Objective:To construct GM-CSF and EB virus LMP2A fusion gene recombinant adenovirus and identificate it , using recombinant adenovirus to do immunologic study.Methods:With RT-PCR and Western blot to detect GC 2A gene expression of the recombinant adenovirus , with ELISA to analysis antibodies of vAd-GC2A stimulation in murine body , lactate dehydrogenase assay to detect the mouse cellular immunity effect , thereby the recombinant adenovirus immune effect was initially analysis.T-test method was used to do preliminary analysis and evaluation of the immune effect of recombinant adenovirus .Results:GC2A fragment amplified by PCR, the size was 1 961 bp and it was the expected.The results showed, the recombinant adenovirus vAd-GC2A proteins can be recognized by sera antibody of GM-CSF ( LMP2A); the results of ELISA showed that vAd-GC2A can stimulate the generation of antibodies.EBV-positive tumor cell killing rate (%) by CTL induced recombinant adenovirus was (66.7 ±6.9)%, at the same time the adenovirus type 5 wild strains and PBS were (24.3 ±2.5)%and(32.4 ±3.1)%only(P≤0.05).Conclusion:Fusion gene has been successfully constructed , vAd-GC2A in mice have humoral and cellular immune function.The data suggest that the vAd-GC2A should play a potent role in preventing the positive EB virus tumor.

0.05), the leukopenia rates 78.13%(25/32)vs 50.00%(17/34)(P

Adult ; Endocarditis ; microbiology ; Humans ; Male ; Mycoses ; Recurrence

Carcinoma, Squamous Cell ; radiotherapy ; Head and Neck Neoplasms ; radiotherapy ; Humans

Objective To investigate the clinical value of PET-CT in aiding CT guided percutaneous puncture biopsy for lung mas-ses.Methods The retrospective data were analyzed including 5 7 patients with lung masses who underwent CT guided percutaneous puncture biopsy combined with PET-CT (regarded as a research group)and other 75 similar patients with single CT-guided biopsy (control group).In two groups,it was divided with malignant,suspicious malignant,can not be diagnosed,inflammatory,benign by pathological findings,and was contrasted with the surgical pathology or final clinical results.To calculate sensitivity,specificity and accuracy of diagnosis for cancer through two puncture method.Recorded complication for pneumothorax and bleeding in two groups .Be compared analysis using the 2 test.Results In study group,it was higher for the diagnosis of malignancy sensitivity, specificity,accuracy.There were 96.3% (52/54),100.0%(3/3)and 96.5%(55/57)in research group,while those were 88.4%(61/69),83.3% (5/6)and 88.0% (66/75)in the control group,respectively.And no statistically significant.Between research and the control group,there was a significantly statistical difference in the rate of twice punctures in one mass (χ2= 10.122,P=0.001).Between the two groups,there was no statistically significant difference in the pneumothorax rate and the hemorrhage rate. Conclusion PET-CT in aiding CT-guided lung biopsy improves the sensitivity,specificity and accuracy of diagnosis.

Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P

Background Researches demonstrated that CpG ODN,a immunostimulatory sequences,has preventing and treating effect on allergic conjunctivitis caused by protein allergen.However,its effect on allergic conjunctivitis caused by fungal allergen is unclear. Objective This study aimed to investigate into whether the Th1-Th2 switching immunostimulatory CpG ODN could reverse the response in the murine allergic conjunctivitis model caused by aspergillus fumigatus. Methods A mixture of spores and hyphae of aspergillus fumigatus strain was used to induce allergic conjunctivitis in male BALB/C mice aged 6-8 weeks.This experiment was designed into preventive or therapeutical treatment program.Under both settings,allergic conjunctivitis of the animals were treated with CpG ODN,nonstimulatory GpC ODN or PBS.After the last challenge with the allergen,the clinical symptoms of the animals were scored based on the criteria of Magone.The animals were sacrificed and the histopathological examination of conjunctiva was performed.Expression of TLR4 mRNA in conjunctiva was analyzed by real-time PCR assay.The responsiveness and populations of lymphocytes in spleen and draining lymph nodes were analyzed by flow cytometry.The use complied with the Standard of Association for Research in Vision and Ophthalmology. Results In the prevention mode.CpG ODN decreased subconjunctival infiltration compared with GpC ODN and PBS groups with the average neutrophil count index(21.25 ±11.59/section,30.75 ±11.44 section and 69.00±9.90/section,respectively).Expression of TLR4 mRNA was up-regulated significantly by CpG ODN.The clinical scores for CpG ODN group were insignificantly lower than those in GpC ODN group and PBS group(P＞0.05).In the therapeutic mode,compared with GpC ODN and PBS groups,the allergic symptom score in CpG ODN group manifested significantly lower(t=4.000.t=2.750,P＜0.01)and showed fewer cellular infiltration(t=4.870,t=3.829,P＜0.01)and higher expression of TLR4 mRNA(P＜0.01).In cultured splenic and draining lymph node cells,increased percentages of CD4+ CD25+ and CD4+ CD25+ CD69+ in CpG ODN group were observed compared with control groups(|P＜0.05). Conclusion CpG ODN can relieve aspergillus fumigatus-induced allergic conjunctivitis via either subconjunctival injection or topical application by upregulating expression of TLR4 and activating Treg lymphocytes.

