Prevalence and Factors Associated with Positivity of Antinuclear Antibodies (ANA) Patterns, Native Anti-DNA and Extractable Nuclear Antigens (ENA) Antibodies: Experience from a Laboratory in Dakar

Background: Diagnosis of autoimmune diseases (AID) is challenging, due to overlapping features with other non-immune disorders. Anti-nuclear antibodies (ANA) are sensitive screening tests but anti-deoxyribonucleic acid-antibody (anti-DNA), and anti-extractable nuclear antigens (anti-ENA) are specific for AIDs. We aimed to look at ANA patterns in our patients and correlated them with anti-ENA for proper interpretation and better patient management cost-effectively. Methods: A retrospective study was conducted over 1 year from January to December 2022 who were tested for ANA at biology medical laboratory of Pasteur Institute of Dakar. Anti-ENA and anti-DNA results were also analyzed for ANA-positive patients. Statistical analysis was performed using STATA 14.0, p Results: 216 patients were analyzed. Women predominated at 79.2% and mean age was 48 years [CI 95%, 46 - 50], with extremes of 10 and 89. Most represented age group was [41 - 60] with 38%. ANA was positive in 27 (12.5%) of patients, 59.2% of whom were strongly positive (titer of 1/1000, 1/3200 or 1/6400). The most common pattern was nuclear speckled, which was found in 77.8% of samples. Anti-ENA and anti-DNA positivity in ANA-positive patients was found respectively in 63% (17/27) and 1.4% (3/27) of the samples analyzed. Most commonly identified anti-ENA was anti-Sm 29.6%, anti-SSA 29.6%, anti-Ro-52 25.9%, anti-RNP 18.5% and anti-SSB 14.8% which was associated with speckled pattern. Association results indicated a significant relationship between both tests and between ANA titer in the anti-ENA- and ANA-positive patients (p 0.001). Conclusions: ANA, Anti-ENA and anti-DNA antibodies are essential for AIDS diagnosis. However, the testing repertoire should follow an algorithm comprising of clinical features, followed by ANA results with nuclear, mitotic, and cytoplasmic patterns, anti-ENA, and anti-DNA for a more meaningful, and cost-effective diagnostic approach.

Peri-nuclear antibodies correlate with survival in Greek primary biliary cirrhosis patients

AIM:To investigate possible associations of anti-nuclear envelope antibody(ANEA)with disease severity and survival in Greek primary biliary cirrhosis(PBC)patients.METHODS:Serum samples were collected at diagnosis from 147 PBC patients(85%female),who were followed-up for a median 89.5 mo(range 1-240).ANEA were detected with indirect immunofluorescence on 1% formaldehyde fixed Hep2 cells,and anti-gp210 antibodies were detected using an enzyme linked immunosorbent assay.Findings were correlated with clinical data,histology,and survival.RESULTS:ANEA were detected in 69/147(46.9%) patients and 31/147(21%)were also anti-gp210 positive.The ANEA positive patients were at a more advanced histological stage(Ⅰ-Ⅱ/Ⅲ-Ⅳ56.5%/43.5% vs 74.4%/25.6%,P=0.005)compared to the ANEA negative ones.They had a higher antimitochondrial antibodies(AMA)titer(≤1:160/>1:160 50.7%/49.3%vs 71.8%/28.2%,P=0.001)and a lower survival time(91.7 ±50.7 mo vs 101.8±55 mo,P=0.043).Moreover,they had more advanced fibrosis,portal inflammation,interface hepatitis,and proliferation of bile ductules(P =0.008,P=0.008,P=0.019,and P=0.027,respectively).They also died more frequently of hepatic failure and/or hepatocellular carcinoma(P=0.016).ANEA positive,anti-gp210 positive patients had a difference in stage(Ⅰ-Ⅱ/Ⅲ-Ⅳ54.8%/45.2%vs 74.4%/25.6%,P= 0.006),AMA titer(≤1:160/>1:160 51.6%/48.4%vs 71.8%/28.2%,P=0.009),survival(91.1±52.9 mo vs 101.8±55 mo,P=0.009),and Mayo risk score(5.5 ±1.9 vs 5.04±1.3,P=0.04)compared to the ANEA negative patients.ANEA positive,anti-gp210 negative patients had a difference in AMA titer(≤1:160/>1:160 50%/50%vs 71.8%/28.2%,P=0.002),stage(Ⅰ-Ⅱ/Ⅲ -Ⅳ57.9%/42.1%vs 74.4%/25.6%,P=0.033),fibrosis(P=0.009),portal inflammation(P=0.018),interface hepatitis(P=0.032),and proliferation of bile ductules(P=0.031).Anti-gp210 positive patients had a worse Mayo risk score(5.5±1.9 vs 4.9±1.7,P=0.038)than the anti-gp210 negative ones.CONCLUSION:The presence of ANEA and anti-gp210 identifies a subgroup of PBC patients with advanced disease severity and poor prognosis.

