Objective To explore the effect of the association of low molecular heparin and Galectin-3 on the cell migration and cell proliferation of vascular endothelial cell from mesenchymal stem cells.Methods Depending on the administration, this study was divided into four groups: low molecular weight heparin group, adding 20 mg/L low molecular weight heparin into the cells;Galectin-3 group, adding 5 mg/L Galectin-3 into the cells;combination group, adding 20 mg/L low molecular weight heparin and 5 μg/ml of Galectin-3 into the cells;control group, equal volume of PBS buffer into the cells.The proliferation of vascular endothelial cell was to be detected by EdU incorporation,the cell growth cycle of vascular endothelial cell was to be detected by flow cytometry,and the cell migration of vascular endothelial cell was to be detected by scrath test.Then to investigate the effect of different conditions on the migration and proliferation of vascular endothelial cell.Results The OD490 of MWH group, Galectin-3 group, combined group and control group were (0.285±0.018), (0.297±0.041), (0.351±0.016), and (0.233±0.005) respectively, which indicates that the combined group could increase the cell proliferation significantly(P<0.05).Cultured for 24 hours, the cell migration rate was(42.02±7.62), (45.82±3.96), (68.53±11.22),and (34.21±3.99), suggesting that combined group have the largest cell migration(P<0.05).Conclusion The association of low molecular heparin and Galectin-3 could improve the cell migration and cell proliferation of vascular endothelial cell from esenchymal stemcells significantly.

Objective To construct a qseC-deleted mutant strain of E.coli by Red recombination and to study the effect of qseC gene on biofilm formation in the mutants.Methods The chloramphenicolresistant gene flanked by homologues of target genes was amplified by PCR and electro-transformed into E.coli MC1000.When induced by L-arabinose,the plasmid pKD46 could express three recombinant proteins of λ-prophage,which led to the replacement of target gene(qseC) with chloramphenicol-resistant gene.Then the chloramphenicol-resistant gene was eliminated by FLP-promoted recombination events.The biofilm formation of wild-type and mutant strain was detected by crystal violet staining.Results The qseC-deleted mutant of E.coli was confirmed by various PCR and DNA sequencing.Gene qseC was completely deleted.There was no significant difference in growth ability between the qseC mutant strain and the wild-type strain MC1000.The biofilm formation of wild-type and mutant strain was quantified by crystal violet staining.The absorbance determined with a plate reader at 570 nm was 1.00±0.15 and 0.47±0.10 respectively.Conclusion The qseC-deleted mutant of E.coli was constructed successfully.And the qseC gene plays an important role in regulation of biofilm formation in E.coli.

BACKGROUND:The limb ischemia model in rats has been widely applied in China to study the pathological process and treatment method of limb ischemia, but there are some controversies on the model construction and evaluation, so a reliable, convenient and economical disease model is needed for the related research. OBJECTIVE:To compare limb ischemia degree, duration and changing rule of acute hind limb ischemia models prepared by different ways in Sprague-Dawley rats, and find a method to prepare models that have moderate and stable limb ischemia and maintain a longer time. METHODS: 72 Sprague-Dawley rats were randomly divided into four groups, with 18 rats in each group. Group A: sham operation group, abdominal aorta below renal artery and iliolumbar artery, right superficial femoral artery, popliteal artery, saphenous artery were separated. Group B: the right superficial femoral artery, popliteal artery, saphenous artery were cut off, and the right femoral artery was resected to establish acute hind limb ischemic model. Group C: abdominal aorta and bilateral abdominal perineal artery were ligated to establish acute hind limb ischemic model. Group D: abdominal aorta, iliolumbar artery and lumbar artery were ligated to establish acute hind limb ischemic model. RESULTS AND CONCLUSION:Right hind limb muscle strengths of rats in groups B, C, D were weaker than that in group A at weeks 2, 4, 6 after operation; muscle strengths in group D was weaker than that in groups B, C at week 4 after operation; muscle strengths in groups B, D was stil weaker than that in group C at week 6 after operation. The partial venous oxygen pressure of right hind limbs in groups B, C, D was lower than that in group A at weeks 2, 4, 6 after operation; the partial venous oxygen pressure in group D was lower than that in groups B and C at weeks 2, 4 after operation; and group D was stil lower than group C at week 6 after operation. At weeks 2, 4 after operation, some muscle cels in the right hind limb muscle tissue were disrupted, fibrous connective tissue hyperplasia and capilary hyperplasia were detected, inflammatory cels infiltrated in the groups B, C, D. These pathological findings were more apparent in group D. Fiber connective tissue hyperplasia was attenuated, capilary hyperplasia, expansion and congestion were found in each group at week 6 postoperatively. The method of ligating abdominal aorta, iliolumbar artery and lumbar artery below renal artery for hind limb ischemia model can obtain moderate and stable ischemia conditions, maintain a long time, and is convenient to prepare.

