Quantitative determination of erlotinib in human serum using competitive enzyme-linked immunosorbent assay

A selective and sensitive competitive enzyme-linked immunosorbent assay(ELISA) method was developed and validated for the quantification of erlotinib in 50 mL of samples of human serum. Anti-erlotinib serum was obtained by immunizing mice with an antigen conjugated with bovine serum albumin and 3,4-bis(2-methoxyethoxy)benzoic acid using the N-succinimidyl ester method. Enzyme labeling of erlotinib with horseradish peroxidase was similarly performed using 3,4-bis(2-methoxyethoxy)benzoic acid. A simple competitive ELISA for erlotinib was developed using the principle of direct competition between erlotinib and the enzyme marker for anti-erlotinib antibody, which had been immobilized on the plastic surface of a microtiter plate. Serum erlotinib concentrations lower than 40 ng/mL were reproducibly measurable using the ELISA. This ELISA was specific to erlotinib and showed very slight cross-reactivity(6.7%) with a major metabolite, O-desmethyl erlotinib. Using this assay, drug levels were easily measured in the blood of mice after oral administration of erlotinib at a single dose of 30 mg/kg. ELISA should be used as a valuable tool for therapeutic drug monitoring and in pharmacokinetic studies of erlotinib.

Seropositivity rates of water channel protein 4 antibodies compared between a cell-based immunofluorescence assay and an enzyme-linked immunosorbent assay in neuromyelitis optica patients

A total of 66 samples （from 27 cases with neuromyelitis optica, 26 cases with multiple sclerosis, aa 13 cases with optic neuritis） were tested for aquaporin-4 antibody by a cell-based immunofluorescence assay and an enzyme-linked immunosorbent assay. The sensitivities and specificities of the two assays were similar. We further analyzed an additional 68 patients and 93 healthy controls using the enzyme-linked immunosorbent assay. A Kappa test showed good consistency between the two methods in terms of detection of anti-aquaporin-4 antibody in the se of neuromyelitis optica patients. No significant correlations were identified with onset age or disea duration, suggesting that aquaporin-4 antibody is a good marker for neuromyelitis optica. The enzyme-linked immunosorbent assay can be used for quantifying aquaporin-4 antibody concentrations and may be useful to dynamically monitor changes in the levels of aquaporin-4 antibody during disease duration.

Development of a competitive enzyme-linked immunosorbent assay for therapeutic drug monitoring of afatinib

Afatinib is an oral tyrosine kinase inhibitor(TKI) approved for treating advanced non-small cell lung cancer. It is necessary to develop a simple quantification method for TKIs in order to facilitate therapeutic drug monitoring(TDM) in clinical settings. This study sought to develop a simple and sensitive competitive enzyme-linked immunosorbent assay(ELISA) to quantify afatinib in plasma for routine pharmacokinetic applications. An anti-afatinib antibody was obtained using(S)-N-4-(3-chloro-4-fluorophenyl)-7-(tetrahydrofuran-3-yloxy)-quinazoline-4,6-diamine(CTQD), which has the same substructure as afatinib, as a hapten. Enzyme labeling of afatinib with horseradish peroxidase was similarly performed using CTQD. A simple competitive ELISA for afatinib was developed based on the principle of direct competition between afatinib and the enzyme marker for the anti-afatinib antibody, which had been immobilized on the plastic surface of a microtiter plate. Plasma afatinib concentrations below the limit of quantification of 30 pg/mL were reproducibly measurable. Also, the values of plasma afatinib levels measured from 20 patients were comparable with those measured by high-performance liquid chromatography, and there was a strong correlation between the values determined by both methods(Y=0.976 X – 0.207, r=0.975). As indicated by its specificity and sensitivity, this newly developed ELISA for afatinib is an important tool for TDM and studies of the pharmacokinetics of afatinib.

