OBJECTIVETo investigate the effects of dendritic cells (DCs) infected with adenovirus vector encoding xenogeneic alpha-fetoprotein (AFP) on breaking the immune tolerance and induction of immunity against hepatocellular carcinomas.

METHODSHuman and mouse alpha-fetoprotein full-length cDNA were cloned from human HepG2 and mouse Hepa 1 - 6 hepatoma cell lines, respectively, using RT-PCR, and then inserted into adenoviral shuttle vectors to construct Ad hAFP and Ad mAFP. Mice were immunized with Ad hAFP-infected DC and in vitro CTL activity against Hepa 1 - 6 cells was examined by standard (51)Cr release assay. Survival was studied of the immunized mice, with or without depletion of CD8+ or CD4+ T cells, inoculated with Hepa 1 - 6 mouse hepatoma cells.

RESULTSThe lytic activity of CTL elicited by the Ad hAFP-infected DCs were much stronger than that by Ad mAFP-infected DCs. 80% of the Ad hAFP/DCs-immunized mice of the inoculated with 5 x 10(6) Hepa 1 - 6 hepatoma cells were still alive two months after inoculation. However, the Ad mAFP/DCs-immunized mice inoculated with 1 x 10(6) Hepa 1 - 6 cells were just 20% surviving two months later. Depletion of CD8+ or CD4+ T cells abolished such an antigen-specific immunity elicited by the DCs infected with Ad hAFP.

CONCLUSIONAdenovirus vector-mediated xenogeneic AFP-infected DCs can effectively break the immune tolerance to hepatocellular carcinomas in an animal model and induce strong antigen-specific T cell response, which are dependent on CD8+ and CD4+ T cells.

Adenoviridae ; genetics ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Carcinoma, Hepatocellular ; immunology ; pathology ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Female ; Genetic Vectors ; Humans ; Immunization ; Liver Neoplasms ; immunology ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; alpha-Fetoproteins ; immunology

OBJECTIVETo investigate the influence of pyrrolidine dithiocarbamate (PDTC) on the expression of transforming growth factor beta(1) (TGF-beta(1)), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor-1 of metalloproteinase (TIMP-1) in rats with pulmonary damage induced by paraquat (PQ).

METHODSFifty-four healthy male SD rats were randomly assigned into the control group (normal saline), the PQ-treatment groups (4 groups) and the PDTC treatment groups (4 groups). Except the rats in the control group, the rats in the PQ group were gavaged only with 40 mg/kg PQ, and PDTC group with 40 mg/kg PQ plus immediate injection 120 mg/kg PDTC (i.p). On the 3rd, the 7th, the 14th and 28th day after treatments, one group rats of each treatments were sacrificed and lung and blood samples were collected. The level of TGF-beta(1) protein in the plasma, the mRNA expression of TGF-beta(1), MMP-2 and TIMP-1 were evaluated using RT-PCR and real-time quantitative PCR, while pathological changes of lung were examined under optical microscope and electrical microscope.

RESULTSThe TGF-beta(1) protein, TGF-beta(1) and MMP-2 mRNA expression were increased significantly in the earlier stage and then decreased after PQ administration (P < 0.05 or P < 0.01), while the mRNA level of TIMP-1 was augmented continuously (P < 0.01) throughout the study compared to the control group. In comparison with the PQ group, in the PDTC treatment group, the TGF-beta(1) mRNA expression on the 3rd and the 14th day, 0.54 +/- 0.08 and 0.72 +/- 0.04 respectively, the MMP-2 mRNA expression on the 7th and 14th day, 1.62 +/- 0.50 and 1.97 +/- 0.34 respective-ly, and the TIMP-1 mRNA on the 7th and 21st day, 1.79 +/- 0.21 and 2.00 +/- 0.34 respectively, were significantly decreased (P < 0.05 or P < 0.01).