Relationship and significance between anti-b2-glycoproteinⅠantibodies and platelet activation state in patients with ulcerative colitis

AIM： To study the relationship between anti-β2- glycoprotein Ⅰ （aβ2GPⅠ） antibodies and platelet activation state in patients with ulcerative colitis （UC） and its significance. METHODS： Peripheral blood samples were collected from 56 UC patients （34 males and 22 females, aged 43.5 years, range 21-66 years）, including 36 at active stage and 20 at remission stage, and 25 sex-and age-matched controls. The level of aβ2GP Ⅰ was measured by ELISA. The platelet activation markers, platelet activation complex- Ⅰ （PAC- Ⅰ ） and P-selectin （CD62P） were detected by flow cytometry. RESULTS： The A value for IgG aβ2GP Ⅰ in the active UC group was 0.61 ± 0.13, significantly higher than that in the remittent UC and control groups （0.50 ± 0.13 and 0.22 ± 0.14, P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. The A value for IgM aβ2GP Ⅰ in the active and remittent UC groups was 0.43 ± 0.13 and 0.38 ± 0.12, significantly higher than that in the control group （0.20 ± 0.12, P 〈 0.01）. However, there was no significant difference between the two groups （P 〉 0.05）. The PAC- Ⅰ positive rate for the active and remittent UC groups was 30.6% ± 7.6% and 19.6% ± 7.8% respectively, significantly higher than that for the control group （6.3% ± 1.7%,P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. The CD62P positive rate for the active and remittent UC groups was 45.0% ± 8.8% and 31.9% ± 7.8% respectively, significantly higher than that for the control group （9.2% ± 2.7%, P 〈 0.01）. There was a significant difference between the two groups （P 〈 0.01）. In the active UC group, the more severe the state of illness was, the higher the A value for IgG aβ2GP Ⅰ was, and the positive rate for PAC-Ⅰ and CD62P was positively correlated with the state of illness （Faβ2GP Ⅰ = 3.679, P 〈 0.05; FPAC-Ⅰ （%） = 5.346, P 〈 0.01; and FCD62P （%） = 5. 418, P 〈 0.01）. Meanwhile, in the same state of illness, the A value for IgG aβ2GP Ⅰ was positively correlated to the positive rates for PAC-Ⅰ and CD62P. CONCLUSION： aβ2GP Ⅰ level, platelet activation state and their relationship of them are closely correlated with the pathogenesis and development of UC.

Prevalence of Anti-Cardiolipin and Anti-β2 Glycoprotein Antibodies in Indian Systemic Lupus Erythematosus Patients

Anti-phospholipid antibodies (APA) like anti-cardiolipin antibodies (ACA) and anti-β2glycoprotien (anti-β2GP) are important cause of venous and arterial thrombosis and other occlusive vascular diseases. The prevalence of these antibodies in SLE patients at the time of diagnosis is not known in Indian SLE patients. This study was conducted to evaluate the prevalence of ACA and anti-β2GP autoantibodies in SLE patients and to correlate them with disease activity and immune parameters such as C3, C4 and CRP levels. where 85 SLE patients referred from Rheumatology Department, KEM hospital, Mumbai were studied. SLE disease activity was evaluated by SLE Disease Activity Index (SLEDAI) score at the time of evaluation. All patients studied were in an active stage of disease of which 37.6% patients had renal disorders, which were categorized as Lupus Nephritis (LN) and 62.3% patients did not show any renal manifestations (non-LN). ACA and anti-β2GP autoantibodies, to IgG and IgM subclasses were tested by ELISA. C3, C4 and CRP levels were detected by nephelometer. It was observed that 12.9% patients were IgG-ACA and IgM-ACA positive and ACA positivity was noted more among LN group Anti-β2GP autoantibody positivity was 27.1% for IgG and 31.8% for IgM., IgG-anti-β2GP antibodies were slightly higher in non-LN patients, whereas a higher incidence of IgM-anti-β2GP antibodies were detected in LN patients. Hence detection both ACA and anti-β2GP antibodies along with associated immune parameters were helpful to evaluate their possible association with disease severity in SLE patients. A long term follow up of patients having ACA and anti-β2GP antibodies without thrombotic event is also needed to detect their possible thrombotic event in future along with their clinical presentation.

