Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three t...Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannanbinding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP.Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays.Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05).Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.展开更多
Background: Sugar moiety of macromolecules is today very well known for its implications in many biological recognition mechanisms including cell-cell, extracellular matrix-cell and/or bacteria-cell interactions. In t...Background: Sugar moiety of macromolecules is today very well known for its implications in many biological recognition mechanisms including cell-cell, extracellular matrix-cell and/or bacteria-cell interactions. In this context lectins, which are carbohydrate-binding proteins displaying a high affinity for sugar groups of other molecules, are of a great importance, notably in immune response involving bacteria, viruses and fungi. As protein-carbohydrate interactions are often mediated by ions such as calcium, zinc or magnesium, we were prompted to study the effect of a thermal spring water (which contains this type of component) on interactions existing between: 1) osidic receptors of human normal keratinocytes and 2) two lectins greatly implicated in the immune response mechanisms (i.e. the dectin-1 and the langerin), and their ligands. Materials and Methods: In a first series of experiments, we studied the effect of increasing concentrations of a thermal spring water on interactions existing between glycosylated molecules and the osidic receptors expressed at the normal human keratinocytes surface. In a second step, and in order to better understand the putative effect of our thermal spring water on the immune response, we analyzed its effect on the interactions existing between the dectin-1 (implicated in the recognition of bacteria, viruses and fungi) and the langerin (expressed by Langerhans cells, the immune cells of the cutaneous tissue), and their ligands in a model using recombinant human lectins and appropriate binding molecules. Results: We showed here that our thermal spring water was able to reinforce interactions between keratinocytes osidic receptors and some of their ligands, in a dose-related manner: From 8% to 55% of increase with 10% to 30% (v/v) of thermal spring water. In the second part of our studies, we also showed that our thermal spring water was able to modulate interactions between dectin-1 and langerin and their ligands through a biphasic effect: Interactions were enhanced by more than 40% and 20% respectively with 10% of thermal spring water, and return to their basal level or lower for higher concentrations. Conclusion: The tested thermal spring water, probably due to its ionic composition, could significantly affect interactions of osidic receptors with their ligands. This property could be of a great interest to help immune system to maintain an appropriate “vigilance state” by using the thermal water at up to a concentration of 10%, and by avoiding any runaway reaction in case of aggression, by using concentrations higher than 10%. .展开更多
This paper report the binding pattern of 12 different kinds of lectin in human breast lesions. Of the 12 kinds of lectins, WGA showed the highest binding activity to the cells of the breast tissue derived; the binding...This paper report the binding pattern of 12 different kinds of lectin in human breast lesions. Of the 12 kinds of lectins, WGA showed the highest binding activity to the cells of the breast tissue derived; the binding of BSL, SBA and DBA were localized to membrane or cytoplasm of cancer cells and to the lumina membrane border of the normal and benign lesions; PNA receptor is related with the differentiation of the breast; cancer; we didn't find any relationship between the lectin receptor and the tendency of metastasis of breast cancer.展开更多
AIM: To evaluate the early expression of mannose-binding lectin 2(MBL2) in human corneal epithelial cells(HCECs) infected by Aspergillus fumigatus(AF).METHODS: HCECs cultured in vitro with AF antigens and sampled at 0...AIM: To evaluate the early expression of mannose-binding lectin 2(MBL2) in human corneal epithelial cells(HCECs) infected by Aspergillus fumigatus(AF).METHODS: HCECs cultured in vitro with AF antigens and sampled at 0, 0.5, 1, 2, 4, 6 and 8h. The expression of MBL2 m RNA was evaluated by semiquantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression of MBL2 protein in supernatant fluid was shown by enzyme linked immunosorbent assay(ELISA). MBL2 protein in HCECs was detected by immunocytochemistry at 0 and 24 h.RESULTS: MBL2 m RNA and protein are expressed in normal HCECs. The expression of MBL2 m RNA and protein in supernatant fluid begin to increase after being stimulated with AF antigens. The most significantly peak of MBL2 m RNA is in 2h. The protein of MBL2 in supernatant fluid decrease gradually after 0.5h. The protein in HCECs expression increase after stimulation of24 h.· CONCLUSION: MBL2 receptor expressed in normal HCECs in vitro. The stimulation by AF antigens can increase the early expression of it.展开更多
