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Objective To understand the capacity of Plasmodium detection and helminth detection and the mastery degree of basic knowledge of parasitic diseases. Methods Three professionals from each city were selected as contestants. The content of the competition contained three parts. The first part included making blood slides and Giemsa staining of Plasmodium,and identifi-cation of species and number with microscopy,the second part included making stool slides with Kato-Katz technique and identifi-cation of common helminth eggs with microscopy,and the third part was basic knowledge of parasitic diseases. Results Totally 51 contestants took part in the competition. The average score of the test of making blood slides was 14.8±3.3,and the passing rate was 82.4%. The average score of the identification of species and number of Plasmodium with microscopy was 19.2 ± 9.3,and the passing rate was 29.4%. The average score of the test of making stool slides was 9.3±0.7,and the passing rate was 100%. The aver-age score of the identification of common helminth eggs with microscopy was 28.0±2.6,and the passing rate was 100%. The aver-age score of basic knowledge of parasitic diseases was 76.3±11.9,and the passing rate was 88.2%. The average score of the test of making blood slides in the female was higher than that in the male(15.7vs.13.5,P<0.05),and the average score of the test of making blood slides in the intermediate title contestants was higher than that in the junior title contestants(16.1vs.14.1,P <0.05). The average score of the basic knowledge of parasitic diseases in the contestants from cities was higher than that in the con-testants from towns(83.2vs.72.6,P<0.05),and the average score of basic knowledge of parasitic diseases in the contestants from high economic level cities was higher than that in the contestants from low economic level cities(82.4vs.71.5,P<0.01). Conclu-sions For professionals in Shandong Province,the capacity of helminth detection was strong and the basic knowledge of parasitic diseases was mastered well,while the capacity of Plasmodium detection was weak.
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Objective To understand the status of intestinal parasitic infections and the related knowledge and behavior in residents of Jiaodong area of Shandong Province,so as to provide the evidence for making an appropriate preventive and control strategy. Methods A total of 18 villages from 6 counties in Jiaodong area were selected as investigation sites according to the stratified sampling method. The feces samples of the permanent residents aged above 3 years were collected and examined by Kato-Katz technique to find the intestinal parasite eggs,and the children under 12 years old were examined by the method of cellophane anal swab to detect the Enterobius vermicularis eggs. In addition,50 households in each survey sites were randomly selected to in-vestigate the basic family situation and the condition of awareness on prevention knowledge and formation of correct behavior of res-idents by using a structured questionnaire. Results Totally 6 163 residents involved in the feces examinations,and the total in-fection rate of intestinal parasites was 6.91%. The infection rates of Trichuris trichiura,Ascaris lumbricoides and hookworm were 6.56%,0.62%and 0.21%,respectively. The infection rate of E. vermicularis in children under 12 years old was 0.51%. The eggs of Clonorchis sinensis and Taenia solium were not found in this survey. The awareness rate of knowledge about preventing parasitic diseases was 49.54%. The formation rates of washing hands before eating,washing hands after using the toilet,never eating raw fruit and vegetable without washing clean,never working in the field with bare feet,and never drinking unboiled water were 97.78%,91.95%,88.81%,92.42%and 86.48%respectively. Conclusions The infection rate of intestinal parasites is low in Ji-aodong area,but there is a significant difference among different counties. The awareness rate of knowledge about preventing para-sitic diseases is low,but the formation rate of healthy behavior is high. In the future,the health education and the strategy of tak- ing medicine among the key population should be enhanced,and the project of reconstructing safe water supply and lavatory should be advanced.
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To obtain the functional fusion protein of rhoptry protein 2, compound rhoptry protein2 and surface antigen 1 of Toxoplasma gondii. the ROP2 and P30 genes from genomic DNA of T.gondii RH strain were amplified by PCR, and were inserted into pMD18-T cloning vector. Then the ROP2 fragment was subcloned to pET-30a(+) plasmid digested by EcoRⅠand Hind Ⅲ to construct plasmid pET-ROP2. Furthermore,the P30 fragment was subcloned into pET-ROP2 digested by BglⅡand EcoRⅠto create plasmid pET-ROP2-P30, the resulting recombinant plasmids , transformed into E.coli BL21 (DE3), were induced with IPTG. and the proteins identified by SDS-PAGE were further purified and refolded. The biological activity was analyzed by Western blot with specific antibody. It was found that the sizes of ROP2 and ROP2-P30 were 1212 and 1896bp with corresponding molecular weight 50- kDa and 75-kDa, respectively. The recombinant protein ROP2 (50-kDa) could specifically react with rabbit-polyclonal antiserum, and complex fusion protein ROP2-P30 (75- kDa) could react with P30 monoclonal antibody.
