Kinetics of fetal cellular and cell-free DNA in the maternal circulation during and after pregnancy: implications for noninvasive prenatal diagnosis

BACKGROUND: Fetal genetic material is detectable in the maternal circulation and has been used for noninvasive prenatal diagnosis. However, few data are available concerning its quantity and natural history during gestation. STUDY DESIGN AND METHODS: This study prospectively characterized the kinetics of cellular and cell-free fetal DNA in the circulation of 25 healthy women during and after uncomplicated pregnancy. Real-time kinetic PCR was used to quantitate human Y-chromosome sequences, and liquid oligomer hybridization with (32)P-labeled probes was used to verify the identity of amplified products. RESULTS: In all male pregnancies, but no female pregnancies, low-level fetal Y-chromosome DNA was detected in both cellular and cell-free compartments beginning at 7 to 16 weeks but increasing steadily after 24 weeks and reaching a peak at parturition. The fetal DNA decreased rapidly after birth. CONCLUSION: Fetal genetic material can be detected throughout pregnancy, and its quantity is a function of gestational age and of whether the plasma or cellular compartment is examined. Both the absolute quantity of fetal DNA and its ratio to total DNA (maternal + fetal) are greater in the plasma than in the cellular compartment. Fetal DNA is cleared rapidly from both compartments after parturition, which suggests that turnover is dynamic. Because they provide prospective and quantitative data concerning fetal DNA levels, these observations and kinetic PCR methods may have implications for noninvasive prenatal diagnosis. Further studies will be needed to determine the immunologic implications of fetal-maternal DNA exchange and cellular microchimerism.     

Increased plasma/serum levels of prolactin in systemic lupus erythematosus: a systematic review and meta-analysis

Background: Prolactin (PRL), a polypeptide hormone produced by the pituitary gland, is involved in the regulation of humoral and cell mediated immune responses. PRL levels have been investigated in several autoimmune diseases including systemic lupus erythematosus (SLE), however, yielded different and inconsistent results. This study aims to derive a more precise evaluation on plasma/serum PRL levels in SLE patients, as well as the potential influential factors.

Methods: Studies published from 1 January 1987 to 31 December 2015 in English, which comparing plasma/serum PRL levels between SLE group and control group were searched in PubMed, EMBASE and The Cochrane Library databases. Pooled standard mean difference (SMD) with 95% confidence interval (CI) was calculated by fixed-effects or random-effect model analysis. Heterogeneity test was performed by the Q statistic and quantified using I², publication bias was evaluated using a funnel plot and Egger’s linear regression test.

Results: Five-hundred and forty-seven articles were obtained after searching databases, and 12 studies with 429 SLE patients and 326 controls were finally included. Meta-analysis revealed that, compared with the control group, the SLE group had significantly higher plasma/serum PRL levels (P < 0.001), with the SMD of 1.26 and 95%CI (0.70,1.82). Subgroup analyses showed that SLE patients from Asia and Europe had higher plasma/serum PRL levels. However, no significant change in plasma/serum PRL levels was observed in SLE patients from America (P > 0.05).

Conclusions: Overall, our study suggests that SLE patients have higher plasma/serum PRL level, but with a regional difference.     

利用母体血浆游离胎儿DNA进行胎儿SRY基因定量检测

目的探讨胎儿游离DNA在无创性产前诊断中的应用价值。方法行产前诊断的孕妇87例,分为中期妊娠(16～24周)61例(中孕组)和早期妊娠(11～14周)26例(早孕组),采用实时荧光定量PCR方法,利用Taqman探针对妊娠早、中期孕妇血浆游离的胎儿Y染色体性别决定基因(sex-determining region of Y-chromosome,SRY)进行扩增和定量分析。结果孕13周后的样本均得到特异性扩增,特异度和灵敏度均为100%;中孕组男性胎儿SRY检测的平均DNA浓度为155.46copies/mL血浆,实测范围为43.6～326.8copies/mL血浆;早孕组平均浓度为26.42copies/mL血浆,实测范围为14.3～73.3copies/mL血浆。结论采用荧光实时定量PCR在妊娠早、中期可稳定检测到胎儿游离DNA,可作为孕期性连锁遗传性疾病研究或产前诊断的有效补充手段。     

Microplate ELISA for detection of antibodies to DNA in patients with systemic lupus erythematosus: specificity and correlation with Farr radioimmunoassay

