Application of Clinical Nursing Pathway in the Peri-Treatment Period of Immunoadsorption Therapy for Rheumatic Immune Diseases

Objective: The paper aims to investigate the clinical nursing pathway (CNP) in the application of immunosorption therapy in patients with rheumatic immune disease. Methods: Convenience sampling method was used to select inpatients who received immunoadsorption therapy from January 2020 to December 2022 in the rheumatology and Immunology department of a 3A hospital in Jingzhou City. 30 patients from January 2020 to June 2021 were selected as control group, and 30 patients from July 2021 to December 2022 were selected as observation group. The control group was given routine nursing. On the basis of the control group, the observation group used a clinical nursing pathway for intervention during the perioperative period of immunosorbent therapy. The incidence of adverse reactions, patient satisfaction, and nurse satisfaction during immunosorbent therapy between the control group and the observation group were compared. Results: After intervention, the incidence of adverse reactions in the observation group was significantly lower than that in the control group, while patient satisfaction and nurse satisfaction in the observation group were significantly higher than those in the control group. The results are all statistically significant (P Conclusion: Clinical nursing pathway is beneficial to reduce the incidence of adverse reactions in patients with immunoadsorption during peri-treatment and improve the satisfaction of patients and nurses.

DNA immunoadsorption for childhood-onset systemic lupus erythematosus

We present a retrospective review of DNA immunoadsorption (DNA-IA) therapy on clinical symptoms as well as indicators in pediatric cases with systemic lupus erythematosus (SLE), and follow up the short-term curative effects. 16 SLE cases were treated by DNA-IA for 3 times every other day. We observed the changes on clinical manifestations and immunological indicators, in order to compare the alteration of these indicators including clinical manifestations, Systemic Lupus Erythematosus Disease Active Index (SLEDAI) scores, 24 hurinary protein excretion, autoantibodies, serum IgG and complement C3. 13 cases were followed up regularly, within 3 months after DNA-IA therapy, 12 cases of clinical manifestations improved (92.3%). SLEDAI scores in 10 cases decreased from (16.20 ± 12.54) to less than 5 (76.9%), 8 cases of ANA, anti-DNA antibodies were negative (61.5%), 13 cases with IgG level in serum recovered to normal (10.39 ± 4.38) g/L, C3 level rose to normal (1.06 ± 0.23) g/L. 3 to 6 months after IA, clinical manifestations and laboratory examinations in all cases got maximum improved. 9 months after IA, SLEDAI score in 2 cases (15.4%) rose to more than 5, anti-DNA antibody in 2 cases (15.4%) became positive, and 1case (7.7%) with serum C3 decreased again. 2 cases died from multiple organs dysfunction within 3 to 6 months after IA. No serious complications were found during DNA-IA. We recommend that DNA immunoadsorption is a safe and effective therapy for active childhood-onset SLE, which could improve clinical symptoms, eliminate ANA and anti-DNA antibodies. Combining with corticosteroids and immunosuppressive drugs, DNA-IA could significantly reduce the activity of disease and protect vital organs function in the short term.

Detection of Atrazine Residue in Food Samples by a Monoclonal Antibody-based Enzyme-linked Immunosorbent Assay

Atrazine（AT,2-chloro-4-ethylamino-6-isopropyl-amino-s-triazine）has been detected in ground water in several areas of the United States for many years,as well as in China,wherein the growth rate of its gross

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.

Comparison between the protective effects of removing circulatory TNF by specific immunoadsorption and by nonspecific Amberlite XAD-7 adsorption on experimental endotoxin shock

Objective: To compare the protective effects of removingcirculatory TNF by specific immunoadsorption and by nonspeciflc Amberlite XAD-7 adsorption on experimental endotoxin shock. Methods: New Zealand white rabbits receiving a lethal dose of endotoxin underwent hemoperfusion through immunoadsorbent or Amberlite BAD-7. Plasma TNF levels. efficiency of the adsorbents and the survival rate were observed. Results: After 2 h of hemopcrfusion through immunoadsorhent or Amberlite XAD-7. plasma TNF levels were significantly lower than those in the control group(P<0.01). and the best result was shown by immunoadsorption (P<0. 01). The 12 h and 18 h survival rates were 70% and 30% in the immunoadsorption group, and 30% and 5% in the Amberlite XAD-7 group respectively (P<0. 05). Conclusion: Compared with the nonspecific Amberlite XAD-7 adsorption,the specific imrnunoadsorption might be a more effective method of removing circulatory TNF and improve the survival rate in endotoxin shock.

