Reflection of neuroblastoma intratumor heterogeneity in the new OHC-NB1 disease model

Accurate modeling of intratumor heterogeneity presents a bottleneck against drug testing. Flexibility in a preclinical platform is also desirable to support assessment of different endpoints. We established the model system, OHC-NB1, from a bone marrow metastasis from a patient diagnosed with MYCN-amplified neuroblastoma and performed whole-exome sequencing on the source metastasis and the different models and passages during model development (monolayer cell line, 3D spheroid culture and subcutaneous xenograft tumors propagated in mice). OHC-NB1 harbors a MYCN amplification in double minutes, 1p deletion, 17q gain and diploid karyotype, which persisted in all models. A total of 80–540 single-nucleotide variants (SNVs) was detected in each sample, and comparisons between the source metastasis and models identified 34 of 80 somatic SNVs to be propagated in the models. Clonal reconstruction using the combined copy number and SNV data revealed marked clonal heterogeneity in the originating metastasis, with four clones being reflected in the model systems. The set of OHC-NB1 models represents 43% of somatic SNVs and 23% of the cellularity in the originating metastasis with varying clonal compositions, indicating that heterogeneity is partially preserved in our model system.     

NRP-1单克隆抗体对乳腺癌裸鼠移植瘤生长的影响

 目的 探讨NRP-1单克隆抗体（NRP-1 MAb）的特异性，以及不同剂量的NRP-1 MAb治疗乳腺癌裸鼠移植瘤的疗效。方法 Western blot和共聚焦免疫荧光法检测NRP-1 MAb是否识别MCF7细胞上NRP-1蛋白。将MCF7细胞接种于BALB/c裸鼠皮下建立乳腺癌细胞移植瘤模型，并进行瘤组织传代。传代的肿瘤体积生长至300~500 mm3时，随机分为对照组、NRP-1 MAb低剂量组、中剂量组和高剂量组，每组6只，给药7次。观察荷瘤裸鼠一般状况，测量瘤体大小及裸鼠体重。实验结束时剥离瘤体称重，提取组织蛋白，Western blot检测组织中VEGF蛋白和NRP-1蛋白的表达量。结果 NRP-1MAb成功识别MCF7细胞上的NRP-1蛋白；NRP-1 MAb能够有效抑制MCF7细胞裸鼠移植瘤的生长，低剂量组（1 mg/kg）抑瘤率为47.01%，中剂量组（5 mg/kg）抑瘤率为65.70%，高剂量组（10 mg/kg）抑瘤率为69.19%。。结论 NRP-1 MAb能够识别并有效结合MCF7细胞膜上的NRP-1蛋白，且可抑制MCF7细胞移植瘤的生长，NRP-1 MAb抑制移植瘤的增长可能与下调NRP-1和VEGF表达有关。     

Incidence of T790M in Patients With NSCLC Progressed to Gefitinib,Erlotinib, and Afatinib: A Study on Circulating Cell-free DNA

BackgroundInsights into the mechanism of resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) could provide important information for further patient management, including the choice of second-line treatment. The EGFR T790M mutation is the most common mechanism of resistance to first- and second-generation EGFR TKIs. Owing to its biologic relevance in the response of non–small-cell lung cancer (NSCLC) to the selective pressure of treatment, the present study investigated whether the occurrence of T790M at progression differed among patients receiving gefitinib, erlotinib, or afatinib.Patients and MethodsThe present retrospective study included patients with NSCLC with an EGFR activating mutation, who had received gefitinib, erlotinib, or afatinib as first-line treatment. Plasma samples for the analysis of cell-free DNA were taken at disease progression and analyzed using a digital droplet polymerase chain reaction EGFR mutation assay.ResultsA total of 83 patients were enrolled; 42 had received gefitinib or erlotinib and 41afatinib. The patient characteristics were comparable across the 2 groups. The median time to progression (TTP) was 14.4 months for the gefitinib and erlotinib group and 10.2 months for the afatinib group (P = .09). Of the 83 patients, 47 (56.6%) were positive for the T790M in plasma. A greater incidence of T790M was observed in patients with progression during gefitinib or erlotinib therapy compared with patients treated with afatinib (33 [79%] vs. 14 [34%], respectively; odds ratio, 7.1; 95% confidence interval, 2.7-18.5; P = .0001).ConclusionsAlthough gefitinib, erlotinib, and afatinib showed a comparable TTP in patients receiving first-line therapy, the incidence of T790M differed among them, as demonstrated by the present study, which could have implications for the choice of second-line treatment.     

