Bactericidal/permeability-increasing protein （BPI） is an endogenous antibiotic protein with activity against gram-negative bacteria. In the present study, we examined the expression of BPI in postnatal mouse testes ...Bactericidal/permeability-increasing protein （BPI） is an endogenous antibiotic protein with activity against gram-negative bacteria. In the present study, we examined the expression of BPI in postnatal mouse testes and epididymides as well as the subcellular localization within epididymal spermatozoa. Our results showed that, BPI mRNA was expressed in testis and epididymis independently. Throughout the epididymis, the BPI protein level gradually decreased in the epididymal epithelium in a spatial manner, specialized within the cytoplasm of clear cells in the cauda part. We detected BPI proteins in intact acrosome, implying its testicular origin; on the other hand, after the acrosome reaction, BPI proteins were observed dispersed across the entire sperm head, especially enriched at the equatorial segment. Our findings suggested a dual origin of the BPI that generated both in the testis and epididymis, and associated with mouse spermatozoa. BPI protein might be involved in the dynamics modification of the sperm plasma membrane and also the fertilization process.展开更多
objective: To investigate the protective effects of bactericidal/permeability-increa protein (BPIP) on rats after endotoxic shock as to provide more experimental evidence for studies on its clinical use. Methods:E. co...objective: To investigate the protective effects of bactericidal/permeability-increa protein (BPIP) on rats after endotoxic shock as to provide more experimental evidence for studies on its clinical use. Methods:E. coli 026:B6 LPS was injected at a dosage of 12. 5 mg/kg through the artery to reproduce endo toxic shock. BPIP at a dosage of 5 mg/kg (BPIP-treated group) or equal volume of normal saline (control group) were injected immediately after the injection of LPS. Results: ①Survival time of the shocked animals was prolonged and the 24 h survival rate was also significantly increased in BPIP-treated group as compared with the control group. ②The mean arterial pressure, left intraventricular systolic pressure, isovolemic ven tricular pressure and ±dp/dtmax. were significantly higher in BPIP-treated group than in control group. ③ Plasma levels of glutamic-pyruvic transaminase and urea nitrogen were markedly higher but those of endotox in and TNFα were lower in BPIP-treated group than in control group. Conclusion: BPIP can exert significant protective effects on cardiac, hepatic and renal functions in rats after endotoxic shock, indicating that BPIP might be a good choice in treatment of sepsis/septic shock.展开更多
The fusion gene of BPI 23 and human Fcγ1 was obtained by PCR method,and the expression plasmid was constructed to express recombinant BPI 23 \|Fcγ1 fusion protein in CHO cells.After transfection with the plasmid and...The fusion gene of BPI 23 and human Fcγ1 was obtained by PCR method,and the expression plasmid was constructed to express recombinant BPI 23 \|Fcγ1 fusion protein in CHO cells.After transfection with the plasmid and selection by methotrexate,the cell lines expressing the fusion protein were obtained.The recombinant protein was purified using cation\|exchange chromatography and its bioactivity was proved with bactericidal assays.展开更多
目的:提高重组抗菌蛋白BPI23在E.coli D H 5α中的表达水平及提纯后蛋白的复性率。方法:通过降低目的基因5'端序列中G C含量和构建双拷贝重组表达载体pBV 220-(synBPI6002),减少低频密码子数目和增加目的基因拷贝数,并采用不同复性...目的:提高重组抗菌蛋白BPI23在E.coli D H 5α中的表达水平及提纯后蛋白的复性率。方法:通过降低目的基因5'端序列中G C含量和构建双拷贝重组表达载体pBV 220-(synBPI6002),减少低频密码子数目和增加目的基因拷贝数,并采用不同复性方法对提纯后的蛋白进行复性。结果:降低目的基因5'端序列中GC含量后,目的蛋白表达量占菌体总蛋白的14.3%;单双拷贝重组表达载体转化E.coli DH 5α后,目的蛋白的表达水平无明显差异;在复性液中加入PEG 4000后,目的蛋白产生聚集沉淀较少。结论:减少目的基因中GC含量可显著提高BPI23的表达水平,增加表达载体中目的基因的拷贝数对其表达水平无影响,在蛋白复性液中加入PEG 4000可明显抑制蛋白聚集沉淀。展开更多
文摘Bactericidal/permeability-increasing protein （BPI） is an endogenous antibiotic protein with activity against gram-negative bacteria. In the present study, we examined the expression of BPI in postnatal mouse testes and epididymides as well as the subcellular localization within epididymal spermatozoa. Our results showed that, BPI mRNA was expressed in testis and epididymis independently. Throughout the epididymis, the BPI protein level gradually decreased in the epididymal epithelium in a spatial manner, specialized within the cytoplasm of clear cells in the cauda part. We detected BPI proteins in intact acrosome, implying its testicular origin; on the other hand, after the acrosome reaction, BPI proteins were observed dispersed across the entire sperm head, especially enriched at the equatorial segment. Our findings suggested a dual origin of the BPI that generated both in the testis and epididymis, and associated with mouse spermatozoa. BPI protein might be involved in the dynamics modification of the sperm plasma membrane and also the fertilization process.
文摘objective: To investigate the protective effects of bactericidal/permeability-increa protein (BPIP) on rats after endotoxic shock as to provide more experimental evidence for studies on its clinical use. Methods:E. coli 026:B6 LPS was injected at a dosage of 12. 5 mg/kg through the artery to reproduce endo toxic shock. BPIP at a dosage of 5 mg/kg (BPIP-treated group) or equal volume of normal saline (control group) were injected immediately after the injection of LPS. Results: ①Survival time of the shocked animals was prolonged and the 24 h survival rate was also significantly increased in BPIP-treated group as compared with the control group. ②The mean arterial pressure, left intraventricular systolic pressure, isovolemic ven tricular pressure and ±dp/dtmax. were significantly higher in BPIP-treated group than in control group. ③ Plasma levels of glutamic-pyruvic transaminase and urea nitrogen were markedly higher but those of endotox in and TNFα were lower in BPIP-treated group than in control group. Conclusion: BPIP can exert significant protective effects on cardiac, hepatic and renal functions in rats after endotoxic shock, indicating that BPIP might be a good choice in treatment of sepsis/septic shock.
文摘The fusion gene of BPI 23 and human Fcγ1 was obtained by PCR method,and the expression plasmid was constructed to express recombinant BPI 23 \|Fcγ1 fusion protein in CHO cells.After transfection with the plasmid and selection by methotrexate,the cell lines expressing the fusion protein were obtained.The recombinant protein was purified using cation\|exchange chromatography and its bioactivity was proved with bactericidal assays.
文摘目的:提高重组抗菌蛋白BPI23在E.coli D H 5α中的表达水平及提纯后蛋白的复性率。方法:通过降低目的基因5'端序列中G C含量和构建双拷贝重组表达载体pBV 220-(synBPI6002),减少低频密码子数目和增加目的基因拷贝数,并采用不同复性方法对提纯后的蛋白进行复性。结果:降低目的基因5'端序列中GC含量后,目的蛋白表达量占菌体总蛋白的14.3%;单双拷贝重组表达载体转化E.coli DH 5α后,目的蛋白的表达水平无明显差异;在复性液中加入PEG 4000后,目的蛋白产生聚集沉淀较少。结论:减少目的基因中GC含量可显著提高BPI23的表达水平,增加表达载体中目的基因的拷贝数对其表达水平无影响,在蛋白复性液中加入PEG 4000可明显抑制蛋白聚集沉淀。
