关于自身免疫性抗体的实验诊断与临床研究

抗核抗体阳性提示患者结缔组织病的可能,但仍要结合患者的临床表现及各结缔组织病的诊断标准。结缔组织病的易感人群一般为中青年女性,老年患者的自身抗体的滴度会升高,但一般为低水平。抗核抗体为一大类自身抗体的总称,ANA阳性进一步分型为针对多种核抗原成份特异性的自身抗体,根据间接免疫荧光的荧光模型(抗原底物为HEP-2和肝片)可区分不同的荧光核型,不同核型也有鉴别意义。不同的结缔组织病可出现特征性抗体:如SLE相关的自身抗体有Sm抗体、ds-DNA抗体、皮肌炎相关的JO-1抗体、MCTD相关的u1RNP抗体等等。     

多价EZH2肿瘤相关抗原肽的制备及其抗原性

目的人工设计制备多价EZH2肿瘤相关抗原肽,并鉴定其抗原特性。方法设计合成编码多价EZH2肿瘤相关抗原肽的DNA序列,克隆入原核表达载体p ET-30a(+)中,构建含4个EZH2肿瘤相关抗原肽段(Aa120-128、Aa165-174、Aa291-299、Aa735-742)的重组多价抗原肽原核表达质粒p ET-30a(+)-EZH2-antigen,转化E.coli BL21(DE3),经IPTG诱导表达后,用Ni-NTA树脂在变性条件下对重组多肽进行亲和层析纯化,Western blot分析重组多价EZH2肿瘤相关抗原肽的抗原特性。用重组多价EZH2肿瘤相关抗原肽免疫大耳白兔,制备多克隆抗体,间接ELISA法检测血清中抗体效价,Western bolt分析多抗的特异性。结果重组原核表达质粒p ET-30a(+)-EZH2-antigen经酶切及测序证实构建正确;表达的重组多价EZH2肿瘤相关抗原肽表观相对分子质量约4 300,主要以包涵体形式存在;纯化的多肽纯度不低于90%,浓度为0.5 mg/ml,且能被兔抗EZH2多抗和兔抗His标签多抗有效识别和结合;纯化的多价EZH2肿瘤相关抗原肽能在动物体内高效诱导抗体生成,抗血清效价为1∶64×104,且能特异性识别和结合PC3人前列腺癌细胞中表达的EZH2蛋白。结论成功制备了一种多价EZH2肿瘤相关抗原肽,该多价抗原肽相对分子质量虽小,但抗原肽密度高,可能具有更强的肿瘤相关抗原的活性,具有重要的应用前景。     

鼠抗GM-CSF单克隆抗体的研究

应用基因工程GM－CSF抗原，通过杂交瘤技术获得3株稳定分泌抗GM－CSF的特异性单抗细胞株，分别命名为LD2、4和6。经分析，3种单抗识别抗原分子上的两个不完全相关的位区，为GM－CSF的亲和层析纯化及抗原的免疫学测定提供了必要的条件。     

肺炎球菌菌种复苏用无动物源性固体培养基的筛选

目的 筛选一种可复苏肺炎球菌(Streptococcus pneumoniae,Spn)菌种的无动物源性物料的固体培养基。方法 首先选取19F血清型Spn对无动物源材料的9种配方培养基进行初步筛选,再对其他23个疫苗血清型Spn进行培养测试验证。通过对传代15代的培养物进行菌种检定与荚膜多糖抗原基因序列分析,判断筛选的配方培养基对各型菌种传代稳定性的影响。用此培养基复苏的菌种按生产程序进行发酵培养,分析培养情况及荚膜多糖抗原产量及质量,确定筛选培养基的生产适用性。结果 在筛选的配方固体培养基上,各型Spn培养11～15h时活菌数量较高,15代终末代次的菌种检定、抗原基因序列与对照(羊血固体培养基培养的菌种)相比无差异。用其复苏的菌种发酵培养后,菌体生长与荚膜多糖产量略有提高,且荚膜多糖相关检定指标均符合《中国药典》三部(2020版)标准。结论 筛选的无动物源性固体培养基具有良好的适用性,可取代血液来源培养基用于Spn疫苗生产菌种复苏。     

免疫原性细胞死亡及其在肿瘤治疗应用中的研究进展

免疫原性细胞死亡(immunogenic cell death,ICD)是细胞死亡的一种形式,通过释放肿瘤相关抗原(tumor asso-ciated antigen,TAA)和肿瘤特异性抗原(tumor specific antigen,TSA),暴露"危险信号"以刺激机体免疫系统产生免疫应答,其特点是释放和/或增加...     