A case of Bickerstaff brainstem encephalitis successfully treated with intravenous immunoglobulin and methylprednisolone after unsuccessful immunoadsorption plasmapheresis

Bickerstaff brainstem encephalitis (BBE) is a rare post-infectious neurological syndrome for which an effective treatment strategy has not been established. Here, we report a case of a 71-year-old male who suffered from an upper respiratory tract infection, and 7 days later, developed numbness of the bilateral upper and lower limbs, unsteady gait and dysarthria. Brain magnetic resonance imaging was normal, nerve conduction study and cerebral spinal fluid analysis were nonspecific. Based on the clinical features, we tentatively diagnosed Guillain-Barré syndrome and started immunoadsorption plasmapheresis. However, consciousness progressively declined to coma level within 10 days. Electroencephalogram showed diffuse slowing, and auditory evoked brainstem response (ABR) demonstrated absence of waves II, III and V. Serum anti-GQ1b IgG autoantibody and anti-GM1b IgG autoantibody were negative. Subsequently, we diagnosed BBE, and clinical symptoms resolved after treatment with intravenous immunoglobulin and methyllprednisolone. On day 62, neurological symptoms were remarkably alleviated with an improvement in ABR. Our observations suggest that immunoadsorption plasmapheresis should be used only when anti-ganglioside antibodies are detected. Combination therapy with intravenous immunoglobulin and methylprednisolone or plasma exchange?is recommended as initial therapy.

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.

Immunoadsorption therapy for Klinefelter syndrome with antiphospholipid syndrome in a patient:A case report

BACKGROUND Klinefelter syndrome(KS) is a genetic disease of male sex chromosome malformations that affects sperm production and reduces testosterone production. It has been reported that there is currently more than 10 cases of KS combined with antiphospholipid syndrome(APS).CASE SUMMARY Here, we describe a 31-year-old male patient with chromosome 47, XXY type, who suffered deep vein thrombosis of the lower limbs accompanied by abnormal antiphospholipid antibody, lupus anticoagulant and factor VⅢ. After treatment with immunoadsorption therapy, glucocorticoids, cyclophosphamide, intravenous immunoglobulin and anticoagulant therapy, the patient showed dramatic symptomatic improvement. During the follow-up, the patient did not develop any new thrombotic events.CONCLUSION Immunoadsorption combined with glucocorticoid and cyclophosphamide shock comprehensive treatment has achieved significant results for patients with KS combined with antiphospholipid syndrome.

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 PRELIMINARY STUDY OF SERUM GLYCOCONIUGATES IN PATIENTS WITH CANCER USING THE ENZYME-LINKED LECTIN ASSAY

After primary analyses on the serum glycocon-jugates of lung cancer and normal individul using the enzyme-linked lectin assay (ELLA) with 12 kinds of lectins, PHA and LCA were selected to further study in the sera of 8 kinds of cancers, 4 kinds of non-malignant disease and 1 kind of postoperative cancer. It was found that the test values of 7 kinds of cancers with PHA or LCA were significantly higher than that of the normal (P<0.01); the values of 4 kinds of non-malignant diseases with PHA were not higher (P>0.05); the values of the postoperative cancer with PHA were obviously lower than that of the preoperative (P<0.05). The results showed that the serum glycoconjugates which can bind to PHA seemed related to the cancerous existence in human bodies. The significance of the findings was discussed.