The electron microscopic topology of Con A and WGA receptors was investi-gated in human peritoneal macrophagcs by utilizing avidin-gold colloids.Our observa-tions confirmed that both the receptors were distributed on ...The electron microscopic topology of Con A and WGA receptors was investi-gated in human peritoneal macrophagcs by utilizing avidin-gold colloids.Our observa-tions confirmed that both the receptors were distributed on the cell membranes,yet thereceptor expression showed variations among the individual cells.Some cells had abun-dant receptors,while others had only few receptors.The data obtained in the presentstudy indicate that human peritoneal macrophages may be a functionally andbiochemically heterogenous population,and the study may also serve as a theorcticalreference for future clinical exploitation of human peritoneal macrophages.展开更多
目的探讨人转运甲状腺素蛋白(transthyretin,TTR)、人凝聚素(human clusterin,CLU)、白细胞介素-6(interleukin-6,IL-6)与脑卒中后继发认知功能障碍的相关性及预后预测价值。方法选取高邮市中医医院收治的500例脑卒中患者,根据简易智能...目的探讨人转运甲状腺素蛋白(transthyretin,TTR)、人凝聚素(human clusterin,CLU)、白细胞介素-6(interleukin-6,IL-6)与脑卒中后继发认知功能障碍的相关性及预后预测价值。方法选取高邮市中医医院收治的500例脑卒中患者,根据简易智能精神状态检查量表(mini-mental state examination,MMSE)得分,分为认知功能障碍组180例和非认知功能障碍组320例,另选取200名健康志愿者作为对照组,对比3组受检者TTR、CLU、IL-6表达水平,并分析与脑卒中后继发认知功能障碍的相关性。随访患者预后情况,根据180例脑卒中后继发认知功能障碍患者治疗后的MMSE得分分为预后良好组110例和预后不良组70例,对比2组患者临床一般情况与TTR、CLU、IL-6表达水平,并用Logistic回归分析分析这些指标对卒中后继发认知功能障碍的预后预测价值。结果认知功能障碍组TTR、IL-6高于非认知功能障碍组和对照组,CLU低于非认知功能障碍组和对照组(P<0.05);Spearman相关分析结果显示,TTR、IL-6与脑卒中后继发认知功能障碍呈正相关,与CLU呈负相关(P>0.05);预后良好组与预后不良组患者体质量指数,是否首发,TTR、CLU、IL-6水平对比差异有统计学意义(P<0.05);Logistic回归分析显示,TTR、CLU、IL-6为脑卒中后继发认知功能障碍的预后独立影响因素(P<0.05)。结论血清TTR、CLU、IL-6水平不仅与脑卒中后继发认知功能障碍具有明显相关性,还为其预后独立影响因素。因此,临床上可考虑将TTR、CLU、IL-6三者作为脑卒中后认知功能障碍的诊断及预后预测指标。展开更多
基金funded by the National Key R&D Program of China[2021YFC2301102]National Natural Science Foundation of China[82202593]Key R&D Program of Hebei Province[223777100D].
文摘Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannanbinding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP.Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays.Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05).Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.
文摘Background: Sugar moiety of macromolecules is today very well known for its implications in many biological recognition mechanisms including cell-cell, extracellular matrix-cell and/or bacteria-cell interactions. In this context lectins, which are carbohydrate-binding proteins displaying a high affinity for sugar groups of other molecules, are of a great importance, notably in immune response involving bacteria, viruses and fungi. As protein-carbohydrate interactions are often mediated by ions such as calcium, zinc or magnesium, we were prompted to study the effect of a thermal spring water (which contains this type of component) on interactions existing between: 1) osidic receptors of human normal keratinocytes and 2) two lectins greatly implicated in the immune response mechanisms (i.e. the dectin-1 and the langerin), and their ligands. Materials and Methods: In a first series of experiments, we studied the effect of increasing concentrations of a thermal spring water on interactions existing between glycosylated molecules and the osidic receptors expressed at the normal human keratinocytes surface. In a second step, and in order to better understand the putative effect of our thermal spring water on the immune response, we analyzed its effect on the interactions existing between the dectin-1 (implicated in the recognition of bacteria, viruses and fungi) and the langerin (expressed by Langerhans cells, the immune cells of the cutaneous tissue), and their ligands in a model using recombinant human lectins and appropriate binding molecules. Results: We showed here that our thermal spring water was able to reinforce interactions between keratinocytes osidic receptors and some of their ligands, in a dose-related manner: From 8% to 55% of increase with 10% to 30% (v/v) of thermal spring water. In the second part of our studies, we also showed that our thermal spring water was able to modulate interactions between dectin-1 and langerin and their ligands through a biphasic effect: Interactions were enhanced by more than 40% and 20% respectively with 10% of thermal spring water, and return to their basal level or lower for higher concentrations. Conclusion: The tested thermal spring water, probably due to its ionic composition, could significantly affect interactions of osidic receptors with their ligands. This property could be of a great interest to help immune system to maintain an appropriate “vigilance state” by using the thermal water at up to a concentration of 10%, and by avoiding any runaway reaction in case of aggression, by using concentrations higher than 10%. .
