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        In vitro immunoregulatory role of recombinant Ancylostoma ceylanicum calreticulin

        Tingting Zhuang,Asmaa M.I. Abuzeid,Xiaoyu Chen,Shilan Zhu,Guoqing Li 대한기생충학ㆍ열대의학회 2024 The Korean Journal of Parasitology Vol.62 No.1

        Ancylostoma ceylanicum is a zoonotic soil-derived nematode that parasitizes the intestines of humans and animals (dogs and cats), leading to malnutrition and iron-deficiencyanemia. Helminth parasites secrete calreticulin (CRT), which regulates or blocks thehost’s immune response. However, no data on A. ceylanicum calreticulin (Ace-CRT) areavailable. We investigated the biological function of recombinant Ace-CRT (rAce-CRT). rAce-CRT showed reliable antigenicity and stimulated the proliferation of mouse splenocytes and canine peripheral blood mononuclear cells. Quantitative reverse-transcriptionPCR assays revealed that rAce-CRT primarily promoted the expression of T helper 2 cytokines, particularly IL-13, in canine peripheral blood lymphocytes. rAce-CRT inhibitedcomplement-mediated sheep erythrocyte hemolysis in vitro. Our findings indicate thatAce-CRT plays an immunomodulatory role and may be a promising candidate moleculefor a hookworm vaccine.

      • KCI등재

        Mitochondrial Genome Sequence of Echinostoma revolutum from Red-Crowned Crane (Grus japonensis)

        Rongkun Ran,Qi Zhao,Asmaa M.I. Abuzeid,Yue Huang,Yunqiu Liu,Yongxiang Sun,Long He,Xiu Li,Jumei Liu,Guoqing Li 대한기생충학ㆍ열대의학회 2020 The Korean Journal of Parasitology Vol.58 No.1

        Echinostoma revolutum is a zoonotic food-borne intestinal trematode that can cause intestinal bleeding, enteritis, and diarrhea in human and birds. To identify a suspected E. revolutum trematode from a red-crowned crane (Grus japonensis) and to reveal the genetic characteristics of its mitochondrial (mt) genome, the internal transcribed spacer (ITS) and complete mt genome sequence of this trematode were amplified. The results identified the trematode as E. revolutum. Its entire mt genome sequence was 15,714 bp in length, including 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and one non-coding region (NCR), with 61.73% A+T base content and a significant AT preference. The length of the 22 tRNA genes ranged from 59 bp to 70 bp, and their secondary structure showed the typical cloverleaf and D-loop structure. The length of the large subunit of rRNA (rrnL) and the small subunit of rRNA (rrnS) gene was 1,011 bp and 742 bp, respectively. Phylogenetic trees showed that E. revolutum and E. miyagawai clustered together, belonging to Echinostomatidae with Hypoderaeum conoideum. This study may enrich the mitochondrial gene database of Echinostoma trematodes and provide valuable data for studying the molecular identification and phylogeny of some digenean trematodes.

      • KCI등재

        Establishment of a Tm-shi Method for Detection of Cat-Derived Hookworms

        Yeqi Fu,Yunqiu Liu,Asmaa M.I. Abuzeid,Yue Huang,Xue Zhou,Long He,Qi Zhao,Xiu Li,Jumei Liu,Rongkun Ran,Guoqing Li 대한기생충학ㆍ열대의학회 2019 The Korean Journal of Parasitology Vol.57 No.1

        Melting temperature shift (Tm-shift) is a new detection method that analyze the melting curve on real-time PCR thermocycler using SYBR Green I fluorescent dye. To establish a Tm-shift method for the detection of Ancylostoma ceyl- anicum and A. tubaeforme in cats, specific primers, with GC tail of unequal length attached to their 5 ́ end, were designed based on 2 SNP loci (ITS101 and ITS296) of the internal transcribed spacer 1 (ITS1) sequences. The standard curve of Tm-shift was established using the standard plasmids of A. ceylanicum (AceP) and A. tubaeforme (AtuP). The Tm-shift method stability, sensitivity, and accuracy were tested with reference to the standard curve, and clinical fecal samples were also examined. The results demonstrated that the 2 sets of primers based on the 2 SNPs could accurately distin- guish between A. ceylanicum and A. tubaeforme. The coefficient of variation (CV) of Tm- values of AceP and AtuP was 0.07% and 0.06% in ITS101 and was 0.06% and 0.08% in ITS296, respectively. The minimum detectable DNA concen- tration was 5.22×10-6 and 5.28×10-6 ng/μl samples of AceP and AtuP, respectively. The accuracy of Tm-shift method reached 100% based on examination of 10 hookworm DNA samples with known species. In the clinical detection of hookworm in 69 stray cat fecal sample, the Tm-shift detection results were consistent with the microscopic examination and successfully differentiated between the 2-hookworm species. In conclusion, the developed method is a rapid, sensi- tive and accurate technique and can provide a promising tool for clinical detection and epidemiological investigation of cat-derived hookworms.

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