Objective: To explore whether acupuncture can improve sleep disturbance, cognitive impairment and emotional disorders caused by sleep deprivation, and its association with the attenuation of oxidative stress injury in prefrontal cortex. Methods: Fifty-two male Sprague-Dawley rats were randomly divided into a control group (n=10), a model group (n=14), a manual acupuncture (MA) group (n=14), and a sham-MA group (n=14). All the groups were established as sleep deprivation models via the modified multiple platform method, except for the control group. Rats in both the MA group and the sham-MA group received corresponding intervention, respectively. After modeling and intervention, the four groups received three behavioral tests, namely sleep monitoring, by comprehensive lab animal monitoring system (CLAMS), Morris water maze (MWM) test and open-field test (OFT), followed by oxygen free radical level test and Western blot (WB) detection for the expression levels of Bax and Bcl-2. Results: The MA group derived more sleep time within 24 h than either the model group or the sham-MA group (both P<0.05). On MWM orientation navigation test day 1, there were no significant differences in escape latency among the control, MA and sham-MA groups (P>0.05), and the escape latency was significantly shorter in these three groups than that in the model group (all P<0.05). On test day 4, the escape latency was markedly shorter in the MA group than that in either the model group or the sham-MA group (both P<0.05); meanwhile, the MA group showed significantly better performance compared with these two groups in space probe test (both P<0.05). In OFT, compared with the control group, there was a significant decline in the horizontal movement score in the other three groups (all P<0.05), and the decrease was more significant in the model group and the sham-MA group than that in the MA group (both P<0.05). The superoxide dismutase (SOD) content was markedly higher and the malondialdehyde (MDA) content was markedly lower in the MA group than those in the model group and the sham-MA group (all P<0.05). Compared with the model group and the sham-MA group, the expression of Bax was significantly lower and the expression of Bcl-2 was significantly higher in the MA group (all P<0.05). Conclusion: MA therapy can lengthen the sleep time in sleep-deprived rats and improve learning and memory impairments induced by sleep deprivation, and the underlying mechanism may be associated with the enhancement of antioxidant capacity in the prefrontal cortex and the inhibition of hippocampal neuronal apoptosis.

According to the super-position principle of the reinforcement of biological activities, a series of novel E-substituted 2, 3-diaryl propenoic acyloxy phosphonate derivatives were designed and synthesized. And the structures of the target compounds were confirmed by IR, 1H NMR, 13C NMR and elemental analysis. Furthermore, the cytotoxicities of all compounds on A-549, SGC-7901 and EC-109 in vitro were evaluated by MTT assay, and some of them showed good antitumor activity. Among the active compounds, especially, the IC50 value of compound 3e was (12.7 ± 1.9) μmol x L(-1) against A-549 cells, similar to cisplatin [IC50 = (8.0 ± 1.5) μmol x L(-1)], compounds 3g and 3k had better inhibition effect on EC-109 cells growth, with the IC50 values of (9.5 ± 1.8) μmol x L(-1) and (11.5 ± 0.9) μmol x L(-1) respectively, and compounds 3i and 3k exhibited good cytotoxic property on A-549, SGC-7901 and EC-109, which were worth further investigation.
Antineoplastic Agents ; chemical synthesis ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Drug Design ; Humans ; Organophosphonates ; chemical synthesis ; pharmacology

OBJECTIVETo observe the effects of Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine on vascular endothelial growth factor (VEGF) in growth retardation induced by Decamethasone and observe its mechanisms.

METHODSThirty one-month-old New Zealand white rabbits were randomly divided into normal group, dexamethasone-treated group and Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine-treated group. The rabbits in dexamethasone group and Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine-treated group received dexamethasone (5 mg/kg x d). The rabbits were sacrificed at the 6th and 12th week after administration, and then rabbit tibia articular was removed. (1) Using TUNEL stain to detect apoptotic index. (2) Using immunohistochemical stain to detect the positive index of the expression of vascular endothelial growth factor (VEGF) in the epiphyseal cartilage of growth. (3) Using fluorescent quantitative PCR to detect the expression intensity of VEGF mRNA in each group.

RESULTSAt the 6th and 12th week after administration, there were significant difference in apoptotic index and cell proliferation index between dexamethasone group and normal group (P<0.01, dexamethasone group more than normal group). Immunohistochemical stain and fluorescent quantitative PCR indicated that the expression of VEGF and VEGF mRNA in dexamethasone group was significantly decreased as compared with that in normal group (P<0.01), and also obviously lower than Chinese herbal medicine-treated group (P<0.01).

CONCLUSIONVEGF has 2. an important role during the growth retardation induced by Dexamethasone. Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine can reduce the growth retardation induced by Dexamethasone through increasing the VEGF expression in growth plate chondrocytes and then increase angiogenesis.

Animals ; Apoptosis ; Cell Proliferation ; Dexamethasone ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression ; drug effects ; Growth Plate ; cytology ; drug effects ; growth & development ; metabolism ; Male ; Rabbits ; Random Allocation ; Vascular Endothelial Growth Factor A ; genetics ; metabolism