Enzyme-linked Immunosorbent Assay for Detection of Anti-idiotype Antibodies to Antibodies to Ligand of Nicotinic Acetylcholine Receptor in Sera of Patients with Myasthenia Gravis

Anti-bungarotoxin anti-serum,which has the internal image of nicotinicacetylcholine receptor,was used as a tool to measure anti-idiotypic antibodies toantibodies to Iigand of nicotinic acctylcholine receptor in scra from 81 patients withmyasthenia gravis.Enzyme-linked immunosorbcnt assay was adopted.Thc positive ratewas 46.9%(38/81).The specific cross inhibitory test with nicotinic acetylcholinereceptor was positive.Anti-idiotype antibodies to antibodies to ligand of nicotinicacetylcholine receptor in sera of different types of myasthenia gravis patients classified ac-cording to modified Osserman’s standard and myasthenia gravis patients with or withoutthymoma were comparcd in this study and the role of anti-idiotype antibodies toantibodies to Iigand of nicotinic acctylcholinc receptor in the immunity of myasthcniagravis and the possibility of thcrapeutic use of anti-idiotype antibodies arc discussed.

Development of an Indirect Enzyme-Linked Immunosorbent Assay for Seromonitoring Contagious Bovine Pleuropneumonia Using Recombinant Lipoprotein LppQ of Mycoplasma mycoides subsp mycoides SC as Antigen

Mycoplasma mycoides subsp mycoides SC （MmmSC） is the etiological agent of contagious bovine pleuropneumonia （CBPP）. The lipoprotein LppQ encoded by lppQ gene is specific to MmmSC and is found in the type strain and in field strains isolated in Europe, Africa, and Australia, as well as in vaccine strains. No serological cross-reactions were observed with the related mycoplasmas of the Mycoplasma mycoides cluster. The N-terminal domain of the mature lipoprotein LppQ is hydrophilic, and it induces a strong, specific, early, and persistent immune response in naturally and experimentally infected animals. Mycoplasma-specific TGA （Trp） codons are utilized as stop codons in most other organisms. The lppQ N-terminal fragment from MmmSC HVRI X strain, the Chinese strain for CF antigen production, was mutated with one-step overlapping extension PCR. Sequence analysis confirmed the successful mutation from A to G in codon 198 in the lppQ gene. The fragment containing the mutation site was subcloned into the pET32a expression vector. The recombinant protein with molecular weight of 42 kDa was purified using the Ni-NTA His.Bind purification kit, with a purity of up to 95%. Western blot indicated that the standard positive serum of CBPP could react with the recombinant protein. The purified protein was diluted to 0.35 μg mL^-1, and coated to microtiter enzyme-linked immunosorbent assay （ELISA） plates. Indirect ELISA reaction conditions were optimized. The value of P/N was determined to be 4.8 （0.934/0.193）, the sensitivity to be 95.8% （46/48）, and the specificity to be 98.9% （161/163）. 3 817 cattle serum samples from three different provinces were detected by the indirect ELISA and CFT. The Kappa value is 0.63, which is middle or high agreement between the two methods.

A Comparison of Enzyme-Linked Immunosorbent Assay versus Multiplex Methodology Using an <i>in Vitro</i>Model of Pulmonary Hypertension and Inflammation

Enzyme-linked immunosorbent assay (ELISA) is the most widely used method for measuring a single cytokine. Recent developments in cytokine quantification such as multiple arrays measure multiple cytokines simultaneously. Although good correlations between ELISA and multiplex methods have been observed, side by side comparisons are limited. In the present study we hypothesized that ELISA and Luminex techniques are comparable in detecting cytokines in culture medium when pulmonary artery smooth muscle cells (PASMC) are exposed to stress. Primary human PASMC were cultured in modular chambers and exposed to 21% FiO2 and peak inspiratory and positive end expiratory pressure of 24 and 8 cmH2O respectively, and 95% FiO2. At 24 hours, culture medium was collected and assayed for interleukin-6 (IL-6) and IL-8 by quantitative ELISA and by Human Cytokine 25-Plex Panel using a Luminex 200 analyzer. A comparative analysis of agreement between our ELISA and Luminex data was detailed for control and stress conditions using the Bland-Altman plot analysis. Each assay resulted in comparable increased (p < 0.001) levels of IL-6 and IL-8 as compared to control in response to oxidative and biophysical stress. The Bland-Altman analysis demonstrated that 95% of the differences between ELISA and Luminex values were within ±1.96 SD from the mean difference indicated by the 95% limits of agreement for the measurements of IL-6 and IL-8. There was no systematic bias as a function of inflammation level. We conclude that in this cell culture model, ELISA and Luminex are comparable in detecting the levels of IL-6 and IL-8 in the culture medium. If measurements of multiple cytokines are demanded and the amount of sample is limited, Luminex multi-analyte profiling technology is accurate and sensitive.