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To provide the basis for preparation of diagnostic kits and vaccines in Toxoplasma gondii infection, the gene coding for the qualified recombinant p30 protein (SAG1) of this parasite was amplified by PCR, and the amplified gene was cloned into prokaryotic expression vector pET-30a(+) to construct the recombinant plasmid, and then transformed to E.coli DH5α. The positive recombinant plasmid was screened by PCR and double enzymes digestion, and the nucleotide sequence of p30 gene was determined by automated DNA sequencing. Meanwhile, the identified recombinant plasmid was transformed to E.coli BL21(DE3) with the expression of p30 on bacteria induced by IPTG and the expressed protein was identified by SDS-PAGE. The protein obtained was then further purified and refolded, and its biological activity was checked by Western blotting. It was shown that the size of the amplified gene was 750 bp with molecular weight of 30 ku, and this protein could specifically react with monoclonal antibody against p30 protein.
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Objective To study the protective effect of ROP2 nuclei acid vaccine in mice.Methods Forty-two BALB/c mice were divided into three groups.Each mouse in experiment group was injected with 50 ?g recombinant plasmid pc-DNA3-ROP2 through musculus quadriceps fexoris.In control groups,each mouse was injected with 50 ?g blank plasmid pc-DNA3 and with 50 ?l PBS respectively.All mice were immunized for three times with an interval of three weeks.The volume was doubled for the final injection in the two plasmid groups.Blood,spleens and lymph nodes of 4 mice in each group were taken for the detection of CD4+,CD8+ T cells and cytokines 2 weeks after the final immunization.The rest mice in 3 groups were challenged with 500 tachyzoites of Toxoplasm gondii RH strain for further observation.Results The vaccine induced strong cellular and humoral immune response.The titer of antibody in serum was high after inoculation and recognized ROP2 protein antigen expressed in vitro.The lymphocyte phenotype was analyzed.CD4+ T cells proliferated sharply(69.5?3.4)%,and the ratio of CD4+/CD8+ increased considerably by(4.69?1.32)%(P
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Objective To investigate the relationship between the different embolization agent and the spinal cord ischemic injury. Methods Bilateral of intercostal arteries were embolized with gelatin sponge and lipiodol to block the blood supply of spinal cord. To detect the somatosensory evoked potentials(SEPs), 6 canines in both groups of gelatin sponge and lipiodol were treated by bilateral intercostal arteries embolization. The median nerve and fibular nerve were recorded and the ratio of lower extremity to upper extremity SEPs amplitude (LE/UE ratio) were calculated; the results were compared with the pathologic findings and the motor function. Results The baseline, LE/UE, evoked potential amplitude ratios in all dogs before operation ranged from 0.22 to 0.92 with a combined total mean of 0.49?0.07. The SEPs changes at 30 minutes post operation were more obvious than pre operation in gelatin sponge group and lipiodol group with statistic significance. The LE/UE ratio of gelatin sponge group recovered one hour after operation, but this was absent in lipiodol group. So did the latency SEPs changes. Conclusion The SEPs could not be used to evaluate the motor function of spinal cord comprehensively.
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Over a 2 year period,53 transbrachial approachs were selectively performed in 42 cases.5～6F catheters were insertend directly over a guide-wire vie a puncture site 1～3cm over the antecubital fossa of left arm,where the artery would be less mobile.Selective cathetenization of the hepatic artery,superior mesenterie artery,infradiaphragmatic artery,renal artery,internalilliac artery,femoral artery and their branches were suessesfully achieved.No significant complications were encountered.This method proved to be a safe,reliable and well-tolerated ahernetive,though the exemination technique was more in demanding.The indications,techniques and complications of the method were discussed.