Diagnostic utility of programmed cell death ligand 1 (clone SP142) immunohistochemistry for malignant lymphoma and lymphoproliferative disorders: A brief review

The programmed cell death 1 (PD1)/PD1 ligand (PD-L1) axis plays an important role in tumor cell escape from immune control and has been most extensively investigated for therapeutic purposes. However, PD-L1 immunohistochemistry is still not used widely for diagnosis. We review the diagnostic utility of PD-L1 (by clone SP142) immunohistochemistry in large-cell lymphomas, mainly consisting of classic Hodgkin lymphoma (CHL) and diffuse large B-cell lymphoma (DLBCL). Neoplastic PD-L1 (nPD-L1) expression on Hodgkin and Reed-Sternberg cells is well-established among prototypic CHL. Of note, EBV+ CHL often poses a challenge for differential diagnosis from peripheral T-cell lymphoma with EBV+ non-malignant large B-cells; their distinction is based on the lack of PD-L1 expression on large B-cells in the latter. The nPD-L1 expression further provides a good diagnostic consensus for CHL with primary extranodal disease conceivably characterized by a combined pathogenesis of immune escape of tumor cells and immunodeficiency. Compared with CHL, the nPD-L1 expression rate is much lower in DLBCL, highlighting some specific subgroups of intravascular large B-cell lymphoma, primary mediastinal large B-cell lymphoma, and EBV+ DLBCL. They consist of nPD-L1-positive and -negative subgroups, but their clinicopathological significance remains to be elucidated. Microenvironmental PD-L1 positivity on immune cells may be associated with a favorable prognosis in extranodal DLBCL. PD-L1 (by SP142) immunohistochemistry has helped us to understand the immune biology of lymphoid neoplasms possibly related by immune escape and/or immunodeficiency. However, knowledge of these issues remains limited and should be clarified for diagnostic consensus in the future.     

银屑病患者皮损间充质干细胞的培养鉴定及其表达HES1和CXCL6的研究

【摘要】 目的 培养鉴定银屑病患者皮损处真皮间充质干细胞（DMSC）,并研究DMSC的发状分裂相关增强子1（HES1）和趋化因子配体6（CXCL6）表达情况。方法 分离培养15例银屑病患者皮损及18例正常人皮肤的DMSC,利用流式细胞仪进行表型鉴定,实时荧光定量聚合酶链反应（RT-PCR）及免疫印迹（Western blot）检测HES1和CXCL6的mRNA及蛋白表达水平,两组间比较采用t检验。结果 银屑病组DMSC形态与正常对照组无差异,但HES1 和CXCL6基因的mRNA水平分别是对照组的3.56和3.44倍,且差异有统计学意义（P < 0.05）。在蛋白水平,银屑病组DMSC中HES1和CXCL6的表达明显高于对照组（P < 0.05）。结论 银屑病患者皮肤DMSC HES1及CXCL6表达升高可能参与银屑病的发病。     

Fas基因沉默对帕金森病大鼠脑纹状体多巴胺能神经元凋亡的影响及机制研究

目的 探讨脂肪酸合成酶(Fatty acid synthase,Fas)基因沉默对帕金森病大鼠脑纹状体多巴胺能神经元凋亡的影响及机制.方法 取40只大鼠,30只大鼠脑纹状体注射4μL 6-羟基多巴胺(6-Hydroxydopamine Hydrobromide,6-OHDA)建立帕金森病大鼠模型,剩余10只为假手术组,...     

Electrophysiological findings in immune checkpoint inhibitor-related peripheral neuropathy