CONCLUSIONPDTC could attenuate paraquat-induced up-regulation of TGF-beta(1) and its mRNA expression, MMP-2 and TIMP-1 mRNA levels, which indicates that PDTC may exert its protective effects on paraquat-induced pulmonary damage by alleviating the earlier inflammation damage and adjust-ing the balance between MMPs and TIMPs. However, further studies are still warranted to investigate and clarify the underlying mechanisms involved in this complicated process.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Paraquat ; poisoning ; Pyrrolidines ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Thiocarbamates ; pharmacology ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism

Objective To investigate the effects of Apollon antisense oligonucleotide (ASODN) on proliferation,apoptosis and drug resistance of human leukemia(K562) cells.Methods Specific phosphorothioate ASODN and missense oligonucleotide (MSODN) of Apollon mRNA were synthesized and transfected into K562 cells following cationic liposome.The proliferation inhibition of K562 cells was assessed by MTT.The apoptosis rate was detected by Annexin V-FITC.The sensitivity of K562 cells to etoposide and vincristine was detected by MTT.Results Apollon antisense oligonucleotide inhibition of K562 cells with the concentration and time increased.ASODN at a final concentration of 600 nmol/L could significantly inhibit the K562 cell proliferation.The apoptosis rate was apparently increased (P ＜ 0.01).Conclusions Apollon ASODN may decrease Apollon gene expression,suppress K562 cells proliferation effectively,and induce significant apoptosis of K562 cells.Apollon ASODN is able to reverse the drug resistance via inhibition of Apollon expression and inducement of apoptosis.

Adult ; Diagnosis, Differential ; Female ; Humans ; Ovarian Neoplasms ; pathology ; Sertoli-Leydig Cell Tumor ; pathology

OBJECTIVETo investigate the mechanism of erectile dysfunction (ED) in rat models with chronic renal failure (CRF).

METHODSChronic renal failure was induced by adult male Sprague-Dawley rats, which were subjected to an excisional 5/6 nephrectomy. The rats in NCRF group and CRF group were divided into three groups randomly. Injected with apomorphine(APO, 80 microg/kg), penile erections of three groups were observed and noted by the 2nd week, 4th week respectively and 6th week from the 14th day of 5/6 nephrectomy and experimental models of ED with CRF were selected; NOS activity was examined and the microstructures of penile were observed under optical microscope with computer configuration image analysis system in selected rat models.

RESULTSCompared with the controls, the areas of smooth muscle and NOS activity in the penile cavernous tissue of ED rats with CRF decreased significantly (P < 0.01 or P < 0.05) , and collagen fibers slightly increased, and these alterations had close correlations with the duration of CRF. The

CONCLUSIONSPenile erection is seriously affected in rats with CRF. The decreases of areas of blood sinus had no obvious changes. the number of smooth muscles and NOS activity might be the most important factors.

Animals ; Disease Models, Animal ; Erectile Dysfunction ; enzymology ; pathology ; Kidney Failure, Chronic ; complications ; pathology ; Male ; Muscle, Smooth ; diagnostic imaging ; Nephrectomy ; Nitric Oxide Synthase ; metabolism ; Penis ; diagnostic imaging ; enzymology ; Rats ; Rats, Sprague-Dawley ; Ultrasonography

OBJECTIVETo observe the effect of dopamine receptor (DR2) activation on hypoxia/reperfusion injury (HRI) in the neonatal rat cardiomyocytes, and to explore its mechanism.

METHODSThe hypoxia/reperfusion (H/R) injury model was established in primarily cultured neonatal rat cardiomyocytes, and randomly assigned: control, H/R, bromocriptine (Bro) and haloperidol (Hal) groups. The cell apoptosis was detected using inverted microscope, transmission electron microscope and flow cytometry (FCM). The lactate dehydrogenase(LDH) and superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cell medium were analyzed. The expression of mRNA and protein of caspase-3, caspase-8, caspase-9, Fas, Fas-L, Cyt C and Bcl-2 were detected by RT-PCR and Western blot, respectively.

