Objectve To detect whether the CTP(phosphocholine cytidylyltransferase) gene was expressed in the asexual erythrocytic stages of Plasmodium falciparum (FCC 1/HN )by using the RT - PCR and to construct eukaryotic expression vector of CTP. Method The erythrocytic stage parasites of Plasmodium falciparum were cultured as described by Trager and Jensen. RNA from erythrocytic stage parasite was extracted by using Trizol reagent. The complete genes coding for CTP gene isolates FCCI/HN were amplified by reverse transcriptase -polymerase chain reaction(RT- PCR). CTP gene was cloned into eukaryotic expression vector pcDNA3. Results CTP encoding gene was amplified from the erythrocytic stages of Plasmodiumfalciparum (FCC 1/HN) and eukaryotic expression vector of CTP was constructed. Conclusion CTP gene was expressed in the erythrocytic stages of Plasmodium falciparum (FCC 1/HN) and eukaryotic expression vector of CTP was successfully constructed.

Adenoviridae ; Adenovirus Vaccines ; China ; Humans ; Vaccination ; Vaccines

[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 construct the eukaryotic expression plasmid for the expression of human Connexin26 in COS-7 cells.Methods Total RNA was isolated from human peripheral blood lymphocytes and used as template for the PCR cloning of the human Connexin26 gene.The human Cx26 cDNA containing the 678 bp whole coding region of the human Connexin26 gene was amplified by PCR using specific primers and cloned into the pCI-neo vector to construct the recombinant eukaryotic expression plasmid,pCI-Cx26.The recombinant plasmid was identified by restriction endonuclease digestion,and transfected into COS-7 cells by liposome.The expression of Cx26 mRNA and the protein were analyzed by RT-PCR and SDS-PAGE,respectively.Results Restriction endonuclease digestion analysis verified successful construction of the recombinant plasmid,pCI-Cx26.The expression of Cx26 mRNA and protein in the transfected COS-7 cells were detected by RT-PCR and SDS-PAGE,respectively.Conclusion The eukaryotic expression plasmid for human Cx26 has been constructed successfully with the capability of expression in COS-7 cells.

Objective:To evaluate the immunogenicity of an experimental SARS-CoV inactivated vaccine.Methods:The virus suspension of F69 strain was inactivated with 0.4% formaldehyde and purified,then used as the immune antigen combined with Freund′s adjuvant.Eight adult New Zealand rabbits were immunized 4 times with this vaccine.12 sets of rabbit serum were sampled from the third day to 74th day after first immunization.Titers of specific IgG antibody and neutralizing antibody were determined by indirect ELISA and micro-cytopathic effect neutralizing test.Results:Rapid and potent humoral immune responses were induced by F69 inactivated vaccine in all eight immunized rabbits.Both specific IgG antibody and neutralizing antibody all peaked just with 2 vaccinations,with the maximum titer of 1∶81 920 and 1∶20 480 respectively about 6 weeks after first immunization.Across neutralizing reaction existed between F69 and Z2-Y3 strains.Conclusion:F69 inactivated vaccine owns strong immunogenicity.Similar antigenic epitopes exist between the F69 strain and Z2-Y3 strain,which ensured the cross neutralizing reaction.

China ; Humans ; Public Health ; Research

AIM: To study the pathological change in mouse organs immunitied by inactivated SARS-CoV vaccine. METHODS: Inactivated SARS-CoV vaccine was injected into BALB/c and C57BL/6 mice. Anti-SARS antibody was analyzed by ELISA. After 8 weeks, the immunitied mice were killed and those organs were analyzed by pathological methods. RESULTS: Anti-SARS antibody in mice was positive after 8 days. Only minimal injury was observed in a few lungs and livers, but the other organs were not. CONCLUSIONS: Inactivated SARS-CoV vaccine induced mice to create antibody, whereas they did not cause severe injury. This result will be valuable for vaccine into clinical research. [

OBJECTIVESTo investigate a survey about acceptance of central slaughtering of live poultry in residents of Guangzhou.

METHODSWe conducted a telephone survey by sampling residents with fixed-line telephone and with normal hearing, whose age is more than 15 years, by Mitofsky-Waksberg two-stage method during Jan 6(th) to 8(th), 2014. 358 residents finished the telephone questionnaire by 12 320 health hot line. We investigated the acceptance rate of city-wide central slaughtering permanently. We compared the difference between the respondents and the 2010 Guangzhou census data by Cohen's effect sizes (w) and weighted by population age and sex. We used χ(2) test to compare the acceptance rate of central slaughtering in residents with different characteristic. We used multiple logistic regression analysis to analyze the factors.

RESULTSThe difference in gender and age was small between respondents and the 2010 Guangzhou census data (w value was 0.13, 0.28, respectively), but that in education and marital status was large (w value was 0.52, 0.31, respectively). 49.0% (95% CI: 43.7%-54.3%) accept city-wide central slaughtering permanently. The acceptance rate of city-wide central slaughtering permanently in those who bought fresh, chilled and frozen poultry in their family in previous year was 54.3% (133/245), 60.0% (57/95) and 59.8% (49/82), respectively. It was more than those who didn't buy fresh, chilled and frozen poultry (38.1% (43/113), 44.9% (118/263) and 45.7% (126/276); χ(2) values were 8.15, 6.40 and 5.03; P values were 0.004, 0.011 and 0.025, respectively). The acceptance rate of city-wide central slaughtering permanently in those who deem fresh poultry taste better than live poultry was 64.9% (24/38). It more than those who deem not (47.0%, 151/320) (χ(2) = 4.22, 6.02, P = 0.040, 0.014, respectively). The acceptance rate of city-wide central slaughtering permanently in the male (OR = 2.68, 95% CI: 1.64-4.37) and those who deem getting sick due to buying live birds from LPM (OR = 1.72, 95% CI: 1.05-2.82), who can accept only fresh poultry carcass supply (OR = 2.39, 95% CI: 1.33-4.30), Who bought live poultry in their family in previous year (OR = 0.29, 95% CI: 0.11-0.74), who will decrease the consumption after ban on live poultry sale (OR = 0.50, 95% CI: 0.30-0.83) was 58.6% (109/186), 59.0% (92/156), 60.7% (139/230), 44.9% (132/295), 36.6% (68/186), respectively.

CONCLUSIONIn the early stage of avian influenza A(H7N9) epidemic in Guangzhou, the rate of acceptance of central slaughtering permanently in residents was not so high. Who deem getting sick due to buying live birds from LPM, who could accept only fresh poultry carcass supply and the male more accept city-wide central slaughtering permanently.

Animals ; Attitude to Health ; Birds ; Epidemics ; Humans ; Influenza A Virus, H7N9 Subtype ; Influenza in Birds ; Influenza, Human ; Male ; Meat-Packing Industry ; Poultry ; Surveys and Questionnaires

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

Antibiotic resistance affects the development of the world economy and threats public health. China is one of the countries with the most severe abuse of antibiotics. Here, we describe the causes of antibiotic resistance in the environment, human and animals as well as the status of antibiotic resistance. Based on the concept of One Health, we propose the promotion of the scientific use of antibiotics, the development of new types of antibiotics, establishment of the antibiotics stereoscopic monitoring network system, the promotion of knowledge education of antibiotic resistance, prevention of infection and other measures. We call for the establishment of interdisciplinary, cross-sectoral, trans-regional communication and cooperation to promote the development of antibiotic resistance prevention and control in China to protect environment and the health of humans and animals.