基于抗体-适配子建立的高尔基体蛋白73检测方法的研究

目的建立基于抗体-适配子双夹心的高尔基体蛋白73(GP73)ELISA检测方法,并用于血清GP73的检测。方法以抗体-适配子双夹心的模式,通过正交试验和方阵滴定试验确定GP73特异性抗体和适配子最佳工作水平、反应的温度和时间等。以GP73重组蛋白建立标准曲线并评价该方法灵敏度、精确度、线性及准确度等。利用该方法对59例对照组患者及77例肝癌患者血清进行GP73水平检测,并同步进行电化学发光定量测定甲胎蛋白(AFP)水平。结果该方法标准曲线的批内变异系数(CV)为3.87%,批间CV平均为4.44%;平均回收率为95.8%,灵敏度达12.0ng/mL。肝癌患者血清GP73水平均明显高于对照组、肝炎组和肝硬化组(P0.05)。GP73对原发性肝癌诊断灵敏度为85.7%,而AFP为62.3%;而早期肝癌中GP73和AFP灵敏度分别为76.7%、36.7%。结论成功建立基于抗体-适配子夹心的GP73ELISA检测方法,该方法可用于临床检测GP73水平。     

Swiss students and young physicians want a flexible goal-oriented GP training curriculum

Background: A growing shortage of general practitioners (GPs), in Switzerland and around the world, has forced countries to find new ways to attract young physicians to the specialty. In 2017, Switzerland began to fund hundreds of new study places for medical students. This wave of young physicians will soon finish University and be ready for postgraduate training. We hypothesized that an attractive postgraduate training program would encourage interested young physicians to pursue a GP career.

Methods: This is a cross-sectional survey of young physicians from the Swiss Young General Practitioners Association (JHaS), members of Cursus Romand de médecine de famille (CRMF), and all current medical students (5^th or 6^th years) (n?=?554) in Switzerland, excluding students indicating definitely not to become GPs. We asked all if they were likely to become a GP (Likert: 1-10), and then asked them to score general features of a GP training curriculum, and likely effects of the curriculum on their career choice (Likert scale). They then rated our model curriculum (GO-GP) for attractiveness and effect (Likert Scales, open questions).

Results: Most participants thought they would become GPs (Likert: 8 of 10). Over 90% identified the same features as an important part of a curriculum (“yes” or “likely yes”): Our respondents thought the GO-GP curriculum was attractive (7.3 of 10). It was most attractive to those highly motivated to become GPs. After reviewing the curriculum, most respondents (58%) felt GO-GP would make them more likely to become a GP. Almost 80% of respondents thought an attractive postgraduate training program like GO-GP could motivate more young physicians to become GPs.

Conclusions: Overall, medical students and young physicians found similar features attractive in the general and GO-GP curriculum, regardless of region or gender, and thought an attractive curriculum would attract more young doctors to the GP specialty.

• Key points

• An attractive postgraduate training program in general practice can attract more young physicians to become GPs.

• In this study cross-sectional survey including medical students (n?=?242) and young physicians (n?=?312) we presented general features for a curriculum and a model curriculum for general practice training, for evaluation of attractiveness to our study population.

• General practice training curriculum provides flexibility in choice of rotations, access to short rotations in a wide variety of medical specialties, training in specialty practices as well, mentoring and career guidance by GPs and guidance in choosing courses/certificate programs necessary for general practice.

• These findings help building attractive postgraduate training programs in general practice and fight GP shortage.

     

An evaluation of a GP out-of-hours service: meeting patient expectations of care

BACKGROUND: The reorganized out-of-hours general practitioner (GP) service, resulting in the creation of out-of-hours cooperatives has been widely welcomed by the medical profession. However, GP satisfaction remains only one aspect of this reorganized service and patients' views and levels of satisfaction must have a contribution to make to the organization and delivery of the service. AIM: To assess patient satisfaction at two out-of-hours cooperatives in Northern Ireland. METHOD: A sample of 4466 patients contacting the out-of-hours service was surveyed by postal questionnaires using a previously validated patient satisfaction instrument. RESULTS: Patients who initially requested to be seen at the out-of-hours centre were more likely to receive the contact they requested than those who requested telephone advice or a home visit. Only 41.8% of patients requesting a home visit actually received one. Patients were generally satisfied with the service provided and most satisfied with the 'doctor's manner' and the 'explanation and advice' received. Patients who received the contact they initially requested were more satisfied with all aspects of the service than other patients. The type of contact actually received had little effect on the satisfaction levels reported by patients who received the contact they initially requested. CONCLUSION: The population should be made fully aware of the services provided by out-of-hours cooperatives to enable them to have realistic expectations. With realistic expectations, patients are more likely to receive the medical contact they request and consequently will be more satisfied with the service provided. High satisfaction level is an important outcome measure of any out-of-hours service as it increases patient confidence and compliance and ultimately clinical outcome.     

