前列腺PI-RADS V2.1评分联合血清PSA相关指标对灰区前列腺癌的诊断价值研究

摘要 目的：探讨前列腺影像报告和数据系统第2.1版（PI-RADS V2.1）评分联合血清前列腺特异抗原（PSA）相关指标对灰区前列腺癌的诊断价值。方法：回顾性分析2016年1月至2019年12月的187例经病理证实且PSA为灰区（4-10 ng/mL）的前列腺癌或前列腺增生患者资料。根据病理结果分为前列腺癌（PCa）组与前列腺增生组（BPH）组。由两名经验丰富的MRI诊断医师通过盲法对所有患者MRI图像进行PI-RADS V2.1评分,统计并计算血清PSA相关指标：总前列腺特异抗原（t-PSA）、游离前列腺特异抗原（f-PSA）、游离前列腺特异抗原与总前列腺特异抗原比值（f-PSA/t-PSA）、前列腺特异抗原密度（PSAD）。采用t检验比较各项指标在两组间的差异性,并使用受试者工作曲线（ROC）分析各项指标对灰区前列腺癌的诊断效能。结果：PI-RADS V2.1评分与PSAD在PCa与BPH组之间的差异具有统计学意义（P＜0.05）,而t-PSA、f-PSA、f-PSA/t-PSA在PCa与BPH组之间的差异均无统计学意义（P＞0.05）。根据ROC曲线分析,PI-RADS V2.1评分、PSAD、PI-RADS V2.1评分联合PSAD诊断灰区前列腺癌的曲线下面积（AUC）分别为0.814、0.671及0.838,且PI-RADS V2.1评分联合PSAD的AUC显著高于单独应用PI-RADS V2.1评分（Z=1.989,P＜0.05）与PSAD（Z=3.174,P＜0.05）。结论：PI-RADS V2.1评分与PSAD对诊断灰区前列腺癌具有较高诊断效能,且联合PI-RADS V2.1评分与PSAD能进一步提高诊断效能。     

血糖与前列腺癌患者的关系

目的:探讨前列腺活检患者的血糖与前列腺癌患者的关系。方法:前瞻性收集416例初次经直肠超声引导下前列腺穿刺活检患者的血糖、前列腺特异性抗原(PSA)和Gleason评分等临床资料,所有患者以血糖浓度6.1 mmol/L为界分成两组,比较高血糖组和正常血糖组前列腺癌检出率和Gleason评分的差异。结果:416例前列腺活检患者中,检出前列腺癌165例,高血糖组38例(40.00%),正常血糖组127例(39.56%),差异无统计学意义(P0.05);低级别前列腺癌(Gleason7分)患者的构成比分别为0.184、0.071,差异有统计学意义(P0.05),Spearman等级相关分析显示前列腺癌患者的血糖值与Gleason评分呈负相关(r=-0.228,P0.05)。结论:血糖对前列腺活检患者中前列腺癌检出率没有影响,但提高了低级别前列腺癌患者的构成比,血糖是影响前列腺癌Gleason评分的一个独立因素。     

血清PSA含量与前列腺癌临床分期、病理分级的相关性

目的:探讨血清前列腺特异性抗原(PSA)与前列腺癌(PCa)临床分期、病理分级的相关性.方法:对自2004年7月～2009年12月南京市13692例50岁以上的男性在健康体检时行血清PSA检测.以PSA≥4.0ng/ml定为前列腺癌可疑病例.建议行前列腺穿刺活检以确诊.共筛查出PCa患者140例,比较不同PSA值PCa患者的Gleason评分及临床分期.结果:随着PSA值的升高,前列腺癌筛查阳性率亦随之升高,低分化前列腺癌患者血清PSA含量明显高于高分化前列腺癌和中分化前列腺癌患者(P＜0.05),晚期前列腺癌患者血清PSA含量明显高于早期PCa患者(P＜0.01).血清PSA含量≥20ng/ml的前列腺癌人群中低分化前列腺癌及晚期前列腺癌的比例高于血清PSA含量＜20ng/ml的前列腺癌人群(P＜0.01).结论:血清PSA可以为前列腺癌患者的诊断、治疗及预后判断提供重要依据.     

