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Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis is a group of systemic small vasculitis characterized by the detection of ANCA in serum. Bactericidal permeability enhancing protein (BPI) is one of the target antigens of ANCA. BPI-ANCA-associated vasculitis is not common clinically, and the combination of bronchiectasis is not accidental. The paper reported a case of BPI-ANCA-associated vasculitis with renal damage combined with bronchiectasis. We reviewed relevant literature to explore the characteristics of BPI-ANCA-associated vasculitis and the correlation between bronchiectasis and ANCA-associated vasculitis, so as to improve the clinician's understanding on this disease.
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Objective To study the antibacterial and tissue reparative effect of BPI-BD3 gene-modified mesenchymal stem cells in a mouse model of wound infection. MethodsC3H10T1/2 cells were transfected with recombinant adenovirus vector pAdxsi-BPI-BD3, the expression of BPI-BD3 fusion protein was verified by RT-PCR and Western blotting. Excision wound with a diameter of 1cm was inoculated with Staphylococcus aureus was made on the back of 30 mice. The mice were randomly divided into 3 groups (10 each). Mice in group T were injected with BPI-BD3 gene-modified C3H10T1/2 cells through caudal vein, those in group C were injected with unmodified C3H10T1/2 cells, and in group N were injected with PBS as control. The wound repair result was evaluated by estimation of the percentage of remaining wound area and the amount of wound bacteria under the scar, followed by observation of pathological changes. Inflammatory reactions of the wounds were assessed accordingly. Results The amount of bacteria under the scar was less in group T than in the other two groups (P<0.05). It was also found that the wound healing process was faster in group T than in group C and group N. Pathological observation showed that the inflammatory reaction in group T was also significantly milder than in the other two groups. Conclusion BPI-BD3 gene-modified mesenchymal stem cells may enhance wound repair by controlling infection and promoting tissue regeneration, thus it may be promising in clinical application.
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Objective:To establish an experimental model for intracellular antibacteria and endotoxin neutralization in vitro to detect the antibacterial and endotoxin neutralization activity of the muBPI_(25) protein.Methods: RAW264.7 cells were transfected with pcDNA3.1(+)muBPI_(36-259), and then were infected with intracellular bacterial of either G ~+/G~-to establish the experimental model of intracellalar antibacteria.The RAW264.7 cells were co-transfected with the pSecTag2B-muBPI_(36-259) and dual-luciferase reporter gene plasmids for establishment of the experimental model of endotoxin neutralization.Results:The experimental model of intracellular antibacteria confirmed that the muBPI_(25) protein could inhibit/kill Salmonella typhi.The experimental model of endotoxin neutralization indicated that the muBPI_(25) protein could neutralize endotoxin.Conelusion: We firstly demonstrate that murine BPI N-terminal functional fragment(muBPI_(25) protein)can inhibit/kill Salmonella typhi,and can neutralize, its lysating product, endotoxin.
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Objective To explore the reparative effect of bactericidal/permeability-increasing protein(BPI) on the damaged mucosa of rats with otitis media with effusion (OME),and state the pathogenesis of OME.Methods Wistar rats(40 ears) were randomly divided into six groups:normal control group (n=4),BPI control group(n=4),eustachian tube obstruction (ETO) group (n=8),lipopolysaccharide (LPS) injection group (n=8),ETO+LPS group (n=8),ETO+LPS+BPI group (n=8).The experimental OME model was made through eustachian tube obstruction and LPS injection.The rats were killed after 1,2 and 4 weeks and the changes of mucosa of middle ear were observed under light and scanning electron microscope.Results The rats in normal control group and BPI control group had the normal mucosa in the tympanic orifice of the eustachian tube.It consisted of pseudostratified ciliated cubical or columnar epithelium which contained an abundant number of ciliated cells and a few goblet cells,these were the mucociliary clearance system of the middle ear.The hypotympanum consisted of thin,squamous epithelium with few microvillus.Middle ear mucosa was obviouly thickened in LPS injection,ETO and ETO+LPS groups.An increase in goblet cells and a decrease in ciliated cells were observed in the tympanic orifice of the eustachian tube.The epithelial layer in the hypotympanum had become more pseudostratified ciliated cubical epithelium.In ETO+LPS+BPI group,there was thin squamous epithelium in the hypotympanum near normal,which was not thickened and contained few microvillus. Conclusion LPS and ETO can result in the occurrence and protracted courses of OME by mimosa's inflammatory reaction which can reduce the activity of ciliary cells and weaken the function of mucociliary clearance system.BPI could bind avidly to LPS,reduce inflammatory reaction,and break the inflammatory cycle and reestablish an effective mucocillary clearance system.The results suggest that BPI treatment is a potential effective drug for prevention and therapy of chronic OME.