A 96-well microplate ELISA for the detection of antibodies to DNA is described. A number of buffers and precoating treatments were used to evaluate the optimal method for coating the plate with DNA. These included pretreatment of the plates with poly-L-lysine or protamine sulfate, and posttreatment with glutaraldehyde, none of which improved the performance of the assay. Whereas bicarbonate and borate coating buffers gave equivalent and satisfactory results, TRIS buffer resulted in very high binding of immunoglobulin to wells not coated with antigen. Sera from groups of patients with autoimmune disease as well as normal sera were tested against plates optimally coated with native E. coli DNA, calf thymus DNA, and heat-denatured DNA. Using native E. coli DNA, virtually none of 35 normal sera had any detectable antibody. With this antigen, as well as with native calf thymus DNA, significant levels of DNA antibody were found only in SLE patients. Most patients with SLE or drug-induced lupus, as well as some patients with rheumatoid arthritis and normal individuals had antibodies that bound to heat-denatured (single-stranded) DNA. Using either native E. coli or calf thymus DNA, a good correlation was found between the amount of DNA antibody detected by ELISA and the Farr-type radioimmunoassay.     

高效液相色谱-荧光法测定SLE患者血清色氨酸及其代谢产物

目的建立一种同时测定血清色氨酸(tryptophan,Trp)、犬尿氨酸(kynurenine,Kyn)和犬尿喹啉酸(kynurenic acid,Kyna)的高效液相色谱-荧光检测法(HPLC-FD),并用此法检测SLE患者血清中Trp及其代谢产物Kyn、Kyna的含量。方法血清标本经0.624 mol/L的高氯酸溶液去除蛋白质后取上清液20μl直接进样分析测定。色谱柱为Hypersil C18柱,流动相为0.20mol/L醋酸锌、8.3 mmol/L醋酸和2.5%的乙腈;流速为1.5 ml/min;荧光检测激发波长和发射波长在0~11 min分别为365nm和480 nm,11~15.5 min变换为344 nm和404 nm,15.5~20 min为254 nm和404 nm。采用建立的方法测定体检健康者和SLE患者血清Trp、Kyn和Kyna的含量。结果Trp的线性范围为0.610~196μmol/L,最低检出浓度为0.005μmol/L,平均回收率为103.71%;Kyn线性范围为0.049~98μmol/L,最低检出浓度为0.025μmol/L,平均回收率为97.45%;Kyna线性范围为1.05...     

BioPlex 2200全自动免疫分析仪检测血清中抗双链DNA抗体的临床应用

目的分析比较BioPlex 2200全自动免疫分析仪和酶联免疫吸附测定(ELISA)检测血清中抗双链DNA(dsDNA)抗体的灵敏度、特异度及其优缺点。方法采用BioPlex 2200和ELISA检测101例纳入研究者的血清抗dsDNA抗体。纳入研究者中49例为临床确诊的系统性红斑狼疮患者,52例作为对照(包括32例其他结缔组织病患者和20例体检健康者)。结果BioPlex 2200检测抗dsDNA抗体的灵敏度为57.14%,特异度为94.23%;ELISA检测抗dsDNA抗体的灵敏度为53.06%,特异度为96.15%。两种方法总体符合率为95.05%,χ2检验分析,差异无统计学意义(χ2=78.69,P>0.05),Kappa值评价一致性较好(Kappa=0.880)。BioPlex 2200检测抗dsDNA抗体,3组抗dsDNA抗体水平比较,差异有统计学意义(F=61.52,P<0.05)。结论BioPlex 2200和ELISA检测血清抗dsDNA抗体结果有较好的一致性,检测迅速,值得临床推广应用。     

SLE患者血清IL-2、IL-10的表达与双链DNA相关性研究

目的 探讨系统性红斑狼疮(SLE)患者血清中IL—2、IL—10的表达与双链DNA(ds—DNA)的关系。方法 用酶联免疫吸附试验(ELISA)检测了22例SLE患者血清中的白细胞介素2(IL—2)、IL—10和抗ds—DNA抗体水平。结果 SLE患者血清中IL—10和IL—10／IL—2比值均与抗ds—DNA抗体呈现较好的正相关性，IL—2与抗ds—DNA抗体呈现低度负相关性或不相关。结论 血清中IL—10、IL—10／IL—2比值与抗ds—DNA抗体的相关性可作为SLE患者病情的诊断、监测和疗效观察的指标。     