程序性死亡蛋白-1抗体治疗晚期胃癌的临床研究新进展

目前化疗仍为晚期胃癌的标准治疗手段,而多数患者诊断时已接近晚期。近年新兴的免疫治疗手段程序性死亡蛋白-1(PD-1,programmed death l)抗体,通过阻断肿瘤细胞逃避自身免疫机制,重新激活自身免疫对肿瘤的杀伤作用,从而达到肿瘤的治疗作用。针对晚期胃癌的PD-1抗体治疗的临床研究已在国内外开展,大部分均有较好效果。本文对近年免疫治疗中的PD-1抗体在晚期胃癌的重要临床研究进展进行介绍。     

脂联素对高脂饮食诱导的胰岛素抵抗子宫内膜癌裸鼠移植瘤生长的影响

目的 探讨脂联素对高脂饮食诱导裸鼠胰岛素抵抗子宫内膜癌移植瘤生长的影响。方法 40只裸鼠随机分成高脂组和普食组，每组20只，分别喂养高脂饲料（high-fat diet，HFD）和普通饲料（normal diet，ND），10周后测定裸鼠空腹血糖（fasting blood glucose，FBG）和空腹血胰岛素（fasting serum insulin，FINS）水平，计算胰岛素抵抗指数（HOMA-IR），建立胰岛素抵抗裸鼠模型。第11周接种子宫内膜癌HEC-1B细胞，待瘤体长至0.5 cm时，两组随机选取10只裸鼠腹腔注射脂联素（adiponectin，APN）分为HFD+APN组、ND+APN组；其余裸鼠分别注射0.9%氯化钠分为HFD组、ND组；14周后，测定各组血糖和血脂的代谢情况。结果 高脂组裸鼠10周后的平均体重、体长、FBG、FINS、HOMA-IR均大于普食组，糖耐量试验和胰岛素耐量试验结果显示血糖水平亦高于普食组（均P＜0.05）；接种子宫内膜癌HEC-1B细胞后均成瘤，建模成功。HFD+APN组和ND+APN组移植瘤平均重量和体积增长速度均分别低于HFD组和ND组（均P<0.05）。HFD组和HFD+APN组14周后的FINS、HOMA-IR、TC和TG均较ND组和ND+APN明显升高（均P<0.01），且HFD组明显高于HFD+APN组（均P<0.01）；HFD+APN组的血脂联素水平低于ND+APN组（P<0.01），HFD组则低于ND组（P<0.05）。结论 脂联素可改善高脂饮食导致的胰岛素抵抗，抑制子宫内膜癌移植瘤生长。     

基于频数分析法的骨伤科中成药用药规律研究

目的：探讨骨伤科中成药的用药规律，为临床合理用药及新药开发提供参考。方法：以《中国药典》2015年版一部收载的骨伤科类中成药共149个为研究对象，运用频数分析的方法研究其主治病症、所用剂型、各类方剂高频药物、核心药物组成，分析骨伤科中成药用药规律。结果：常用骨伤科中成药中治疗骨关节痹症的最多，其次为跌打损伤和骨关节退行性疾病的中成药；剂型以胶囊剂、片剂、丸剂为主；以活血化瘀药、祛风湿药、补虚药为核心药物组合，各类方剂中三者比例偏重不同。结论：通过对研究结果进行分析，提出了新药研发的方向。     

Therapeutics and delivery vehicles for local treatment of osteomyelitis

Osteomyelitis, or the infection of the bone, presents a major complication in orthopedics and may lead to prolonged hospital visits, implant failure, and in more extreme cases, amputation of affected limbs. Typical treatment for this disease involves surgical debridement followed by long-term, systemic antibiotic administration, which contributes to the development of antibiotic-resistant bacteria and has limited ability to eradicate challenging biofilm-forming pathogens including Staphylococcus aureus—the most common cause of osteomyelitis. Local delivery of high doses of antibiotics via traditional bone cement can reduce systemic side effects of an antibiotic. Nonetheless, growing concerns over burst release (then subtherapeutic dose) of antibiotics, along with microbial colonization of the nondegradable cement biomaterial, further exacerbate antibiotic resistance and highlight the need to engineer alternative antimicrobial therapeutics and local delivery vehicles with increased efficacy against, in particular, biofilm-forming, antibiotic-resistant bacteria. Furthermore, limited guidance exists regarding both standardized formulation protocols and validated assays to predict efficacy of a therapeutic against multiple strains of bacteria. Ideally, antimicrobial strategies would be highly specific while exhibiting a broad spectrum of bactericidal activity. With a focus on S. aureus infection, this review addresses the efficacy of novel therapeutics and local delivery vehicles, as alternatives to the traditional antibiotic regimens. The aim of this review is to discuss these components with regards to long bone osteomyelitis and to encourage positive directions for future research efforts.     

Academic–Industrial Collaboration: Toward the Consilience of Two Solitudes

Current major advances in drug discovery can be traced back to pioneering contributions originating from academics over a century ago. Living in a symbiotic yet noninvasive coexistence, the academic community and the pharmaceutical industry have strived, each in their own way, to develop the modern medicines that benefit humankind today. The subject is presented from a historical and personal perspective.     

宫颈癌顺铂耐药研究进展

顺铂（DDP）作为第一个被发现的金属抗癌药物，是目前最有潜力和应用最广泛的抗肿瘤药物。顺铂在宫颈癌的治疗中尤为重要，目前已被推荐为同步放化疗的首选药物。但随着广泛的使用，顺铂的耐药性逐渐显露出来，并成为限制临床疗效和部分患者肿瘤治疗进展的主要原因之一。顺铂的耐药机制复杂，发生环节较多，但具体耐药机制尚不明确，目前按照顺铂耐药发生的环节可分为:①顺铂在血液循环过程中产生耐药；②顺铂通过细胞膜的流入或流出产生耐药；③顺铂在胞质中产生耐药；④顺铂与DNA结合后产生耐药。本文综述了宫颈癌顺铂耐药可能发生的四个环节及克服耐药常用的手段，为提高顺铂对宫颈癌的疗效提供依据。