10.3969/j.issn.1000-8179.2013.12.014

Objective:To study the vaccine potency of gene-modified tumor cells. Methods:The EL-4 lymphoma was transduced with recombinant retrovirus containing the murine B7-1 gene. The effect of gene transduction on antitumor immunity was investigated.Results:The appearance, growth rate and surface marker of MHCⅠand MHCⅡ molecules of EL-4 cells transduced with B7-1 gene were the same with control cells except for CD80 positive in B7-1 gene transduced cells. B7-1 gene transduced EL-4 cells resulted in remarkable loss of tumorigenicity in syngenetic mice. EL-4/B7-1 cells could induce system protective immunity. Therapeutic vaccine of EL-4/B7-1 cells could retard the growth of established early-stage EL-4/Wt tumor significantly, but not retard the growth of late-stage EL-4/Wt tumor. Irradiated EL-4/B7-1 vaccine showed weak effect against challenged EL-4 cells.Conclusion:B7-1 gene transduced EL-4 cells can induce system protective immunity. It suggested that this vaccine have a potential application value in human cancer treatment.

BACKGROUND: Muscle-derived stem cells have some problems in pudflcation, amplification and directional differentiation during in vitro culture. Basic fibroblast growth factor as a membrane of bloactive factor has been paid great attention, due to its biological activity.OBJECTIVE: To explore effects of basic flbroblast growth factor on/n vitro proliferation of rat musde-derived stem cells.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Shenyang Medical College from April to July 2008.MATERIALS: Five healthy male Wistar rats were supplied by Experimental Animal Center, Shenyang Medical College. Basic fibroblast growth factor was produced by Zhuhai Biological Product Co., Ltd.METHODS: Left anterior limb triceps brachii was sterilely obtained from rats. Muscle-derived stem cells were harvested by isolation with enzyme digestion, pudfied by density gradient centrifugation and differential attachment. At the second passage,musde-derived stem cells were treated with basic fibroblast growth factor at final concentration of 6.25, 12.50, 25.00, 50.00,100.00 pg/L. Cells received 200 uL growth medium as blank control group, and those treated with 200 μL growth medium +musde-derived stem celts as negative controls. MTT was added at 24, 48, 72 and 96 hours of culture.MAIN OUTCOME MEASURES: Immunocytochomical method was used to determine rat musde-dedved stem cells. MTT assay was utilized to detect cell proliferation.RESULTS: Musde-denved stem cells were mostly positive for Sca-1, but few was positive for Desmin. Compared with the negative control group, vanous concentrations of basic fibroblast growth factor had significant proliferation effects on musde-derived cells (P < 0.05). At 6.25-50.00 μg/L, the proliferation effect became strong with the increased concentration (P <0.01). At 50.00 μg/L, the proliferation promotion effect nearly reached a peak. Musde-derived stem cells significantly proliferated over time. Compared with the negative control group, significant proliferation promotion effect was detected in the basic fibroblast growth factor groups at 96 hours (P < 0.01).CONCLUSION: Basic fibroblast growth factor can promote proliferation of rat muscle-derived stem cells/n vitro in a dose-time dependent fashion. 50.00 μ g/L and 96 hours are separately good concentration and time for in vitro culture.

Objective:To study the morphologic and phenotypic characters of a macrophage-tumor vaccine,and to observe the effect of macrophage-tumor vaccine on inducing CTL respose.Methods:The super-structure and the expression of CD14,CD68,CD80,CD86,MHC Ⅱ molecules of macrophage-tumor cells were detected with electron microscope,immunofluorescence staining and flow cytometry respectively.Meanwhile,H22 tumor cells were transplanted to the mice that had been immunized with different tumor vaccines.The weight and volume of tumors,the tumor cell injure rate and the level of LDH in culture supernatant were detected with direct measurement,MTT and selection methods.Results:The macrophage tumor vaccine cells were large cells with an irregular outline,and generally displayed pseudopodium,membrane folding,and vesicles on the cell surface.The predominant cytoplasmic organelles were lysosomes,secondary lysosomes and residual bodies.The percentage of CD14,CD68,CD80,CD86 and MHC Ⅱ positive cells within the differentiated population were 53.90%,98.60%,26.50%,90.20% and 25.40% respectively.The results of experiment in vivo revealed that the tumor forming rate,volume and weight of the group immunized with macrophage-tumor vaccine were much lower than that of control group and the group that were immunized with the macrophages that were induced by liquid paraffin (P0.05),the tumor weight and volume of the group immunized with the macrophage-tumor vaccine were lower than those of the group immunized with inactivated tumor cells(P