宫颈癌化疗敏感性研究进展

许多研究表明,宫颈癌细胞内在的抗原成分、基因、蛋白与化疗的敏感性相关。可以通过检测患者体内的相关标志物来判断患者的病情,为做出综合治疗方案提供依据。     

人源性抗甲型肝炎病毒Fab噬菌体抗体分子的初步筛选

目的从自建的大容量非免疫Fab噬菌体抗体库中筛选具有抗甲型肝炎病毒(HAV)抗原活性的噬菌体抗体,并进行鉴定。方法采用纯化的甲型肝炎病毒作为包被抗原,对自建的容量为1.26×1011pfu/ml的Fab噬菌体抗体库进行筛选和富集,分别对原始库和筛选后的抗体库进行Phage-ELISA和活性检测,经酶切鉴定和DNA测序分析后进行诱导表达。结果经2轮筛选,抗-HAV抗原的噬菌体抗体得到了明显富集,ELISA检测证实其具有良好的抗甲肝病毒抗原的特异性,抑制率为38.61%。酶切鉴定表明抗体库重组较稳定。SDS-PAGE检测显示Fab抗体蛋白在E.coliDH5α中得到了表达。结论从自建的非免疫Fab噬菌体抗体库中成功获得了抗甲肝病毒噬菌体抗体,所构建的非免疫抗体库可用于人源性抗体的筛选。     

脊髓灰质炎病毒Ⅲ型Sabin株C抗原和D抗原的分离及特性分析

目的分离脊髓灰质炎病毒Ⅲ型Sabin株(sPVⅢ)C抗原和D抗原,并分析其部分特性。方法采用CsCl密度梯度离心法分离sPVⅢ的C抗原和D抗原,电镜观察病毒形态;SDS-PAGE检测病毒结构蛋白组成;微量BCA法检测蛋白浓度,计算每种抗原所占比例;细胞病变法检测病毒滴度。同时检测上样浓度及超离次数对C抗原纯度的影响。结果sPVⅢC抗原和D抗原在透射电镜下均呈球形,C抗原病毒颗粒密度较小,外壳大多较为松散,D抗原病毒颗粒密度较大,结构更稳固;C抗原由VP0、VP1、VP3蛋白组成,D抗原由VP1、VP2、VP3和VP4蛋白组成;C抗原和D抗原分别约占总蛋白含量的10%和90%,部分C抗原在分离前及分离过程中易产生聚集;D抗原的病毒滴度高于C抗原。降低上样浓度及进行二次超离可提高C抗原纯度。结论采用CsCl密度梯度离心法成功分离了sPVⅢ的C抗原和D抗原,C抗原滴度远低于D抗原,且部分易产生聚集,较难达到较高纯度。     

猪血凝性脑脊髓炎病毒HE基因的克隆及原核表达

目的构建猪血凝性脑脊髓炎病毒株(67N)血凝素酯酶蛋白(HE蛋白)原核表达系统,为建立特异性免疫学诊断方法提供备选材料。方法克隆猪血凝性脑脊髓炎病毒(67N)HE蛋白抗原表位富集区基因,构建重组表达质粒pET-HE,并在大肠杆菌中诱导表达。经包涵体的初步纯化,金属螯合层析进一步纯化HE蛋白,SDS-PAGE和Western blot进行检测。结果大肠杆菌可高效表达HE蛋白,目的蛋白表达量可占菌体总蛋白的42.2%。纯化后蛋白浓度为0.6 mg/ml,纯度为85.4%。所表达的蛋白可被兔抗猪血凝性脑脊髓炎阳性血清所识别。结论已成功构建猪血凝性脑脊髓炎病毒HE蛋白原核表达载体,重组HE蛋白抗原具有良好的特异性,可作为检测猪血凝性脑脊髓炎病毒血清抗体的候选抗原。     