前列腺特异性抗原在前列腺癌诊断中的应用研究

前列腺特异性抗原(prostate specific antigenPSA)是1979年Wang等首先从前列腺组织内分离和提纯出来的一种特异性糖蛋白,PSA在前列腺腺上皮细胞合成和分泌,其产生和分泌受雄激素的调节,具有组织特异性。正常状态下,PSA存在于     

uPCA3mRNA、tPSA、fPSA/tPSA、PSAD对PSA灰区前列腺癌诊断的临床意义

目的：探讨uPCA3mRNA、tPSA、fPSA/tPSA、PSAD对PSA 灰区前列腺癌的诊断价值。方法：经前列腺穿刺活检诊断为前列 腺增生(BPH) 111 例和前列腺癌(PCa) 89 例,测定血清tPSA、fPSA 与前列腺体积,所有病例均计算fPSA／tPSA、PSAD。用 RT-PCR 的方法检测PSA mRNA、PCA3 mRNA 的含量,以PCA3 mRNA/PSA mRNA 表示PCA3 mRNA 的含量。比较两组间 tPSA、fPSA/tPSA、PSAD 及PCA3mRNA各指标的差异,分析各指标在ROC 曲线下的面积、各指标的诊断敏感性和特异性。结果： PCa 组的fPSA／tPSA较BPH 组降低（P<0.01）,PCa 组的tPSA、PSAD及PCA3 mRNA较BPH 组均升高（P<0.01）。tPSA、fPSA／ tPSA、PSAD及PCA3 mRNA在ROC 曲线下的面积从大到小依次是PCA3 mRNA>PSAD > fPSA ／tPSA>tPSA。当PCA3 mRNA 和PSAD 临界值分别为0.27 和0.15 时,诊断PCa 的敏感性和特异性分别是86.5%和77.5%,80.9%和55.9%。结论：uPCA3 mRNA、fPSA／tPSA、PSAD 的测定能显著提高灰区PCa 诊断的敏感性和特异性,且uPCA3 mRNA 诊断效能最高。     

PSA及其相关检测指标在早期前列腺癌诊断中的临床价值

前列腺癌(PCa)是影响老年男性的恶性肿瘤之一,具有较高的发病率和死亡率,我国PCa患者人数逐年升高,因此进行早期的诊断和治疗具有重要意义。前列腺特异性抗原(PSA)对于早期PCa的诊断、治疗和预后具有重要作用,本文分别对PSA相关指标(F/TPSA、CPSA、PSAV、PSAD、PSATZ)应用于早期PCa诊断中的价值进行综述,旨在为临床诊断PCa提供理论依据。     

根治性前列腺切除术后大体病理较术前穿刺病理升级的危险因素分析

目的:前列腺穿刺病理Gleason评分(GS)和根治性前列腺切除术后病理Gleason评分经常出现差异.本文主要研究肿瘤病理升级的可能影响因素.方法:选择1999-01至2007-11在本院行前列腺穿刺活检确诊并行根治性前列腺切除术的95例前列腺癌患者,考察的临床资料包括患者确诊时的年龄,前列腺特异性抗原水平(PSA),前列腺体积(PV),前列腺特异性抗原密度(PSAD),术前是否接受新辅助内分泌治疗(NHT),穿刺病理GS,手术后病理GS及肿瘤体积(TIV).使用t-检验或卡方检验比较不用组别之间的变量,分别使用单因素和多因素Logistical回归分析引起GS升级的相关因素.结果:患者的平均年龄是67岁,平均PSA水平为24.3 ng/ml,平均前列腺体积是33.1ml.将前列腺体积分为≤25ml(25例),25-50ml(59例),≥50ml(11例)三组,将穿刺病理GS分为4-5(13例),6(35例),7(32例),8-10(15例)四组.前列腺体积较大组(≥50m1)比体积较小组(≤25ml,25-50m1)的肿瘤升级比率明显较低(48% vs 24%,18%,p＜0.05).穿刺病理GS较高组(8-10)比较低组(4-5,6,7)的肿瘤升级比率明显减低(46% vs 34%,25%,13%,p＜0.05).多因素Logistic回归分析显示,PV、穿刺GS及内分泌治疗与病理升级呈负相关(p＜0.05),而肿瘤体积及PSAD与其呈正相关(p＜0.05).结论:较大的前列腺体积,较高的穿刺病理GS,接受内分泌治疗以及较低的PSAD均可降低其肿瘤升级的可能.泌尿外科医师在决定由穿刺活检确诊的前列腺癌患者的治疗方案时应想到上述结论.     

