Objective To observe the effect and mechanism of imiquimod on T helper (Th) cell subsets in the Parabronchial lymph node (PBLN) cell cultures from ovalbumin (OVA)-sensitized rats.Methods PBLN were isolated and cultured. PBLN cells were divided into A~F, according to different concentrations of intervention. Cultured for 0, 3, 6, 12, 24, 48 hours, the expressions of IL-4 and IFN-? in supernatants were determined by ELISA. The mRNA expressions of the cytokines in cells were detected by RT-PCR.Results In the group A, only low concentrations of IFN-? were detected. Based on the cultured time, the concentrations of IFN-? were increased significantly if imiquimod concentration was between 1 and 10 ?g/ml. Levels of IL-4 were increased slowly compared with those in the group B (P0.05).Conclusion Imiquimod show the best effect on antigen-specific Th cell subsets when cultured for 12h. The results suggest that imiquimod have benefit in atopic diseases such as the late inflammation reaction of asthma.

Objective To evaluate the relationship between Helicobacter pylori antigen in saliva and the activity of gastritis and like intestinal metaplasia(IM)or atypical hyperplasia(AH).Methods From June 2004 to June 2005 Helicobacter pylori antigen were detected in saliva of 246 persons by enzyme-linked immunosorbent assay(ELISA),and the Hp-positive rates in saliva in patients with different gastritis were compared.Results The Hp-positive rate in saliva in patients with active chronic gastritis was 74.29%(26/35),which was significantly higher(P

Transcatheter aortic valve replacement is a new method for the treatment of patients with symptomatic aortic stenosis and high surgical risk.With the development of technology, this method has been widely used in the world.The current research shows that the short and medium-term curative effect is worthy of affirmation.Due to clinical experience, more and more surgical approaches have been proposed.This article reviews the advantages and disadvantages, clinical research, application progress and problems of transcardiac and transcatheter aortic valve replacement.

Ski, as an evolutionary conserved protein, is widely involoved in the proliferation and differentiation of many kinds of cells in different species. Ski also plays an irreplaceable role in many physiological and pathological processes of nervous system, including em-bryonic nervous system development, central and peripheral nervous system diseases, and so on, which may be assiciated with the signal pathways of transforming growth factor-beta and another family member bone morphogenetic protein.

Aim To explore the role of cytoplasmic FKBP52 in AAV-mediated transduction.Methods Murine embryo fibroblasts(MEFs)cultures from FKBP52 wild-type(WT),heterozygous(HE),and knockout(KO)mice were established.The role of FKBP52 in intracellular trafficking of AAV was analyzed by fluorescence-activated cell sorting(FACS)analyses,electrophoretic mobility shift assays(EMSA),southern blot,immunoprecipitations and western blot analyses.Results Conventional AAV vectors failed to transduce WT MEFs efficiently,and the transduction efficiency was not significantly increased in HE or KO MEFs.AAV vectors failed to traffick efficiently to the nucleus in these cells.Treatment with hydroxyurea(HU)increased the transduction efficiency of conventional AAV vectors by～25-fold in WT MEFs,but only by～4-fold in KO MEFs.The use of self-complementary AAV(scAAV)vectors,which bypass the requirement of viral second-strand DNA synthesis,revealed that HU treatment increased the transduction efficiency～23-fold in WT MEFs,but only～4-fold in KO MEFs,indicating that the lack of HU treatment-mediated increase in KO MEFs was not due to failure of AAV to undergo viral second-strand DNA synthesis.Following HU treatment,～59% of AAV genomes were present in the nuclear fraction from WT MEFs,but only ～28% in KO MEFs,indicating that the pathway by which HU treatment mediates nuclear transport of AAV was impaired in KO MEFs.When KO MEFs were stably transfected with an FKBP52 expression plasmid,HU treatment-mediated increased in the transduction efficiency was restored in these cells,which correlated directly with improved intracellular trafficking.Intact AAV particles were also shown to interact with FKBP52 as well as with dynein,a known cellular protein involved in AAV trafficking.Conclusion These studies suggest that FKBP52,being a cellular chaperone protein,facilitates intracellular trafficking of AAV,which has implications in the optimal use of recombinant AAV vectors in human gene therapy.