RESULTSCompared with the control group, apoptosis rate, LDH activity, MDA content and the expression of pro-apoptotic factors and anti-apoptotic factors were increased, but SOD activity was decreased in H/R group. Compared with the H/R group, all index above-mentioned were down-regulated or reversed in Bro-group, and had no obvious differences in Hal-group.

CONCLUSIONThe neonatal rat cardiomyocytes injury and apoptosis caused by hypoxia/reperfusion can be inhibited with DR2 activation, which mechanism is related to scavenging oxygen radical.

Animals ; Animals, Newborn ; Apoptosis ; Cell Hypoxia ; Myocardial Reperfusion Injury ; etiology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; Rats ; Rats, Wistar ; Receptors, Dopamine D2 ; metabolism

OBJECTIVETo investigate the spatial learning and exploration along with the CNS excitatory amino acid neurotransmitters profiles in adult rats subchronically exposed to the anticholinesterase organophosphorus insecticide dimethoate.

METHODSRats were gavaged daily with dimethoate (0, 5, 10 or 20 mg/kg via oral) in NS. for 90 days. Morris water maze tasks were used to test the spatial learning and memory in the rats after the dimethoate exposure. Simultaneously, rats were decapitated for the determination of brain cholinesterase AChE activities, glutamate concentrations, and the NMDA receptor NMDA-R densities and affinities in hippocampus.

RESULTSLatencies to find a hidden escape platform were significantly longer in dimethoate dosed groups than that of the control group in the place navigation tests. Subsequently, the times of crossing the location of platform which had been removed obviously decreased in the highest dose group compared with that of the control in the spatial probe tests (P < 0.05). AChE activity was significantly reduced 42% approximately 78% by all three doses of dimethoate (P < 0.05). Glutamate concentrations were increased significantly 132.9% approximately 134.5% by the two highest doses of dimethoate (P < 0.05). In addition, the NMDA receptor bindings were reduced 21.2% approximately 23.2% with the statistical significance at the same two highest doses (P < 0.05). Furthermore, the receptor affinities was reduced 33.1% by the highest dose group (P < 0.05). The lesions of spatial memory were statistically corrected with the decrease of the NMDA-R affinities (P < 0.05).

CONCLUSIONThe cholinergic lesion as well as the excitatory amino acid system alteration might attribute to the inferior ability in spatial learning and memory in dimethoate subchronically exposed rats.

Acetylcholinesterase ; metabolism ; Animals ; Chronic Disease ; Dimethoate ; toxicity ; Disease Models, Animal ; Glutamic Acid ; metabolism ; Insecticides ; toxicity ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Toxicity Tests, Subchronic

OBJECTIVETo explore the clustering character of overweight and obesity with multiple cardiovascular disease risk factors such as blood pressure, serum lipids and glucose, and to provide evidence for intervention in childhood.

METHODSA cross-sectional study was conducted on 913 children aged 7-13 years in urban area of Shijiazhuang. Measurements included height, weight, systolic blood pressure (SBP), diastolic blood pressure (DBP) and fasting serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and serum glucose (GLU).

RESULTSOverall prevalence rate of overweight and obesity was 29.24% (boys: 34.98% vs. girls: 24.23%). When compared with normal children, children with overweight or obesity had significantly higher levels of SBP, DBP, TC, TG but low HDL-C. The prevalence rates of high SBP, DBP, TC, TG, GLU and low HDL-C in overweight and obese children were higher than in normal children. After adjusted for gender and age, the odds ratios of overweight and obese children for high SBP, DBP, TC, TG, GLU and low HDL-C were 6.77, 3.22, 2.55, 6.42, 3.85 and 2.94 (95% CI: 4.15-11.04, 1.38-7.49, 1.59-4.11, 3.46-11.92, 1.69-8.78 and 1.83-4.73), respectively. Odds ratios of overweight and obese children holding any one, two or three of the selected six risk factors appeared to be 2.74, 13.15 and 15.33 (95% CI : 1.92-3.92, 6.69-25.87 and 4.17-56.39), respectively.