Periodontitis and Inflammation: Plasma High Titer Naturally Occurring Anti-Glucan Antibodies Form Immune Complex with Streptococcus mutans Antigen

Atheromatous plaques usually contain antigens of the periodontitis-causing bacteria Streptococcus mutans though molecular mechanism of this incorporation remains unknown. Since vascular adhesion and inflammatory potential of Immune Complexes (IC) are known we investigated the naturally occurring plasma antibodies that recognize major antigens from S. mutans. S. mutans-binding plasma proteins (SMBP) prepared by affinity chromatography on a column of heat-killed S. mutans could recognize α- and β-linked glucose in dextran and yeast respectively but not galactose in glycoproteins. SMBP contained only three proteins, each corresponding in electrophoretic mobility to standard plasma IgG, IgA or IgM. The major positively and negatively charged protein antigens (PSMAg and NSMAg) isolated from S. mutans by electrophoresis and ion exchange chromatography respectively were recognized sugar-reversibly by the anti-β-glucan antibody (ABG) and though less avidly, by the dextran-binding immunoglobulin (DIg) in normal plasma. NSMAg addition resulted in near doubling of IC-bound immunoglobulins in immunoglobulin-rich fraction of plasma. IC isolated from above fraction after NSMAg addition had substantially more IgA and IgM content than total plasma immunoglobulins. IC formation by NSMAg was significantly inhibited by ABG- and DIg-specific sugars or by selective withdrawal of ABG or DIg from plasma. ABG and DIg being relatively high titer plasma antibodies IC formation with them suggested a possible route for vascular adhesion and damage by S. mutans and its antigens. Further, high IgA content of these ICs indicated their susceptibility to tissue uptake through cell surface galectin-1 for which IgA is the lone immunoglobulin ligand.

孕妇抗核抗体谱与胎儿发育异常的关系

目的:探讨孕妇抗核抗体谱与胎儿发育异常的关系。方法:选择2020年1月至2023年12月在郑州大学第三附属医院行抗核抗体谱检测的孕妇2490例,通过胎儿四维彩超和超声心动图检测胎儿发育情况。使用免疫印迹法检测孕妇血清抗核抗体谱(抗双链DNA、核小体、组蛋白、核糖体蛋白P、SM、nRNP、SSA/Ro60、SSA/Ro52、SSB/La、着丝点、Scl-70和Jo-1抗体)。比较胎儿发育正常和发育异常孕妇血清抗核抗体谱阳性检出情况,胎儿心脏发育正常和发育异常孕妇血清抗着丝点抗体、抗SSA/Ro52抗体、抗着丝点或SSA/Ro52抗体和抗SSA/Ro60+SSA/Ro52+SSB/La抗体阳性检出情况。分析抗核抗体谱在胎儿心脏异常中的诊断价值。结果:2490例孕妇抗核抗体谱阳性检出率为5.38%。胎儿发育异常孕妇抗核抗体谱阳性检出率为7.2%(37/514),高于胎儿发育正常孕妇的4.9%(97/1976)(P=0.040)。胎儿发育正常和发育异常孕妇抗SSA/Ro60、SSA/Ro52、SSB/La和着丝点抗体阳性检出率比较,差异无统计学意义(P>0.05);78例胎儿心脏异常,其中心血管畸形51例、心律失常27例。胎儿心脏发育正常和发育异常孕妇抗着丝点抗体、抗SSA/Ro52抗体、抗着丝点或SSA/Ro52抗体、抗SSA/Ro60+SSA/Ro52+SSB/La抗体阳性检出率比较,差异有统计学意义(P<0.05)。孕妇抗着丝点或SSA/Ro52抗体预测胎儿心脏发育异常的敏感度和阴性预测值分别为0.090和0.971;抗着丝点抗体预测的特异度和阳性预测值分别为0.999和0.400。结论:孕妇血清抗着丝点抗体、抗SSA/Ro52抗体以及抗SSA/Ro60+SSA/Ro52+SSB/La抗体阳性,可能提示胎儿心脏发育异常。

血清抗核抗体阳性对大动脉炎患者冠状动脉病变风险的影响

目的探讨大动脉炎患者中抗核抗体阳性的临床意义及对患者冠状动脉病变风险的影响。方法纳入首都医科大学附属北京安贞医院2014年1月至2022年2月住院的大动脉炎且无其他免疫疾病的患者158例。根据抗核抗体检测结果,分为抗核抗体阳性组(46例)和抗核抗体阴性组(112例)。收集研究对象的一般资料、实验室检查和影像学资料,并进行组间相关分析。采用Logistic回归方法分析相关危险因素,并进行预测模型的建立;采用受试者工作特征(ROC)曲线进行不同预测模型曲线下面积(AUC)的比较,分析大动脉炎患者抗核抗体阳性的临床意义。结果抗核抗体阳性组的高脂血症病史发生率高于抗核抗体阴性组[23.9%(11/46)比7.1%(8/112)](P=0.003)。抗核抗体阳性组冠状动脉病变发生率明显高于抗核抗体阴性组[50.0%(23/46)比22.3%(25/112)](P=0.001)。多因素Logistic回归分析显示,抗核抗体阳性和高脂血症是大动脉炎患者冠状动脉病变的独立危险因素(均P<0.05)。建立大动脉炎冠状动脉病变的预测模型,ROC曲线分析显示,以发病年龄、性别、病程、体重指数、美国国立卫生研究院(NIH)评分和高脂血症病史为基础的模型AUC为0.713(P=0.001),而加入抗核抗体阳性作为危险因素后预测模型AUC提高至0.761(P<0.001)。针对无高脂血症病史的患者(139例),基于多因素Logistic回归分析构建预测模型,包括发病年龄、性别、病程、体重指数、NIH评分的基线模型AUC为0.645(P=0.046),而加入抗核抗体阳性作为危险因素后,AUC提高至0.709(P=0.004)。结论大动脉炎患者合并抗核抗体阳性时需警惕冠状动脉病变,尤其是在既往无高脂血症的患者中,更具有临床提示意义。

