Pei-Liang Yang

Nanfang Hospital, Shengcheng, Guangdong, China

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Publications (7)2.45 Total impact

  • Pei-Liang Yang, Xiao-Guang Chen
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    ABSTRACT: Toxoplasma gondii undergoes a complex life cycle that involves multiple development stages, hosts and environments. The ability to transform from one stage to another and adapt to changing environments demands precise regulation of gene expression. Bioinformatic surveys of the sequenced genomes of T. gondii revealed a peculiar absence of DNA-binding transcription factors that are well-conserved from yeast through humans, but a wealth of epigenetic machinery present in T. gondii. Evidence from reports demonstrates that remodeling of the chromatin structure particularly through post-translational modifications of histones, such as acetylation, methylation, phosphorylation, ubiquitination, and sumoylation, is potentially a major process that coordinates regulation of its gene expression. In addition, no-coding RNAs may play an important role in modulating gene expression of T. gondii. These results provide reliable foundations for prevention of toxoplasmosis by revealing its pathogenic mechanism.
    Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases 06/2012; 30(3):228-32.
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    ABSTRACT: The goal of the present study is to develop an enzyme-linked immunosorbent assay (ELISA) using a truncated surface antigen 1 (SAG1) gene of Toxoplasma gondii for the diagnosis of human toxoplasmosis. The truncated SAG1 gene was highly expressed in Escherichia coli. An ELISA kit based on the purified recombinant truncated SAG1 (rtSAG1) was developed, which was used to detect antibodies against T. gondii in human sera. The results showed that the infection of T. gondii could be detected sensitively and specifically by this serologic method. The positive concordance between rtSAG1-ELISA and Western blot, the gold standard, was 93.9% (31/33). However, the positive concordance between the commercial available ELISA Kit 1 (Haitai, Zhuhai, China) and ELISA Kit 2 (DiaSorin ETI-TOXOK-M reverse Plus, Italy) with Western blot was 79.5% (31/39) and 91.2% (31/34), respectively. Comparatively, the positive concordance of ELISA Kit 1 and 2 with Western blot was lower than rtSAG1-ELISA, in particular, the ELISA Kit 1 (P < 0.01), which indicated that the rtSAG1 protein could be used as the diagnostic antigen for human toxoplasmosis.
    Diagnostic microbiology and infectious disease 05/2009; 64(3):261-6. · 2.45 Impact Factor
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    ABSTRACT: To establish a colloidal gold immunochromatographic assay (GICA) for detecting Schistosome japonicum (Sj). Eight monoclonal antibodies (mAbs) against Sj p38 antigen (1A6, 3C4, 3D12, 6F10, 6G12, 9H6, 9G7 and A5H) prepared previously were purified by protein-G affinity chromatography. The affinity constant (K(aff)) was determined by indirect enzyme-linked immunosorbent assay. All the mAbs were labeled with horseradish peroxidase by sodium oxidation method and the mAb pairs with high affinity and stability were identified according to their optical density at 450 nm (OD450). Four mAbs (1A6, 6G12, 9H6 and 9G7) were chosen for colloidal gold labeling. GICA was then performed by further optimization of the labeling and the conditions were determined. The sera of mice at different infection stages were examined with GICA dipstick, with the sera collected before infection and those of Toxoplasma gondii-infected mice as negative controls. The purity of the 8 mAbs was higher than 95% with K(aff) ranging from 2.8x10(-10) to 1x10(-8) mol/L. 9G7 coating (2.5 mg/ml) as the capture antibody and detection with 1A6 (diluted at 1:4) as the labeling antibody was determined as the best reaction model. With this combination, the positivity rates of the detection were 40%, 50%, 60% and 80% for mouse sera collected at 3, 4, 5 and 6 weeks after Schistosome japonicum infection, respectively, without positive results for the negative control samples. GICA established in this study is characterized by simplicity, rapidity and good sensitivity, and the prepared rSjP38 dipstick can test the circulating antigen SjP38 in early stage of infection.
    Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA 06/2005; 25(5):538-41.
