Polarization variable-angle synchronous fluorescence spectrometry was proposed to determine samples in turbid solution. A mixture of fluorescein, rhodamine 6G and rhodamine B was used to evaluate the technique. The ba...Polarization variable-angle synchronous fluorescence spectrometry was proposed to determine samples in turbid solution. A mixture of fluorescein, rhodamine 6G and rhodamine B was used to evaluate the technique. The background caused by scattering light was decreased remarkably. The limits of detection were 0.6 ng/ml for fluorescein, 2.3 ng/ml for rhodamine 6G and 4.1 ng/ml for rhodamine B, respectively.展开更多
Caulerpa lentillifera is a green algae that distributes worldwide and is cultivated for food. We assessed vegetative propagation of C. lentillifera by measuring the specific growth rate （SGR） and chlorophyll fluores...Caulerpa lentillifera is a green algae that distributes worldwide and is cultivated for food. We assessed vegetative propagation of C. lentillifera by measuring the specific growth rate （SGR） and chlorophyll fluorescence of the green algae cultured at different salinities and nutrient levels. The results indicated that C. lentillifera can survive in salinities ranging from 20 to 50, and can develop at salinities of 30 to 40. The maximum SGR for C. lentillifera occurred at a salinity of 35. Both chlorophyll content and the ratio of variable to maximum fluorescence （F_v/F_m） were also at a maximum at a salinity of 35. Photosynthesis was inhibited in salinities greater than 45 and less than 25. Both the maximum SGR and maximum chlorophyll content were found in algae treated with a concentration of 0.5 mmol/L of NO3-N and 0.1 mmol/L of PO_4-P. The photosynthetic capacity of photosystem Ⅱ （PSⅡ） was inhibited in cultures of C. lentillifera at high nutrient levels. This occurred when NO_3-N concentrations were greater than 1.0 mmol/L and when PO4-P concentrations were at 0.4 mmol/L. As there is strong need for large-scale cultivation of C. lentillifera, these data contribute important information to ensure optimal results.展开更多
Total RNA was isolated from the hybridoma cell line （LC- 1 ）, which secretes anti-lung adenocarcinoma monoclonal antibody, and was transferred into cDNA. Based on the FRl （framework region l） and FR4 conserved reg...Total RNA was isolated from the hybridoma cell line （LC- 1 ）, which secretes anti-lung adenocarcinoma monoclonal antibody, and was transferred into cDNA. Based on the FRl （framework region l） and FR4 conserved regions of LC-1 gene, the variable regions of heavy chain （Vh） and light chain （Vl） were amplified, and the Vh and modified Vl were connected to single chain Fv （ScFv） by SOE-PCR （splice overlap extension PCR）. The modified ScFv was fused with green fluorescent protein （GFP） and introduced into E. coli JM109. The fusion protein induced by lPTG （Isopropylthiogalactoside） was about 57000 on a 10% SDS-PAGE gel （10% Sds Polyacrylamide Gel Electrophoresis）, and primarily manifested as inclusion bodies. The renatured protein purified by Ni-NTA Superflow resins showed ability to bind to antigen on SPC-A-l lung adenocarcinoma. In addition, the induced host cells fluoresced bright green under 395 nm wavelength, which indicated that the expected protein with dual activity was expressed in the prokaryotic system. The ScFv with GFP tag used in this research can be applied as a new reagent to detect immunological dye, and provide a feasible way to detect adenocarcinoma in a clinical setting.展开更多
目的:研究荧光磁性纳米粒子与前列腺癌特异性抗原(prostate cancer specific antigen,PSA)单链抗体片段结合的复合纳米探针作为前列腺癌核磁共振靶向显像剂与治疗剂的可行性。方法:荧光磁性纳米粒子(FMCNPs)与前列腺癌特异性单链抗体片...目的:研究荧光磁性纳米粒子与前列腺癌特异性抗原(prostate cancer specific antigen,PSA)单链抗体片段结合的复合纳米探针作为前列腺癌核磁共振靶向显像剂与治疗剂的可行性。方法:荧光磁性纳米粒子(FMCNPs)与前列腺癌特异性单链抗体片段(ScFv)桥联制备成复合纳米探针(FMCNPs-ScFv),应用高分辨电镜、荧光光谱仪与磁强度计进行鉴定。将FMCNPs-ScFv与前列腺癌LNCaP细胞共培养,观察其进入癌细胞的靶向性;采用MTT法评价复合纳米探针的细胞毒性。建立裸鼠前列腺癌细胞移植模型,进行免疫组化和病理学鉴定;荷前列腺癌裸鼠尾静脉注射复合纳米探针,采用荧光成像系统观察纳米探针在裸鼠体内的分布消除过程;利用核磁共振观察复合纳米探针肿瘤靶向显像效果;给予体外磁场照射(100W功率)30min,观察肿瘤体积的变化。结果:成功制备复合纳米探针FMCNPs-ScFv;细胞培养实验显示复合纳米探针能够进入前列腺癌细胞质;在显像浓度范围内复合纳米探针细胞毒性很低,不影响细胞增殖。成功制备裸鼠前列腺癌模型。荧光成像系统显示复合纳米探针快速地在裸鼠各重要器官分布,并逐渐集中于肿瘤部位;核磁共振显示纳米探针在24h内能够清晰显示前列腺癌图像;体外磁场照射4d后,注射复合纳米探针的裸鼠肿瘤组织生长显著慢于对照裸鼠的肿瘤组织(P<0.05)。结论:制备的复合纳米探针FMCNPs-ScFv能有效地靶向前列腺癌组织,可用于前列腺癌的核磁共振成像,也能用于体外磁场作用下的肿瘤治疗。展开更多
文摘Polarization variable-angle synchronous fluorescence spectrometry was proposed to determine samples in turbid solution. A mixture of fluorescein, rhodamine 6G and rhodamine B was used to evaluate the technique. The background caused by scattering light was decreased remarkably. The limits of detection were 0.6 ng/ml for fluorescein, 2.3 ng/ml for rhodamine 6G and 4.1 ng/ml for rhodamine B, respectively.