Determnation of ochratoxin A in grain by monoclonal antibody－based enzyme－linked immunosorbent assay

The simple rapid and sensitive enzyme-linked immunosorbent assay (ELISA) methods, di-rect and indirect ELISA, for quantitation of ochratoxin A in cereal had been developed by theutilization of monoclonal antibody on immunomicroplate. Direct FLIAS was found to be less timeconsuming than indirect ELISA. For direct FLISA, recovery of 1 -500 ppb OA added to wheat was78.9-100.0% and rice was 88.9- 120.0%. For indirect EI.IAS, recovery of 1-500 ppb OA addedto wheat was 79.0- 110.0% and rice was 82.0 120.0%. The minimal detection level for OA was Ippb. Analyses of 31 samples that caused humanintoxicant for OA showed that the ELISA resultsagreed wtll with those obtained by thin-layer chromatogrdphy.

SERODIAGNOSIS OF CLONORCHIASIS BY ENZYME—LINKED IMMUNOSORBENT ASSAY WITH HRP—SPA

In thes paper the authors used the Horseradish peroxidase labelledstaphylococcal protein A(HRP—SPA)in ELISA,for the detection of Clo-norchis sinensis infection.Serum tests were made on 116 confirmed cases ofclonorchiasis,103(88.8%)of them showed positive,while only 6(4.4%)werepositive among 138 healthy people.Samples were collected on filter paperstrips,111(95.7%)cases were positive among 116 comfirmed cases tested,but only 2(1.5%)were positive out of 138 healthy persons.The resultswere similar to those obtained by sheep antihuman IgG.Animal experimentalso showed that the SPA—ELISA can be used for the diagnosis ofclonorchiasis.In an endemic area,stool egg positive rate was 8.8%(62/703).whenchecked with SPA—ELISA,the rate of conformity in both filter paperstrips and stool examinations was 90.3(56/62).Among 641 serum testsfrom individuals negative in stool examinations,only 35(5.5%)reactedpositively.The authors suggested—that SPA—ELISA with soluble Clo-norchis antigens could be used in a large scale seroepidemiological surveyin endemic areas.

S-烯丙基巯基半胱氨酸促进CD8+T细胞杀伤功能的机制研究

目的探究S-烯丙基巯基半胱氨酸(S-allylmercaptocysteine,SAMC)对CD8^(+)T细胞杀伤鼻咽癌细胞功能的影响及机制。方法将人鼻咽癌细胞HK-1和C666-1与SAMC共培养,分为0、25、50、100μmol/L SAMC组,Western blot检测肿瘤细胞程序性死亡配体-1(programmed cell death-ligand 1,PD-L1)表达。CD8^(+)T细胞分别与HK-1和C666-1细胞以10∶1的比例共培养并加入0、25、50、100μmol/L SAMC,检测CD8^(+)T对HK-1和C666-1的杀伤能力,酶联免疫法(ELISA)检测干扰素(INF-γ)和肿瘤坏死因子-α(TNF-α)浓度,构建鼻咽细胞HK-1的小鼠皮下移植瘤模型,分为对照组、CD8^(+)T细胞组、SAMC组、SAMC+CD8^(+)T细胞组,各组小鼠治疗期间测量瘤体积,治疗结束后取小鼠肿瘤组织,Western blot检测肿瘤组织中PD-L1表达,ELISA检测小鼠血清INF-γ和TNF-α浓度。结果相比于0μmol/L SAMC组,25、50、100μmol/L SAMC组HK-1和C666-1细胞PD-L1表达均显著下调(P<0.05),相比于0μmol/L SAMC+CD8^(+)T细胞组,25、50、100μmol/L SAMC+CD8^(+)T细胞组HK-1和C666-1细胞培养上清中INF-γ和TNF-α浓度均能显著增加,HK-1和C666-1细胞裂解率显著增加(P<0.01)。相比于对照组,CD8^(+)T细胞组和SAMC+CD8^(+)T细胞组小鼠瘤体积和瘤重显著下降(P<0.05),小鼠血清INF-γ和TNF-α浓度显著增加,相比于CD8^(+)T细胞组,SAMC+CD8^(+)T细胞组小鼠瘤体积和瘤重显著下降(P<0.05),小鼠血清INF-γ和TNF-α浓度显著增加(P<0.05),肿瘤组织PD-L1表达显著下调(P<0.01)。结论SAMC可通过抑制人鼻咽癌细胞PD-L1表达而促进CD8^(+)T细胞的杀伤功能。