高尔基体蛋白73对中国人群肝癌诊价值的Meta分析

目的 评价高尔基体蛋白73（golgi protein 73,GP73）检测在肝癌诊断中的价值.方法 检索万方、中国知网、NCBI PubmMed,EMBSASE数据库中2000-2013年符合纳入标准的国内外公开发表的有关以血清GP73诊断中国人肝癌的文献8篇,采用meta-disc1.4软件进行统计软件分析.结果 GP73诊断肝癌敏感度为78％,95％可信区间为0.74-0.81;特异度为75％,95％可信区间为0.73-0.78;阳性预测值为5.07,95％可信区间为2.95-8.72;阴性预测值为0.29,95％可信区间为0.23-0.37;诊断优势比为18.98,95％可信区间为8.46-42.54;综合SROC曲线下面积（AUC）为0.86,Q值为0.79.结论 GP73作为肝癌肿瘤标志物在肝癌的诊断中有一定价值.     

GP35 ANOAL, an abundant acidic glycoprotein of female Anopheles albimanus saliva

Salivary glands of female mosquitoes produce proteins, not completely described yet, that participate in carbohydrate and blood feeding. Here, we report an acidic glycoprotein of 35 kDa (GP35 ANOAL) secreted in the saliva of the malaria vector mosquito Anopheles albimanus. GP35 ANOAL is produced exclusively in the distal lateral lobes of adult female salivary glands, it has a pI of 4.45 and is negatively stained by regular silver stain. An 888 bp cDNA clone encoding a predicted product of 240 amino acids has a signal peptide, potential post-translational modification sites, and a disintegrin signature RGD. The GP35 ANOAL sequence depicts high similarities with the 30 kDa saliva allergen of Aedes aegypti, 30 kDa allergen-like hypothetical proteins, and GE-rich proteins present in several Anopheles species, as well as in Ae. albopictus and Culex pipiens quinquefasciatus. The function of this protein family is still unknown.     

GP2015 as a promising therapy for rheumatoid arthritis

Introduction: Rheumatoid arthritis is a common inflammatory joint disease with a myriad of systemic manifestations. Over the last 20 years its treatment has been revolutionised by the introduction of a number of different biologic drugs, including the TNF-receptor Fc fusion protein, Etanercept. However, these drugs are expensive and their widespread use puts a financial burden on healthcare systems. As many biologic treatments begin to come off patent new ‘biosimilar’ versions are being developed which can lead to significant cost savings. GP2015 (Erelzi®) is the second biosimilar version of Etanercept which is licensed for the treatment of rheumatoid arthritis.

Areas covered: We discuss the Chemistry, pharmacokinetics and pharmacodynamics of GP2015 in relation to reference Etanercept. Preclinical trials have shown pharmacokinetic equivalence between GP2015 and the reference drug. The recently completed Phase III, randomised, double blind EQUIRA study has shown equivalent efficacy and safety between GP2015 and Etanercept in patients with rheumatoid arthritis.

Expert opinion: GP2015 has shown equivalent efficacy and safety to reference Etanercept. With a growing number of biosimilar medications becoming available and another biosimilar Etanercept already being widely prescribed it is likely to be the cost of the drug that will determine if it is used widely.     

GP2017, an adalimumab biosimilar: pharmacokinetic similarity to its reference medicine and pharmacokinetics comparison of different administration methods

ABSTRACT

Background: To compare the pharmacokinetics of Sandoz biosimilar adalimumab (GP2017) with reference adalimumab (Humira) in healthy volunteers (PK similarity study) and to compare the pharmacokinetics of GP2017 administered by autoinjector (AI) or prefilled syringe (PFS; delivery study).

Methods: Healthy male subjects were randomized to receive a single 40 mg subcutaneous injection of GP2017, US-licensed or EU-authorized reference adalimumab (US/EU-Humira; PK similarity study) or a single 40 mg subcutaneous injection of GP2017 via AI or PFS (delivery study). Pharmacokinetics, safety, and immunogenicity were assessed over 72 days post-injection.

Results: The geometric mean ratios (90% confidence intervals) for C_max and AUC_0–inf were 1.05 (0.99–1.11) and 1.04 (0.96–1.13) for GP2017/EU-Humira and 1.00 (0.94–1.06) and 1.08 (1.00–1.18) for GP2017/US-Humira, all within the prespecified margin of 0.80–1.25 (PK similarity study). Pharmacokinetic parameters of GP2017 matched between AI and PFS (delivery study). Safety and immunogenicity were similar across groups in both studies.

