Primary Study of the Structure and Antigenic Epitopes of Antigen Pf332 / 热带医学杂志

Objective To understand the primary structure and potential antigenic epitopes of antigen Pf332(Ag332) of P.falciparum iso late FCC1/HN.Methods　Based on the published Pf332 gene sequence , nine pairs of primers were designed for the PCR amplification of the Pf332 gen e fragments from genomic DNA of P.falciparum isolate FCC1/HN. The amplified gene fragments were subcloned into pMD-18T vectors and sequenced. The sequences were aligned using DNAstar software to obtain the full-length sequence of the gene Pf332. The primary structure and sequence homology of Ag332 were analyzed by SAPS, Tmpred, SingalP and Blastn programs. Three fragments, R0, R1 and R2, cor responding to nt#9595-10083, nt#10339-10767 and nt#10855-11247 of Pf332 gene were subcloned into the eukaryotic expression vector pcDNA3-S separately. The Balb/c mice were immunized with pcDNA3-S-R0, pcDNA3-S-R1 and pcDNA3- S-R2 separately, and the expressions of the recombinant proteins were detected by immunohistochemistry assay. The protective immune responses elicited by DNA I mmunization were analyzed by ELISA and parasite growth inhibition tests in vitro .Results　Nine Pf332 gene fragments were specifically amplif ied, subcloned into pMD-18T vectors and sequenced. Pf332 gene of the P.falci parum isolate FCC1/HN was 16,377 bp in length, encoding a protein of 5,458 ami no acids, about 615.28kDa. The Ag332 contains 17 regions of highly degenerated Glu-rich repeats, with 30.18% Glu in total amino acids of Ag332. Ag332 of P.falciparum isolate FCC1/HN and 3D7 exhibited 94.55 % homology in amino acid residues. The results of immunohischemistry assay showed that R0, R1 and R2 were expressed in mice muscle tissue. The amount of IgG antibody of the groups immu nized with pcDNA3-S-R0, pcDNA3-S-R1 and pcDNA3-S-R2 were higher than those of blank and pcDNA3 groups (P＜0.05). The result of parasite growth inhibition test showed that the immunized sera at 1∶5 dilution of groups of pcDNA3-S-R0, pcDNA3-S-R1 and pcDNA3-S-R2 had an incomplete inhibitor y effect on P.falciparum growth. Conclusion　The antigen Pf332 is an large protein containing highly degenerated Glu-rich repeats. Pf332 gene fragments, R1 and R2 encoding potent antigenic epitope repeats.

Generation and analysis of 113 adult stage Schistosoma japonicum (Chinese strain) expressed sequence tags / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To rapidly and economically obtain knowledge about adult stage Schistosoma japonicum (Chinese strain) expressed genes using expressed sequence tag (EST).</p><p><b>METHODS</b>A directional cDNA library constructed from Schistosoma japonicum (Chinese strain) adult stage RNA was used to generate expressed sequence tags (ESTs). These were compared against an EMBL-parasites database and GENBANK database by BLASTn and BLASTx.</p><p><b>RESULTS</b>A total of 314 phage clones were randomly selected for generating expressed sequence tags (ESTs). From these clones, 132 EST-quality sequence were obtained. Among these EST-quality sequences, 113 ESTs were successfully submitted to the dbEST at GenBanK. A total of 7.6% of these EST-quality sequences were previously identified sequence of Schistosoma japonicum, while 4.5% were putatively identified sequences of Schistosoma japonicum. A total of 23.5% of these EST-quality sequences were putatively identified sequence of Schistosoma mansoni or other organisms. 57.6% had no matches in the database and were classified as unknown sequences. Most ESTs with the putative protein identified belonged to housekeeping proteins. Information about several interesting genes was found.</p><p><b>CONCLUSION</b>Partial cDNA sequencing to generate expressed sequence tags (ESTs) has the potential to rapidly and economically increase our knowledge about adult stage Schistosoma japonicum (Chinese strain) expressed genes.</p>

Effects of Alcohol Extract of Herba Epimedii on the Release of NO from Human Umbilical Vein Endothelial Cells in Vitro / 中药新药与临床药理

To investigate the mechanism of Herba Epimedii alcoho l extract for imp otence, Giess reagent was used to observe the effect of the alcohol extract of H erba Epimedii in mice and its effect on release of NO fro m human umbilical vein endothelial cells in vitro was also studied. The results showed that alcoh ol extract of Herba Epimdii had no effect on the release of NO. But the serum co llected from the mice after the medication of alcohol extract of Herba Epimedii for 120min and 180min promoted the production of NO, and the reaction reached th e peak at 120 min. It is indicated that the promotion of NO release from endothe lial cells may be one of the mechanisms of Herba Epimedii for impotence.

Effect of Xiao Chaihu Decoction on the Release of NO from Mouse Peritoneal Microphage in Vitro / 中药新药与临床药理

To explore the antihepatotoxic mechanism of alcohol_extract of Xiao Chaihu Decoction (XCD),Giress reagent was used to observe the effect of the sero_alcohol_extract of XCD and alcohol_extract of XCD on the release of NO from mouse peritoneal macrophage in vitro.The results showed that alcohol_extract of XCD had no effect on the release of NO while the production of NO was improved in the cultured peritoneal macrophage after the medication of sero_alcohol_extract of XCD for 120min,180min and 240min.It is indicated that improving the release of NO from macrophage was one of the mechanisms of XCD in counteracting hepatic damage.

Effects of Alcohol Extract of Herba Epimedii on the Release of NO from Human Umbilical Vein Endothelial Cells in Vitro / 中药新药与临床药理

To investigate the mechanism of Herba Epimedii alcohol extract for impotence, Giess reagent was used to observe the effect of the alcohol extract of Herba Epimedii in mice and its effect on release of NO from human umbilical vein endothelial cells in vitro was also studied. The results showed that alcohol extract of Herba Epimdii had no effect on the release of NO. But the serum collected from the mice after the medication of alcohol extract of Herba Epimedii for 120min and 180min promoted the production of NO, and the reaction reached the peak at 120 min. It is indicated that the promotion of NO release from endothelial cells may be one of the mechanisms of Herba Epimedii for impotence.