不同品牌ELISA试剂盒检测细胞培养上清液中IL-8对比研究

目的 对比研究2种不同品牌ELISA试剂盒检测细胞培养上清液中人白细胞介素-8(IL-8)水平情况。方法 不同浓度聚肌胞苷酸(Poly I:C)刺激A549细胞10、24 h,收集细胞上清液,共获得40份样本,分为1~10组,每组4份。采用2种ELISA试剂盒(试剂盒A、试剂盒B)检测细胞培养上清液中IL-8水平,对比2个试剂盒检测得到的组间数据或同组数据的一致性或差异性。结果 2种试剂盒拟合曲线中相关系数值分别为:0.999 600、0.999 602,检测标准品结果相关性良好。试剂盒B检测样品1、2组中IL-8水平比较,差异有统计学意义(P<0.01),但试剂盒A检测样品1、2组中IL-8水平比较,差异无统计学意义(P>0.05);2种试剂盒检测样品8、9组及样品9、10组中IL-8水平比较,差异均有统计学意义(P<0.01)。仅试剂盒B检测样品1、5组中IL-8水平明显低于试剂盒A,差异有统计学意义(P<0.01)。结论 不同品牌ELISA试剂盒均可用于检测IL-8水平,但检测结果仍在一定程度上存在差异。为避免实验误差,研究中应尽量选择同一个厂家生产的同品牌试剂盒。

人C-反应蛋白磁微粒化学发光酶免疫测定法的建立

人C-反应蛋白(C-reactive protein,CRP)是炎症以及各种相关疾病如病毒感染、心血管疾病等诊断、治疗和预后的临床检测指标。为了建立一种快速、准确的CRP定量免疫测定方法,将表达纯化的重组CRP作为抗原免疫小鼠,获得了5株稳定分泌抗体的单克隆抗体细胞株,采用双抗体夹心酶联免疫吸附测定法(double antibody sandwich enzyme-linked immunosorbent assay,DAS-ELISA)初步鉴定筛选的抗人CRP单克隆抗体,并分别选择mAb 9D6和mAb 9G4作为捕获抗体与检测抗体,建立用于人CRP检测的化学发光酶免疫测定法(chemiluminescence enzyme immunoassay,CLEIA),最后通过测定分析临床血清CRP样本,评价CLEIA的性能。结果显示,基于9D6/9G4-AP单克隆抗体对的CLEIA测定范围为0.1767~500μg/L(可扩展至100 mg/L);所建立的CLEIA与医院采用的免疫散射比浊法(R^(2):0.9496,P<0.0001)表现出良好的相关性,且Bland-Altman分析中96.36%(106/110)的点在95%一致性界限范围内显示两种检测方法具有较好的一致性。结果初步表明,建立的分析方法在临床诊断中具有较好的应用前景。

2016-2020年云南省部分猪场猪繁殖与呼吸综合征免疫抗体检测分析

为了解2016年-2020年云南省部分地区猪场猪繁殖与呼吸综合征疫苗免疫情况,采集了云南省9个州市188个健康猪场的4179份血清样品,应用ELISA方法进行猪繁殖与呼吸综合征病毒抗体检测,分析不同州市、不同年份的猪繁殖与呼吸综合征病毒抗体阳性率。结果显示,4179份血清样品中,阳性样品2932份、阴性样品1247份,总体抗体阳性率为70.16%;2016-2018年云南省部分地区抗体阳性率分别为76.97%、76.32%、75.75%,达到我国农业农村部要求的猪繁殖与呼吸综合征病毒抗体阳性率70%的标准;2019-2020年抗体阳性率分别为63.68%、56.50%,未达到农业农村部规定的标准。9个州市中,保山、楚雄、红河、普洱、版纳、玉溪的抗体阳性率分别为84.85%、70.75%、76.46%、80.67%、76.82%、81.39%,达到农业农村部规定标准;大理、昆明、曲靖抗体阳性率分别为61.89%、67.42%、57.88%,未达到农业部规定标准。结果表明,云南省部分地区总体猪繁殖与呼吸综合征病毒抗体阳性率呈现下降趋势,各州市抗体阳性率也存在差异性。为了降低猪繁殖与呼吸综合征疫情发生风险,建议定期做好猪繁殖与呼吸综合征病毒血清学、病原学监测,优化猪繁殖与呼吸综合征疫苗免疫策略,制定更加科学合理的综合防控方案,从而进一步保障云南省生猪养殖业高质量发展。

