot.Laser scanning confocal microscopy was used to detect Nrf2 nuclear translocation at the end of incubation,and Nrf2 activity was recorded in the nucleus of myocardial cells.Results Compared with group C,the mitochondrial function score was significantly increased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was down-regulated,and Nrf2 activity in the nucleus was enhanced in A/R and EI groups.Compared with group A/R,the mitochondrial function score was significantly decreased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was up-regulated,and Nrf2 activity in the nucleus was enhanced in group EI.Conclusion The mechanism by which emulsified isoflurane postconditioning mitigates A/R-induced damage to rat cardiomyocytes is related to induced nuclear translocation of Nrf2 and activated Nrf2-ARE signaling pathway.