影像引导下前列腺定向穿刺活检术的研究进展

确诊前列腺癌的金标准无疑是病理组织学检查,影像引导下的前列腺定向穿刺活检又是这当中最有效、最便捷的方式。而基于前列腺特异抗原(prostate specific antigen,PSA)筛查以及6分活检结果的前列腺癌诊断,已经由于其对低度肿瘤的过度诊断和对临床上有意义前列腺癌(格林森评分≥7)的诊断不足遭到批评。所以纵观国内外,影像引导已经增加到先用多参数MRI(multi-parametric magnetic resonance imaging,mpMRI)扫描检出病变组织再进行前列腺定向穿刺活检,包括,in-bornMRI引导,认知融合引导,以及MRI/TRUS融合引导下的活检。本文就一些影像引导下前列腺定向穿刺活检术的研究进展进行综述。     

一个以前列腺特异抗原密度和穿刺评分为基础的中国人群中前列腺癌根治术后病理升级预测模型的建立

目的：分析前列腺癌根治术后病理得分较穿刺得分增加的原因，并建立一个可以预测中国人群中前列腺癌根治术后病理升 级的模型。方法：以2008 年8 月至2013 年12 月在我院泌尿科行前列腺癌根治性切除术的264例患者的临床资料为基础，根据 术前和术后患者病理得分的变化将其分为升级组和未升级组。运用单因素和多因素logistic 回归分析病理升级的原因，并通过多 因素回归系数建立预测病理升级的诺模图。结果：264 例患者中，共238 例最终纳入统计分析，多因素logistic 回归分析显示前列 腺特异抗原密度(0R=3.854，P=0.001 )和穿刺Gleason(≤ 6)评分是中国人群中前列腺癌根治术后病理升级的独立危险因素。前列腺 特异抗原密度和穿刺得分的ROC 最佳截断取值为0.37 ng/ml 2和8 分。运用上述两个变量建立了一个可用于预测病理升级的诺 模图。结论：前列腺特异抗原密度和穿刺Gleason 评分是预测中国人群中前列腺癌根治术后病理升级的独立危险因素，本研究所 得的诺模图可以很好地预测前列腺癌根治术后的病理升级。     

PSCA与前列腺癌研究进展

前列腺干细胞抗原(prostate stem cell antigen,PSCA)是一种前列腺癌相关肿瘤抗原,也是一种GPI(gly-cosyl phosphatidylinositol)锚定蛋白,通过其C端的GPI锚定结构锚定到细胞膜表面.PSCA在正常前列腺组织中的表达较低,提高的PSCA表达伴随着增加的肿瘤分期、分级以及雄激素非依赖性和转移癌的形成,且不随癌症进展而降低,是前列腺癌诊断和治疗的理想靶抗原.动物实验显示,PSCA抗体和疫苗可能在前列腺癌免疫靶向治疗中具有重要价值.     

Targeted treatment of prostate cancer

Over a half century ago, Charles Huggins demonstrated the response of prostate cancer to androgen deprivation therapy. Subsequently, many discoveries and evolving findings continued to support a research rationale focused on the androgen receptor (AR) as a key target for prostate cancer. More recently, preliminary trials have suggested that other targets could also be useful in the treatment of prostate cancer, and the proposed strategies for treatment have ranged from targeted toxins to immunotherapeutic agents. We provide an overview of some of these approaches, with an emphasis on those that employ prostate specific membrane antigen (PSMA) as a target.     

用亲和层析技术鉴别布氏菌强弱毒差异抗原成分

采用CDI和CCA两种方法活化Sepharose4B,制备亲和层析柱,并对这两种方法的优劣进行了比较。分别用超声液破碎和Ribi机器压榨制备布氏菌强毒M28株和疫苗M25株的可溶性抗原,作为亲和层析柱的配基。抗M28的血清经过M28和M5柱,洗脱液中的抗体再分别和M28、M5抗原反应经电转印检查证实,M28抗原中的一条蛋白带只与M28柱洗脱液反应,不与M5柱洗脱液反应。这种只与M28柱洗脱液反应的蛋白可能即是布氏菌强毒M28株所特有的抗原成分,也就是强毒株和弱毒株的差异抗原成分。     