ENZYME-LINKED IMMUNOSORBENT ASSAY OF HUMAN PLACENTA TYPE GLUTATHIONE S-TRANSFERASE AND ITS APPLICATION IN THE DIAGNOSIS OF HEPATOCARCINOMA

GST-π was purified from human placenta and its antiserum was raised in rabbits. The antibody IgC was purified and degraded into Fab' fragment which was conjugated with horseradish peroxidase (HRP) using N-succinimidyl-4-(N-maleimido-methyl) cyclo-hexane-1-carboxylate (SMCC) as crosslinking reagent to produce Fab'-HRP conjugate. A sandwich ELISA was established for the microquantitative determination of GST-π. The sensitivity was 11 pg/tube, which was far more sensitive than the radioimmunoassay so far reported. Using this method, the serum GST-π of 41 cases normal adult was found to be 1.06±0.94 ng/ml. The upper limit of the normal value was 2.6 ng/ml. In 30 cases of primary hepatocarcinoma, the level of serum GST-π was 24.4± 17.4 ng/ml, which was 23 times higher than the normal average value (P<0.01). The positive rate was 90%. In contrast, serum GST-π in 25 cases of chronic hepatitis was determined to be 1.74±1.16 ng/ml, which was not significantly different from the normal value (P>0.05). The pseudo-positive rate was 12.0%.

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.

Is it feasible to remove circulating TNF specifically by immunoadsorption

In order to evaluate the feasibility of specific removal of circulating TNF by imrnunoadsorption,the efficacy and safety of the anti-TNF monoclonal antibody immobilized immunosorbent were studied fromdifferent points of view. The results showed: (l ) The physical characteristics of the immunosorbent wereperfect; (2) The maximum of antibody 1 ml was about 8. 12 mg; (3) Leakage of the antibody from the immunosorbent was not detectable; (4 ) The immunosorbent could remove circulating TNF effectively; (5 )Small amounts of Plasma proteins were adhered nonspecifically, but most of them could be washed out easily;(6 ) The sorbent capacity was not affected obviously when it was disinfected with ethanol; (7 ) The immunosorbent was of good biocompatibility. These findings suggest that the immunosorbent might be used forremoving circulatory TNF in endotoxic shock in the

HA280免疫吸附在重度活动性系统性红斑狼疮治疗中的作用

目的:探究HA280免疫吸附配合激素、环磷酰胺、羟氯喹治疗重度活动性系统性红斑狼疮的疗效及安全性。方法:选取32例重度活动性系统性红斑狼疮患者为研究对象,根据治疗方法不同将患者分为吸附组与对照组,每组各16例。对照组进行激素、环磷酰胺、羟氯喹等常规治疗;吸附组在此基础上应用HA280免疫吸附治疗,两组患者疗程均为6个月。对比分析两组患者的免疫球蛋白、补体、疾病活动度、激素累积剂量的变化情况。结果:治疗6个月后,吸附组IgG、IgA及总球蛋白下降水平、C3、C4回升及SLE活动度降低水平均高于对照组(P<0.05);治疗后,吸附组的达标用时小于对照组(P<0.05);激素累积量小于对照组(P<0.05)。结论:HA280免疫吸附联合激素、环磷酰胺、羟氯喹治疗重度活动性系统性红斑狼疮的疗效显著,更能快速控制病情活动度、及早达标,并可减少激素的累积剂量,减少相关副作用。

2023年美国血液分离学会“治疗性血液分离技术临床实践指南”(第9版)解读

2023年4月美国血液分离学会(American Society for Apheresis,ASFA)发布了第9版《治疗性血液分离技术临床实践指南》。该指南基于目前已发表的文献,确定治疗性血液分离技术治疗疾病的证据质量及推荐强度,讨论了该技术在91种疾病(166种适应证)中的临床应用。本文重点解读血浆净化技术,即血浆置换和免疫吸附技术在临床多学科中的应用。

抗体清除血液净化技术治疗抗中性粒细胞胞质抗体相关血管炎的研究进展

抗中性粒细胞胞质抗体相关血管炎(AAV)是一类累及多系统的自身免疫性疾病,有较高的病死率和进展至终末期肾病的风险。体外循环血液净化技术已成为除糖皮质激素和免疫抑制剂之外治疗AAV的重要辅助手段,包括血浆置换、双重滤过血浆置换和免疫吸附。本文系统综述了抗体清除血液净化技术治疗AAV的研究进展以及指南推荐的变化,为临床应用提供参考。