系统性红斑狼疮合并心力衰竭患者心肌肌钙蛋白I水平的监测及临床意义

目的探讨心肌肌钙蛋白I（cTnI）变化对诊断系统性红斑狼疮（SLE）心脏损伤的临床意义及其与病情活动的关系。方法应用Beckman Coulter Access Immunoassay System 2对SLE合并心力衰竭患者、SLE无心力衰竭患者和健康人血清cTnI水平进行测定,用SLE活动指数积分评价SLE患者病情活动情况。结果SLE心力衰竭组血清cTnI增高,与无心力衰竭组的差异有统计学意义;无心力衰竭组血清cTnI与健康对照组的差异无统计学意义。血清cTnI的变化与SLE病情活动指数无相关性;其变化与病程有相关性。结论SLE合并心力衰竭时有心肌损伤存在,提示预后较差,治疗时应加强保护心肌。     

Effective plasma concentrations of mycophenolic acid and its glucuronide in systemic lupus erythematosus patients in the remission-maintenance phase

What is known and Objective: Mycophenolate mofetil (MMF) has been reported recently to be effective in the treatment of systemic lupus erythematosus (SLE). The therapeutic range of mycophenolic acid (MPA) in SLE in the remission‐maintenance phase remains to be clarified. The aim of this study was to evaluate the therapeutic efficacy of MMF and predose plasma concentrations of MPA and its phenolic glucuronide (MPAG) in patients with SLE in the remission‐maintenance phase. Methods: Thirty‐one patients with SLE receiving a fixed dosage regimen of MMF (median and interquartile range, 1500 and 1000–2000 mg/day) for at least 1 month and who had not experienced any adverse drug reactions for more than 3 months were enrolled. Results: Significant improvement was observed after MMF administration in total haemolytic complement CH₅₀ and its fractions C3 and C4, immunoglobulins IgG, IgA and IgM, anti‐dsDNA antibody, serum concentration of albumin and red blood cell count, even though the mean daily dose of prednisolone was significantly reduced (P = 0·02). Median predose plasma concentrations of MPA and MPAG were 1·95 and 26·2 μg/mL (interquartile ranges, 0·94–2·96 and 18·6–53·7 μg/mL). Predose plasma concentrations of MPA and MPAG correlated significantly with MMF dose (r = 0·64, P < 0·01 and r = 0·39, P = 0·03). What is new and Conclusions: MMF improved clinical laboratory markers and reduced prednisolone dosage in SLE patients with predose plasma concentration of MPA and MPAG in the interquartile ranges of 0·94–2·96 and 18·6–53·7 μg/mL, respectively.     

β2-GPI-dependent and independent binding of anticardiolipin antibodies in patients with recurrent spontaneous abortions

We measured anticardiolipin antibodies (aCL) in plasma samples from 214 women with a history of recurrent spontaneous abortions by an enzyme-linked immunosorbent assay (ELISA) utilizing solid phase cardiolipin (CL) and β₂-glycoprotein I (β₂-GPI). Sixteen patients (7.5%) were positive for β₂-GPI-dependent aCL. Though β₂-GPI appeared to enhance the binding of aCL, β₂-GPI-independent aCL were also observed in these patients (4.7%). The patients were classified into three groups on the basis of their medical history, and analysis of data of individual groups revealed that the incidence of β₂-GPI-dependent aCL was significantly higher in patients who had experienced at least one fetal loss in the second or third trimester. © 1994 Wiley-Liss, Inc.     

The predictive value of fibrinogen‐to‐albumin ratio in the active,severe active,and poor prognosis of systemic lupus erythematosus: A single‐center retrospective study

ObjectiveTo evaluate the prediction and effect of fibrinogen‐to‐albumin ratio (FAR) on active, severe active, and poor prognosis of systemic lupus erythematosus (SLE).MethodsOne hundred and sixty‐eight patients with SLE who were treated in our hospital were enrolled, the clinical data, laboratory indexes, and disease prognosis of all patients were collected and analyzed.ResultsTriglyceride (TG), FAR, ESR, and anti‐dsDNA (+) were the influencing factors, while complement 3 (C3) was the protective factor of active SLE, the odds ratio (OR) values were 2.968, 3.698, 2.114, 2.727, and 0.652, respectively (p < 0.05). FAR, ESR, and anti‐dsDNA (+) were the influencing factors, while C3 was the protective factor of severe active SLE, the OR values were 3.791, 1.953, 2.187, and 0.742, respectively (p < 0.05). SLE disease activity index (SLEDAI), TG, FAR, and anti‐dsDNA (+) were the influencing factors, while C3 was the protective factor of poor prognosis SLE, the OR values were 3.024, 2.293, 3.012, 2.323, and 0.801, respectively (p < 0.05). FAR and FIB were positively correlated with SLEDAI, while ALB was negatively correlated with SLEDAI, the related coefficient (r) were 0.398, 0.267, −0.270, respectively. The receiver operating curve (ROC) analysis showed that the predictive values of FAR for active, severe active and poor prognosis SLE were 0.769, 0.769, and 0.734, respectively, were significant higher than FIB and ALB (p < 0.05).ConclusionFibrinogen‐to‐albumin ratio was an influencing factor of active, severe active, and poor prognosis SLE had higher predictive value than FIB and ALB for the activity and prognosis of SLE.     