Coronary Sinus ; abnormalities ; Humans ; Male ; Middle Aged ; Subclavian Vein ; Vena Cava, Superior

Objective To study the effect of epinephrine on biofilm formation of the qseC-deleted mutant of Escherichia coli on biomaterial.Methods The strains used in this study are Escherichia coli MC1000 and MC1000AqseC.LB was used for all the experiments.To determine the effect of epinephrine on motility,halos were measured in LB medium at 37℃ in the presence of epinephrine(50 μmol/L).LB with epinephrine and without epinephrine were used,and then the experiment of bacterial biofilm formation on PVC material was taken.The relative amount of biofilm was estimated.The thickness of bacterial community and bacterial community quantity in the unit area on PVC materials were measured by confocal laser scanning microscope( CLSM),and the surface structure of biofilm formation was observed by scanning electron microscope(SEM).Results The mutant strain formed less biofilm than the wild-type strain in LB.The increment in motility of wild-type strain due to epinephrine addition was shown,but mutant strain is unaffected.Similarly,biofilm formation of the wild-type strain was increased by epinephrine,but epinephrine did not affect the biofilm formation of the qseC mutant.The CLSM and SEM showed that epinephrine stimulated biofilm formation of wild-type strain on PVC materials,but had no effect on qseC-deleted mutant strain.Conclusion Epinephrine increases Escherichia coli biofilms on biomaterials through qseC.

Objective To investigate the role of quorum sensing Escherichia coli regulator C (qseC) in intestinal bacterial translocation in rats subjected to hemorrhagic shock.Methods Thirty Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 5 groups (n =6 each):control group (group C),MC1000-sham shock group (group M-SS),MC1000qseC-sham shock group (group △-SS),MC1000-hemorrhagic shock group (group M-HS),and MC1000△ qseC-hemorrhagic shock group (group △-HS).The rats drank 150 μg/ml of disinfect water containing streptomycin in 3 consecutive days to inhibit the autochthonous flora in the intestinal tract.From 4th day,the rats were fed with Escherichia Coli MC1000 or MC1000△ qseC 1 ml/100 g by gastric perfusion once a day for another 3 consecutive days in the other 4 groups,while the rats were fed with normal saline instead in group C.Hemorrhagic shock was induced by blood-letting.The mesenteric lymph node (MLN),spleen and liver specimens were obtained at 24 h after operation for bacterial culture and the bacteria were identified.Bacterial translocation from gut to MLN,spleen and liver was observed and the number of bacteria in MLN,spleen and liver tissues were counted.Results The rate of bacterial translocation was significantly higher,and the number of bacterial colonies in MLN,spleen and liver tissues and the total number of bacterial colonies were significantly larger in groups M-HS and △-HS than in group C,and in group M-HS than in groups M-SS and △-SS (P ＜ 0.05).The rate of bacterial translocation was significantly lower,and the number of bacterial colonies in MLN,spleen and liver tissues and the total number of bacterial colonies were significantly smaller in group △-HS than in group MHS.Conclusion QseC is involved in the intestinal bacterial translocation following hemorrhagic shock in rats.

Objective To compare surgical gastrojejunostomy and endoscopic stenting in palliation of malignant gastric outlet obstruction.Methods This retrospective study investigated patients treated for malignant gastric outlet obstruction from January 2007 to January 2014 in the first affiliated hospital of Nanjing Medical University.Endoscopic stenting was placed in 29 patients and surgical gastrojejunostomy was performed in 42 patients.The outcomes assessed included diet scores,time to diet,length of hospital stay,treatments fees and complications.Results Both endoscopic stenting and surgical gastrojejunostomy can relieve patients' syndrome with significant higher GOOSS score compared with that before treatment (P ＜0.05),but score improves faster in stenting group.Clinical success for endoscopic stenting and surgical gastrojejunostomy was 96.6％ and 92.9％ respectively,and technical success was 100％ for both of them.Endoscopic stenting group was found to have lower early complication rate(3.4％ VS 23.8％,P ＜0.05),higher late complication rate(24.1％ VS 6.9％,P ＜0.05),less time to diet,hospital stay and treatment fees(all P value ＜ 0.05)than surgical gastrojejunostomy group.The major complication after endoscopic stenting is re-obstruction while it is infection and leak of anastomotic site for surgical group.There were no significant differences in complication between two groups (27.6％ VS 11.9％,P ＞ 0.05).Conclusion Both endoscopic stenting and surgical gastrojejunostomy can relieve patients' syndrome effectively and safely,but endoscopic stenting improves GOOSS scores more rapid with less time to diet,less early complication rate and easy-dealing late complications,also it needs less hospital stay and fees.It's a better choice for patients with less survival expectation.