ObjectiveTo report the electrodiagnostic features of immune checkpoint inhibitor (ICI)-related neuropathy.MethodsWe retrospectively reviewed clinical presentations and electrodiagnostic features of 23 patients studied after receiving immune checkpoint inhibitors (ICIs). The presentations for electrodiagnostic evaluation included an acute neuropathy or neuromuscular junction disorder. We applied established electrodiagnostic criteria for polyneuropathy and acute demyelinating neuropathy.ResultsWe identified acute demyelinating neuropathy (13 cases), axonal sensory motor neuropathy (5), pure sensory neuropathy (4) and mononeuropathy (1). 13 patients had acute demyelinating neuropathy confirmed by demonstrating demyelination in 2 or more nerves; 3 additional patients had demyelination in only one nerve. Analysis of motor nerve conduction parameters revealed demyelination involving median and ulnar nerve distal motor latencies as well as median, ulnar and peroneal nerve conduction velocities. Conduction block was found in median, ulnar and peroneal nerves. The remaining one-third patients without demyelination had acute painful axonal neuropathy. Coexisting myopathic changes (6) and neuromuscular junction dysfunction (4) were also identified.ConclusionsOur findings suggest that, while immune-mediated motor nerve demyelination is the primary underlying mechanism of ICI-related neuropathy, axonal painful neuropathy can also be an important presentation. Early recognition and effective intervention may reduce morbidity and permanent disability.SignificanceElectrophysiological studies might be useful in the evaluation of ICI-related neuropathy.     

CD40激发对恶性B淋巴细胞体外生物学行为的影响

目的 观察重组人可溶性CD40配体（rhsCD40L)及CD40L基因转染细胞（CD40L-TC）对恶性B淋巴细胞体外生物学行为的影响，探讨rhsCD40L在肿瘤生物治疗中的可能作用。方法 利用基因工程技术获得了rhsCD40L和CD40L-TC，分别与恶性B淋巴细胞株XG2，XG7，U266，8226，Raji及Daudi共同作用，分析了CD40激发对肿瘤细胞的体外增殖（锥虫蓝计数法），细胞周期（碘化丙锭掺入法），细胞表面共刺激分子的表达（免疫荧光标记法）以及细胞凋亡（Anexin-V-ETTC法）的效应。结果 （1）恶性B淋巴细胞株CD40表达呈异质性，XG2高表达CD40，8266，Raji和Daudi中度表达，而XG7和U266不表达CD40。显微镜下观察发现，rhsCD40L(5μg/ml）可引起XG2或Daudi细胞的同型聚集，该效应在作用6-8h后即可出现；与CD40L-TC细胞共育后（肿瘤细胞；CD40L-TC=5：1），XG2，Raji和Daudi细胞可粘附于CD40L-TC表面；（2）rhsCD40L和CD40L-TC均可显著抑制XG2，Raji和Daudi细胞的体外增殖，导致XG2细胞呈现G1期阻滞，而Raji和Daudi细胞阻滞于G2期，并诱导XG2，Raji,Daudi细胞的凋亡，凋亡率分别为：XG2细胞23.3%和18.8%,Raji细胞阻滞于G2期，并诱导XG2，Raji,Daudi细胞的凋亡，凋亡率分别为：XG2细胞23.3%和18.8%,Raji细胞11.6%和8.9%,Daudi14.2%和15.9%;(3)表型分析显示；rhsCD40L/CD40L-TC可明显上调XG2,Raji和Daudi细胞CD95的表达水平以及Raji细胞CD80的表达，而下调Raji细胞CD18的表达。结论 rhsCD40L能直接并显著地抑制恶性B淋巴瘤细胞株Raji,Daudi和多发性骨髓瘤细胞株XG2的体外增殖，诱导其凋亡，并上调Raji细胞免疫共刺激株Raji,Daudi和多发性骨髓瘤细胞株XG2的体外增殖，诱导其凋亡，并上调Raji细胞免疫共刺激分子CD80的表达，具有膜型CD40L的同样生物学功能，故rhsCD40L具有潜在的抗恶性B淋巴细胞肿瘤的作用。     

Reversible dougong structured receptor–ligand recognition for building dynamic extracellular matrix mimics

Dynamic biomaterials excel at recapitulating the reversible interlocking and remoldable structure of the extracellular matrix (ECM), particularly in manipulating cell behaviors and adapting to tissue morphogenesis. While strategies based on dynamic chemistries have been extensively studied for ECM-mimicking dynamic biomaterials, biocompatible molecular means with biogenicity are still rare. Here, we report a nature-derived strategy for fabrication of dynamic biointerface as well as a three-dimensional (3D) hydrogel structure based on reversible receptor–ligand interaction between the glycopeptide antibiotic vancomycin and dipeptide d-Ala-d-Ala. We demonstrate the reversible regulation of multiple cell types with the dynamic biointerface and successfully implement the dynamic hydrogel as a functional antibacterial 3D scaffold to treat tissue repair. In view of the biogenicity and high applicability, this nature-derived reversible molecular strategy will bring opportunities for malleable biomaterial design with great potential in biomedicine.

