Objective] To determine the nucleotide sequence of the 3′\|termal of the RESA gene Plasmodium falciparum isolate FCC1/HN, and find out the differences of the sequences of RESA gene among isolate FCC1/HN,FC27,NF7 and Palo Alto. [Methods] 3′\|terminal fragment of RESA gene of P.falciparum isolate FCC1/HN was amplified by PCR method, then was cloned into pMD18\|T vector. The recombinant was screened and identified by BamHI+XhoI and PCR technique. The nucleotide sequence of the 3′\|terminal of the RESA gene was determined by the dideoxy chain termination method. DNASTAR and BLAST software were used to compare and analyze the RESA gene sequences among the different isolates. [Results] The 3′\|termal fragment of the RESA gene with about 846 bp was specifically amplified by PCR, the recombinant pMD18\|T\|RESA was successfully constructed. Different degrees of diversity of the RESA gene sequences were found among P.falciparum isolates FCC1/HN、FC27,NF7 and Palo Alto. [Conclusion] There were differences in the sequences of RESA gene among the P.falciparum isolate FCC1/HN and three other isolates (FC27,NF7 and Palto alto).

Objective To evaluate the ability to show anatomical structures of the normal middle ears with helical CT virtual endoscopy(CTVE).Methods CTVE were performed to observe anatomical structures of bilateral middle ears in 100 healthy volunteers.Results Anatomical structures of middle ears were displayed stereoscopically by CTVE from multiple views.But superficial and ting structures were showed pooly.Conclusion CTVE is a new noninvasive imaging tool for observe stereoscopically auditory ossicudar chains.

[Objective] To construct a eukaryotic expression plasmid containing a gene encoding a 41-3 kilodalton blood stage antigen (p41-3) of Plasmodiu falciparum isolate FCC1/HN, and to determine the sequence of p41-3 gene and analyze the homology of the sequences of 341-3 gene of different P. falciparum isolates. [ Methods] Two pairs of primers were designed according to the known sequence of p41-3 gene. Using PCR technique, the p41-3 gene was obtained by amplification from genomic DNA of isolate FCC1/HN. By cloning target gene into a eukaryotic expression vector, pcDNA3, a recombinant plasmid pcDNA3-p41-3 was con structed and trarsferred into E. coli DH5α. The positive clones were screened and identified by agarose gel electrophoresis, endonu clease digestion and PCR technique. The correct recombinant plasmid pcDNA3-p41-3 was used as template, and the nucleotide se quence of p41-3 gene was determined by the dideoxy chain termination method. Using softwares to analyze the structure and sequence homology of p41-3 gene between isolate FCC1/HN and FCBR. [Results] The p41-3 gene was specifically amplified from genomic DNA of Plasmodiumm falciparum isolate FCC1/HN, and the correct recombinant plasmid pcDNA3-p41-3 was screened and identi fied. The result of sequence determination showed that the p41-3 gene of isolate FCC1/HN was 2 137 base pairs in full length, encod ing 375 amino acids. Isolate FCC1/HN and isolate FCBR exhibited 98.98 % homology in the nucleiotide sequences and 99.73 % ho mology in the encoded anino acids of p41-3 gene. [Conclusion] The eukaryotic expression plasmid pcDNA3-p41-3 is successfully con structed and nucleotide sequence of p41-3 gene of isolate FCC1/HN is determined. The p41-3 genes of isolate FCC1/HN and isolate FCBR share quite high homology.

Objective To observe the effects of hyperbaric oxygen (HBO) on the expression of FasL mRNA and caspase-3 protein in renal tissue after renal ischemia-reperfusion injury (IRI) in order to elucidate the underlying mechanisms.Methods Rats were randomly divided into thrcc groups: sham group(n=8),IRI group(n=8) and IRI+HBO group(n=8).The IRI group and the IRI+HBO group recieved 45 minutes hibateral renal ischima and the IRI +HBO group received additional HBO therapy at the 1st,24th and 48th hour after ischemia.The kidneys were removed at the end of HBO therapy.Malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were measured to determine the extent of oxidative stress.The expression of FasL mRNA and caspase-3 protein was detected by quantitative real-time PCR and immunohistochemical staining in renal tissue respectively.Results Compared with the sham group,MDA level increased markedly and SOD activity decreased markedly after ischemia.After HBO treatment,MDA level decreased and SOD activity increased significantly (P ＜0.05).In IRI group,the expression of FasL mRNA and caspase-3 protein were higher than those in the sham group (P＜0.01),which were reduced significantly by HBO treatment (P＜0.01).Conclusion The expression of FasL mRNA and caspase-3 protein increases along with the lasting of reperfusion and HBO exhibites protection against cell apoptosis through improving the antioxidant-oxidant balance and reducing IRI in acute stage of IRI.

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.

OBJECTIVETo evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases.

METHODSVero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells.

RESULTSFive pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect.

CONCLUSIONsiRNA may be effective in inhibiting SARS-associated coronavirus replication.

Animals ; Cercopithecus aethiops ; RNA, Small Interfering ; pharmacology ; SARS Virus ; drug effects ; Transfection ; Vero Cells ; Virus Replication ; drug effects

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