Protozoans,the natural host of the facultative intracellular pathogen Legionella species,play an important role in survival,proliferation,virulence and stress resistance of Legionella species. By repeating transformation and selection,a spontaneous mutant of plasmid over expressing green fluorescent protein was obtained. This mutant replicates and is maintained stably in Legionella cells. The colonies of L. pneumophila harbouring the mutated plasmid were intense green in colour even under the daylight. After feeding BF1 strain of Tetrahymena thermophila with transformed L. pneumophila,the intracellular dynamic of changing of bacterial shape,bacterial proliferation and lysis of the host cell due to the bacterial proliferation were observed clearly under fluorescent microscopy. Thus,the present paper provides a simple and intuitionistic strategy for investigating the ecological and cellular relationship between L. pneumophila and its host.

Objective To explore the effects of single rectangle cage interbody fusion combined with pedicle screw fixa?tion for isthmic spondylolisthesis. Methods Data of the 34 cases with isthmic spondylolisthesis hospitalized at our department from March 2012 to February 2015 were retrospectively analyzed. All cases were operated by posterior pedicle screw fixation com?bined with single rectangle cage interbody fusion. There were 21 males and 13 females, aging from 18 to 63 years old (with an aver?age age of 41.6±8.2 years). Lesion segments were L4 in 15 cases and L5 in 19 cases;there were 12 cases in Meyerding I degree, 16 in II degree and 6 in III degree;28 patients had lower back pain associated with lower limb pain;19 patients had lower back pain associated with numbness of lower limb;18 patients had lower back pain associated with lower limb muscle weakness and 2 pa?tients had urination and defecation function disturbance. The surgical operation time, bleeding, postoperative drainage flow, graft fusion rate, complications were measured, and the intervertebral disc height, Japanese Orthopaedic Association (JOA), Oswestry disability indes (ODI), visual analogue scale (VAS) score were compared before and after treatment. Results The operation of 34 patients was successfully completed with the average operation time of 1.8 ± 0.8 h, and the bleeding in operation was 362.4 ± 93.7 ml, while postoperative drainage flow was 116.3±54.2 ml. All patients were effectively followed up for 12 months to 46 months (av?erage, one year and 10 months). Two weeks after operation, the intervertebral disc height were obviously increased (9.85±1.11 mm, 9.63 ± 1.04 mm), and there were significant difference compared with preoperative results (6.53 ± 0.98 mm, 6.24 ± 1.07 mm), but there were no significant difference compared with the latest follow?up results (9.23±1.33 mm, 9.30±1.16 mm). At the latest follow?up, JOA score was 13.19±1.08, ODI score 12.34±7.52, VAS score 2.23±1.51, and there were significantly different in each score compared with preoperative results. The symptoms of low back pain, lower limb nerve dysfunction and quality of life were signifi?cantly improved compared with preoperative situation. At the 16 month follow?up, all postoperative patients with vertebral body bone graft were judged to be fused. There was no pedicle screw loosening, fracture and so on. Conclusion Single rectangle cage interbody fusion combined with pedicle screw fixation for the treatment of isthmic spondylolisthesis is a kind of mini?invasive, less bleeding, safe and effective surgical method.

Objective To investigate the changes of circulaling progenitor cells and endothelial progenitor cells(EPCs)in non-septic and septic shock patients using flow cytometry.Method A total of 27 sepsis patients hospitalized in emergency department of Zhongshan hospital during August 2007 to February 2008 were enrolled in this study.The patients were dividedinto septic shock group(n=12)and non-septic shock group(n=15).Ten healthy individuals and ten non-sepsis ICU patients were collected as controls.Peripheral blood mononuclear cells(PBMCs) were isolated by Ficoll density gradient centrifugation,and EPCs labelled with antibodies against CDl33,CD34 and VEGFR-2 were identified and isolated by three-color fluorescence flow cytometry.Differences within the groups were analyzed using One way ANOVA.Results The percentages ofprogenitor cells and EPCs in the PBMC fraction in healthy controls were(0.25%4-0.14%),(0.09%-I-0.02%),respectively,and those in ICU controls were(O.38%.4-0.29%),(0.12%.4-0.02%).The percentages of progenitor cells and EPCs were significantly higher in栅sel如c shock patients(0.57%±0.12%),(0.22%.4-0.10%)than in heathy and non-sepsis ICU controls(P<0.05).However.the percentages of progenitor cells and EPC8 in septic shock pa.tienta(O.20%±0.12%,0.04%-t-O.01%)was obviousely lower than those in healthy,ICU controls and ilionseptic shock patients(P<0.05).Sptic shock survivors had significantly higher numbers of cEPCs than nonsur.vivors(P<0.05).Conclusions The level of progenitor cells and EPC8 in peipheral blood of sepsis patients might be the valuable markers to as.se88 the severity and outcome ofthese ptienS.