Conclusion: PK similarity between GP2017, EU- and US-Humira was demonstrated. The safety pro?le of GP2017 was consistent with previous reports for Humira. These results contribute to the ‘totality-of-the-evidence’ supporting biosimilarity of GP2017 to Humira. PK and tolerability were equivalent for GP2017 dosed by AI or PFS.

Trial registration: PK similarity study EudraCT no. 2015-000579-28; Delivery study: EudraCT no. 2014–002879-29     

血小板输注无效患者血小板同种抗体及血小板特异性糖蛋白抗体表达的研究

目的 探讨血小板输注无效与血小板同种抗原或血小板特异性抗原的相关性.方法 选择65例临床确诊血小板输注无效患者作为研究对象,应用酶联免疫吸附实验(ELISA)方法检测血清、血小板洗脱液中血小板特异性抗体;应用HLA抗体特异性检测试剂盒,对组合反应性抗体(PRA)阳性的患者进行HLA抗体特异性分析;用HPA分型试剂盒检测8个血小板同种抗原系统HPA-1、2、3、4、5、6、9、15;用HLA分型试剂盒对HLA-A/B抗原进行基因分型.结果 65例患者HLA-A/B抗原,HPA-1、2、4、5、6、9、15抗原的基因频率分布与健康献血员比较差异无统计学意义.HPA-3a、3b抗原频率分别为0.65、0.35,与健康献血员比较差异有统计学意义(P＜0.05).65例患者中HLA抗体单独阳性24例(36.9%),HLA抗体和血小板特异性糖蛋白抗体共同阳性14例(21.5%);HLA抗体和血小板洗脱液特异性糖蛋白抗体共同阳性6例(9.2%),血小板洗脱液特异性糖蛋白抗体阳性13例(20%),HLA抗体、血小板特异性糖蛋白抗体及血小板洗脱液特异性糖蛋白抗体共同阳性4例(6.2%);HLA-A/B特异性抗体中,HLA-A*9抗体占全部抗体的46.2%,HLA-B*40抗体占33.6%.血清血小板特异性抗体以GPⅡb/Ⅲa为主(26.2%),其次为GP Ⅰa/Ⅱa(21.5%),血小板洗脱液中,血小板特异性抗体以GPⅡb/Ⅲa和GP Ⅰb/Ⅸ为主(41.5%).对2例患者进行了遗传学调查,发现产生的血小板特异性糖蛋白抗体和HLA抗体与父母血小板抗原及HLA抗原不相合呈密切相关.结论 血小板输注无效患者中,HLA抗体占主要地位,其次为血小板特异性糖蛋白抗体.     

Comparative functional and pharmacological characterization of Sandoz proposed biosimilar adalimumab (GP2017): rationale for extrapolation across indications

Background: Biosimilars are approved biologics that match reference medicine in quality, safety, and efficacy. The development of Sandoz proposed biosimilar adalimumab (SPBA; GP2017) involved a target-directed, iterative state-of-the-art quality-by-design development program. Here, we describe the functional and pharmacological characterization of SPBA and its proposed mechanism of action in immune-mediated inflammatory diseases.

Methods: Sensitive in vitro binding and functional characterization studies, and nonclinical evaluations (pharmacokinetics, pharmacodynamics, and safety/toxicology) were performed as part of a stepwise approach to confirm the biosimilarity of SPBA with reference adalimumab.

Results: Matching values were reported for SPBA and reference adalimumab in binding assays involving tumor necrosis factor (TNF)-α, complement 1q and human immune effector cell Fcγ receptor subtypes in cell-based bioassays for Fc receptor function (complement- and antibody-dependent cytotoxicity), and in apoptosis inhibition. Furthermore, SPBA and reference adalimumab were equivalent in terms of membrane TNF binding and induction of reverse signaling. Pharmacokinetics of SPBA and reference adalimumab were comparable in rabbits, and the two biologics were equally effective in a human TNF transgenic mouse model of polyarthritis.

Conclusion: SPBA matches reference adalimumab with regards to target binding, functional, pharmacokinetic, and pharmacodynamic properties at the nonclinical level supporting its approval in all indications of the reference adalimumab.     