More than 2,500 y ago, the Chinese built their nail- and glue-less architectures (e.g., from Beijing''s Forbidden City to Sichuan Province''s Bao''en Temple) with a “dougong” structure, which is part of the network of wooden supports essential to the timber frame structure of the building. A typical dougong consists of a flat block of wood (dou), the top of which is fixed with an interlocking set of curved bows (gong), without the aid of nails or glue, to provide mechanical support to hold the three-dimensional (3D) network structure. Intriguingly, the microstructure of extracellular matrix (ECM) also demonstrates such reversible interlocking structure, which supports the integrity of tissues and organs. In natural ECM, the dougong structure occurs at the cell–ECM interfaces accompanied by a constant remodeling of the ECM network, giving rise to specific cell signaling, intracellular cascades, and subsequently, all relevant cell behaviors (1–3). Biomaterial designs based on the reversible interactions mimicking the cell–ECM interfaces are believed to boast distinct advantages, including the capability to modulate cell–biomaterial interactions, adapt to the development of cellular processes (1, 4–6), and facilitate the morphogenesis of tissues and organs (7, 8). Although the dynamic design of biomaterials is relatively complicated and challenging, this field attracts significant attention in building dynamic ECM mimics for regenerative medicine (1, 9–11).To faithfully reproduce the dynamics of ECM in artificial matrices, various strategies, including congenitally reversible noncovalent interactions (e.g., hydrogen bonds, coordinate bonds, hydrophobic forces, π–π interactions, van der Waals forces, and electrostatic effects) and dynamic covalent bonds (e.g., reversible boronic esters and benzoic–imine bonds and photosensitive nitrophenyl and azobenzene groups), have been exploited. Currently, biomaterial interfaces with dynamically functionalized bioligands are mainly designed through reversible covalent phenylboronic esters or benzoic–imine bonds (12–16), deformable azobenzene bonds (17), DNA and peptide molecular assemblies (18, 19), cyclodextrins/cucurbiturils-based macrocycle host–guest supermolecules (20–22), metal–ligand coordination (23, 24), and other multiple noncovalent interactions (25–29). They can elicit controllable and reversible cell behaviors (e.g., adhesion, migration, differentiation, and apoptosis) on the biomaterial interfaces (1, 4, 8). Unfortunately, these dynamic ECM-mimicking strategies carry critical problems (13). First, most dynamic strategies are based on nonbiogenic chemical molecules, which are usually nonbiocompatible and probably harmful. Second, the dynamics of these strategies commonly rely on the nonbiological stimuli (e.g., ultraviolet [UV] light or toxic chemicals), which are potentially invasive to cells. Third, current studies on mimicking dynamic ECM are usually limited to either reversible bioligand presentation or remoldable network fabrication; few works focus on both. In this context, the exploration of biocompatible molecular means for recapitulation of both dynamic bioactivity and dynamic structure in ECM is highly anticipated.Here, we present a nature-derived reversible strategy inspired by the receptor–ligand molecular recognition for design of dynamic ECM-mimicking biomaterial. The receptor–ligand molecular recognition relies on multi-noncovalent interaction between two or more molecules with exquisite complementarity in their chemical groups and geometries (30). With this in mind, we focus our attention on a typical simple yet elegant receptor–ligand system (i.e., the glycopeptide antibiotic vancomycin [Van] and the dipeptide d-Ala-d-Ala [AA]). Produced by a bacterial species named Amycolatopsis orientalis, Van exhibits strong bactericidal effect by inhibiting cell wall biosynthesis via the specific binding (K_d = 1.6 μM) (31) toward the terminal AA dipeptide of the bacterial cell wall precursors (Fig. 1A). As a proof of concept, we employ the reversible Van–AA interaction for building both a reversible dynamic biointerface and a 3D hydrogel network (Fig. 1 B and C). Due to the specific but reversible AA–Van molecular recognition, the dynamic biointerface demonstrates excellent reversibility in binding to cell-adhesive tripeptide arginylglycylaspartic acid (RGD) and modulating adhesion of multiple cells, demonstrating our strategy’s general applicability. In addition, the 3D hydrogel network based on the reversible AA–Van molecular recognition demonstrates self-recovery and injectability. The inherently antibacterial activity of the Van–AA hydrogel well equips the 3D hydrogel network for treating infected open skin wounds; the hydrogel could adapt to the shape of wound sites, resist self-fragmentation, and inhibit proliferation of pathogenic bacteria while continuously supporting wound healing. We believe that the specific but reversible Van–AA molecular recognition would be a strategy for dynamic biomaterial fabrication, and the easy-handling merit, ECM-like remoldability, and inherently antibacterial activity involved in this dynamic system will bring insights to biomaterial scaffold design in tissue engineering and regenerative medicine.Open in a separate windowFig. 1.Schematics showing the mechanism of dynamic biointerface and 3D ECM mimics based on a reversible dougong-structured natural receptor–ligand recognition. (A) The Van–AA molecular recognition on bacterial cell wall in nature. (B) Schematic illustration of the dynamic biointerface based on the reversible Van–AA interaction. Reversible bioligand presentation and controllable cell behaviors could be readily realized through the Van–AA interaction. (C) Schematic illustration of the dynamic hydrogels with remoldable network structure and its application in tissue repair.     