Objective The present study was to explore association of PvuⅡand XbaⅠpolymorphism in ER-?gene with genetic susceptibility for breast cancer without BRCAl/2 gene mutation. Methods 113 BRCA1/2 negative hereditary breast cancer patients from independent families and 113 agematched healthy control subjects were analyzed. Genotype analysis was conducted by polymerase chain reaction (PCR) and then DNA direct sequencing. The odd-ratios (OR) and 95% confidence intervals (CI) was calculated by unconditional logistic regression model. Results The frequency of PvuⅡpolymorphism CC(PP) ,CT(Pp) ,TT(pp) genotype in patients was found in 16 cases(14.2% ), 58 cases(51. 3% ) , and 39 cases (34. 5% ). The distribution of AA (xx) , AG (Xx) , GG (XX) genotype of XbaⅠpolymorphism were found in 76 cases ( 67. 2% ) , 34 cases ( 30. 1% ), and 3 cases ( 2. 7% ) among patients. Among premenopausal women, CT genotype of PvuⅡconfered a significantly increased risk for breast cancer compared with CC genotype ( adjusted OR = 2. 07; 95% CI, 0. 68 - 6. 30) ; Carriers of GG of XbaⅠhad a decreased risk for breast cancer (adjusted OR =0. 11; 95 % CI, 0. 01 - 1. 27) compared with AA genotype. Furthermore, combined analysis of two polymorphisms indicated individuals carrying PvuⅡCT and XbaⅠAA genotype were at increased risk for breast cancer as compared with those with PvuⅡCC and XbaⅠGG genotype (Oft = 11.43, 95% CI, 1.12-116.7) among premenopausal women. Conclusions PvuⅡand XbaⅠpolymorphisms in ER-?gene could be a candidate locus for low penetrance breast cancer susceptibility in Chinese population, especially among premenopausal women.

Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

0.05); the levels of LL, LI, RRS in CBA group and CBA+IBT group were significantly lower than those in control group(P

Objective To study the effects of MF59 in combination with heat-killed BCG ( hBCG) as adjuvant on the immunogenicity of Mycobacterium tuberculosis fusion protein PstS1-LEP.Methods BALB/c mice were divided into six groups from group 1 through group 6.They were immunized with PstS1-LEP+MF59 ( group 1 ) , PstS1-LEP+MF59/hBCG ( group 2 ) , PstS1-LEP+hBCG ( group 3 ) , MF59 ( group 4 ) , PstS1-LEP (group 5) and hBCG (group 6) for three times at intervals of two weeks , respectively.The mice were sac-rificed two weeks after the last immunization .The serum samples were collected for antibodies detection .The splenic lymphocytes and peritoneal macrophages were isolated and cultured with PstS 1-LEP.Indirect ELISA and sandwich ELISA were used to detect PstS 1-LEP-specific antibodies and cytokines in the supernatants of culture , respectively.Results The level of IFN-γ, IL-1β, IgG, IgG1 and IgG2a in group 1 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 and IL-4 in group 1 were higher than those in groups 4 and 6 (P<0.05).The level of IFN-γ, IL-1β, IL-12, IgG, IgG1 and IgG2a in group 2 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 was higher in group 2 than that in groups 4 and 6 (P<0.05). The level of IL-4 in group 3 was higher than that in group 4 ( P=0.05 ) .The level of IL-1βin group 3 were higher than that in groups 4 and 5 ( P<0.05 ) .The level of IgG was higher in group 3 than that in groups 4 and 6 (P<0.05).IgG1 level in group C was up-regulated in comparison with that in groups 4, 5 and 6 (P<0.05 ) .Conclusion hBCG as PstS1-LEP adjuvant induces a shift towards Th 2-type immune response , while MF59 induces Th1/Th2-type immune response.The combination of MF59 and hBCG inhibits the secretion of IL-4 by spleen lymphocytes , but enhances the secretion of IL-12 by macrophage .