老年系统性红斑狼疮患者抗核抗体谱检测阳性率观察

目的探讨ANAs检测在老年SLE患者中的阳性率及不同年龄段抗体分布特点与临床应用。方法用IFA检测217例老年SLE患者和211例非老年SLE患者ANA和抗ds-DNA抗体,免疫印迹法检测ANAs。对SLE患者不同年龄段ANAs阳性率进行统计分析,比较阳性率特点。结果ANA在老年组、非老年组SLE患者中阳性率分别为98.58%、99.54%;ANAs在老年组、非老年组SLE患者中阳性率分别为98.58%、92.17%,其中抗ds-DNA抗体、nRNP、抗Sm抗体、ANuA、r-RNP和PCNA的阳性率,老年组低于非老年组,差异有统计学意义(P<0.05);nRNP、AMA M2的阳性率,在老年组SLE患者不同年龄段间差异有统计学意义(P<0.05)。结论ANA检测在老年组、非老年组SLE患者中均有较高的阳性率,ANAs老年组阳性率低于非老年组;抗nRNP抗体和抗AMA M2抗体随年龄发生不同的改变可能是ANAs检测SLE老年组有别于非老年组的特点,总之ANAs检测对老年SLE诊断具有重要意义。

自身免疫性肝炎临床误诊分析

目的探讨自身免疫性肝炎(AIH)的诊治措施及误诊原因、防范措施。方法回顾性分析2020年1月—2022年5月收治的AIH误诊为病毒性肝炎21例的临床资料。结果21例主要症状为食欲缺乏、乏力、黄疸、发热,伴腹胀12例,恶心5例,胸闷和胸痛4例,呕吐及关节痛各3例。体形消瘦,巩膜及皮肤黏膜黄染明显。查血丙氨酸转氨酶和天冬氨酸转氨酶升高;7例γ-谷氨酰转肽酶升高,碱性磷酸酶和总胆红素升高各6例。腹部B超检查示肝大18例,肝内回声不均。初期外院诊断为病毒性肝炎,予相应治疗15 d无好转,遂转我院。查血抗平滑肌抗体(SMA)、抗核抗体(ANA)阳性,血γ-球蛋白、IgG升高,结合肝炎病毒血清学检测阴性及相关病史,遂明确诊断为AIH。误诊时间18~21 d。确诊后,18例予泼尼松单独治疗,3例予泼尼松联合硫唑嘌呤治疗。治疗1年后随访,患者病情稳定,无复发。结论AIH发病较隐匿,以年轻女性高发,临床表现多样且无特异性,易误诊为病毒性肝炎,行肝炎病毒血清学检查及ANA、SMA、抗肝肾微粒体、免疫球蛋白或肝组织病理检查可区分二者,确诊后应及时予有效治疗,以改善患者预后。

骨保护素/核因子κB受体活化因子/核因子κB受体活化因子配体调节骨代谢及其靶向治疗在口腔领域的应用

背景:骨保护素/核因子κB受体活化因子/核因子κB受体活化因子配体是偶联破骨细胞、成骨细胞分化和活化的重要细胞因子,是调节骨代谢的关键因子,影响着免疫系统、骨的再生和重塑,与牙槽骨的生理性及病理性改建密切相关。目的:分析总结骨保护素/核因子κB受体活化因子/核因子κB受体活化因子配体信号通路对牙槽骨改建的影响及其靶向治疗在口腔领域应用研究中的进展。方法:检索中国知网及PubMed数据库收录的相关文献。中文检索词为“骨保护素,抗RANKL抗体,核因子κB受体活化因子配体,牙周炎,正畸牙移动,种植,牙齿萌出,根尖周病变,牙槽骨吸收”,英文检索词为“OPG,anti-RANKL antibody,RANKL,periodontitis,orthodontic tooth movement,implant,tooth eruption,periapical lesion,alveolar bone resorption”,最终纳入63篇文献进行归纳总结。结果与结论:①抗核因子κB受体活化因子配体通过靶向抑制破骨细胞形成和牙槽骨吸收来治疗口腔疾病;②局部和全身抗核因子κB受体活化因子配体治疗可以抑制牙周炎、种植体周围炎、根尖周病变的进展,且其在预防正畸后复发、增强正畸支抗和种植体骨结合方面也发挥重要作用;③核因子κB受体活化因子配体通过靶向促进破骨细胞分化来治疗口腔疾病;④核因子κB受体活化因子配体治疗可以加速正畸牙移动、缩短治疗周期、减少正畸并发症的发生;⑤虽然抗核因子κB受体活化因子配体治疗存在局限性,但是可以通过合理应用如应用前排除危险因素,应用期间定期口腔维护、避免创伤性牙槽手术等措施来规避。