Myth and reality: practical test system for the measurement of anti‐DNA antibodies in the diagnosis of systemic lupus erythematosus (SLE)

The myth persists that only the labor intensive Farr radioimmunoassay and Crithidia luciliae immunofluorescence (CL‐IFA) are systemic lupus erythematosus (SLE)‐specific tests. We compared them to ELISA with bacteriophage λ DNA (EL‐dsDNA) and denatured calf thymus DNA (EL‐ssDNA). By percentile ranking, the specificity cut‐off level was set both out of clinical context (SOCC) on 100 blood bank donors, and in clinical context (SICC) on 100 patients with either rheumatoid arthritis or scleroderma (50/50). Clinical sensitivity was calculated on 100 random SLE patients. At 95% SICC, the sensitivity of Farr, CL‐IFA, EL‐dsDNA, and EL‐ssDNA was similar (95%CI): 76% (66–84), 76% (66–84), 63% (53–72), and 75% (65–83), respectively; 87% of the patients were positive by at least one method and 55%by all methods. At 99% SICC, the sensitivity was also similar (95% CI): 57% (47–67), 47% (37–57), 58% (47–67), and 43% (33–53), respectively. The areas under ROC curve were similar (95% CI) when patients were used as controls for specificity. At 99% SOCC, EL‐ssDNA identified 89% positive, 2 negative but positive by another method at 95% SICC, and 9 negative (i.e. 89/2/9), followed by CL‐IFA (80/6/14), Farr (76/12/12), and EL‐dsDNA (64/23/13). Thus, at relatively low cost and easy automation, under the same conditions of specificity, the two ELISA tests combined were at least as good, if not superior, to CL‐IFA or Farr: they showed similar clinical sensitivity and also identified more patients with anti‐DNA antibodies. J. Clin. Lab. Anal. 24:77–84, 2010. © 2010 Wiley‐Liss, Inc.     

104例系统性红斑狼疮患者血清指标的变化及C1q抗体的表达

目的检测系统性红斑狼疮(SLE)患者血清指标及C1q抗体(Anti-C1q)的变化,并探讨其临床意义。方法选取104例SLE患者(SLE组)及100例体检健康者(健康对照组)作为研究对象,检测其血清中Anti-C1q、双链DNA(dsDNA)抗体、其他相关自身抗体及生化指标,并对结果进行分析。结果与健康对照组相比,SLE组的三酰甘油(TG)水平升高[(3.89±0.65)mmol/L vs.(1.12±0.48)mmol/L,P0.05],高密度脂蛋白(HDL)水平降低[(0.8±0.2)mmol/L vs.(1.2±0.3)mmol/L,P0.05],超敏C反应蛋白(hs-CRP)水平升高[(5.5±1.4)mg/L vs.(0.9±0.6)mg/L,P0.05],补体C3水平降低[(87.6±14.7)mg/L vs.(128.2±20.3)mg/L,P0.05],补体C4水平降低[(16.2±4.8)mg/L vs.(29.1±7.4)mg/L,P0.05]。SLE组Anti-C1q水平较健康对照组明显升高[(44±20)U/mL vs.(6.5±1.8)U/mL,P0.05],其中活动期SLE患者Anti-C1q水平明显高于非活动期患者(P0.05),狼疮性肾炎(LN)患者Anti-C1q水平明显高于非LN患者(P0.05),dsDNA抗体阳性者Anti-C1q水平明显高于dsDNA抗体阴性者(P0.05)。结论 SLE患者外周血中各种生化指标和自身抗体的异常为患者疾病诊断提供了一定的理论依据,其中Anti-C1q的异常表达更有助于对活动期SLE和LN的诊断。     

Immunotherapies application in active stage of systemic lupus erythematosus in pregnancy: A case report and review of literature