Objective To observe the clinical efficacy of bamboo-circled salt-partitioned moxibustion in treating arthritis of temporomandibular joint.Method Eighty patients were randomized into two groups. Forty cases in the bamboo-circled salt-partitioned moxibustion group received bamboo-circled salt-partitioned moxibustion at temporomandibular joint; forty cases in the warm needling group were intervened by selecting Xiaguan (ST7), Ashi point, etc. at the affected side. For the two groups, 3-day treatment was taken as a treatment course, and the therapeutic efficacy was analyzed after 2 treatment courses. The improvements in pain and mouth opening were observed before and after the treatment, and the treatment efficacy was evaluated by a 1-month follow-up study.Result The bamboo-circled salt-partitioned moxibustion group was superior to the warm needling group in comparing the real-time analgesic effect (P<0.05) and in the improvement of mouth opening (P<0.05); the comprehensive markedly effective rate was respectively 67.5% and 45.0% in the bamboo-circled salt-partitioned moxibustion group and warm needling group, and the between-group difference was statistically significant (P<0.05), indicating that bamboo-circled salt-partitioned moxibustion is better than warm needling in treating arthritis of temporomandibular joint; the follow-up study revealed satisfactory therapeutic efficacies in both groups: the effective rate was 92.5% in the bamboo-circled salt-partitioned moxibustion group versus 87.5% in the warm needling group, and the difference was statistically insignificant (P>0.05).Conclusion Bamboo-circled salt-partitioned moxibustion can produce a real-time analgesic effect and improve mouth opening; it's especially suitable to treat the patients who are afraid of needling, as it's significantly effective, safe, non-invasive,and easy-to-operate.

Aim To identify paclitaxel differential binding protein in the cells of cell line MCF-7 and MCF-7/Taxol and to find new target for antitumor agents.Methods Synthesis and activity assay of biotinylated paclitaxel was used to gain paclitaxel binding protein by semiautomatic in vitro selection using the affinity magnetic beads method.LC MS/MS analysis and Western immunoblot analysis were used to identify the differential binding protein.Results The experimentation identified paclitaxel binding protein in the cells of MCF-7 and MCF-7/Taxol.An absent strap in MCF-7/Taxol was discovered by comparing the straps of two cell lines, which contained 25 kinds of proteins, among which 3 proteins were identified by western blot techniques: Heat shock protein HSP 90,Dermcidin Precursor,Actinin.Conclusions By comparing the straps of two cell lines, the differential protein in the cells of MCF-7 and MCF-7/Taxol are discovered, implying that they may be the novel mechanism of taxane resistance and may lead to find a new approach to finding a new target for oncotherapy drugs.

Objective To identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.

Objective To explore the expression and the changes of ski with time in the injured spinal cord in rats. Methods Sixty adult female Sprague-Dawley rats were randomly divided into sham group (n=30) and injury group (n=30), each group were further divided into 1 week, 2 weeks, 4 weeks, 8 weeks and 12 weeks subgroups, with 6 rats in each subgroup. Spinal cord injury at T10 was established with modi-fied Allen's technique (10 g × 25 mm) in the injury group. The hindlimbs behavior of rats was rated with Basso-Beattie-Bresnahan (BBB) scores 1 day, 3 days, 1 week, 2 weeks, 4 weeks, 8 weeks and 12 weeks after spinal cord injury. Three rats in each subgroup were stained with HE staining to observe the pathological changes of the spinal cord and the formation of cavity. The other 3 rats were analyzed with im-munofluorescence staining of ski and semi quantitative analysis. Results The BBB scores of each time point were less in the injury group than in the sham group (P<0.05). Necrosis was the major pathological change in the injury groups 1 and 2 weeks after injury;cystic cavity completely formed 4 weeks after injury, with dense scar tissue around it;there was no significant change in the cavity and scar 8 and 12 weeks after injury, however, the adjacent spinal cord was obviously thinner. Ski expressed little in the normal spinal cord, and expressed more and more after injury, peaked at 8 weeks and decreased then. Ski was mainly observed in white matter in the sham group and 12 weeks injury subgroup, which was in gray matter 2, 4 and 8 weeks after injury. Ski was highly expressed around the cavity in injury center and formed high expression band. Conclusion Ski expresses after spinal cord injury in rats, that may be associated with the activation and prolif-eration of astrocytes and the formation of glial scar.

AIM:To explore the time-dependent change of Ski protein expression in normal and activated astrocytes in rats.METHODS:The astrocytes were obtained from rat cerebral cortex and cultured in vitro.The astrocytes were treated with LPS and scratch injury for activation.Western blot analysis was used to determine glial fibrillary acidic protein (GFAP) and Ski protein levels in activated astrocytes at a series of time points.The indirect immunofluorescence staining method was performed to detect the location of Ski protein in the astrocytes.RESULTS:The protein of GFAP was naturally expressed in the astrocytes, beginning to increase after treated with LPS and scratch injury.Little protein expression of Ski in the normal astrocytes was observed.The Ski protein expression began to increase after treated with 1 mg/L LPS, peaked at 4 d (P<0.05) and then deceased, but was stills higher than that in the normal cells.The protein expression level of Ski after scratch injury was highly consistent with above mentioned.Ski was mainly observed in the nucleus of the normal cells and the cells treated with LPS for 6 d, while it was observed in the cytoplasm 2 and 4 d after treated with LPS.CONCLUSION:The protein of Ski is expressed in the astrocytes, and the expression level is increased in activated astrocytes,mainly located in the nucelus.Ski may plays an essential roles in the processes of activation and proliferation of astrocytes.