药物性肝损伤患者血清抗核抗体阳性临床意义探讨

目的分析不同临床类型和血清抗核抗体(ANA)阳性的药物性肝损伤(DILI)患者临床特征和转归。方法2019年12月~2023年4月安徽医科大学第一附属医院感染病科诊治的DILI患者64例,均行肝活检术和常规血清检测,排除自身免疫性肝炎(AIH)。其中血清ANA阳性31例,ANA阴性33例。结果在本组64例DILI患者中,导致疾病的药物为中草药占59.4%,抗生素占10.9%和非甾体类药物占4.7%;肝细胞型42例(65.6%),胆汁淤积型11例(17.2%)和混合型11例(17.2%);ANA阳性组肝细胞型、胆汁淤积型和混合型分别占77.4%、6.5%和16.1%,而ANA阴性组分别为54.5%、5.2%和30.3%,无统计学差异(P>0.05);ANA阳性组血清球蛋白和Ig G水平分别为26.1(22.2,29.9)g/L和13.6(12.1,17.7)g/L,显著高于ANA阴性组【分别为22.9(21.6,25.2)g/L和11.5(9.4,12.9)g/L,P<0.05】;两组所有患者均获得肝功能指标恢复正常,其中在31例ANA阳性DILI患者中,1例最终导致药物诱自身免疫性肝炎(DI-AIH),2例考虑为免疫介导的药物性肝损伤(IM-DILI),这3例患者均加用糖皮质激素治疗后,血清ALT和AST水平恢复正常。结论大多DILI患者预后良好,但血清ANA阳性者可能发展至DI-AIH或IM-DILI,需加用皮质激素治疗,值得注意。

局部注射不同剂量抗RANKL抗体对大鼠牙槽骨骨质疏松的影响

目的:探究局部注射抗核因子κB受体活化因子配体(receptor activator of nuclear factor kappa-B ligand,RANKL)抗体对大鼠牙槽骨骨质疏松的影响及不同剂量间的比较研究。方法:采用SD大鼠30只(对照组9只,实验组21只),通过皮下注射地塞米松建立大鼠牙槽骨骨质疏松模型,建模5周后进行模型验证,模型验证成功后,将实验组随机分为3组;实验组Ⅱ、Ⅲ于1、3、7 d在大鼠右上第一磨牙腭侧黏骨膜下注射兔抗大鼠RANKL抗体0.1μg/位点和1μg/位点,实验组Ⅰ注射等体积生理盐水。1周后处死取材,采用微计算机断层扫描技术(micro computed tomography,Micro-CT)测量右上第一磨牙区牙槽骨骨密度(bone mineral density,BMD)、骨体积分数(bone volume fraction,BV/TV);牙槽骨组织病理学染色观察组织形态学改变及破骨细胞数目变化;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测牙龈中RANKL/骨保护素(osteoprotegerin,OPG)比值变化。结果:成功建立大鼠牙槽骨骨质疏松模型。Micro-CT和组织病理学结果显示,注射RANKL抗体后,牙槽骨BMD、BV/TV增加,破骨细胞数目减少,成骨细胞活跃,骨髓腔减小,但高、低剂量抗体相比变化不显著;ELISA结果显示,注射高剂量抗体后牙龈中RANKL/OPG比值显著降低,注射低剂量抗体后RANKL/OPG比值虽降低,但变化不显著。结论:局部注射抗RANKL抗体可以改善大鼠牙槽骨骨质疏松,低剂量即可达到改善效果。