荧光定量对肺炎支原体IgM抗体诊断效能的影响

目的:探讨荧光定量对肺炎支原体IgM抗体诊断效能的影响。方法:选取186例我院2020年4月至2022年4月就诊于我院的呼吸道感染疑似肺炎支原体感染患儿作为研究对象。入院后均采用荧光定量聚合酶链式反应(Polymerase chain reaction,PCR)法、酶联免疫吸附(Enzyme linked immunosorbent assay,ELISA)法对肺炎支原体IgM抗体进行检测。以间接荧光免疫法检测结果为“金标准”,比较荧光定量PCR、ELISA法对肺炎支原体IgM抗体诊断结果。结果:经间接荧光免疫法检测结果显示,186例呼吸道感染患儿中,肺炎支原体IgM抗体阳性102例,经荧光定量PCR检测结果显示,肺炎支原体IgM抗体阳性94例,经ELISA法检测结果显示,肺炎支原体IgM抗体阳性85例;与ELISA法比较,荧光定量PCR法对于IgM抗体阳性诊断灵敏度、准确度较高,漏诊率较低(P<0.05)。结论:荧光定量PCR可有效提高肺炎支原体感染诊断准确效能,为临床早期诊断提供参考。

蛋白A免疫吸附治疗重症ANCA相关性血管炎肾损伤的临床效果

目的:抗中性粒细胞胞质抗体(anti-neutrophil cytoplasmic antibody,ANCA)相关性血管炎(ANCAassociated vasculitis,AAV)是一组累及多系统及器官的疾病,肾是最常受累的靶器官之一,部分患者短期内迅速进展至终末期肾病。既往标准诱导缓解方案治疗后仍有部分患者的缓解情况及肾脏预后较差。蛋白A免疫吸附(protein A immunoadsorption,PAIA)是一种选择性血液净化方式,在多种重症自身免疫系统疾病治疗方面表现出巨大的优势。本研究旨在探讨PAIA联合激素及免疫抑制剂在重症AAV肾损伤中的短期疗效。方法:回顾性纳入中南大学湘雅二医院于2019至2020年间收治的10例重症AAV肾损伤患者。在诱导缓解期对患者在使用标准激素联合免疫抑制剂治疗的基础上联合PAIA治疗,于初次治疗前、PAIA治疗后、PAIA治疗后1个月及3个月时分别收集患者的人口学特征,比较治疗前后患者的免疫学指标、伯明翰血管炎活动性评分(Birmingham Vasculitis Aactivity Score,BVAS)及相关临床指标等变化,并记录其不良反应情况,评估该疗法的短期疗效。结果:10例患者中男性6例,女性4例,年龄为(61.5±11.4)岁,从发病到确诊治疗的时长为(2.8±1.8)个月,均为抗髓过氧化物酶抗体阳性,2例合并抗蛋白酶3(anti-proteinase 3,PR3)抗体阳性(≥2.3 U/mL)。10例患者普遍表现为多器官受累,肾损伤严重,估算肾小球滤过率(estimated glomerular filtration rate,eGFR)均<30 mL/(min·1.73 m2)。随访3个月期间,10例患者BVAS、红细胞沉降率(erythrocyte sedimentation rate,ESR)及血红蛋白较治疗前的水平均可见持续改善(均P<0.05)。与治疗前相比,治疗3个月后患者的ANCA滴度、血清肌酐及尿红细胞水平均下降,eGFR水平回升(均P<0.05);患者的血清白蛋白、C反应蛋白及补体水平在治疗前后的差异均无统计学意义(均P>0.05)。1例患者纳入时已进入肾脏替代治疗阶段的患者治疗后未脱离维持性透析;1例患者在治疗过程中出现一过性低血压,对症处理后好转;余患者治疗过程中耐受性良好。结论:PAIA联合激素和环磷酰胺治疗可快速降低血清ANCA水平,有效改善AAV合并肾损伤患者疾病活动情况,提示较好的短期肾脏预后,且整体安全性良好。