Objective To investigate the application of of Gaussian 09/GuassView 5.0 in spectra teaching in Analytical Chemistry.Method Undergraduates of Pharmaceutical Science in 2014 grade of Kunming Medical University were selected to teach with a method with the help of Gaussian 09/GuassView 5.0 soft.Calculations of UV spectra,IR spectra and NMR spectra of compounds were introduced to make better understanding in the learning of relative knowledge points.The teaching effect was evaluated by the comparison of theoretical exam results of 2013 grade which didn't use soft.Result The theoretical test results showed that the scoring averages of the 2014 grade in UV,IR and NMR spectra were significantly higher than that in 2013 grade (P＜0.05).Conclusion Gaussian 09/GuassView 5.0 can visualize the nonobjective knowledge,and imporve the students' interesting,thus improving the efficiency of teaching and learning.

BACKGROUNDThere were some papers published in the Jonrnal of Science, December 2000 suggesting that one or more important susceptibility genes for late onset Alzheimer's disease (LOAD) were located on the long arm of chromosome 10. Linkage analysis showed maximum lod score close to D10S1225 loci, which indicated the loci might contribute to the etiology of Alzheimer's disease (AD).

METHODSFifty-nine LOAD patients and 107 controls were recruited. Apolipoprotein E (ApoE) genotypes were determined by reverse dot blotting hybridization assay. The D10S1225 was genotyped by 12% nondenaturing polyacrylamide gels electrophoresis and analyzed by silver staining. Statistical analysis was used to compare genotype and allele distributions between LOAD group and control group for ApoE and D10S1225 polymorphisms.

RESULTSApoE epsilon 4 was significantly higher in LOAD group in comparison with the control group (chi(2) = 6.530, P = 0.011). Seven different alleles of D10S1225 have been identified. The length of these gene fragments were 178 bp, 181 bp, 184 bp, 187 bp, 190 bp, 193 bp, and 196 bp, respectively. A total of 21 different genotypes were observed. There was no relationship between D10S1225 polymorphism and LOAD (chi(2) = 4.488, P > 0.05). Conclusion This study suggests that ApoE epsilon 4 is a risk factor for LOAD, however, the results indicated that there is not any possible linkage for disequilibria with a nearby AD risk gene near D10S1225.

Aged ; Aged, 80 and over ; Alleles ; Alzheimer Disease ; genetics ; Apolipoproteins E ; genetics ; Female ; Genotype ; Humans ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymorphism, Genetic

Nine morphologic traits, plant height, ground diameter, leaf length, leaf width, leaf area of plant, leaf fresh weight, blades, length/width ratio, plant fresh weight of Anoectochilus roxburghii from 13 different areas were determined for correlation analysis, path analysis and principal components analysis. Different source of morphological trait variation coefficient of A. roxburghii was 2.96% -12.59%, plant fresh weight was significant positively correlated with ground diameter, plant height and leaf number, and positively correlated with leaf fresh weight. Path analysis showed that plant height had the largest positive direct effect on plant fresh weight, the leaf fresh weight and blades number had indirect effects on the plant fresh weight. Through principal component analysis, morphological traits of A. roxburghii can be divided into "Determinants of high-yielding morphology" and "Determinants of leaf production". In the actual process of production and breeding of A. roxburghii, we should pay attention to plant height, leaf fresh weight, blades numbers and other traits.
China ; Orchidaceae ; anatomy & histology ; chemistry ; classification ; growth & development ; Phenotype ; Plant Leaves ; anatomy & histology ; chemistry ; growth & development