Progress of Legumain in malignant tumors / 国际肿瘤学杂志

Legumain, also called asparaginyl endopeptidase, is one new member of the cysteine protease C13 family and plays various roles in human. In recent years, studies have displayed that Legumain is over expressed in a variety of tumors types and promotes the proliferation of tumor cells, tumor invasion and metastasis as well as angiogenesis through varied mechanisms. Further investigation of Legumain will be helpful to elucidate the pathogenesis and progression mechanism of malignant tumors and provide new molecular for targeted therapy of tumors.

Recent Incidence of Paragonimus westermani Metacercariae in Freshwater Crayfish, Cambaroides similis, from Two Enzootic Sites in Jeollanam-do, Korea

An epidemiological study was performed to know the recent infection status of Paragonimus westermani metacercariae (PwMc) in freshwater crayfish, Cambaroides similis, from 2 streams in Jeollanam-do, Republic of Korea. Crayfish were collected from creeks in Bogil-do (Island), Wando-gun, and in a creek near Daeheung Temple in Haenam-gun. The infection rate of crayfish with PwMc in Bogil-do was 89.8%, and the metacercarial burden was 37 PwMc per the infected crayfish. Crayfish in a creek near Daeheung Temple were larger and twice heavier than those in Bogil-do. Of them, 96.5% were infected with PwMc. An average of 140 metacercariae was found in the infected crayfish, almost quadruple to those of Bogil-do. There was a strong correlation between the number of PwMc and body weight of the crayfish. These results suggest that P. westermani metacercariae are still prevalent in crayfish of the 2 regions in Jeollanam-do, Korea.

Effect of modified Huangqi-Jianzhong decoction on the quality of ulcer healing in rats with chronic acetous gastric ulcer / 国际中医中药杂志

Objective To investigate the effect of modified Huangqi-Jianzhong decoction on healing quality of gastric ulcer in rats and to explore its mechanism. Methods 40 Wistar rats were randomly divided into a normal group, a model group, a modified Huangqi-Jianzhong decoction group and a omeprazole group (n=10) according to the random number table method. These rats were used to model of chronic gastric ulcer induced by acetic acid 100%. On third day after surgery, the rats in the modified Huangqi-Jianzhong decoction group were treated with 10% of the modified Huangqi-Jianzhong decoction according to the dose of 20 ml per kilogram of body weight. The rats in the omeprazole group were treated with a 3% omeprazole suspension according to the of 20 ml per kilogram of body weight. The rats in the normal group and the model group were treated with the same volume of saline. After treated with corresponding treatment for 16 days, the gastric mucosa pathology, ulcer index, regenerative mucosal thickness, mucosal surface mucus thickness, muscularis mucosal layer width of the defect, cystically dilated glands number and level of serum prostaglandin (6-K-PGF1α), epidermal growth factor (EGF), nitric oxide (NO) were observed. Results Compared with the model group, the rat ulcer index (8.95 ± 2.78 mm2 vs. 20.82 ± 5.12 mm2), mucosal muscularis defect width (123.56 ± 32.89 μm vs. 229.32 ± 49.69 μm), cystic glandular expansion (1.65 ± 0.76 vs. 6.12 ± 1.23) were lower in the modified Huangqi-Jianzhong decoction group (P<0.01). Thickness of regenerated mucosa (329.55 ± 32.22 μm vs. 198.22 ± 5.83 μm), the surface thickness of mucus (44.3 ± 2.8 μm vs. 24.5 ± 7.8 μm), serum 6-K-PGF1α (93.7 ± 8.9 pg/ml vs. 58.5 ± 5.8 pg/ml), EGF (2.11 ± 0.29 ng/L vs. 0.82 ± 0.20 ng/L) , NO (0.51 ± 0.03 μmol/L vs. 0.22 ± 0.05 μmol/L) was higher (P<0.01). In addition to the ulcer index, other each target improvement flavored were better in the modified Huangqi-Jianzhong decoction group than in the omeprazole group (P<0.05). Conclusion Modified Huangqi-Jianzhong decoction can improve the quality of ulcer healing by promoting gastric mucosa ulcer repair, reducimg ulcer index, mucous muscularis layer defect width and cystic glandular expansion and improving the thickness of regenerative mucosa, mucosal surface mucus thickness and cyst, EGF, NO level and thus improve the experimental gastric ulcer healing quality.

Identification and Molecular Characterization of Parkin in Clonorchis sinensis

Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn2+ were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

Identification and Molecular Characterization of Parkin in Clonorchis sinensis

Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn2+ were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

Expression and identification of the multiple gene ROP2-P30 of Toxoplasma gondii in E.coli BL21 / 中国人兽共患病学报

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.

Studies on the Immuno-Protection of ROP2 Nuclei Acid Vaccine in Toxoplasma gondii Infection / 中国寄生虫学与寄生虫病杂志

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