酿酒酵母钙通道膜蛋白单克隆抗体制备及鉴定

以酿酒酵母电压门控钙通道膜蛋白（Cch1p）、牵张敏感性钙通道膜蛋白（Mid1p）和瞬时受体电位钙通道膜蛋白（Yvc1p）为研究材料，制备其单克隆抗体。采用生物信息学方法确定3种膜蛋白抗原表位，根据分析结果克隆抗原基因，并进行原核表达和表达产物分析鉴定，通过Ni²⁺-NTA树脂亲和层析技术获得重组抗原蛋白，免疫小鼠后细胞融合技术制备单克隆抗体，酶联免疫吸附测定（ELISA）检测抗体效价，免疫印迹技术检测单克隆抗体对重组纯化抗原和天然酿酒酵母钙通道膜蛋白的反应性和特异性。生物信息学分析结果表明，Cch1p、Mid1p和Yvc1p抗原表位可能分别位于1~300位氨基酸残基、359~548位氨基酸残基、1~236位氨基酸残基；克隆目的基因条带大小分别为926bp、570bp和708bp，与预期结果一致；原核表达抗原蛋白分子量分别为60000、25000和30000，Western blot检测条带正确；重组纯化抗原免疫BALB/c小鼠，细胞融合技术制备单克隆抗体，ELISA检测显示单克隆抗体效价分别高达1∶256000、1∶128000和1∶64000，Western blot检测到3种重组纯化抗原和天然酿酒酵母钙通道膜蛋白Cch1p、Mid1p和Yvc1p。这些结果说明本文制备的单克隆抗体可以成功用于检测酿酒酵母钙通道膜蛋白Cch1p、Mid1p和Yvc1p表达的相关研究。     

The Extraction and Separation of Ho,Y, and Er(III) with the Mixtures of Cyanex 302 and Another Organic Extractant

Abstract

The extraction and separation of Ho, Y, and Er(III) with the mixtures of bis(2,4,4‐trimetylpentyl)monothiophosphinic acid (Cyanex 302) and another organic extractant, such as acidic organic extractant (di‐2‐ethylhexyl phosphoric acid P204, 2‐ethylhexyl phosphoric acid mono‐2‐ethylhexyl ester P507, di‐2‐ethylhexyl phosphinic acid P229, and sec‐nonylphenoxy acetic acid CA‐100), neutral organic extractant (tri‐n‐butyl phosphate TBP, di‐(1‐metylheptyl)metyl phosphate P350, and branched trialkylphosphinic oxide Cyanex 925) or primary amine N1923, has been investigated in this paper. The extractability and separation ability for the Ho, Y, and Er with the mixtures of Cyanex 302 and organic extractants has been compared. The synergistic effect of the Ho, Y, and Er extraction with the mixtures of Cyanex 302 and P229, Cyanex 925, CA‐100, or N1923 has been explored and the synergistic enhancement coefficients have been calculated. At last, the Y³⁺ synergistic extraction with the mixtures of Cyanex 302 and CA‐100 has been determined and the extracted complex has been deduced.     

Fe-EDTA-Bisamide and Fe-ADR-925, The Iron-Bound Hydrolysis Product of the Cardioprotective Agent Dexrazoxane, Cleave DNA Via the Hydroxyl Radical

Use of the antitumor drug doxorubicin is limited by cardiomyopathic side-effects which are believed to be due to iron-mediated hydroxyl radical generation. Dexrazoxane reduces this cardiotoxicity, possibly by removal of iron from doxorubicin by the EDTA-like hydrolysis product of dexrazoxane, ADR-925. However, EDTA-diimides like dexrazoxane, previously used as antitumor agents, are themselves carcinogenic, and recent studies have found that Fe-ADR-925 can also promote hydroxyl radical production. This study demonstrates that, like Fe-EDTA, Fe-ADR-925 and a related desmethyl complex can cleave plasmid DNA under Fenton conditions, and suggests by radical scavenger study that this cleavage is probably via the hydroxyl radical. Differences in DNA cleavage dependence upon concentrations of Fe-EDTA, Fe-ADR-925 and Fe-EDTA-bisamide can be explained by differences in the solution chemistry of the complexes.     

Separation of scandium(III) from lanthanides(III) with room temperature ionic liquid based extraction containing Cyanex 925

The separation of Sc(III) from Y(III), La(III) and Yb(III) in [C₈mim][PF₆] containing Cyanex 925 has been investigated, and is reported in this paper. A cation exchange mechanism of Sc(III) in [C₈mim][PF₆] and Cyanex 925 is proposed by study of the influence of anionic and cationic species on the extraction. The coefficient of the equilibrium equation of Sc(III) was confirmed by slope analysis of log D_Sc vs log [Cyanex 925], and the loading capacity also confirmed the stoichiometry of Cyanex 925 to Sc(III) was close to 3:1. Infrared data for Cyanex 925 saturated with Sc(III) in [C₈mim][PF₆] indicated strong interaction between P?O of Cyanex 925 and Sc(III). In addition, the relationship between log D_Sc and temperature showed that temperature had little influence on the extraction process, and the resulting thermodynamic parameters indicated that an exothermic process was involved. Copyright © 2007 Society of Chemical Industry     

Solvent extraction of lanthanides and yttrium from nitrate medium with CYANEX 925 in heptane