肺炎支原体感染鼠P1特异抗原的动态检测

通过实验动物模型探讨肺炎支原体感染动物肺泡灌洗液中特异抗原检出率的动态变化,为肺炎支原体感染的临床诊断提供理论依据。小鼠经鼻自然感染肺炎支原体,分别采集感染后不同时间点小鼠的支气管灌洗液,应用量子点标记肺炎支原体P1蛋白抗体,直接免疫荧光法检测感染鼠肺泡灌洗液中肺炎支原体P1特异抗原,同时通过PCR检测肺组织肺炎支原体DNA及肺组织病理切片观察肺部炎性变化确定小鼠感染。结果显示,感染鼠肺炎支原体特异抗原在感染后第3天检出阳性率为75%,第7天达高峰为83%,之后随病程延长,抗原检测的阳性率逐渐下降,在感染后第14、21天检出阳性率分别为58%和25%。肺炎支原体特异抗原在感染早期检出率高。应用量子点标记肺炎支原体P1蛋白抗体,直接免疫荧光法检测肺炎支原体特异抗原可应用于肺炎支原体感染的早期诊断。     

Differential responsiveness of prostatic acid phosphatase and prostate-specific antigen mRNA to androgen in prostate cancer cells

Androgens regulate the expression of both human prostatic acid phosphatase (PAcP) and prostate-specific antigen (PSA), two major prostate epithelium-specific differentiation antigens. Due to the important role of these two enzymes as prostate epithelium differentiation markers, we investigated their regulation of expression at the mRNA level in LNCaP human prostate carcinoma cells. Interestingly, phenol red, a pH indicator in the culture medium, promoted cell growth. To eliminate this non-specific effect, a phenol red-free, steroid-reduced medium was utilized. When high-density cells were grown in that medium, 5alpha-dihydrotestosterone (DHT) suppressed PAcP but stimulated PSA. However, tumor promoter phorbol ester 12-o-tetradecanoyl phorbol-13-acetate (TPA) functioned as a potent inhibitor of both PAcP and PSA expression. Prolonged treatment with DHT as well as TPA resulted in a similar down-regulation of protein kinase C and cellular PAcP activities. Thus, the levels of PAcP and PSA mRNA are differentially regulated by androgens in LNCaP cells.     

Control of immunologically crossreactive leptospiral infection by administration of lipopolysaccharides from a nonpathogenic strain of Leptospira biflexa

In our previous paper (Matsuo, K., Isogai, E., and Araki, Y., Carbohydr. Res., 328: 517-524, 2000), antigenic polysaccharides obtained from the lipopolysaccharide (LPS) fraction of a nonpathogenic leptospira, Leptospira biflexa patoc Patoc I, are shown to be broadly crossreactable with most rabbit antisera elicited by immunization with various pathogenic leptospires. The result led us to test a protective effect of the same LPS in a hamster model system by heterologously challenging with a pathogenic leptospira, L. interrogans manilae UP-MMG. Firstly, a similarity in the antigenic epitopes of L. biflexa and L. interrogans was confirmed by the following assays. In the microscopic agglutination test (MAT), a hamster antiserum elicited by immunization with the L. biflexa-LPS preparation was shown to agglutinate cells of L. interrogans. Contrarily, in the enzyme-linked immunosorbent assay (ELISA), the L. biflexa-LPS preparation was shown to crossreact with a hamster antiserum elicited by immunization with whole cells of L. interrogans. These results suggest that the same or closely related antigens may be present on the cell surfaces of both L. biflexa patoc Patoc I and L. interrogans manilae UP-MMG. Furthermore, in a protective assay, the prior administration of a L. biflexa-LPS preparation resulted in raising a protective response in hamsters against challenge by L. interrogans without any side effect. The protective effect was strongly dependent on the dose amounts and/or administration times of L. biflexa-LPS. Thus, L. biflexa-LPS preparations can use as a potent vaccine against leptospirosis caused by various leptospires.     