以脊柱关节痛为首发症状的布氏杆菌病患者的血清学检测分析

目的:探讨与分析以脊柱关节痛为首发症状的布氏杆菌病患者的血清学检测价值.方法:选择2020年4月—2023年2月青海省西宁市大通回族土族自治县疾病预防控制中心收治的86例布氏杆菌病患者作为研究对象,其中以脊柱关节痛为首发症状的36例患者为关节痛组,非以脊柱关节痛为首发症状的50例患者为对照组,检测2组的血清学指标,判定患者的脊柱关节疼痛情况并进行相关性分析.结果:2组的白细胞、红细胞、血红蛋白、血小板计数等对比,组间差异无统计学意义(P>0.05).关节痛组的血清抗核抗体、抗角蛋白抗体、抗滑膜环瓜氨酸肽抗体、基质金属蛋白酶-3含量均高于对照组,血清1,25-二羟基维生素D3含量均低于对照组,差异有统计学意义(P<0.05).关节痛组的脊柱关节疼痛VAS评分明显高于对照组,差异均有统计学意义(P<0.05).在86例患者中,Pearson分析显示,血清抗核抗体、抗角蛋白抗体、抗滑膜环瓜氨酸肽抗体、基质金属蛋白酶-3、1,25-二羟基维生素D3含量与脊柱关节视觉模拟量表(VAS)评分存在相关性,差异均有统计学意义(P<0.05).二元logistic回归分析显示,血清抗核抗体、抗角蛋白抗体、抗滑膜环瓜氨酸肽抗体、基质金属蛋白酶-3、1,25-二羟基维生素D3含量为影响脊柱关节疼痛VAS评分的重要因素,差异均有统计学意义(P<0.05).结论:以脊柱关节痛为首发症状的布氏杆菌病患者比较常见,多表现为血清抗核抗体、抗角蛋白抗体、抗滑膜环瓜氨酸肽抗体、基质金属蛋白酶-3含量增高与血清1,25-二羟基维生素D3含量降低,加强血清学检测具有重要的临床意义.

抗CCL20单克隆抗体对血清淀粉样蛋白A相关的结节病中促炎性细胞因子的抑制作用

目的探讨抗CCL20单克隆抗体是否可抑制高水平血清淀粉样蛋白A(SAA)相关的结节病外周血单个核细胞(PBMC)中促炎性细胞因子的表达。方法提取健康人和结节病患者的PBMC,并进行分组。除空白对照组(正常人PBMC)和结节病PBMC对照组(患者PBMC)外,另将结节病组PBMC中分别加入SAA、SAA+抗CCL20单克隆抗体、SAA+NF-κB抑制剂BAY11-7082或地塞米松,经培养48 h后收集各组细胞,分别作为PBMC+SAA刺激组、PBMC+SAA+NF-κB抑制剂BAY11-7082组、PBMC+SAA+抗CCL20单抗组、PBMC+地塞米松干预组,使用ELISA分别测定各组细胞上清液中细胞因子IL-17A、IL-23、IL-6、CCL20和TGF-β1的表达,使用RT-PCR测定各组细胞中CCR6、IL-17、ROR-γt和Foxp3的mRNA表达水平,使用Western blot检测各组细胞中p-NF-κB和NF-κB蛋白的表达水平。结果PBMC+SAA+抗CCL20单抗组与PBMC+SAA组相比,细胞上清液中的IL-17A、IL-23和IL-6的水平显著降低(P<0.01),而TGF-β1的含量显著升高(P<0.01)。PBMC+SAA+抗CCL20单抗组相较于PBMC+SAA组,细胞中CCR6、IL-17A、ROR-γtmRNA水平显著降低(P<0.01),而Foxp3 mRNA差异无统计学意义(P>0.05)。PBMC+SAA+抗CCL20单抗组中p-NF-κB蛋白表达水平较PBMC+SAA组明显降低(P<0.01)。结论抗CCL20单克隆抗体可抑制高水平SAA相关的结节病PBMC中促炎性细胞因子的表达。

抗核抗体联合T淋巴细胞亚群检测对复发性流产妊娠结局的预测价值

目的:探究抗核抗体(ANA)联合T淋巴细胞亚群检测对复发性流产(RSA)妊娠结局的预测价值。方法:选择2020年2月—2022年7月赣州市妇幼保健院收治的50例RSA患者入研究组,另选择50例同期于本院行产前检查的正常妊娠孕妇入对照组。采集所有参检者清晨空腹肘静脉血5 mL,检测ANA、T淋巴细胞亚群(CD4^(+)、CD8^(+)、CD16^(+)CD56^(+))水平,并计算CD4^(+)/CD8^(+)值。结果:研究组29例患者于孕10周前发生流产,流产率为58.00%(29/50);对照组于孕10周前发生流产共13例,流产率为26.00%(13/50),两组流产率对比,差异有统计学意义(χ^(2)=10.509,P=0.001)。相比于对照组,研究组ANA阳性检出率较高,CD4^(+)、CD16^(+)CD56^(+)、CD4^(+)/CD8^(+)水平均较高,CD8^(+)水平较低,差异均有统计学意义(P<0.05)。联合检测在RSA中的预测价值最高,AUC为0.872,敏感度为0.875,特异度为0.393,其次为CD16^(+)CD56^(+)、CD4^(+)、CD4^(+)/CD8^(+)、CD8^(+)、ANA。结论:ANA联合T淋巴细胞亚群检测在RSA诊断中应用价值较高,能够有效预测RSA发生风险,为临床尽早干预提供可靠的参考。