Comparison of chemiluminescence enzyme immunoassay based on magnetic microparticles with traditional colorimetric ELISA for the detection of serum α-fetoprotein

A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELISA).A systematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer dosages of immunoreagents,less total assay time,and better linearity,recovery,precision,sensitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results were compared with commercial electrochemiluminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R 2 0.6703;however,the correlation between MPs-CLEIA and ECLIA (R 2 0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R 2 0.6866).

120例无偿献血者HIV-ELISA单试剂反应性结果的分析

目的分析研究无偿献血者人类免疫缺陷病毒抗原抗体酶联免疫吸附法(human immunodeficiency virus antigen antibody enzyme-linked immunosorbent assay,HIV-ELISA)单试剂反应性结果的检测性能,为提高实验室的检测能力作参考。方法采用伯乐酶联免疫吸附法(BR-HIV)试剂对笔者血站2023年2月份的7069例标本进行检测,其中有120例BR-HIV单试剂反应性,再对其该单试剂反应性标本采用另外3家不同厂家HIV-ELISA试剂、化学发光免疫分析(chemiluminescence immunoassay,CLIA)、核酸检测技术(nucleic acid detection technology,NAT)、Western blot法进行检测,统计分析ELISA、CLIA、NAT、Western blot法其检测结果关系。结果BR-HIV单试剂反应性阳性率出现了阶段性的增加,差异有统计学意义(χ^(2)=4.058,P<0.05)。通过电话追踪其120例标本信息,将BR-HIV单试剂反应性的病毒感染者与未感染者进行比较,S/CO值感染者明显高于未感染者;假阳性率1.69%(120/7069)比2019年2月份假阳性率0.22%(8/3619)明显增加,差异有统计学意义(χ^(2)=44.099,P<0.05)。CLIA、NAT、另外3家HIV-ELISA检测试剂具有一致性(Kappa=1),具有等效性。Western blot法检测结果均为阴性。结论曾经感染过病毒的无偿献血者的标本BR-HIV单试剂假阳性率增高,可以用CLIA、NAT以及其他酶免试剂暂时替代检测,减少血液的报废,提高对捐献血液的检测能力。

Monoclonal Antibody-Based Enzyme Linked Immunosorbent Assay for the Analysis of Jasmonates in Plants

Methyl jasmonate （MeJA） and its free-acid form, jasmonic acid （JA） are naturally occurring plant growth regulators widely distributed in higher plants. In order to improve the sensitivity for the analysis of MeJA at low levels in small amounts of plant samples, a monoclonal antibody （MAb） （designated as MAb 3E5D7C4B6） against MeJA was derived from a JA- bovine serum albumin （BSA） conjugate as an immunogen. The antibody belongs to the IgG1 subclass with a κ type light chain and has a dissociation constant of approximately 6.07 ×10^-9 M. MAb3E5D7C4B6 is very specific to MeJA. It was used to develop a direct competitive enzyme-linked immunosorbent assay （dcELISA）, conventional and simplified indirect competitive ELISAs （icELISA）. JA was derivatized into MeJA for the ELISA analysis. The IC50 value and detection range for MeJA were, respectively, 34 and 4-257 nglmL by the conventional icELISA, 21 and 3-226 nglmL by the simplified icELISA and 5.0 and 0.7-97.0 nglmL by the dcELISA. The dcELISA was more sensitive than either the conventional or simplified icELISA. The assays were used to measure the content of jasmonates as MeJA in tobacco leaves under drought stress or inoculated with tobacco mosaic virus and tomato leaves inoculated with tomato mosaic virus or Lirioinyza sativae Blanchard as compared with the corresponding healthy leaves. The increased jasmonates content indicated its role in response to the drought stress and pathogens.