Dual‐labeled chemiluminescence enzyme immunoassay for simultaneous measurement of total prostate specific antigen (TPSA) and free prostate specific antigen (FPSA)

The specificity for early diagnostic of prostate‐specific antigen (PSA) is low because the current technology mostly allows the detection of only one biomarker at one time. In this work, a dual‐labeled chemiluminescence enzyme immunoassay (CLEIA) for simultaneous measurement of total PSA (TPSA) and free PSA (FPSA) was proposed. Anti‐PSA McAb (Mab1) was immobilized on a microplate as the solid phase, horseradish peroxidase (HRP)‐labeled anti‐TPSA monoclonal antibody (McAb2) and alkaline phosphatase (ALP)‐labeled anti‐FPSA McAb3 were used as detection antibodies. Two chemiluminescence reactions of HRP with luminol and ALP with 4‐methoxy‐4‐(3‐phosphate‐phenyl)‐spiro‐(1,2‐dioxetane‐3,2′‐adamantane) (AMPPD) were used as the signal detecting system. Based on a sandwich model, the amount of FPSA and TPSA could be determined simultaneously. The effects of several physico‐chemical parameters were studied and optimized. Cross‐reactivities of six common tumor markers in serum were studied. The proposed method presented the sensitivity of 0.03 ng ml^?1 and 0.05 ng ml^?1 for FPSA and TPSA respectively, with low cross‐reactivities. Compared with the results from commercial chemiluminescent kits there was good correlation, indicating that this established method could be used to simultaneously to measure the concentrations of FPSA and TPSA in one serum sample and also could greatly facilitate the early diagnosis for PCa in clinical practice.     

狂犬病病毒糖蛋白表达及纯化及其记忆性B细胞结合能力的分析

表达纯化不同标签、不同大小3个狂犬病病毒糖蛋白,分析其结合功能后,得到具备高亲和力的、可特异性结合记忆性B细胞的狂犬病病毒糖蛋白。本实验通过基因工程的方法,采用不同的原核表达系统分别表达带有不同标签的、全长和膜外区的RVG,纯化蛋白并分析比较其结合功能,荧光标记候选蛋白,结合CD19及CD27的抗体,流式细胞术检测狂犬疫苗免疫后PBMCs中抗狂犬病病毒特异性记忆性B细胞的情况,确认候选蛋白与抗狂犬病毒特异性记忆性B细胞的结合功能。本实验成功构建了3个表达载体pGEX-5X-1-RVG、pET28a-RVG和pET30a-G,优化表达纯化条件成功获得了糖蛋白GST-RVG、His-RVG和His-G。纯化后的GST-RVG、His-RVG和His-G经Western blotting和ELISA鉴定均有抗原特异性;由竞争ELISA法测得3个纯化后糖蛋白与抗狂犬病病毒抗体的亲和力。通过流式细胞术可以检测到狂犬疫苗免疫后阳性志愿者PBMCs中的抗狂犬病病毒特异性记忆性B细胞,从而获得了高亲和力、可用于分选抗原特异性的记忆性B细胞的狂犬病病毒糖蛋白。     

Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer

Adoptive transfer of T cells expressing chimeric antigen receptors (CARs) is considered to be a novel anticancer therapy. To date, in most cases, single-chain variable fragments (scFvs) of murine origin have been used in CARs. However, this structure has limitations relating to the potential immunogenicity of mouse antigens in humans and the relatively large size of scFvs. For the first time, we used camelid nanobody (VHH) to construct CAR T cells against prostate specific membrane antigen (PSMA). The nanobody against PSMA (NBP) was used to show the feasibility of CAR T cells against prostate cancer cells. T cells were transfected, and then the surface expression of the CAR T cells was confirmed. Then, the functions of VHH-CAR T cell were evaluated upon coculture with prostate cancer cells. At the end, the cytotoxicity potential of NBPII-CAR in T cells was approximated by determining the cell surface expression of CD107a after encountering PSMA. Our data show the specificity of VHH-CAR T cells against PSMA⁺ cells (LNCaP), not only by increasing the interleukin 2 (IL-2) cytokine (about 400 pg/mL), but also the expression of CD69 by almost 38%. In addition, VHH-CAR T cells were proliferated by nearly 60% when cocultured with LNCaP, as compared with PSMA negative prostate cancer cell (DU-145), which led to the upregulation of CD107a in T cells upto 31%. These results clearly show the possibility of using VHH-based CAR T cells for targeted immunotherapy, which may be developed to target virtually any tumor-associated antigen for adoptive T-cell immunotherapy of solid tumors.