基于胃黏膜MyD88、ATF2、NF-κB蛋白表达情况分析化浊解毒方加减治疗慢性萎缩性胃炎的机制

目的:基于胃黏膜髓样分化因子88(MyD88)、活化复制因子2抗体(ATF2)、核因子-kB(NF-κB)蛋白表达情况分析化浊解毒方加减治疗慢性萎缩性胃炎(CAG)的机制。方法:采用随机数字表法将于2021年1月~2023年5月在上海中医药大学深圳医院接受治疗的120例CAG患者分为A组和B组,每组各60例。A组给予幽门螺杆菌根除三联疗法,B组在A组基础上给予化浊解毒方加减治疗,两组均持续治疗2周。比较两组治疗2周后的临床疗效,治疗前、治疗2周后的胃镜黏膜征象积分、成纤维细胞生长因子23(FGF-23)、白细胞介素-32(IL-32)、降钙素基因相关肽(CGRP)、人表皮生长因子(EGF)、胃黏膜MyD88、ATF2、NF-κB蛋白表达情况,治疗期间的不良反应。结果:B组治疗2周后的总有效率高于A组(88.33%vs 71.67%,P<0.05)。两组治疗2周后的胃镜黏膜征象各项评分均比治疗前低,且与A组比较,B组较低(P<0.05)。两组治疗2周后的血清FGF-23、IL-32、EGF水平均比治疗前低,且与A组比较,B组较低;两组血清CGRP水平比治疗前升高,且与A组比较,B组较高(P<0.05)。两组治疗2周后的胃黏膜MyD88、ATF2、NF-κB蛋白表达均比治疗前低,且与A组比较,B组较低(P<0.05)。B组和A组治疗期间的不良反应发生率比较,差异无统计学意义(5.00%vs 11.67%,P>0.05)。结论:化浊解毒方加减治疗可有效减轻CAG患者胃黏膜炎症反应及胃黏膜损伤程度,分析其机制可能与其能调节胃黏膜MyD88、ATF2、NF-κB蛋白的表达有关,有助于提高治疗效果,且安全性良好。

Appearance of an inhibitory cell nuclear antigen in rat and human serum during variable degrees of hepatic regenerative activity

AIM To determine whether proliferating cell nuclear antigen (PCNA) is present in the peripheral circulation and whether PCNA levels correlate with enhanced regenerative activity.METHODS In animal studies, adult male Sprague-Dawley rats (n=3-4/ group) were sacrificed at 0, 12, 24, 36, 48, 72 and 96 hours following 70% partial hepatectomy. At each interval, sera were analyzed by Western blot for PCNA by two monoclonal antibodies (PC-10 and 19F-4). In human studies, sera from 4 patients with liver cirrhosis and 4 healthy controls were tested in a similar manner.RESULTS The PC-10 monoclonal antibody identified a protein with a molecular mass of 120 KD which remained stable in rat sera for 24 hours following partial hepatectomy, then increased 1.5-fold at 48 hours prior to returning to baseline at 96 hours after partial hepatectomy. However, it was not detected in the sera of patients with or without liver disease. In the 19F-4 monoclonal antibody, a protein with a molecular mass of approximately 46 KD was found. which was present in rat sera prior to partial hepatectomy and for 12 hours after surgery. Thereafter, levels fell by approximately 50% at 24 hours, 65% at 36 hours and 75% at 48 hours where they remained until 96 hours after partial hepatectomy. The decrease in levels correlated with the extent of partial hepatectomy. In human sera, the appearance of this inhibitory cell nuclear antigen (ICNA) was higher in the sera of patients with cirrhosis than in healthy controls.CONCLUSION The PC-10 monoclonal antibody can detect a protein in the circulation when active hepatic regenerative activity is taking place. The 19F-4 monoclonal antibody, however, identifies a protein in both rat and human sera that inversely correlates with hepatic regenerative activity. This protein which is tentatively referred to as inhibitory cell nuclear antigen (ICNA) may be used in documenting the extent of suppression of hepatic regeneration.

