Protein kinase B (Akt) is a Ser/Thr kinase,which in mammals comprise three highly homologous members known as PKB?/Akt1,PKB?/Akt2 and PKB?/Akt3. PKB is activated by hormones?growth factor and extra cellular matrix. The activation occurs downstream of PI3K. PKB phosphorylates and regulates the function of many cellular protein involved in processes that include survival,apoptosis, proliferation,glycogen metabolism and cancer progression. Although many mechanisms remains to be fully characterized,the research of PKB is thought to have a useful profect.

The family of HSP60 belongs to heat shock proteins with highly species conservatism and some important biologic functions. It can help other proteins for their assembling, folding and translocating, and plays a role in protecting cells against injuries and other types of stress. In addition, HSP60 is frequently recognized by the immune system as predominant antigens during infections and the progression of certain autoimmune diseases and might provide a novel strategy for the development of immunotherapeutics. This review focuses on distribution, molecular chaperone mechanism, function and gene expression regulation of HSP60. [

After the first recognition occurs between the activated sperm and zona pellucida of the oocyte from the mammalians, ?-1,4-galactosyltransferase I (? 4GalT I) combines the N-GlcNAc terminals by O-ligands on the ZP3 of the zona pellucida, which plays a difunctional role in the fertilization. The G protein signal system on the sperm membrane then is consequently activated by ZP3 via ?4GalT I, contributing to the induction of acrosome reaction. It was proved that in the activation of the G protein system, both the BBXB and BBXXB motifs on the N terminal of the long ? 4GalT I are necessary.

Src is a non-receptor protein tyrosine kinase activated by a number of extracellular signal moleculars. It is recruited to peripheral sites through myristoylation and the SH3 domain. Src initiates intracellular signal trandsduction pathways that influence cell adhesion, migration, growth, differentiation and survival though catalytic domain. Src is normally maintained in an inactive conformation because of carboxy terminal Src kinase, but can be activated transiently during cellular events such as mitosis or constitutively by abnormal events such as mutation and some cancers. In additions, c-Src protein is found to be highly activated and the Src gene is frequently over-expressed in many cancers. These findings suggest that the relationship between c-Src activation/over-expression and cancer progression appears to be significant.

Objective: To investigate the regulation of respiratory syncytial virus CTL epitope fused with G protein antigen fragment G1 to the specific immunoresponses. Methods: The recombinant plasmid pET-DsbA-G1 or pET-DsbA-G1F/M2 was transferred into E.coli BL21(DE3) and the fusion protein DsbA-G1F/M2 or DsbA-G1 was expressed.The expressing product was induced and purified by affinity chromatography. The two proteins were used to immunized BALB/c mice i.p, respectively. Serum and spleen cells were collected regularly. RSV-specific CTL responses were measured by MTT, IgG and IgG1 and IgG2a antibodies by ELISA, neutralizing antibodies by plaque reduction assay. Results: The recombinant proteins were expressed successfully and the purity was over 90% after purified by affinity chromatography. The protein DsbA-G1F/M2 induced significant RSV-specific CTL responses, while DsbA-G1 without CTL epitope did not induce detctable CTL responses. Strong IgG antibody responses were elicited,indicated by both. IgG1 and IgG2a titers induced by DsbA-G1F/M2, while only IgG1 was induced by DsbA-G1.The ratio of IgG1/IgG2a was downregulated significantly. Both antigens induced high level of neutralizing antibodies, but the latter was a little lower. Conclusion: DsbA-G1F/M2, the fusion protein of a CTL epitope and G protein fragment G1 can induce both cellular immunity and humoral immunity. The activation of CTLs downregulates the ratio of IgG1/IgG2a.The more balanced immunoresponse is advantageous for improving safety of the candidate vaccine.

AIM To analyze the toxicity and inhibitory mechanism of extracts from cultured cells (F 4 cell line) of Taxus chinensis on cancer cell lines SMMC 7721 and HEp 2. METHODS MTT assay for cell viability and flow cytometry for cell cycle analysis. RESULTS IC 50 of SMMC 7721 and HEp 2 were 0 161 4 g DCW?L -1 and 0 275 6 g DCW?L -1 respectively,tumor cells in G 2～M stage all increased with higher concentration and longer incubation of extracts from Taxus chinensis cells. CONCLUSION Extracts from cultured cells of Taxus chinensis could have cytotoxic effect on SMMC 7721 and HEp 2 and could induce apoptosis of both two cancer cells.

