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 investigate the influence of aerobic exercise on the vulnerability of atherosclerotic plaques and to observe the concentration of matrix metalloproteinase-9 ( MMP-9 ) and matrix metalloproteinase-14(MMP-14) in carotid atherosclerotic plaques in apolipoprotein E (APoE) gene deficient mice.Methods Eighty male, APoE gene deficient mice were divided into two equal groups: an aerobic exercise group and a limited action control group. Carotid atherosclerotic lesions were induced by perivascular constrictive collars placed on the right common carotid artery. Blood lipid levels in the exercise group were measured after 8 weeks of treadmill running and also in the control group. The morphological characteristics of carotid atherosclerotic plaques were observed in the two groups. Plaque area and fibrous cap thickness were measured. Lipid and collagen positive areas were quantified and the ratios correlated with the entimal areas were calculated. Plaque rupture rate and vulnerable index were calculated.Immunostaining was used to detect MMP-9 and MMP-14 expression in the atherosclerotic plaques. The mRNA levels of MMP-9 and MMP-14 mRNA in the fresh carotid plaques were quantified using a real-time reverse-transcriptase polymerase chain reaction (RT-PCR). Western blotting was performed for examining MMP-9 and MMP-14 protein expression in the fresh carotid plaques.Results No significant difference in serum lipid profiles or plaque area was found between the exercise and control groups. Compared with the control group, mean fibrous cap thickness, cap/core ratio and collagen content were all significantly higher in the exercise group, and lipid content was significantly lower. Plaque rupture rate and the vulnerable index were both significantly lower in the exercise group. Immunostaining showed that MMP-9 and MMP-14 expression were lower in the exercise group compared with the control group.The mRNA expression of MMP-9 and MMP-14 was also significantly lower.Conclusions Aerobic exercise can decrease the expression of MMP-9 and MMP-14 in carotid atherosclerotic plaques in ApoE gene deficient mice. Aerobic exercise may play a role in forestalling atherosclerosis by increasing the stability of plaque and decreasing plaque vulnerability.

Administration, Inhalation ; Animals ; Disease Models, Animal ; Female ; Fluorine ; blood ; Fluorocarbons ; adverse effects ; pharmacokinetics ; Male ; Rabbits ; Rats

Anti-Inflammatory Agents ; pharmacology ; Brain ; drug effects ; metabolism ; Humans ; NF-kappa B ; metabolism ; Neurons ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; alpha-MSH ; pharmacology

0.05).The degreeⅢ～Ⅳthrombocytopenia was more common in group A than in group B,but the degreeⅢ～Ⅳhypolekocytosis and phlebitis was more serious in group B.Conclusion NC and GC for treating refractory metastatic breast cancer have a high response rate and tolerable side effects.

Objective To observe the correlation of histologicalactivity(HAI) of chronic hepatitis B (CHB) with CCL20 expression, and to investigate the impact of CCL20 expression in CHB infection. Methods On the basis of established competitive quantitative RT-PCR with an internal standard, the expression of the CCL20 in the hepatocytes in different infected patterns of HBV infected cells and liver biopsies were quantified and at the same time its correlation to HAI were explored. Results In the cell levels, the expression quantity of CCL20 in control cells (HepG2), persistent HBV infected hepatocytes( HepG2. 2. 15) are (2. 65 ± 0. 02) pg/106 cells, ( 1.22± 0. 04) pg/106 cells, respectively. There were significantly differences between them ( t = 39. 66, P < 0. 01 ). The expression of CCL20 was enhanced in hepatocytes stimulated by PMA but their expression pauern was not changed. Moreover, CCL20 expression in liver biopsies with CHB was (3.54 ± 0. 65 ) pg/20 mg and CCL20 expression in control groups was ( 8. 74±0. 56) pg/20 mg. The expression of CCL20 between two groups was different (t =30. 09,P <0. 01 ) and correlation lied in between HAI and CCL20 expression in liver biopsies of CHB patients ( r = 0. 675, P =0. 023 ). Conclusion CCL20 expression was down-regulated and it was correlated to HAI of liver biopsies in CHB patients.

OBJECTIVETo explore the effect of Qiling Decoction (QD) combined highly active antiretroviral treatment (HAART) on expression levels of peripheral blood Th17 and Treg cells in HIV/AIDS patients.

METHODSTotally 55 HIV/AIDS patients were randomly assigned to the treatment group (28 cases) and the combination group (27 cases). Besides, 21 HIV negative patients were recruited as the healthy control group. Those in the treatment group received HARRT alone, while those in the combination group received HAART combined QD. The observation lasted for 24 weeks. Meanwhile, according to peripheral blood CD4+ T cell counts before treatment, HIV/AIDS patients were assigned to three subgroups. For patients in subgroup 1, 1 cells/microL < CD4+ T cell counts < or = 100 cells/microL; For patients in subgroup 2, 101 cells/microL < CD4+ T cell counts < or = 200 cells/lL; For patients in subgroup 3, 201 cells/microL < CD4+ T cell counts < or = 350 cells/microL. Expression of peripheral blood Th17 and Treg cells, and number of CD4+ T cell counts were detected using flow cytometry (FCM)in HIV/AIDS patients at the pre-treatment baseline, week 4, 12, and 24, as well as those in the healthy control group.