Prevalence, significance and predictive value of antiphospholipid antibodies in Crohn's disease

AIM: To assess the prevalence and stability of different antiphospholipid antibodies(APLAs) and their association with disease phenotype and progression in inflammatory bowel diseases(IBD) patients.METHODS: About 458 consecutive patients [Crohn's disease(CD): 271 and ulcerative colitis(UC): 187] were enrolled into a follow-up cohort study in a tertiary IBD referral center in Hungary. Detailed clinical phenotypes were determined at enrollment by reviewing the patients' medical charts. Disease activity, medical treatment and data about evolvement of complications or surgical interventions were determined prospectively during the follow-up. Disease course(development f complicated disease phenotype and need for surgery),occurrence of thrombotic events, actual state of diseaseactivity according to clinical, laboratory and endoscopic scores and accurate treatment regime were recorded during the follow-up,(median, 57.4 and 61.6 mo for CD and UC). Sera of IBD patients and 103 healthy controls(HC) were tested on individual anti-β2-Glycoprotein-I(anti-β2-GPI IgA/M/G), anti-cardiolipin(ACA IgA/M/G)and anti-phosphatidylserine/prothrombin(anti-PS/PT IgA/M/G) antibodies and also anti-Saccharomyces cerevisiae antibodies(ASCA IgA/G) by enzyme-linked immunosorbent assay(ELISA). In a subgroup of CD(n = 198) and UC patients(n = 103), obtaining consecutive samples over various arbitrary timepoints during the disease course, we evaluated the intraindividual stability of the APLA status. Additionally,we provide an overview of studies, performed so far, in which significance of APLAs in IBD were assessed.RESULTS: Patients with CD had significantly higher prevalence of both ACA(23.4%) and anti-PS/PT(20.4%) antibodies than UC(4.8%, p < 0.0001 and10.2%, p = 0.004) and HC(2.9%, p < 0.0001 and15.5%, p = NS). No difference was found for the prevalence of anti-β2-GPI between different groups(7.2%-9.7%). In CD, no association was found between APLA and ASCA status of the patients.Occurrence of anti-β2-GPI, ACA and anti-PS/PT was not different between the group of patients with active vs inactive disease state according to appropriate clinical, laboratory and endoscopic scores in CD as well as in UC patients. All subtypes of anti-β2-GPI and ACA IgM status were found to be very stable over time, in contrast ACA IgG and even more ACA IgA status showed significant intraindividual changes.Changes in antibody status were more remarkable in CD than UC(ACA IgA: 49.9% vs 23.3% and ACA IgG:21.2% vs 5.8%). Interestingly, 59.1% and 30.1% of CD patients who received anti-TNF therapy showed significant negative to positive changes in ACA IgA and IgG antibody status respectively. APLA status was not associated with the clinical phenotype at diagnosis or during follow-up, medical therapy, or thrombotic events and it was not associated with the probability of developing complicated disease phenotype or surgery in a Kaplan-Meier analysis.CONCLUSION: The present study demonstrated enhanced formation of APLAs in CD patients. However,presence of different APLAs were not associated with the clinical phenotype or disease course.

Autoimmune liver disease-related autoantibodies in patients with biliary atresia

AIM To investigate the prevalence and clinical significance of autoimmune liver disease(ALD)-related autoantibodies in patients with biliary atresia(BA).METHODS Sera of 124 BA patients and 140 age-matched non-BA controls were assayed for detection of the following autoantibodies: ALD profile and specific anti-nuclear antibodies(ANAs), by line-blot assay; ANA and antineutrophil cytoplasmic antibody(ANCA), by indirect immunofluorescence assay; specific ANCAs and antiM2-3 E, by enzyme linked immunosorbent assay. Associations of these autoantibodies with the clinical features of BA(i.e., cytomegalovirus infection, degree of liver fibrosis, and short-term prognosis of Kasai procedure) were evaluated by Spearman's correlation coefficient.RESULTS The overall positive rate of serum autoantibodies in preoperative BA patients was 56.5%. ALD profile assay showed that the positive reaction to primary biliary cholangitis-related autoantibodies in BA patients was higher than that to autoimmune hepatitis-related autoantibodies. Among these autoantibodies, anti-BPO was detected more frequently in the BA patients than in the controls(14.8% vs 2.2%, P < 0.05). Accordingly, 32(25.8%) of the 124 BA patients also showed a high positive reaction for anti-M2-3 E. By comparison, the controls had a remarkably lower frequency of anti-M2-3 E(P < 0.05), with 6/92(8.6%) of patients with other liver diseases and 2/48(4.2%) of healthy controls. The prevalence of ANA in BA patients was 11.3%, which was higher than that in disease controls(3.3%, P < 0.05), but the reactivity to specific ANAs was only 8.2%. The prevalence of ANCAs(ANCA or specific ANCAs) in BA patients was also remarkably higher than that in the healthy controls(37.9% vs 6.3%, P < 0.05), but showed no difference from that in patients with other cholestasis. ANCA positivity was closely associated with the occurrence of postoperative cholangitis(r = 0.61, P < 0.05), whereas none of the autoantibodies showed a correlation to cytomegalovirus infection or the stages of liver fibrosis.CONCLUSION High prevalence of autoantibodies in the BA developmental process strongly reveals the autoimmunemediated pathogenesis. Serological ANCA positivity may be a useful predictive biomarker of postoperative cholangitis.