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    ABSTRACT: To obtain soluble expression product of immunoreactive recombinant multiepitope antigen of Toxoplasma gondii from E.coli. The gene encoding the multiple epitopes (MEG) of Toxoplasma gondii was amplified by PCR from the original plasmid containing MEG gene and cloned into the prokaryotic soluble expression vector pET32a. After identification by enzyme digestion and sequencing, the positive recombinant plasmid pET32a-MEG was transformed into BL21(DE3), which was induced with IPTG for expression of the target antigen. The relative molecular mass, solubility and antigenicity of the expression products were analyzed by SDS-PAGE and Western blotting. The recombinant expression plasmid pET32a-MEG was successfully constructed and the highly efficient expression of the antigen was achieved after IPTG induction of E.coli. Improvement of the induction condition increased the expression product which accounted for about 28% of the total bacterial protein. The target protein, with good solubility and a relative molecular mass of about 31 000, was purified by immobilized metal affinity chromatography (Ni-NTA resin) and could be well recognized by mouse and rabbit antisera derived by infection of the animals with Toxoplasma gondii B36 and RH, respectively. The recombinant multiepitope antigen has good antigenicity and potential value in diagnosis and vaccine development of toxoplasmosis.
    Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA 06/2005; 25(5):528-30, 544.
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    ABSTRACT: To construct the plant expression vectors containing the multiepitope gene of Toxoplasma gondii (TGMG). 1. TGMG was subcloned into pBAC55 vector to construct the intermediate plasmid pB35MG. The E35S/TGMG/NOS3' fragment was cleaved from pB35MG and ligated into the plant binary vector pCAMBIA2300 to construct the plant expression vector pC35MG. 2. Tomato fruit-specific E81. 1 promoter was introduced to pB35MG to construct pB35E1MG vector. The E35SE81. 1/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pC35E1MG. 3. Tomato fruit-specific E82.2 promoter was inserted to pB35MG to construct pBE2MG vector. The E82.2/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pCE2MG. The insert gene TGMG in the vectors pB35MG, pC35E1MG and pCE2MG were confirmed by sequencing. 4. pC35MG, pC35E1MG and pCE2MG were introduced into Agrobacterium tumefaciens strain LBA4404 competent cell. Digestion with restriction enzymes proved that all recombinant vectors had the inserts with expected length of the target fragments. And the sequencing results were confirmed correct. The TGMG intermediate vectors pB35MG, pB35E1MG and pBE2MG and the plant expression vectors pC35MG, pC35E1MG and pCE2MG were constructed successfully, and the three plant expression vectors were introduced into Agrobacterium tumefaciens.
    Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases 02/2003; 21(2):106-9.
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    ABSTRACT: To study the changes in the constituents of the cercaria antigen of Schistosoma japonicum before and after ultraviolet irradiation. The cercaria of Schistosoma japonicum were exposed to ultraviolet light (UV) irradiation at a dose of 400 mgrW/cm2 for 1 min, and the UV-irradiated cercaria antigen (UVCA) and normal cercaria antigen (NCA) were simultaneously analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. At least 2 antigens with relative molecular mass (Mr) of 212 000 and 82 000 were identified in UVCA but not in NCA by SDS-PAGE analysis, and the concentrations of the antigens with Mr of 116 000, 26 000 and 16 000 in UVCA were significant higher than those in NCA. On the other hand, the antigenic molecule with Mr of 67 000 in NCA was recognized by serum from pigs vaccinated with UV-attenuated cercariae, but not by serum from pigs with Schistosoma japonicum infection. Antigens with Mr of 79 000 and 94 000 were apparently more strongly reactive with the former porcine serum than with the latter. The results suggest that all the novel antigens arising from or increased by UV exposure, or antigens specifically recognized by serum from pigs vaccinated by UV-attenuated carcariae may be the principal factors in the highly protective immunity provoked by irradiated cercariae.
    Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA 09/2002; 22(8):697-9.
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    ABSTRACT: To establish rabbit model of renal allograft transplantation with reduced complications and high survival rate using microsurgical technique. Twelve healthy adult rabbits were randomly divided into 2 groups of equal number, one as donor group and the other recipient. The left kidneys of the donor rabbits were removed followed by immediate reperfusion with 4 degrees celsius H-CA solution, before they were transplanted into the recipient rabbits with their left kidneys excised and end-to-end anastomosis of the renal arteries, veins and ureter respectively performed with microsurgical technique. Another 12 normal rabbits received operations to temporarily block the right renal arteries and veins, serving as control group, in which 11 completed the experiment. No thrombosis or stricture occurred at the site of anastomosis in rabbits with renal allograft transplantation, and the survival rate reached 91.7% (11/12). This rabbit model of renal allograft transplantation has markedly fewer complications with improved survival rate, thus providing a more practical and reliable model for experimental and clinical studies of renal transplantation.
    Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA 09/2002; 22(8):713-5.