Prognostic Value of PD‐L1 in Breast Cancer: A Meta‐Analysis

Programmed cell death 1 ligand 1 (PD‐L1) is a promising therapeutic target for cancer immunotherapy. However, the correlation between PD‐L1 and breast cancer survival remains unclear. Here, we present the first meta‐analysis to investigate the prognostic value of PD‐L1 in breast cancer. We searched Pubmed, Embase, and Cochrane Central Register of Controlled Trials databases for relevant studies evaluating PD‐L1 expression and breast cancer survival. Fixed‐ and random‐effect meta‐analyses were conducted based on heterogeneity of included studies. Publication bias was evaluated by funnel plot and Begg's test. Overall, nine relevant studies with 8583 patients were included. PD‐L1 overexpression was found in 25.8% of breast cancer patients. PD‐L1 (+) associated with several high‐risk prognostic indicators, such as ductal cancer (p = 0.037), high tumor grade (p = 0.000), ER negativity (p = 0.000), PR negativity (p = 0.000), HER2 positivity (p = 0.001) and aggressive molecular subtypes (HER2‐rich and Basal‐like p = 0.000). PD‐L1 overexpression had no significant impact on metastasis‐free survival (HR 0.924, 95% CI = 0.747–1.141, p = 0.462), disease‐free survival (HR 1.122, 95% CI = 0.878–1.434, p = 0.357) and overall specific survival (HR 0.837, 95% CI = 0.640–1.093, p = 0.191), but significantly correlated with shortened overall survival (HR 1.573, 95% CI = 1.010–2.451, p = 0.045). PD‐L1 overexpression in breast cancer associates with multiple clinicopathological parameters that indicated poor outcome, and may increase the risk for mortality. Further standardization of PD‐L1 assessment assay and well‐controlled clinical trials are warranted to clarify its prognostic and therapeutic value.     

促性腺激素释放激素拮抗剂对大鼠睾丸支持细胞雄激素受体和Fas配体基因表达的影响

本实验利用注射促性腺激素释放激素拮抗剂（ＧｎＲＨ Ａ）引起睾丸生精细胞程序化死亡的大鼠模型，用地高辛标记的大鼠雄激素受体和Ｆａｓ配体的ＲＮＡ探针对大鼠睾丸石蜡切片进行原位杂交，观察雄激素受体和Ｆａｓ配体基因在生精细胞凋亡中的变化。结果显示，ＧｎＲＨ Ａ处理后曲细精管有退行性变，原位杂交表明此时Ｓｅｒｔｏｌｉ细胞的雄激素受体表达明显减低，而Ｆａｓ配体表达显著升高。提示Ｆａｓ配体的表达可能与生精上皮细胞程序化死亡关系密切。