Mechanism and application of molecular self-assembly in Sup35 prion domain of Saccharomyces cerevisiae / 生物工程学报

Sup35 in its native state is a translation termination factor in Saccharomyces cerevisiae. The prion domain of Sup35p can form amyloid-like proteinaceous fibrils in vitro and in vivo. Furthermore, the in-register cross beta-sheet structure of Sup35p amyloid fibrils is similar to those formed in other species. Therefore, studies on mechanism of Sup35p self-assembly can be an appropriate model to study protein misfolding-related diseases and prion biology. Because of its ability to self-assemble into nanowires, the prion domain of Sup35p has been widely used in biotechnology and nanotechnology.

Transformation of enhanced green fluorescent protein gene in Paecilomyces lilacinus mediated by Agrobacterium tumefaciens / 生物工程学报

The main aim of this study was to transform the enhanced green fluorescent protein gene (egfp) into biocontrol fungus Paecilomyces lilacinus strain 9410. We constructed the expression vector pUPNGT of the fusion gene nptII-egfp using pcDNA3.1(-) as a helper plasmid. The egfp gene was then transformed into P. lilacinus strain 9410 via Agrobacterium tumefaciens-mediated transformation. PCR and Southern blotting analysis showed that the egfp gene was integrated into the genomes of the tested transformants and the integration manner was single-copy. The transformants could generate green fluorescence when they were excited by 488 nm blue laser. These results indicated that the egfp gene had been successfully transformed into P. lilacinus 9410 and expressed in the tested transformants. Our work may provide a new approach to assess environmental safety and practical biocontrol efficacy ofP. lilacinus under different conditions.

Comparison and clinical significance of different imageological methods in the detection of transitional carcinoma of upper urinary tract:Analysis of 234 cases / 北京大学学报(医学版)

Objective:To determine the diagnostic value of multislice CT urography (MSCTU) in patients with transitional cell carcinoma ( TCG) of upper urinary tract by comparing other imageology methods used. Methods: Two hundred and thirty four cases of transitional cell carcinoma of upper urinary tract, in which 82 cases were diagnosed pathologically with pelvic carcinoma and 152 cases with ureteral carcinoma, between June 2004 and September 2006 in our institute were enrolled in a retrospective study. Most of them underwent urological ultrasound, intravenous urogram (IVU) , retrograde pyelography and MSCTU. We compared the positive rate (PR) and diagnostic rate (DR) of these methods used by chi-square test. Results: Among the 234 cases, 215 patients underwent urologic ultrasound, in which 152 cases were detected to be abnormal, with the PR of 70.1% ;Meanwhile, 58 cased were diagnosed by this examination, with the DR of 27. 0%. IVU was performed in 193 patients and 132 cases were found to be abnormal, and the PR was 68. 4% , 65 cases were diagnosed by IVU and the DR was 33.7%. And 132 patients underwent retrograde pyelography, by which 115 cases of lesion were detected, with the PR of 87. 1% ; In the meantime, 93 cases were diagnosed, with the DR of 70. 5%.MSCTU was performed in 226 cases and 220 cases were found to be abnormal, and the PR was 97.3% ;214 cases were diagnosed by MSCTU, with the DR of 94. 7%. The DR of detecting TCC of retrograde pyelography had statistically significant difference with that of ultrasound and IVU(P＜0.001). As compared with retrograde pyelography, MSCTU had statistically significant superiority (P＜0.001). Conclusion: To shorten the diagnosis time and mitigate the sufferings, patients with hematuria supposed to be TCC of upper urinary tract should be recommended to undergo MSCTU first.

Preparation and activity analysis of hu3D3/core-streptavidin fusion protein as a universal carrier targeting to lung cancer / 中华微生物学和免疫学杂志

Objective To prepare a novel fusion protein of hu3D3 and core-streptavidin(hu3D3/cSA) as a universal carrier targeting to lung cancer,and analyze its activities. MethodscSA gene was acquired by PCR,and inserted into plasmid hu3D3/pET-22b(+)to construct a recombinant plasmid hu3D3/cSA/pET-22b(+)．The fusion gene was expressed in E. Coli BL21(DE3).The fusion protein was purified through nickel-affinity chromatography column and renatured using TEA buffer. The tetrameric hu3D3/cSA complex was analyzed by SDS-PAGE and Western blot.The hu3D3/cSA protein was labeled with FITC,then its antigen-binding activity was analyzed using fluorescence microscopy,and the functional affinity of hu3D3/cSA and hu3D3 were analyzed and compared by flow cytometry. The biotin-binding activity of hu3D3/cSA was tested bv EUSA and Western blot. Results The recombinant plasmid hu3D3/cSA/pET-22b(+)with correct sequence was obtained. The fusion protein was found after expression in E. Coli BL21 (DE3)mainly in the form of inclusion body. After being purified and refolded,tetrameric complex was formed. The purified tetrameric hu3D3/cSA complex retained both antigen-binding activity of hu3D3 moiety and biotin-binding activity of cSA moiety:furthermore,the avidity of the hu3D3/cSA to its target antigen increased by about 14 times as compared with that of monomeric hu3D3. Conclusion The fusion protein hu3D3/cSA with both antigen- and biotin-binding activity is SHCCessfully prepared,and the avidity of hu3D3 moiety to 3D3 antigen is enhanced. Consequently,hu3D3/cSA could be a universal carrier targeting to lung cancer, and could be an alternative,convenient method to realize target therapy to lung cancer by the combination of multiple biotinylated anti-tumor drugs or agents.

A comparison of four methods for laparoscopic hysterectomy / 中国微创外科杂志

0 05). No postoperative pyrexia and severe complications occurred in all the four groups. Conclusions The four procedures are all safe and feasible. The selection of these procedures is based on the patient’s individual conditions.

Expression and characterization of fusion protein RGD/tTF for targeting therapy of cancer / 中国免疫学杂志

Objective:To express and purify a new fusion protein(RGD/tTF) for targeting therapy of cancer, and analyze its activities.Methods:The fused gene RGD/tTF was constructed by PCR, and then was inserted into vector pET22 b(+),expressed in E.coli BL21(DE3).The fusion protein was purified through Nickel-affinity chromatography column. The tTF activity of the fusion protein was analyzed by clotting assay and FⅩ activation assay. The specific binding of RGD/tTF to ?_v?_3 was analyzed by indirect ELISA.Results:The recombinant plasmid pET22 b(+)/RGD/tTF with correct sequence was obtained. The fusion protein was expressed with high yield in E.coli BL21(DE3). The purified fusion protein could activiate FⅩ and cause blood coagulation, and bind to ?_v?_3 specifically.Conclusion:The recombinant plasmid pET22 b(+)/RGD/tTF was constructed.The fusion protein retained TF activity and binding specificity to ?_v?_3, lays the foundation for studying the function of inducing thrombosis in tumor vasculature in vivo.