CONCLUSIONChildhood overweight and obesity increased the clustering of children's risk factors on cardiovascular diseases, as well as increasing the risk of children acquiring multiple cardiovascular disease risk factors. Controlling overweight and obesity would help early prevention on children from getting cardiovascular diseases.

Adolescent ; Blood Glucose ; metabolism ; Blood Pressure ; Cardiovascular Diseases ; prevention & control ; Child ; China ; epidemiology ; Cities ; statistics & numerical data ; Cluster Analysis ; Female ; Humans ; Lipids ; blood ; Male ; Obesity ; blood ; epidemiology ; physiopathology ; Risk Factors ; Schools ; statistics & numerical data

OBJECTIVETo investigate the heterogeneity of human breast cancer cells, their influence on biological behavior of tumor cells and clinical implications.

METHODSThe subpopulations of MCF-7 breast cancer cells were isolated by Percoll gradient centrifugation. DNA content and cell cycle distribution were detected with flow cytometry. Tumor chemosensitivity analysis was performed with MTT assay.

RESULTSHeterogeneity was observed in DNA content and cell cycle distribution among four subpopulations of breast cancer cells, which were related to their proliferation ability and chemosensitivity results.

CONCLUSIONHereditary instability and intrinsic characteristics of most tumor cells, not only lead to tumor progression and heterogeneity but also cause the loss of monoclonality and the generation of subclones. Further study on some profiles of tumor heterogeneity such as DNA content, cell cycle distribution and their influence on tumor proliferation and chemosensitivity may very well improve the clinical treatment.

Breast Neoplasms ; drug therapy ; pathology ; Cell Cycle ; Cell Division ; Cell Line, Tumor ; Centrifugation, Density Gradient ; DNA, Neoplasm ; analysis ; Female ; Humans

OBJECTIVETo develop 22 Chinese Mandarin monosyllable lists with good psychometrical equivalence. This study was to evaluate the test-retest reliability of these lists when it was used in speech recognition test in normal hearing dialectal speakers.

METHODSSeven cities including Dalian, Shanghai, Hangzhou, Wuhan, Guangzhou, Fuzhou and Xiamen were selected as testing centers which contain 6 typical Chinese dialectal regions including north of China, East of China, north of Fujian, south of Fujian, Guangdong and mid-south of China. At each center, 22 local normal hearing people were selected to join this study. Every participant was tested by each recognition test of all 22 lists twice in two sessions and same test order respectively. The second run of testing was carried out within 10 days-1 month since first run of testing.

RESULTSThere was a significant correlations between scores obtained at the two sessions (r = 0.682, P < 0.01). Paired student-t test had shown that a gross score of all dialectal participants was significantly higher than that of initial test to retest (P < 0.01). The mean increment of score was (2.7 +/- 10.1)%. A significant difference of test-retest score in 7 sites was 19.8% and it was equal to 5 test items. A one way ANOVA analysis had indicated that there were statistically significant difference between the score improvement of 7 test sites (P < 0.01). Another analysis had shown that there was no significant correlation between test-retest score improvement and intra-session intervals (P = 0.947).

CONCLUSIONSMandarin monosyllabic recognition test seems to be more stable, and the present study has indicated a systematic differences in Chinese Mandarin monosyllable recognition scores between test and retest. Monosyllable recognition test is not susceptible to memory effect. Pearson's correction analysis is not suitable to evaluation for test-retest reliability.

Adolescent ; Adult ; Analysis of Variance ; Asian Continental Ancestry Group ; Audiometry, Speech ; Female ; Humans ; Language ; Reproducibility of Results ; Speech Discrimination Tests ; Young Adult