Development of Monoclonal Antibody-based Enzyme-linked Immunosorbent Assay to the Estrogen Diethylstilbestrol

Diethylstilbestrol （DES） is a synthetic estrogen that has ever been used worldwide. Polyclonal antibodies （PAbs） were used in immunoassay for detection of DES residues in environmental and agricultural samples in previous paper. In this paper, an indirect competitive enzyme-linked immunosorbent assay （icELISA） was developed based on monoclonal antibody （MAb） for the determination of diethylstilbestrol. Mono-o-carboxypropyldiethylstilbestrol （DES-CP） and mono-o-carboxymethyldiethylstilbestrol （DES-CME） were synthesized to be haptens. DES-CP was coupled to bovine serum albumin （BSA） to be an immunogen in BALB/c female mouse for MAb production. The MAb was characterized for specificity and affinity to DES in icELISA. Under the optimum condition, the icELISA showed an ICs0 of 9.8 ng/mL, the limit of detection （IC20） of 2.3 ng/mL and a working range of 2-42 ng/mL. Hexestrol and dienestrol exhibited cross-reactivity values were 44% and 27%, respectively. Cross-reactivity of natural estrogen 17β-estradiol was less than 0.1%. The influences of some factors such as salt concentration, pH and organic solvent concentration on the assay were evaluated. The concentrations of DES in the fortified water samples determined by the assay were correlated well with the fortification levels. The results were conf＇m＇ned with analysis by HPLC.

Tetra-substituted Amino Aluminum Phthalocyanine as a New Red-region Fluorescent Substrate for Horseradish Peroxidase Based Enzyme-linked Immunosorbent Assay

The use of tetra-substituted ammo aluminum phthalocyanine (TAAIPc) as a new red-region fluorescent substrate for horseradish peroxidase (HRP)-based enzyme-linked immunosorbent assay was investigated. TAAIPc displayed an excitation maximum at 610 nm and emission maximum at 678 nm in a strong acidic medium. In the presence of HRP, trace amounts of H2O2 could rapidly and significantly react with TAAIPc, thus quenching the fluorescence of TAAIPc. The Michaelis-Menten parameters Km and Vmax were measured to be 2.82 × 10?6 mol/L?1 and 6.0 × 10?9 mol·L?1, respectively. In this paper, TAAIPc was used in an HRP-based enzyme-linked immunosorbent assay (ELISA) of α-fetoprotein (AFP) in human serum with satisfactory results. AFP could be determined in the concentration range of 0.5–200 ng/mL with a detection limit of 0.2 ng/mL, which was close to that of radioimmunoassay. The advantage of proposed method was strongly minimizing the interference resulting from background fluorescence or scattering light and had a high analytical sensitivity.

脑脊液D-二聚体对脑胶质瘤的诊断意义

目的测定脑胶质瘤患者术前、术后以及后续放化疗过程中的脑脊液D-二聚体水平,探讨脑脊液D-二聚体水平对脑胶质瘤的诊断分级、治疗效果及评估预后的临床意义。方法纳入2019年3月1日至2022年3月1日于新乡医学院第一附属医院接受手术治疗并后续放化疗的患者68例,分别于术前、术后1周,术后1个月、术后3个月、术后6个月及术后12个月留取脑脊液标本测定D-二聚体水平;分别比较脑胶质瘤与脑膜瘤(对照组)、不同级别脑胶质瘤、脑胶质瘤术前与术后、进展前后脑脊液D-二聚体水平以及治疗过程中的脑脊液D-二聚体水平变化,采用受试者工作特征曲线分析各个级别脑胶质瘤脑脊液D-二聚体水平的截断值,总结其水平变化的临床意义。结果低级别胶质瘤组与脑膜瘤对照组血浆D-二聚体水平相比,差异无统计学意义(P=0.971);Ⅲ级及Ⅳ级胶质瘤组与脑膜瘤对照组血浆D-二聚体水平相比,含量均升高,差异均具有统计学意义(P<0.01);伴随胶质瘤级别的增高,血浆D-二聚体水平升高,各级别之间差异均具有统计学意义(P<0.01);术后脑脊液D-二聚体水平低于术前组(P<0.01),全切患者脑脊液D-二聚体水平下降率高于未全切患者(P<0.01);进展后患者脑脊液D-二聚体水平高于进展前(P<0.01)。低级别、Ⅲ级及Ⅳ级脑胶质瘤患者脑脊液D-二聚体含量受试者工作特征曲线中脑脊液D-二聚体截断值分别为96.84 ng/ml、588.82 ng/ml和8855.24 ng/ml。结论脑脊液D-二聚体可作为脑胶质瘤的诊断分级、治疗效果及评估预后的标志物。