BACKGROUND: Chronic rejection limits the long-term success of cardiac transplantation and the underlying causes of the disease are unknown. Connective tissue growth factor (CTGF) is considered as a mitogenic and chemotactic factor for fibroblasts and is associated with cell proliferation and collagen synthesis.OBJECTIVE: To evaluate the role and significance of expression of CTGF in rat chronic rejection heart aliografta.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Laboratory Animal Center of the Second Xiangya Hospital between April and August 2007.MATERIALS: Twenty Wistar rats serving as donors and twenty Sprague-Dawely (SD) rats serving as recipients were included. An additional 10 Wistar rats were included as controls.METHODS: After intra-abdominal heterotopic heart transplantations, rats received cyclosporine A, mycophenolate, and methylprednisolone immunosuppression. Ten recipient rats were anesthetized and sacrificed for heart harvesting at 2 and 8 weeks postoperation, respectively.MAIN OUTCOME MEASURES: Coronary vessel density, fibrosis grade, and intimal occlusion were observed by hematoxylin-cosin staining and Van Gieson staining. Myocardial fibrosis was semi-quantitatively scored. CTGF expression was detected by immunohistochemistry. The associations between CTGF expression and allograft fibrosis and CAV formation were analyzed.RESULTS: Allografts harvested at 8-week post-surgery showed more obvious coronary intimal proliferation, fibrosis and higher CTGF expression compared with the 2-week allografts and the controls (P < 0.05-0.01 ) while the cardiac artery density was lower than the control group (P < 0.05). However, the control group in our study showed negligible CTGF expression. There were strong negative correlations between the gray value of CTGF protein expression and cardiac fibrosis and coronary intimal occlusion (r = -0.734, -0.713, P < 0.01), demonstrating that CTGF protein expression was positively correlated with cardiac fibrosis and coronary intimal occlusion.CONCLUSION: CTGF is expressed in cardiac myocyte with CAV. The increased expression of CTGF in the cardiac allograft is associated with CAV development and fibrosis formation and is involved in the pathogenesis of cbronic heart rejection

Objective To investigate the mRNA expression of COX-2 and MMP-2 on transitional cell carcinoma of bladder(TCCB) and their relationship with the invasion and metastasis of the bladder neoplasm. Methods Surgical bladder specimens were obtained from 54 TCCB patients and 5 benign prostate hyperplasia (BPH) patients to make paraffin slices, and 8 specimens which against cancers. In situ hybridization (ISH) was used to assay the mRNA expression of COX-2 and MMP-2. Results The mRNA expression of COX-2 and MMP-2 in bladder neoplasm were 59.2 % and 57.4 % respectively. Compared to the control, their expression was higher (P

Objective:To explore social support and personality characteristic of patients with clinically chronic pains to provide a new idea for clinical psycho-intervention.Method:45 patients with clinically chronic pains were evaluated by the Symptom Checklist(SCL-90),EPQ and SSRS,and compared with the control group.Results:Somatization,interpersonal sensitivity,anxiety,fear and psychotic factors have significant differences from those of the control group when being compared(p