文摘This paper report the binding pattern of 12 different kinds of lectin in human breast lesions. Of the 12 kinds of lectins, WGA showed the highest binding activity to the cells of the breast tissue derived; the binding of BSL, SBA and DBA were localized to membrane or cytoplasm of cancer cells and to the lumina membrane border of the normal and benign lesions; PNA receptor is related with the differentiation of the breast; cancer; we didn't find any relationship between the lectin receptor and the tendency of metastasis of breast cancer.
基金Supported by National Natural Science Foundation of China(No.81170825No.81300730)
文摘AIM: To evaluate the early expression of mannose-binding lectin 2(MBL2) in human corneal epithelial cells(HCECs) infected by Aspergillus fumigatus(AF).METHODS: HCECs cultured in vitro with AF antigens and sampled at 0, 0.5, 1, 2, 4, 6 and 8h. The expression of MBL2 m RNA was evaluated by semiquantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression of MBL2 protein in supernatant fluid was shown by enzyme linked immunosorbent assay(ELISA). MBL2 protein in HCECs was detected by immunocytochemistry at 0 and 24 h.RESULTS: MBL2 m RNA and protein are expressed in normal HCECs. The expression of MBL2 m RNA and protein in supernatant fluid begin to increase after being stimulated with AF antigens. The most significantly peak of MBL2 m RNA is in 2h. The protein of MBL2 in supernatant fluid decrease gradually after 0.5h. The protein in HCECs expression increase after stimulation of24 h.· CONCLUSION: MBL2 receptor expressed in normal HCECs in vitro. The stimulation by AF antigens can increase the early expression of it.
文摘The electron microscopic topology of Con A and WGA receptors was investi-gated in human peritoneal macrophagcs by utilizing avidin-gold colloids.Our observa-tions confirmed that both the receptors were distributed on the cell membranes,yet thereceptor expression showed variations among the individual cells.Some cells had abun-dant receptors,while others had only few receptors.The data obtained in the presentstudy indicate that human peritoneal macrophages may be a functionally andbiochemically heterogenous population,and the study may also serve as a theorcticalreference for future clinical exploitation of human peritoneal macrophages.
文摘目的探讨人转运甲状腺素蛋白(transthyretin,TTR)、人凝聚素(human clusterin,CLU)、白细胞介素-6(interleukin-6,IL-6)与脑卒中后继发认知功能障碍的相关性及预后预测价值。方法选取高邮市中医医院收治的500例脑卒中患者,根据简易智能精神状态检查量表(mini-mental state examination,MMSE)得分,分为认知功能障碍组180例和非认知功能障碍组320例,另选取200名健康志愿者作为对照组,对比3组受检者TTR、CLU、IL-6表达水平,并分析与脑卒中后继发认知功能障碍的相关性。随访患者预后情况,根据180例脑卒中后继发认知功能障碍患者治疗后的MMSE得分分为预后良好组110例和预后不良组70例,对比2组患者临床一般情况与TTR、CLU、IL-6表达水平,并用Logistic回归分析分析这些指标对卒中后继发认知功能障碍的预后预测价值。结果认知功能障碍组TTR、IL-6高于非认知功能障碍组和对照组,CLU低于非认知功能障碍组和对照组(P<0.05);Spearman相关分析结果显示,TTR、IL-6与脑卒中后继发认知功能障碍呈正相关,与CLU呈负相关(P>0.05);预后良好组与预后不良组患者体质量指数,是否首发,TTR、CLU、IL-6水平对比差异有统计学意义(P<0.05);Logistic回归分析显示,TTR、CLU、IL-6为脑卒中后继发认知功能障碍的预后独立影响因素(P<0.05)。结论血清TTR、CLU、IL-6水平不仅与脑卒中后继发认知功能障碍具有明显相关性,还为其预后独立影响因素。因此,临床上可考虑将TTR、CLU、IL-6三者作为脑卒中后认知功能障碍的诊断及预后预测指标。