柴芩解热颗粒对发热大鼠解热作用及血清TNF-ɑ、IL-1β、PGE_(2)、AVP表达水平的影响

目的观察柴芩解热颗粒对脂多糖(LPS)致热大鼠的解热作用及血清中肿瘤坏死因子-ɑ(TNF-ɑ)、白介素-1β(IL-1β)、前列腺素E_(2)(PGE_(2))及精氨酸加压素(AVP)表达水平的影响.方法将60只SPF级SD大鼠随机分为正常对照组(生理盐水+灌胃蒸馏水)、模型对照组(LPS造模+灌胃蒸馏水)、小儿柴桂退热颗粒组(LPS造模+灌胃3.6 g/kg小儿柴桂退热颗粒)和柴芩解热颗粒高、中、低剂量组(LPS造模+灌胃2.1 g/kg、1.05 g/kg、0.525 g/kg柴芩解热颗粒),每组10只.直肠测温确定各组大鼠温度变化;腹主动脉取血,分离血清,酶联免疫法(ELISA)检测TNF-ɑ、IL-1β水平;处死大鼠,取下丘脑,检测PGE_(2)、AVP水平.结果与模型对照组比较,柴芩解热颗粒高剂量组有显著的解热效果(P<0.01).柴芩解热颗粒高、中、低剂量组均可降低大鼠血清中TNF-ɑ、IL-1β的含量,与模型组比较差异具有统计学意义(P<0.01或P<0.001);柴芩解热颗粒低剂量组可降低大鼠下丘脑匀浆液中PGE_(2)的含量,与模型组比较差异具有统计学意义(P<0.05);同时还能升高大鼠下丘脑匀浆液中AVP的含量,与模型组比较差异具有统计学意义(P<0.01).结论柴芩解热颗粒具有良好的解热作用,解热机制可能与TNF-ɑ、IL-1β、PGE_(2)及AVP有关.

2016年-2020年云南省部分猪场猪瘟免疫抗体检测与分析

为了解2016年1月-2020年12月云南省猪场猪瘟疫苗免疫效果,采集云南省11个州(市)198个猪场猪瘟疫苗免疫后14 d~21 d血清样本4622份。应用ELISA方法进行猪瘟病毒抗体检测,分析不同州(市)、不同年份的猪瘟病毒抗体阳性率。结果显示,4622份血清样品中,阳性样品4078份,阴性样品544份,总体猪瘟病毒抗体阳性率为88.23%。其中,保山市猪瘟病毒抗体阳性率最高,其次为大理州和西双版纳州,猪瘟病毒抗体阳性率分别为98%、95.57%和95.26%;德宏州猪瘟病毒抗体阳性率最低,为60.81%。不同年份连续5年猪瘟病毒抗体检测中,猪瘟病毒抗体阳性率呈现波浪形变化,其中2017年度猪瘟病毒抗体阳性率最高,全年共检测血清样品1156份,阳性样品1054份,猪瘟病毒抗体阳性率为91.18%;2018年度猪瘟病毒抗体阳性率最低,全年度共检测血清样品845份,阳性样品720份,猪瘟病毒抗体阳性率为85.21%。结果表明,云南省猪场的总体猪瘟病毒抗体阳性率较高,但是各州(市)猪瘟病毒抗体阳性率存在差异,各年度猪瘟病毒抗体阳性率不稳定。为了有效防控猪瘟疫情发生,建议定期做好猪瘟病毒血清学、病原学监测,实时调整猪瘟疫苗免疫程序,对猪瘟病毒抗体阳性率低的猪场加强一次猪瘟疫苗免疫。