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ABSTRACT: To prokaryotically express and purify fusion protein containing extracellular region of human BTLA and prepare the antiserum of it.
Human BTLA (hBTLA) gene was amplified by PCR, digested with enzymes, ligated and subcloned into a his-tagged prokaryotic expression vector to generate a recombinant plasmid named pET28a-hBTLA. Then pET28a-hBTLA was transformed into E.coli BL21 (DE3). The hBTLA fusion protein was obtained upon IPTG induction, purified by Ni-NTA Purification System, and analyzed by SDS-PAGE. Rabbit anti-hBTLA antiserum was prepared and identified.
The pET28a-hBTLA plasmid was confirmed to carry the correct hBTLA gene by sequencing. It expressed a 15.7kD protein that could be purified by Ni-NTA purification system. The titer of the multiclonal antibody was 1:16 detected by double diffusion test, and 1:20 000 by ELISA, respectively. The anti-hBTLA antibodies can bind to hBTLA specifically shown by Western blot.
The prokaryotic expression vector has been constructed successfully, leading to highly purified hBTLA protein. The antiserum of hBTLA has been prepared, with high titer and specificity.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 04/2011; 27(4):419-21.
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ABSTRACT: To study the clinical significance of determination of serum B7-H4 in patients with malignant hematologic diseases.
Serum B7-H4 levels were determined in 65 patients with leucemia, 34 patients with lymphoma, 12 patients with multiple myeloma as well as in 50 healthy controls.
The serum B7-H4 levels in patients with lymphoma [(38.81+/-10.34) kappag/L] were significantly higher than healthy controls [(31.62+/-9.850) kappag/L] (P<0.01). But there are no significant difference of B7-H4 levels in serum among patients with leucemia, patients with multiple myeloma and healthy controls.
These results suggest that the B7-H4 may correlated with lymphoma, but uncorrelated with leucemia and multiple myeloma. Measurement of serum B7-H4 level provide useful information for distinctive diagnosis of different kinds of malignant hematologic diseases.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 09/2010; 26(9):893-4.
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Jun-Fa Xu,
Huan Xiao,
Guo-Yan Hu,
Shu-Hua Zheng, Wei Liu,
Chun-Lei Yuan,
Heng Yang,
Jing Lü,
Fang Zheng,
Cong-Yi Wang,
Fei-Li Gong
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ABSTRACT: Previous studies demonstrate that both membrane B7-H4 and B7-H4-Ig fusion protein could inhibit T-cell responses. In the present study, we explored the potential effect of B7-H4-Ig on liver injury in a hepatitis mouse model induced by concanavalin A (ConA). A B7-H4-Ig construct was introduced into animals by the hydrodynamic gene delivery approach. It was found that ectopic expression of B7-H4-Ig could inhibit ConA-induced elevation of serum levels of ALT and AST, suppress liver necrosis and even mortality of mice. Furthermore, we observed that pretreatment of B7-H4-Ig dramatically decreased serum levels and the expression of mRNA for IL-2, IFN-gamma and IL-4, but increased IL-10 in ConA-treated mice. Our results suggest that B7-H4-Ig may protect animals from liver injury induced by ConA, which could be associated with reduced serum levels for IL-2, IFN-gamma and IL-4 as well as enhanced IL-10 production.
Clinical Immunology 03/2010; 136(1):30-41. · 4.05 Impact Factor
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ABSTRACT: Mineral oil was selected to protect HeLa cells from water evaporation during low-energy ions implantation in the present paper. Then, HeLa cells having been treated with vacuum and low-energy N+ ions implantation were used to collect ultraviolet absorption spectrum by spectrophotometer. Analytical results indicated that HeLa cells had some characteristic absorption peaks near 202 and 260 nm, respectively. And then the study also found: (1) The spectral intensity increased with the vacuum treatment time. In addition, the effect of vacuum on cellular spectrum was greater than that of mineral oil. (2) The influence of low energy N+ ions on absorption spectrum was far more than that of vacuum. (3) The spectral intensity increased with the implantation dose. According to these results, the effect of low-energy N+ ions implantation and vacuum on tumorous cells (HeLa cells), especially on the molecular configuration and component of tumorous cells (HeLa cells) was discussed. In a word, this study provides a basis for further research on the functionary mechanism of low-energy ions implantation on biomaterial.
Guang pu xue yu guang pu fen xi = Guang pu 08/2009; 29(8):2034-7. · 0.84 Impact Factor
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ABSTRACT: On the basis of classical theory about spectral analysis, the present article used the method of principal component analysis to get the specificity of 83 ultraviolet absorption spectra from mammary gland patient pathology pieces of 83 cases. The authors chose 44 principal component data as training samples and the rest 39 as testing samples. After training discrete and continual sensation network, the authors found that the recognition rate of cancer was only 43.3% and the recognition of noncancerous one was 38.7% when using the discrete sensation network However, because fuzzy-mathematics was introduced to the continual sensation network and the output value of this model was expanded to [0,1], the recognition rate of cancer reached 83.6% and that of noncancerous one was 76.3% when using this model.
Guang pu xue yu guang pu fen xi = Guang pu 11/2008; 28(10):2396-400. · 0.84 Impact Factor