Lesion was made individually to the various cortical areas in 14 rats.The cortico- fugal fibers terminating into cochlear nuclei were traced in sections by means of the modified Nauta and Fink-Heimer silver staining method.The results were as follows: 1.After damaging the auditory area or somato-sensory area 1(SI)of the cerebral cortex unilaterally,degenerating fibers were found bilaterally in the ventral and dorsal cochlear nuclei.This result showed that the above cortical areas have direct descending connection with the cochlear nuclei. 2.No degenerating fibers were found in the cochlear nuclei after the ablation of the motor or visual area of the cerebral cortex,which demonstrated that the cochlear nuclei do not receive descending fibers from the above cortical areas. 3.The pathway from the cerebral cortex to the cochlear nuclei was:The descending fibers which originated from the auditory area or SI area ipsilaterally passed through the capsula interna and basis pedunculi,and part of these fibers passed dorsally and downward to the lateral portion of the reticular formation of the midbrain;and from there the fibers passed through the lemniscus lateralis and dorsal acustic stria to the cochlear nuclei of the ipsilateral side.The cochlear nuclei in contralateral side also received the corticofugal fibers by way of the dorsal acustic stria.These fibers might cross to the contralateral side at the region in the posterior commissure,superior colliculus commissure and inferior colliculus commissure.

Objective To establish a quick method to analyze vinorelbine ( NVB) in plasma of beagle dogs and study its pharmacokinetics of long-circulation and thermosensitive liposome loaded vinorelbine bitartrate.Methods The plasma was treated with liquid-liquid extraction after precipitation in methanol.The analysis was perfomed on a Venusil XBP C18 column(2.1 mm ×50 mm, 3 μm) at 35℃,the mobile phase consisted of methanol and water( containing 10 mmol/L ammonium acetate, 1%acetonitrile) 80∶20 and injection volume was 10μl.The type of mass spectrum was multireactive monitoring(MRM) in a positive mode.The monitor transitions were m/z 779.4-765.4 for vinorelbine and m/z 825.4-122 for vincristine.Results The concentration range from 10 to 2500 ng/ml had a good linearity ( r=0.0994).The precision, accuracy and extraction efficiency were acceptable.The plasma samples were stable for 10 days at -20℃ and 24 h at room temperature.Pharmacokinetic study in beagle dogs showed that the main parameters for injection and liposome were Cmax(833.51 ±150.42) and (1397.95 ±443.05)ng/ml, AUC(0-t) (577.16 ±223.57) and (1059.82 ±408.27) ng/ml· h, Cl(3014.64 ±1049.17)and 1633.10 ±551.77 ml/(h· kg), respectively.Conclusion A reliable HPLC-MS/MS method for vinorelbine analysis is established and can be applied to the pharmacokinetics study of liposome.The results show that liposome has a higher AUC, Cmax and longer Cl than injection.Meanwhile, liposome has a lower irritability.

Objective:To investigate whether His-tag to change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.Methods:The G1 and F/M2 gene fragments were amplified by PCR method and then ligated into the expressing vector pET-His or pET-DsbA-His.Each recombinant plasmid was transferred into E.coli BL21(DE3) and the expression was induced by IPTG.The expressed His-G1F/M2 or DsbA-His-G1F/M2 was purified by affinity chromatography.The latter was digested with thrombase and G1F/M2 was purified by affinity chromatography.His-G1F/M2 or G1F/M2 was used to immunize BALB/c mice.Anti-RSV antibody was measured by ELISA and RSV-specific CTL responses by MTT.Results:No significant difference was observed between the level of anti-RSV antibody or RSV-specific CTL response induced by G1F/M2 and that by His-G1F/M2.Conclusion:His-tag does not change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.

Objective To establish a quick method to identify BMSC by fast violet B salt staining and evaluate the clinic value. Methods Smears of separated and cultured BMSC, bone marrow, pleural and ascitic fluids were made, then the staining of fast violet B salt was performed. The BMSC in aplastic anemia (AA), high hyperplasia and normal groups were counted and compared with each other. Meanwhile, the diagnostic value of this method to AA was evaluated. Results The cytoplasm of BMSC presented mauve, while the nucleus were negative, other cells such as myelocytes, nucleated erythrocytes, megakaryocytes, monocytes, macrophages, lymphocytes and plasmacytes were negative. The count of BMSC in AA, high hyperplasia and normal group was 1.07 ± 0. 29, 2. 26 ± 0. 37 and 1.58±0. 33, respectively. Significant differences were found between AA and high hyperplasia groups, AA and normal groups, high hyperplasia and normal groups, respectively (P < 0.01). The sensitivity, specificity, positive likelihood ratio and negative likelihood ratio of this method for diagnosis of AA were 90%, 93%, 12. 86 and 0. 11,respectively. Conclusions The fast violet B salt staining is simple and convenient. It could be used to identify BMSC and play an important role in judging the hyperplasia extent and differentiation of AA.