BACKGROUNDPregnancy in the setting of systemic lupus erythematosus can worsen the condition from the stable to active stage, with quality of life and fertility desire being particular concerns. Pregnancy in the active stage of systemic lupus erythematosus (ASLE), although rare and complicated to manage, can be treated favorably with immunotherapies ifs used properly. Here we report such a success case.CASE SUMMARYA 31-year-old primigravida patient, diagnosed with SLE seven years ago, was induced ASLE after a cold at 21 + weeks. The patient’s vital signs on presentation were normal. Her laboratory exam was remarkable for significant proteinuria, liver and renal dysfunction, and low C3 and C4 levels. Infectious work-up was negative. The patient was diagnosed with ASLE. She was given immunosuppressive agents (methylprednisolone, gamma globulin and azathioprine etc.) and plasma adsorption therapy, monitoring blood pressure every 8 h, fetal heart rate twice a day, and liver and renal function at least twice a week. Successful maternal and fetal outcomes are presented here.CONCLUSIONChild-bearing in ASLE has become more promising, even for this difficult case of ASLE with multiple organ damage. Thorough antepartum counseling, cautious maternal-fetal monitoring, and multi-organ function monitoring by multidisciplinary specialties are keys to favorable pregnancy outcomes.     

联合检测抗核抗体和抗双链DNA抗体在狼疮性肾炎诊断中的作用

目的探讨联合检测抗核抗体(ANA)和抗双链DNA抗体在狼疮性肾炎(LN)诊断中的作用.方法 对406例系统性红斑狼疮(SLE)患者(其中LN 122例)和120例健康体检者采用间接免疫荧光法测定ANA,应用欧蒙印迹法测定抗双链DNA抗体.结果 406例SLE患者,ANA阳性率平均为94.49%;与对照组比较差异有统计学意义(P<0.001).ANA阳性者核型,LN组核均质型和核颗粒型占84.35%,不伴肾炎组占72.12%(P<0.05).ANA滴度:LN组较不伴肾炎组,低滴度结果低,而高滴度结果高(P<0.05).ANA阳性者检测抗双链DNA抗体,其阳性率为:LN组65.22%;不伴肾炎组51.67%(P<0.05).抗双链DNA抗体滴度,低滴度结果LN组64.00%,不伴肾炎组81.30%;高滴度结果LN组36.00%,不伴肾炎组18.70%(P<0.05).两组抗双链DNA抗体高滴度时,其相应的ANA滴度:LN组高滴度结果占绝大多数,不伴肾炎组结果分布较均匀,两组间差异有统计学意义(P<0.05).结论 联合检测ANA及抗双链DNA抗体,并综合分析判断,对提高LN的诊断率及疗效观察、判断预后等方面具有重要意义.     

Concurrent monoclonal gammopathy and systemic lupus erythematosus in a known case of ulcerative colitis: A case report

Our patient had previously been diagnosed with Ulcerative colitis. The clinical manifestations of the patient along with laboratory tests such as anti‐dsDNA and proteinuria were also positive. Therefore, the clinical manifestation was consistent with SLE. In the following work up, monoclonal gammopathy in serum electrophoresis was also detected.     

系统性红斑狼疮患者免疫球蛋白轻链及补体B因子测定的临床意义

目的 探讨血清中免疫球蛋白轻链κ和λ含量、κ/λ比值以及补体B因子(PFB)含量的检测在系统性红斑狼疮(SLE)诊疗中的应用价值.方法 在特定蛋白仪上,采用免疫速率散射比浊法测定SLE患者及对照组血清中轻链κ和轻链λ、PFB含量,并计算κ/λ比值.结果与30例对照组比较,51例SLE患者血清中轻链κ[(16.51±6.27)g/L,P<0.01]和轻链λ[(10.30±4.35)g/L,P<0.01]均升高;κ/λ比值下降(1.60±0.32,P<0.01),PFB含量下降[(0.214±0.063)g/L,P<0.01].结论 血清中免疫球蛋白轻链κ和轻链λ含量、κ/λ比值及PFB含量检测对SLE的临床诊疗具有重要的意义.     

系统性红斑狼疮患者疾病不确定感与生活质量的相关分析

目的探讨系统性红斑狼疮患者疾病不确定感、生活质量状况及两者之间的关系。方法选择60例住院系统性红斑狼疮患者为研究对象,应用疾病不确定感量表和生活质量量表进行测评。结果系统性红斑狼疮患者疾病不确定感总分为（103．79±11．37）分;生活质量状况评分：社会功能（39．24±12．27）分,生理职能（41．67±28．35）分,情感职能（43．36±19．23）分,精神健康（47．21±21．52）分,活力（57．36±18．28）分,总体健康（61．23±15．21）分,生理功能（63．32±16．57）分,身体疼痛（66．29±17．26）分;疾病不确定感总体水平与生理职能、总体健康、社会功能、情感职能、精神健康呈负相关。结论系统性红斑狼疮患者存在较高水平的疾病不确定感,与生活质量存在一定的相关性。应加强临床护理及健康教育,开展科学随访,以降低患者的疾病不确定感,提高患者生活质量,改善其预后。