CCNB1 is involved in bladder cancer pathogenesis and silencing CCNB1 decelerates tumor growth and improves prognosis of bladder cancer.

In search of an effective therapeutic target for bladder urothelial carcinoma (BLCA), the present study aimed to investigate the expression of cyclin B1 (CCNB1) and its putative mechanism in BLCA. BLCA sequencing data from Gene Expression Omnibus and The Cancer Genome Atlas were used to analyze expression of CCNB1 mRNA and high CCNB1 expression had a poorer prognosis compared with those with low expression. Immunohistochemistry (IHC) samples collected from the Human Protein Atlas database were analyzed for CCNB1 protein expression. Short hairpin (sh) CCNB1-transfected BLCA T24 and 5637 cells were used to investigate the effects of CCNB1 and inhibit the proliferation, migration and invasion of BLCA cells, affect the cell cycle distribution and promote apoptosis of 5637 cells. A sh-CCNB1 BLCA chicken embryo chorioallantoic membrane (CAM) transplantation model was established to observe the impacts of sh-CCNB1 on the tumorigenesis of BLCA in vivo. Analysis of sequencing data showed that CCNB1 mRNA was significantly elevated in tumor and BLCA compared with normal tissues [standardized mean difference (SMD)=1.21; 95% CI: 0.26-2.15; I²=95.9%]. IHC indicated that CCNB1 protein was localized in the nucleus and cytoplasm and was significantly increased in BLCA tumor tissues. The in vitro tests demonstrated that proliferation of T24 and 5637 cells transfected with sh-CCNB1 was significantly inhibited and cell migration and invasion ability were significantly decreased. sh-CCNB1 decreased the percentage of T24 cells in G0/G1, 5637 cells in the G0/G1 phase and S phase and increased percentage of 5637 cells in the G2/M phase and increased early apoptosis of 5637 cells. The in vivo experiments demonstrated that the mass of transplanted tumors was significantly decreased compared with the control group following silencing of CCNB1. The present results suggested that CCNB1 was involve in the development and prognosis of BLCA and silencing of CCNB1 may be a promising targeted therapy for BLCA.

[Detection of binding of antibodies with recombinant house dust mite allergen (rDer p2) by surface plasmon resonance technique].

In the present paper, the authors aimed to detect the binding of monoclonal antibody and polyclonal antibodies in sera of patients with recombinant house dust mite allergen (rDer p2) by surface plasmon resonance (SPR) technique. This technique is superior to other methods, like enzyme-linked immunosorbent assays (ELISAs) in studying the interaction between biomolecules because no labeling and sample separation are needed. The allergen rDer p2 was immobilized on carboxymethyldextran-modified sensor chip surface by amine coupling. Surface plasmon resonance measurements of monoclonal antibody and polyclonal antibodies in patients' sera revealed that their bindings diverge widely; the binding of patients' sera was remarkably lower than that of monoclonal antibody. At the same time, the binding of patients' sera with rDer p2 varied among patients allergic to dust mite. This study could provide an easy, fast and real-time way for clinical allergic diseases diagnosis.

[Study on structures of blattela germanica allergen, Bla g 2 expressed by eukaryotic and prokaryotic vectors using fluorescence and CD spectra].

The recombinant allergen, Bla g 2, was expressed by prokaryotic vector E. coli and eukaryotic vector P. Pastoris. The different structures and configurations of the Bla g 2 from E. coli and P. Pastoris were studied by fluorescence and circular dichroism. The secondary structures of Bla g 2 in solution, and the composition besides the type of its tertiary structure were proposed. These studies help understand the differences between prokaryotic and eukaryotic expression systems, reveal the relationship between the structure and the function of Bla g 2, and improve the production of this significant allergen.

[Study of fluorescence spectra of blattela germanica allergen, derenatured and renatured Bla g 2].

Recombinant proteins extracted from inclusion body remain in denaturation status. Renaturation in vitro after initial purification is a key step of downstream processing. A common method of renaturation of recombinant proteins is the dilution method. With Bla g 2 as a model protein, the conformational changes of denatured and renatured Bla g 2 were investigated by applying fluorescence spectra. The effects of different urea concentrations, different SDS concentrations and different pH on the fluorescence intensity of renatured protein were also investigated. The reasons for these were studied with the knowledge of molecular structure.