Objective: To investigate the effects of lycopene on T lymphocyte subpopulations and pulmonary alveolar macrophagic (PAM) functions in rats with acute lung injury (ALI). Methods: Rats were randomly divided into the following groups. (1) Control group, (2) ALI model group, (3) Low dose group, (4) Mid dose group and (5) High dose group. Control group and ALI model group were treated with solvent of lycopene, and the other groups were gastrically incubated with lycopene. Thirty-five days later, control group were given physiological saline, ALI model group and lycopene administrated groups were injected with lipopolysaccharide (LPS) (6.0 mg/kg) to induce ALI. One hour, four hours or six hours after LPS or physiological saline challenged, abdominal aorta blood for measuring lymphocyte subpopulations and bronchoalveolar lavage fluid for measuring function of PAM were gathered respectively. Results: (1) At h 1, the percentages of CD3+,CD4+ and CD8+ of lycopene administrated groups compared with control group were not significantly different. At h 4, the percentage of CD4+ was similar to that at h 1. As for the percentages of CD3+, except high dose group [(28.8?9.9)%] was significantly lower, low dose, mid dose and ALI model group showed no significant difference compared with control group[(39.5?4.5)%]. The percentages of CD8+ of ALI model and lycopene administrated rats, separately (10.2?3.9)%,(10.3?2.8)%,(9.8?2.8)%,(10.1?3.5)%, had been significantly reduced compared with control group[(15.1?2.5)%]; between ALI model and lycopene ad-ministrated groups there was no significant difference. The instance at h 6 was the same as that at h 4. The percentage ratios of CD4+ T-lymphocyte to CD8+ T-lymphocyte of ALI model rats were not significantly different compared with control group or lycopene administrated groups at h 1 and h 6. At h 4, the ratio of the CD4 + and CD8 + in Low dose and Mid dose groups had significant difference and ALI model, high dose hadn’t when they were compared with control group. (2) Lycopene increased the phagocytic function of PAMs significantly at h 1(P

Objective To study genotypes,antibiotic resistance and epidemiology of extendedspectrum β-lactamases (ESBL)-producing Shigella isolated in Tianjin,and to discuss the relationship between ESBL-producing Shigella and drug-resistance plasmid.Methods A total of 136 Shigella spp.were isolated from stool specimens of patients with diarrhea who presented mainly with bloody purulent stool from Tianjin Children's Hospital,Tianjin Medical University No.2 Hospital and Tianjin No.1 Central Hospital between May 2009 and September 2010.Suspicious ESBL-producing isolates were screened by K-B disc diffusion method. The conjugation experiment was performed in the confirmed ESBL-producing strains and antibiotic resistance was compared between clinical strains and transconjugants to confirm the plasmid-mediated resistance. The genotypes of these isolates were detected by polymerase chain reaction (PCR) using universal primers for TEM,SHV,CTX-M-1 group,CTX-M-2 group,CTX-M-9 group,respectively.Intergenic consensus PCR (ERIC-PCR) was employed to understand the molecular homology of the ESBL-producing isolates. The data were analyzed by x2 test.ResultsESBL were identified in 14.7％ (20/136) of Shigella isolates,but no AmpC enzyme were detected.Among all the Shigella isolates,16 strains were genotype CTX-M-14,4 were genotype CTX-M-15.The strains with CTX-M ESBL were resistant to multiple antibiotics,while 100％ sensitive to imipenem.The transconjugant test of 18 ESBL-producing isolates were positive,and these conjugations were only resistant to β-lactamases. Conclusions CTX-M type is the common genotype of ESBL-producing Shigella isolates in Tianjin. ESBL-producing is the main cause of multiple resistance to β-lactams.The transmission of CTX-M producing strains is mainly mediated by plasmids.

Objective To investigate dynamic changes of plasma D-dimer, C-reactive protein (CRP) and fasting plas?ma glucose (FBG) levels in elderly patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) combined with type 2 diabetes mellitus. Methods AECOPD group (n=65) was used as A group, AECOPD combined with type 2 diabetes mellitus group (n=65) was used as B group. Levels of D-dimer, CRP and FBG were observed and compared between two groups in the first day, the 3rd day, the 7th day and the 14th day after hospital admission. The correlation be?tween D-dimer, CRP and FBG was analyzed. Logistic regression analysis was used to estimate the factors affecting increased plasma levels of D-dimer. Results Levels of D-dimer, CRP and FBG were significantly higher in B group than those of A group in the first day, the 3rd day, the 7th after hospital admission (P<0.01). Values of D-dimer and CRP were declined ob?viously in A group in the 7th day after hospital admission, but in B group they were declined obviously until the 14th day. There was no significant difference in FBG of A group. The level of FBG was significantly decreased in B group. There was a positive correlation between D-dimer, CRP and FBG (P<0.05). Logistic regression analysis showed that CRP, FBG, p(O2) and age were the influence factors for the increased levels of D-dimer. Conclusion In elderly patients with AECOPD com?bined with type 2 diabetes mellitus, we should pay attention to anti-inflammatory and controlling blood sugar, and the dy?namic monitoring levels of D-dimer, which helps to improve the prognosis for patients with AECOPD, and reduce mortality.