基金Supported by the Technology Program of Basic Research of Qingdao(No.12-1-4-8-(2)-jch)
文摘Caulerpa lentillifera is a green algae that distributes worldwide and is cultivated for food. We assessed vegetative propagation of C. lentillifera by measuring the specific growth rate （SGR） and chlorophyll fluorescence of the green algae cultured at different salinities and nutrient levels. The results indicated that C. lentillifera can survive in salinities ranging from 20 to 50, and can develop at salinities of 30 to 40. The maximum SGR for C. lentillifera occurred at a salinity of 35. Both chlorophyll content and the ratio of variable to maximum fluorescence （F_v/F_m） were also at a maximum at a salinity of 35. Photosynthesis was inhibited in salinities greater than 45 and less than 25. Both the maximum SGR and maximum chlorophyll content were found in algae treated with a concentration of 0.5 mmol/L of NO3-N and 0.1 mmol/L of PO_4-P. The photosynthetic capacity of photosystem Ⅱ （PSⅡ） was inhibited in cultures of C. lentillifera at high nutrient levels. This occurred when NO_3-N concentrations were greater than 1.0 mmol/L and when PO4-P concentrations were at 0.4 mmol/L. As there is strong need for large-scale cultivation of C. lentillifera, these data contribute important information to ensure optimal results.
基金Project (No. 396007) supported by the National Natural ScienceFoundation of China
文摘Total RNA was isolated from the hybridoma cell line （LC- 1 ）, which secretes anti-lung adenocarcinoma monoclonal antibody, and was transferred into cDNA. Based on the FRl （framework region l） and FR4 conserved regions of LC-1 gene, the variable regions of heavy chain （Vh） and light chain （Vl） were amplified, and the Vh and modified Vl were connected to single chain Fv （ScFv） by SOE-PCR （splice overlap extension PCR）. The modified ScFv was fused with green fluorescent protein （GFP） and introduced into E. coli JM109. The fusion protein induced by lPTG （Isopropylthiogalactoside） was about 57000 on a 10% SDS-PAGE gel （10% Sds Polyacrylamide Gel Electrophoresis）, and primarily manifested as inclusion bodies. The renatured protein purified by Ni-NTA Superflow resins showed ability to bind to antigen on SPC-A-l lung adenocarcinoma. In addition, the induced host cells fluoresced bright green under 395 nm wavelength, which indicated that the expected protein with dual activity was expressed in the prokaryotic system. The ScFv with GFP tag used in this research can be applied as a new reagent to detect immunological dye, and provide a feasible way to detect adenocarcinoma in a clinical setting.
文摘目的:研究荧光磁性纳米粒子与前列腺癌特异性抗原(prostate cancer specific antigen,PSA)单链抗体片段结合的复合纳米探针作为前列腺癌核磁共振靶向显像剂与治疗剂的可行性。方法:荧光磁性纳米粒子(FMCNPs)与前列腺癌特异性单链抗体片段(ScFv)桥联制备成复合纳米探针(FMCNPs-ScFv),应用高分辨电镜、荧光光谱仪与磁强度计进行鉴定。将FMCNPs-ScFv与前列腺癌LNCaP细胞共培养,观察其进入癌细胞的靶向性;采用MTT法评价复合纳米探针的细胞毒性。建立裸鼠前列腺癌细胞移植模型,进行免疫组化和病理学鉴定;荷前列腺癌裸鼠尾静脉注射复合纳米探针,采用荧光成像系统观察纳米探针在裸鼠体内的分布消除过程;利用核磁共振观察复合纳米探针肿瘤靶向显像效果;给予体外磁场照射(100W功率)30min,观察肿瘤体积的变化。结果:成功制备复合纳米探针FMCNPs-ScFv;细胞培养实验显示复合纳米探针能够进入前列腺癌细胞质;在显像浓度范围内复合纳米探针细胞毒性很低,不影响细胞增殖。成功制备裸鼠前列腺癌模型。荧光成像系统显示复合纳米探针快速地在裸鼠各重要器官分布,并逐渐集中于肿瘤部位;核磁共振显示纳米探针在24h内能够清晰显示前列腺癌图像;体外磁场照射4d后,注射复合纳米探针的裸鼠肿瘤组织生长显著慢于对照裸鼠的肿瘤组织(P<0.05)。结论:制备的复合纳米探针FMCNPs-ScFv能有效地靶向前列腺癌组织,可用于前列腺癌的核磁共振成像,也能用于体外磁场作用下的肿瘤治疗。