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OBJECTIVE To investigate systematically the existence of bactericidal/permeability increasing protein(BPI) in normal rat organs and tissues,for providing important information about BPI in clinical application.METHODS The total RNA was extracted from organs and tissues homogenates.Then the first-strand cDNA was synthesized and BPI DNA was amplified by PCR.RESULTS BPI mRNA was found in kidneys,ovary,testes,liver,small intestine,large intestine,thymus,spleen and neutrophils of 21 various organs and tissues.Among them,BPI mRNA content was the richest in testes,relatively rich in liver and large intestine.CONCLUSIONS BPI mRNA presents in many organs and tissues of rat and it indicates that the role of BPI in host defense against bacterial infection is relatively widespread.
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Endotoxins are lipopolysaccharides ( LPS) , major component of out wall in Gram-negative bacteria. Often they are considered as "prime criminals" of triggering systemic inflaming reaction such as sepsis, bacteremia and so on. It was found in recent years that bactericidal/permeability-increasing protein, a 55kDa member of lipid-related protein family, was a kind of natural molecule of anti-infection and it has special endotoxin-neutralising activity and antibacterial activity. Comprehensive phase II/III clinical trials demonstrated the feasibility and safety of recombinant BPI. So pharmaceutical corporations are attracted to try to apply it to therapy.
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@#ObjectiveTo explore changes of lipopolysaccharide (LPS), lipopolysaccharide-binding protein (LBP), bactericidal/permeability-increasing protein (BPI), soluble CD14 (sCD14) in blood of cirrhosis patients and their clinical significance.MethodsSerum samples of 45 cirrhosis patients were detected with chromogenic limulus amebocyte lysate assay for LPS and detected with ELISA for LBP, BPI and sCD14. While, serum samples of 15 normal subjects were used as controls.ResultsLevels of LPS, LBP, BPI and sCD14 in blood of cirrhosis patients with liver function being grade A, B and C were significantly higher than that in normal subjects. Also, those indexes fore mentioned were obviously higher in died cirrhosis patients than that in survived cirrhosis patients.Conclusion High levels of LBP, LPS and relative deficiency of BPI in cirrhosis patients accompanied with intestinal endotoxemia (IETM) may significantly increase the sensitivity of body to endotoxin.
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Objective: To investigate the associations of single nucleotide polymorphisms(SNP) on bactericidal/permeability-increasing protein(BPI) and lipopolysaccharide-binding protein(LBP) with the complication of sepsis. Methods: The SNP genotypes of BPI Lys216Glu,PstⅠin intron 5,G545C,LBP Cys98Gly and Leu436Pro were examined by restriction fragment length polymorphism PCR(PCR-RFLP).Results:The rate of BPI Lys216Glu G/G genotypes and G-alleles in patients with sepsis were significantly higher than that of volunteers.The rate of BPI Lys216Glu G/G genotypes and G-alleles in nonsurviving patients were significantly higher than that of survivor patients. Conclusion: The BPI Lys216Glu G-allele may be associated with the high risk of sepsis.
