The functions of spontaneous suppressor T cells and T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome were compared. In the early stages of disease, decreased activity of spontaneous suppressor T cells was concurrent with increased numbers of CD8+ cells and a reversed CD4:CD8 ratio. These changes were related to abnormalities in serum C3 level and circulating immune complexes. In the recovery stages of the illness, spontaneous suppressor T cell activity and T cell subsets returned to normal levels.

The gene and cDNA sequence encoding PAL from Chinese medicinal plant Rhus chinensis were cloned and analyzed, furthermore the biochemical properties, kinetic parameters, differential expression and key sites were studied. Rhus chinensis is a well-known Chinese medicinal plant. Phenylalanine ammonia-lyase (PAL) is the first enzyme of phenylpropanoid pathway. Several recent studies suggested that PAL also play an important role in plant-aphid interaction. In this study, both the cDNA and the genomic sequence encoding PAL from Rhus chinensis (designated as RcPAL) were cloned and analyzed. The 3,833 bp gene contained a 1,342 bp intron and two extrons. The ORF was 2,124 bp and predicted to encode a 707-amino acid polypeptide. The results of real-time PCR showed that RcPAL expressed in all tested tissues and followed the order: stems > young leaves > petioles > roots > seeds > mature leaves. RcPAL was successfully expressed in E. coli with the pET-28a-RcPAL recombinant vector. The recombinant protein exhibited a high level of PAL activity. Biochemical properties and kinetic parameters of recombinant RcPAL were further studied. The results showed that the optimal temperature and pH for RcPAL activity were 45 °C and 9.0, and the K m and K cat values were 7.90 mM and 52.31 s(-1), respectively. The active sites and substrate selectivity site were also investigated with site-directed mutagenesis methods, suggesting that Phe(126) is responsible for the substrate selectivity. To our knowledge, this was the first full-length PAL gene cloned and characterized from the family Anacardiaceae so far.

The 420 bp upstream regulatory region of rd29A gene from Arabidopsis thaliana genome was amplified by PCR. Sequence analysis showed that it shared 100% identity with the reported rd29A promoter and contained several cisacting elements including dehydration responsive elements (DRE), ABA responsive elements (ABRE) and so on. The plant expression vector prd29A-S65T was constructed with this fragment linked up with green fluorescent protein (GFP) gene controlled by rd29A promoter, which was transferred to sugarcane callus by bombardment. The expressions of gfp in the transformed sugarcane callus and leaves treated with 6g/L PEG and 40%PEG, respectively, were detected by fluorescent microscope. The results showed that the expression vector had been transferred into sugarcane callus and GFP gene could be induced by PEG treatment. Through PCR analysis, 5 plants were identified to be the transgenic. At the same time, plant expression vector rd29a-dreb-hyg was constructed with this fragment linked up with DREB2B gene controlled by inducible promoter rd29A, which will lay the foundation for droughtresistant transgenic breeding in sugarcane. Molecular analysis showed that the rd29a-dreb-hyg has been successfully integrated into corresponding sugarcane variety.

During embryonic development of Artemia sinica, environmental stresses induce the embryo diapause phenomenon, required to resist apoptosis and regulate cell cycle activity. The small ubiquitin-related modifier-1 (SUMO), a reversible post-translational protein modifier, plays an important role in embryo development. SUMO regulates multiple cellular processes, including development and other biological processes. The molecular mechanism of diapause, diapause termination and the role of As-sumo-1 in this processes and in early embryo development of Artemia sinica still remains unknown. In this study, the complete cDNA sequences of the sumo-1 homolog, sumo ligase homolog, caspase-1 homolog and cyclin B homolog from Artemia sinica were cloned. The mRNA expression patterns of As-sumo-1, sumo ligase, caspase-1, cyclin B and the location of As-sumo-1 were investigated. SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E proteins were analyzed during different developmental stages of the embryo of A. sinica. Small interfering RNA (siRNA) was used to verify the function of sumo-1 in A. sinica. The full-length cDNA of As-sumo-1 was 476 bp, encoding a 92 amino acid protein. The As-caspases-1 cDNA was 966 bp, encoding a 245 amino-acid protein. The As-sumo ligase cDNA was 1556 bp encoding, a 343 amino acid protein, and the cyclin B cDNA was 739 bp, encoding a 133 amino acid protein. The expressions of As-sumo-1, As-caspase-1 and As-cyclin B were highest at the 10 h stage of embryonic development, and As-sumo ligase showed its highest expression at 0 h. The expression of As-SUMO-1 showed no tissue or organ specificity. Western blotting showed high expression of As-SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E at the 10 h stage. The siRNA caused abnormal development of the embryo, with increased malformation and mortality. As-SUMO-1 is a crucial regulation and modification protein resumption of embryonic diapause and early embryo development of A. sinica.

The distribution and duration of Hantaan virus (HTNV) in the body fluids of patients were studied by immunofluorescence, reverse passive hemagglutination, and cell culture assays. Virus antigen of hemorrhagic fever with renal syndrome in peripheral blood mononuclear cells (PBMCs) was usually present before day 11 of the disease, especially from days 4-7. Virus isolates were more readily recovered from plasma early in the course of the illness and less frequently after day 7. The use of PBMCs rather than plasma enabled isolates to be recovered at a rate nearly twice that permitted by plasma and allowed the isolation peak of HTNV (days 4-7 after onset of disease) to extend an additional 2 or 3 d, thus prolonging the period of detectable viremia until days 8-11. PBMCs were especially useful in isolating viruses from patients with hemorrhagic fever with renal syndrome in whom antibody titers were generally high during the acute phase of the disease. HTNV was isolated from the cerebrospinal fluid of patients, but was difficult to recover from other body fluids.