AIM:To investigate the inhibitory effect of small interfering RNA ( siRNA) on the expression of protein kinase Cε( PKCε) in human hepatoma SK-Hep-1 cells, and the biological behaviors of the transduced cells , inclu-ding proliferation and invasion , were investigated.METHODS:The cultured SK-Hep-1 cells were divided into 3 groups, including PKCε-siRNA group , negative control ( NC)-siRNA group and control group .MTT assay was used to analyze the proliferation of the SK-Hep-1 cells in the respective groups , while invasion potency was determined by Transwell assay .The protein levels of functional biomarkers such as Ki 67 and matrix metalloproteinase 9 ( MMP-9 ) were measured by Western blotting .The Luciferase reporter gene assay was used to explore the activity of the NF-κB pathway .RESULTS:PKCεex-pression in SK-Hep-1 cells transfected with PKCε-siRNA was significantly down-regulated at both mRNA and protein levels compared with that in the normal SK-Hep-1 cells (P<0.01), with the decreases in the protein levels of Ki67 and MMP-9. The invasion and proliferation of SK-Hep-1 cells were obviously inhibited in PKCε-siRNA group compared with control group (P<0.01).Furthermore, the transcriptional activity of NF-κB was down-regulated when PKCε was effectively in-hibited by PKCε-siRNA (P<0.01).CONCLUSION:Down-regulation of PKCεinhibits the proliferation and invasion of hepatic carcinoma cells , which might be mediated via the NF-κB signaling pathway .

Objective To analyze the associations between particulate air pollution (PM10,PM25) and stroke daily attack or mortality. Methods Meta-analysis method was used to polysynthetically analyze 16 quantitative studies about the associations between particulate air pollution and stroke daily attack or mortality. The relative odds ratio(OR) of stroke attack or mortality associated with per 10 μg/m3 increase of particulate matter concentration was used as effective value,taking a sensitivity analysis for the results. Results A 10 μg/m3 increase in PM10was associated with a 1.09% (95%CI: 0.10%-2.08% ) increase in stroke daily attack (OR=1.011,95%CI: 1.001-1.021)and 0.70% (95%CI: 0.60%-0.80% ) increase in stroke daily mortality (OR= 1.007, 95%CI: 1.006-1.008). The results of sensitivity analysis supported above results. As for PM2.5 OR appeared to be 1.001 (95%CI: 0.992-1.010) with a 10 μg/m3 increase in stroke daily attack and 1.052 (95%CI:0.958-1.154) for daily mortality. Conclusion There are positive associations between PM10 and stroke daily attack and mortality, increase of PM25 was not associated with stroke attack and mortality.

Objective To analyse the changes of metabolic parameters and condition of metabolic syndrome(MS)in females with obstructive sleep apnea-hypopnea syndrome(OSAHS),and explore the interaction between OSAHS and MS.Methods Sixty females undergoing polysomnography were included,and were divided into simple snorer group(n=19),mild OSAHS group(n=21)and moderate to severe OSAHS group(n=20).Epworth sleepiness scale(ESS)scores and metabolic parameters were compared among groups.Forty-one patients with OSAHS were subdivided into group with MS(MS group,n=15)and group without MS(non-MS group,n=26).ESS scores,apnea hypopnea index(AHI),oxygen desaturation index(ODI)and lowest pulse oxygen saturation(LSpO_2)were compared between MS group and non-MS group.Results There was no significant difference in age,body mass index(BMI),total cholesterol(TC),triglyceride(TG),low density cholesterol(LDL),high density cholesterol(HDL),apolipoprotein A-I(apoA-I)and apolipoprotein B(apoB) among simple snorer group,mild OSAHS group and moderate to severe OSAHS group(P>0.05).Correlation analysis revealed ESS was positively related to AHI(r=0.327,P=0.011).ESS scores and proportions of hypertension,fasting plasma glucose(FPG)and MS were significantly higher in moderate to severe OSAHS group than those in simple snorer group(P<0.05).BMI,FPG,TG,proportion of hypertension,ODI and ESS score in MS group were significantly higher than those in non-MS group(P<0.05),while there was no significant difference in AHI and LspO_2 between these two groups(P>0.05). Conclusion ESS score may reflect the severity of OSAHS in females.Females with OSAHS matched for age and BMI have no difference in blood fat.With the increase of severity of OSAHS,glycometabolism can be impaired,the prevalences of hypertension and MS increase.MS can exacerbate the severity of OSAHS,indicating that MS and OSAHS interact with each other.