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 explore the appropriate frequency of ventilator tube replacement by researching the influence of different ventilator circuit change frequency on ventilator-associated pneumonia (VAP).Methods A prospective randomized sampling study was conducted.The patients undergoing invasive mechanical ventilation over or equal to 3 days admitted to emergency intensive care unit (EICU) of the First Affiliated Hospital of Kunming Medical University from December 2012 to December 2015 were enrolled.The patients were divided into 3,7 and 10 days group according to the frequency of ventilator tube replacement.Bacteriology of ventilator tube and the incidence of VAP were compared among the groups.Results Ninety-eight patients were enrolled,mainly with the artificial airway of endotracheal intubation or tracheotomy,with 56 male and 42 female,aging 8 to 86 years with mean of (51.97 ± 17.56) years.There were no statistical differences in gender,age,Glasgow coma scale (GCS) score,cough function and application of glucocorticoid,enteral nutrition,atomization and sedative therapy among three groups,indicating that the risk factors among three groups were consistent.The bacteria detection rates of extension tube,breathe out tube,breathe in tube,and hydrops collection cup were 36.7％,36.7％,33.3％,and 33.3％ respectively in 3 days group,and they were 73.0％,67.6％,62.2％,and 62.2％ in 7 days group respectively,and were all 90.3％ in 10 days group.It was showed that the bacteria detection rate in different pipe parts was almost the same with the same change frequency,and the rate was higher with the longer usage of ventilator tube (x2 values were 20.599,19.879,21.975,21.975,all P =0.000).The longer of the tube used time,the higher incidence of VAP.The incidence of VAP in 3,7,10 days groups were 26.7％,59.5％ and 77.4％,respectively,but there was statistically significant difference among all groups (x2 =30.486,P ＜ 0.001).Based on the value of 3 days group,the incidence of VAP in the 7 days group was 15.950 folds of 3 days group,and the incidence of VAP in the 10 days group was 18.333 folds of the 3 days group (both P ＜ 0.001).Conclusion This study suggests that the longer of pipeline using time,the more serious degree of bacterial contamination of pipeline,the higher incidence of VAP.

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.

Objcetive To evaluate the clinical significance of bone biochemical metabolic markers in osteoporosis( OP),and investigate the clinical application value of the determination.Methods BMD at various skeletal sites was measured by XR-36 dual energy X-ray absorptiometry for 65 patients.Enzyme linked immunosorbent assay (ELISA) were used to determine bone biochemical metabolic markers in sera of 65 patients with osteoporosis and 62healthy controls.Results The levels of serum TRACP5b and 25-OH-VD3/25-OH-VD2 in OP group were higher than that in control group( P ＜0.05),meanwhile the level of BGP,BAP and Ca were varying degrees higher than that in control group,but the level of P was lower than that in control group.Conclusion TRACP5b,BGP,BAP,VD3/2 and other bone metabolic markers can be used as indicators of early detection of osteoporosis,in order to provide valuable data for early treatment of osteoporosis.

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 evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) singling pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by zinc sulfate preconditioning in rats.Methods SPF healthy male Sprague-Dawley rats,weighing 250-280 g,aged 16-20 weeks,were used in this study.After the animals were anesthetized,their hearts were immediately removed and retrogradely perfused with an oxygenated K-H solution at 37 ℃ in a Langendorff apparatus.Forty isolated rat hearts were randomly divided into 4 groups (n=10 each):control group (group C),I/R group,zinc sulfate preconditioning group (group Zn) and zinc sulfate preconditioning plus Nrf2/ARE singling pathway blocker luteolin group (group Zn+Lut).After 20 min of equilibration,the hearts were continuously perfused for 100 min in group C,the hearts were perfused with 4 ℃ St.Thomas′ cardioplegic solution before ischemia and then subjected to 40 min global ischemia followed by 60 min reperfusion to establish the model of I/R in group I/R,200 μmol/L zinc sulfate 1.5 ml/kg was injected intraperitoneally,and 24 h later the model of I/R was established in group Zn,and in group Zn+Lut,the hearts were perfused for 3 min with K-H solution containing luteolin 50 μmol/L starting from the time point immediately before ischemia,and the other treatments were similar to those previously described in group Zn.Heart rate (HR),left ventricular end diabetic pressure (LVEDP),left ventricular developed pressure (LVDP) and the maximum rate of increase of left ventricular pressure (+dp/dtmax) were recorded at the end of equilibration and reperfusion.Coronary effluent was collected at the end of reperfusion to measure the levels of lactate dehydrogenase (LDH) and malondialdehyde (MDA).The expression of heme oxygenase-1 (HO-1),quinone oxidoreductase (NQO1),superoxide dismutase 1 (SOD1) and Nrf2 in myocardial tissues was detected by Western blot.Results Compared with group C,HR,+dp/dtmax and LVDP were significantly decreased,LVEDP was increased,and the levels of LDH and MDA in coronary effluent were increased at the end of reperfusion in I/R and Zn+Lut groups,and the expression of NQO1,HO-1,Nrf2 and SOD1 was up-regulated in I/R,Zn and Zn+Lut groups (P<0.05).Compared with group I/R,the HR,+dp/dtmax and LVDP were significantly increased,LVEDP was decreased,and the levels of LDH and MDA in coronary effluent were decreased at the end of reperfusion,and the expression of NQO1,HO-1,Nrf2 and SOD1 was up-regulated in group Zn (P<0.05).Compared with group Zn,HR,+dp/dtmax and LVDP were significantly decreased,LVEDP was increased,and the levels of LDH and MDA in coronary effluent were increased at the end of reperfusion,and the expression of NQO1,HO-1 and SOD1 was down-regulated(P<0.05),and no significant change was found in Nrf2 expression in group Zn+Lut (P>0.05).Conclusion The mechanism by which zinc sulfate preconditioning reduces myocardial I/R injury is related to activation of Nrf2/ARE singling pathway in rats.