The extraction behavior of lanthanides and yttrium usinsg CYANEX 925 (mixture of branched chain alkylated phosphine oxides) in n‐heptane from nitrate medium has been studied. The effects of aqueous phase ionic strength, CYANEX 925 concentration in the organic phase, and temperature on Sm³⁺, Nd³⁺ and Y³⁺ extraction have been investigated. The extractability of the lanthanides and yttrium increases with increasing nitrate concentration, as well as with increasing CYANEX 925 concentration. An extraction mechanism is proposed based on slope analysis. Furthermore, the infra‐red spectra of CYANEX 925 saturated with lanthanides are employed to provide evidence of the composition of the complex. The relationship between the logarithm of the distribution ratio and lanthanide atomic number is also discussed which indicates that yttrium can be separated from light lanthanides. In addition separation of the light and heavy lanthanide groups is also possible using CYANEX 925. From the temperature dependence data, the thermodynamic parameters values (ΔH, ΔS and ΔG) are calculated. Copyright © 2007 Society of Chemical Industry     

双金属纳米颗粒在硅基底上直接催化生长碳纳米管

以甲烷为碳源,双金属纳米颗粒Fe／Ru（1：1）、Fe／Ru（1：1）、Ru／Fe（1：1）为催化剂,在含有加厚氧化层的硅基底上,于875-950℃制备纳米碳管,并用原子力显微镜（atomic force micmscopr——AFM）和拉曼光谱进行表征,发现随着反应温度的降低,3种催化剂的催化效果具有明显的不同程度的降低,最佳反应温度为925℃;所制备的纳米碳管都是单壁的,并且高度都约是1．25nm;对催化剂进行预处理（925℃,1min）后,其催化效果会有显著的提高,这对实现碳纳米管的定点生长具有重要的意义。     

Extraction and Separation of Zr(IV) and Hf(IV) from Nitrate Medium by Some CYANEX Extractants

Abstract

A solvent extraction study has been carried out to extract and separate zirconium and hafnium from nitrate medium by using some phosphine oxide extractants (CYANEX 921, CYANEX 923, and CYANEX 925) in kerosene. The influence of the different factors affecting the extraction process was studied in detail. Apparently the rate of extraction of Zr(IV) and Hf(IV) in CYANEX 921, CYANEX 923, and CYANEX 925 is reasonably fast. The extraction increases with increasing temperature, suggesting that the reaction is endothermic. The stripping percent of Zr(IV) and Hf(IV) by 0.5 M HNO₃ from the loaded organic phase after two stages reached 97.5% and 10.2%, respectively, which lead to good separation of the two metals. Under the optimum conditions, the extraction of zirconium was about 90, 87.6, and 91.6% and separation factors equal to 17, 21.4, and 40.7 were obtained for CYANEX 921, CYANEX 923, and CYANEX 925, respectively. The results obtained reveal that 2.0 M nitric acid is the optimum acid concentration for the separation of Zr(IV) and Hf(IV) and 0.4 M CYANEX 925 performs more efficient separation compared with other organophosphorus extractants.     

新型高效破乳剂在青霉素生产中的应用

本文介绍了我国青霉素征税中使用的破乳剂现状，对破乳机理进行了比较详细的阐述，新型高效破乳剂Ｄ９２５ｍ采用了混合的表面活性剂体系，可调性好，适应性强，已受到国内主要青霉素生产厂家的关注。     

频率和功率对轮胎微波加热的影响

在双波导入射功率均为5 k W时,分别对(915±25)MHz频率范围和925 MHz频率时的双波导不同功率分配下的微波加热效率以及轮胎温度场分布进行了模拟。结果表明:微波加热效率随微波频率的变化呈类似于正/余弦曲线波动,在微波作用频率为925 MHz时,微波加热效果最好;双波导功率分配越趋近于一致,微波加热效率越高,轮胎温度场分布越均匀。     

Extraction of Lanthanum and Samarium from Nitrate Medium by some Commercial Organophosphorus Extractants

Abstract

The extraction of lanthanum(III) and samarium(III) from nitrate solutions by some phosphine oxide compounds (Cyanex 921, Cyanex 923, and Cyanex 925) in kerosene was investigated. The influence of the different factors affecting the extraction was studied in detail. The extraction of these metals using the above extractants was compared and the sequence of extraction was found to be Cyanex 921>Cyanex 923?Cyanex 925. The stripping percent of La(III) and Sm(III) by 0.75 M HNO₃ from the loaded organic phase after two stages were 72% and 5.2%, respectively, which could enable a good separation between these two lanthanides.