RESULTSCompared with the healthy control group, CD4+ T cell counts and the baseline expression level of Th17 cells in the peripheral blood of HIV/AIDS patients significantly decreased, the expression level of Treg cells significantly increased P < 0.01). Compared with before treatment in the same group, CD4+ T cell counts all increased at week 4, 12, and 24 in the two treatment groups, showing statistical difference (P < 0.05, P < 0.01). There was no statistical difference in the effective rate at various CD4+ T cell levels between the two groups (P > 0.05). There was no statistical difference in expression levels of Th17 and Treg cells between the combination group and the treatment group at any time point (all P >0.05). The Th17/Treg ration significantly increased in the combination group after 24 weeks of treatment, showing statistical difference when compared with the treatment group (U = 2.135, P = 0.038).

CONCLUSIONQD could improve the immune balance of Th17/Treg cells, which might be one of its mechanisms for improving HIV/AIDS patients' immunity.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

Objective To investigate the clinical value of emergent perspective stent implantation for leftside obstructive colonic cancer.Methods The clinical data of 26 patients with obstructive colonic cancer who received emergent perspective stent implantation at the Affiliated Xuzhou Hospital of Southeast University from October 2011 to February 2014 were retrospectively analyzed.A soft and a hard guidewire and a catheter were applied in the operation.The guidewires were put through the hole of the intestinal tumor,and then the metal stent and the conveyor were guided by the guidewires and were pulled through the hole of the intestinal tumor,finally the stent was released and the conveyor was adjusted to ensure that the stent was at the right position.Patients were followed-up through outpatient examination till 4 weeks after tumor resection.Results Of the 26 patients,9 were with rectal cancer,10 with sigmoid colonic cancer,6 with descending colonic cancer,1 with splenic flexure cancer.The median length of stenosis was 4.6 cm (range,2.0-8.0 cm).The surgery of the 26 patients was successful.The mean operation time was 35.2 minutes (range,15.2-72.0 minutes),and the mean time of stent implantation was 5.6 minutes (range,2.6-26.9 minutes).Patients had watery or loose stool for 4-8 times after stent implantation.Ten hours after the operation,all the patients were given liquid diet.The remission rate of clinical symptoms was 100.0％ (26/26).No colonic perforation was detected during the operation.Two patients were complicated with slight bleeding,and was alleviated by medication.Twenty-six patients received stage Ⅰ tumor resection procedure within 7-10 days after the symptoms of intestinal obstruction were remised.The success rate of surgery was 100.0％ (26/26).No infection and other drainage were detected after tumor resection through follow-up.Conclusion Emergent perspective stent implantation for left-side obstructive colonic cancer is safe and effective.

By non-steady method,the effective diffusivity of ferrous sulphate within alginate calcium gel entrapped without bacteria was measured.Meanwhile the oxidation ability of entrapped bacteria was analyzed.Experimental results showed that the effective diffusion coefficient of ferrous sulphate decreased with the increase of alginate concentration,the optimum alginate concentration is 2%(W/V).The effect of calcium chloride on the effective diffusivity was neglectable.The incubation of ferrooxidans would pass through 10 hours,and the diffusion coefficient within gel entrapped Thiobacillus ferrooxidans cells was less remarkably than that of ferrous sulphate without entrapped cells.For the entrapped cells,the absolute oxidation time was shortest and the rate change was fastest with the initial Fe concentration 5g/L.The absolute oxidation time was same when the initial Fe concentration was 8g/L and 10g/L.

Objective To investigate the effects of in vivo electroporation on plasmid mediated reporter gene expression and immunogenicity of DNA vaccine. Methods Luciferase expression plasmid was administered intramuscularly to BALB/c mice at 8μg and 40μg dosage level through injection with or without eletroporation Luciferase expression level in murine muscle was detected by IVIS imaging system 24 h after injection. DNA vaccine plasmid p1.0-gp1455m carrying codon-optimized env gene of CN54 strain ( HIV-1 CRF07_BC) was administered to mice at dosages of 8μg and 40μg through the two approaches mentioned above. Mice were immunized at week 0,2 and 4. Env-specific immune responses were detected at two weeks post the second and the third vaccinations. Env-specific antibody immune responses were determined by ELISA. Euv-specific cellular immune responses were determined by IFN-γ ELISPOT. Results Luciferase expression level in murine muscle was significantly increased as much as 35 folds through in vivo eletroporation. Results of ELISA and ELISPOT revealed that in vivo eletroporation could significantly enhance both the humoral and cellular immune responses induced by DNA vaccination. The responses induced by electrodelivered p1.0-gp1455m at 8 μg dosage were better than those induced by simple intramuscular injection with 40 μg of plasmid DNA. On the other hand, 2 injections followed by electroporation elicited comparable level of humoral and cellular immune responses with those induced by 3 injections without electroporation. Conclusion In vivo electroporation was capable of enhancing both the plasmid-mediated gene expression and immunogenicity of DNA vaccine.