从K562细胞中提取热休克蛋白GP96及其对人树突状细胞分化和功能的影响

本研究旨在建立从K5 6 2细胞系中提取热休克蛋白GP96的方法 ,进而研究GP96对人树突状细胞分化和功能的影响。采用蛋白分离提纯技术 ,将K5 6 2细胞裂解 ,经饱和硫酸铵沉淀、亲和层析、离子交换层析后提纯出热休克蛋白GP96 ,用SDS PAGE凝胶电泳和Westernblot方法对其进行鉴定。将提纯的GP96加入从正常人外周血单个核细胞诱导培养的树突状细胞 (dendriticcell,DC)中 ,通过流式细胞术检测DC细胞表型变化 ,用MTT法测定混合淋巴细胞反应 (mixedlymphocytereaction ,MLR)。结果表明 :从 1× 10 1 0 K5 6 2细胞中能提纯出热休克蛋白GP96 6 0 - 80 μg ;加入GP96制备的HSP DC ,其细胞表面标志CD83、CD86和HLA DR的表达率比常规培养DC明显升高 ,CD1a表达率降低 (P <0 .0 5 ) ;HSP DC比常规培养DC具有更强的刺激混合淋巴细胞反应的能力 (P <0 .0 5 )。结论 :从K5 6 2细胞中可以提取出热休克蛋白GP96 ;热休克蛋白GP96可以促进树突状细胞分化成熟 ,使其具有更强的刺激同种异体人T淋巴细胞增殖的能力     

Differences in immunogenicity associated with non-product related variability: insights from two pharmacokinetic studies using GP2017, an adalimumab biosimilar

ABSTRACT

Introduction: Antidrug antibody (ADA) development is known to occur with adalimumab treatment and impacts adalimumab exposure. Here, we compare the impact of immunogenicity on pharmacokinetics (PK) across two randomized PK studies of GP2017, an approved biosimilar adalimumab, in healthy subjects.

Methods: Healthy male subjects (N= 107 in study GP17-104; N= 90 in study GP17-103) received a single 40 mg subcutaneous injection of the same GP2017 drug product batch. Cross-study PK comparison was performed for log-transformed C_max, AUC_0–360h, AUC_0–last, and AUC_0–inf, using an ANCOVA model.

Results: The proportion of ADA-positive subjects was higher in GP17-103 (in total 71.1%) vs. GP17-104 (57.9%). Comparison of GP2017 PK between studies showed that the exposure was lower in GP17-103 vs GP17-104, with 90% confidence intervals (CIs) for geometric mean ratios of AUC_0–last and AUC_0–inf being outside the range of 0.80–1.25. A subgroup analysis showed that in ADA-negative subjects 90% CIs for all PK parameters were within range, with geometric mean ratios close to 1.00.

Conclusion: The differences in GP2017 PK between the two groups are not considered to be product-related, but may be due to currently unknown factors related to differences between the two study populations.     

Study on the antigens and antibodies of Mur and Mia blood groups in southern China

BackgroundThe frequency of Mur and Mi^a blood group antigens in Asian population is much higher than that in Caucasian population. However, due to the scarcity and high price of commercial detection reagents, there are few studies on antigen and antibody detection and comparative analysis in large samples.ObjectiveTo study the occurrence frequency, antigen correlation and antibody properties of Mur and Mi^a antigens and their corresponding antibodies in southern China.MethodsMur and Mi^a antigens and antibodies in local blood donors and patients were detected by routine serological microplate method. Statistical methods were used to calculate the incidence of two antigens and antibodies and analyze their correlation.ResultsAmong blood donors, the positive rates of Mur and Mi^a antigens were 6.4 % and 6.5 % respectively, with no significant difference (P > 0.05). In this region, the incidence of anti-"Mur" and anti-"Mi^a" was 0.65 % and 0.45 % respectively. But significant difference existed between blood donors and patients (P < 0.05). Among the anti-"Mur" and anti-"Mi^a" positive patients, most of the antibodies were IgM or IgM + IgG mixed type and had saline activity.ConclusionMur and Mi^a antigens and their corresponding antibodies have a high frequency in the population of southern China. In the routine clinical detection of irregular antibodies, Mur or Mi^a positive (GP.Mur) should be added to screen erythrocytes. Moreover, given the high correlation between Mur and Mi^a antigens expression on red blood cells, using monoclonal antibodies against Mi^a could predict the presence of Mur antigen.     

GP stress and patient dissatisfaction with nights on call: an exploratory study. GP stress and patient satisfaction.

OBJECTIVES: To compare the relative effects of being on or off duty at night on general practitioners' (GPs') levels of stress, and the satisfaction of their patients with daytime consultations surrounding these nights. DESIGN: A within-subjects, counterbalanced design was used. Two "on-call" and two "off-duty" nights were studied per GP over 4 weeks. SETTING: Primary health care in the UK. PARTICIPANTS: 26 GPs and their patients seen in consultations either side of nights studied. MAIN OUTCOME MEASURES: GPs completed validated stress questionnaires at the beginning and end of the consultation sessions immediately before and after nights on call and off duty. RESULTS: GPs experienced elevated levels of stress when on call, compared to when they were off duty. Patients seen in the consultation sessions before and after a night on call were less satisfied than patients seen before and after a night off duty. CONCLUSION: Being on call at night raised GP stress levels from at least the start of the afternoon consultation session before until the end of the morning consultation after the night on call. Daytime patient satisfaction is also reduced in periods surrounding nights on call.