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OBJECTIVE To investigate the protective mechanism of recombinant bactericidal permeability-increasing protein 21(rBPI21) in rat endotoxemia.METHODS The different dosage of rBPI21 in endotoxemia rats was injected and the changes in lipopolysaccharide(LPS), lipopolysaccharide binding protein(LBP) and tumor necrosis factor ?(TNF?) contents in blood of different group rats were continuously observed.RESULTS At 6 and 12 hours,the levels of LPS in rBPI21 treatment 1 group(rBPI21 dosage 0.625 mg/kg) were significantly lower than those in endotoxin group at the same time.Serum LBP and TNF? in rBPI21 treatment 1 group were both lower than those in endotoxin group at any time point.Compared with the endotoxin group,the levels of LPS,LBP,and TNF? in rBPI21 treatment 2,3 and 4 groups(rBPI21 dosage 1.25 mg/kg,2.5mg/kg,and 5.0mg/kg,respectively) markedly dropped at any time points,the survival rates were increased from 16.7%(endotoxin group) to 58.3%,91.7% and 100%(rBPI21 treatment 2,3,and 4 groups) individually.CONCLUSIONS The protection of rBPI21 in endotoxemia rats is primarily achieved through neutralizing LPS,decreasing LPS activity in vivo and inhibiting LBP and TNF-? synthesis.
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Human bactericidal/permeability-increasing protein(hBPI)cDNA was amplified by reverse transcription(RT)and touchdown PCR(TD-PCR)from blood stem cells collected from healthy human of Uygur nationality in Xinjiang Uygur Autonomous Region of China, and then was subcloned into pEGFP-N1 vector,hBPI cDNA sequence consists of 1,464bp.Comparison with other 4 hBPI cDNA sequences registered in GenBank identified 99% homology in DNA sequence.However,there were two base substitutions(nucleotide 576G→C,nu- clotide 676A→G),one of which resulted in an amino acid(residue 185 Lys→Glu).
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Objective:To express and secrete rBPI m23 bactericidal protein in Pichia pastoris expression system. Methods:pPICZ? synBPI m600 recombinant expression vector was constructed with prepared vectors (pUC18 synBPI 414 and PBV220 BPI m420 ) according to Molecular Cloning Laboratory Manual. The Pichia pastoris GS115 were electroporated with linearized pPICZ? synBPI m600 vectors , positive clones were screened on low salt LB medium with Zeocin , then the positive clones were identified by PCR and Mut phenotype analysis. rBPI m23 was induced to express in BMMY ,purified by SP Sepharose beads , identified by SDS PAGE and Western blot and measured with BCA method. Results:①pPICZ? synBPI m600 expression vector was successfully constructed.②Pichia pastoris GS115 strains integrated with synBPI m600 stably were obtained.③SDS PAGE showed the molecular weight of the recombinant protein was approximate 23 kD,and Western blot confirmed that the protein could bind to the anti BPI antibody specially.④The supernate protein concentration was about 2 9 mg/L.Conclusion:rBPI m23 was expressed and secreted effectively in Pichia pastoris GS115.
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Objective To investigate the fluctuated pattern of endogenous bactericidal/permeability-increasing protein(BPI)levels,and to elucidate its significance in endotoxin response after major abdominal surgery.Methods Eleven patients undergoing major abdominal surgery were involved in present study.The levels of plasma BPI,lipopolysaccharide-binding protein(LBP)and interleukin-6(IL-6)were measured preoperatively and on postoperative days 1,3,5,7,and 14 by ELISA.Plasma endotoxin concentration was determined with the chromogenic limulus amebocyte lysate(LAL),which was modified by perchloric acid(PCA)pretreatment of samples.The polymorphonuclear neutrophil count was also done at each time point.Results In comparison with normal controls,plasma BPI/LBP ratios [(1.058?0.074)?10-3,P=0.004] were significantly increased on postoperative day 1,maintained at same level on day 3,and declined on day 5.No plasma IL-6 was detected from the blood of all the 11 patients prior to operation,but plasma IL-6 level rose rapidly on day 1 after operation(27.440?12.144pg/ml,P=0.04),maintaining a high level(11.530?5.312pg/ml)on postoperative day 3,and disappeared on day 5.Conclusions Plasma BPI/LBP ratios were markedly elevated during 3 days after a major abdominal surgery,and endogenous elevated BPI may play an important role in prevention from the development of endotoxemia in surgical patients.