乳腺癌（BC）发病率持续上升，已成为严重威胁公共健康的重大挑战，转移仍是导致患者死亡的主要原因。目前，切除肿瘤并辅以放化疗后仍有较大几率发生转移。此外，早期转移隐匿性强，干预措施不完善及耐药发生迅速均是预后不良的重要因素。细胞外囊泡（EVs）是由细胞释放的、具有膜结构的纳米级颗粒，是细胞间通讯的重要介质。EVs通过转运膜性组分、胞质蛋白及核酸等，调控乳腺癌的发生、进展及转移定植，其来源异质性决定了其在乳腺癌转移中发挥独特作用。本文重点总结了不同来源EVs作为液体活检标志物在转移预警中的价值，以及靶向EVs分泌或阻断其通讯网络在克服免疫治疗耐药中的潜在临床应用，以期为临床医生提供有价值的诊疗参考。

     The incidence of breast cancer（BC） continues to rise and has become a major public health challenge．Metastasis remains the principal cause of BC-related mortality．Currently，there remains a high probability of metastasis even after surgical tumor resection followed by radiotherapy and chemotherapy．The insidious nature of early metastasis，inadequate intervention strategies，and rapid development of drug resistance are all critical factors contributing to poor prognosis．Extracellular vesicles（EVs） are membrane-enclosed，nanoscale particles released by cells．As mediators of intercellular communication，EVs transfer membrane components，cytosolic proteins，and nucleic acids to recipient cells，thereby modulating BC initiation，progression，and metastatic dissemination．The heterogeneity of EV origins determines their distinct roles in breast cancer metastasis．This review focuses on summarizing the value of EVs from different sources as liquid biopsy biomarkers for metastasis warning，as well as their potential clinical applications in targeting EV secretion or blocking their communication networks to overcome immunotherapy resistance，aiming to provide valuable diagnostic and therapeutic references for clinicians．

       目的  探讨TAP水平与乳腺癌分子亚型及临床病理参数之间的相关性。方法  以2021年3月—2025年1月期间收治的150例乳腺癌病例为样本，采用静脉采血方式测定TAP凝聚物表面积指标，通过免疫组织化学EnVision双步染色技术，对雌激素受体（ER）、雄激素受体（AR）、孕激素受体（PR）、Ki-67及p53等表达水平进行分析，采用荧光原位杂交（FISH）对人表皮生长因子受体2（HER2）基因扩增状态进行检测。结果  150例患者中，TAP强阳性131例，TAP弱阳性15例，TAP阴性4例，TAP阳性率97.33%。免疫表型：ER阴性43例，ER阳性107例；AR阳性133例，AR阴性17例；PR阴性60例，PR阳性90例；p53阳性73例，p53阴性77例；HER2强阳性41例，HER2弱阳性89例，HER2阴性20例；Ki-67增殖指数≥20% 116例，Ki-67增殖指数＜20% 34例。FISH对65例免疫组织化学检测结果为HER2（2+ ）的乳腺癌病例进行基因扩增状态分析，其中阳性7例，阴性58例。Ki-67高增殖组TAP表达水平显著高于低增殖组（P＜0.05）；不同临床分期患者TAP表达水平存在差异（P＜0.05）；三阴型、HER2阳性型、Luminal A型和Luminal B型的患者之间的TAP表达水平存在差异（P＜0.05），各分子分型（HER2阳性型、三阴型、Luminal A型和Luminal B型）与其对应非分型组的TAP表达均无统计学差异（均P＞0.05）。结论  TAP在乳腺癌中广泛表达，且与Ki-67增殖指数、临床分期呈正相关。虽然不同分子分型间TAP表达存在总体差异，但具体亚型对比未显示显著性，后期需扩大样本量验证。

       Objective  To explore the relationship between tumor abnormal protein（TAP）level and molecular typing and clinicopathological features of breast cancer．Methods  A total of 150 breast cancer cases admitted from March 2021 to January 2025 were enrolled in this study．The surface area of TAP condensates was measured using venous blood samples．The expression levels of estrogen receptor（ER），androgen receptor（AR），progesterone receptor（PR），Ki-67，and P53 were analyzed via immunohistochemistry（IHC）using the EnVision two-step staining technique．The amplification status of the human epidermal growth factor receptor 2（HER2+）gene was determined using fluorescence in situ hybridization（FISH）．Results  Among 150 patients，131 cases were strongly positive，15 cases were weakly positive and 4 cases were negative，with a positive rate of 97.33%．Immunophenotype：ER positive in 107 cases and ER negative in 43 cases，133 cases were  positive for AR and  17 cases were negative，PR was positive in 90 cases and negative in 60 cases，73 cases were positive for p53 and 77 cases were negative．HER2 is strongly positive in 41 cases，weakly positive in 89 cases and negative in 20 cases．There were 116 cases with Ki-67 proliferation index ≥ 20% and 34 cases with Ki-67 proliferation index ＜ 20%．Sixty-five cases of breast cancer HER2（2 ）were detected in the later stage．by FISH，of which 7 cases were positive and 58 cases were negative．The expression level of TAP in patients with high Ki-67 proliferation index was higher than that in patients with low Ki-67 proliferation index（P＜0.05）．The expression level of TAP in patients with different clinical stages was different（P＜0.05）．There were differences in TAP expression levels among patients with triple negative type，HER2 positive type，Luminal A type and Luminal B type（P＜0.05）．There was no statistical difference in TAP expression between each molecular type（triple negative type，HER2 positive type，Luminal A type and Luminal B type）and its corresponding non-typing group（all P＞0.05）．Conclusions  TAP is widely expressed in breast cancer，and it is positively correlated with Ki-67 proliferation index and clinical stage．Although there is a general difference in TAP expression among different molecular typing，the comparison of specific subtypes shows no significance，and it needs to be verified by expanding the sample size 

       目的   了解育龄期乳腺癌患者对子代健康和遗传风险担忧现状及其对癌症诊断前后生育意愿改变的影响。方法   于2019年11月—2020年9月，采用中文版癌症后生育忧虑量表（RCAC）-子女健康亚量表对广州市某三甲医院的230例育龄期乳腺癌患者进行调查，应用SPSS 26.0及PSM插件对结果进行统计分析。在进行子女健康亚量表得分低分组和中高分组患者的生育意愿改变率的比较时，应用倾向性评分匹配法控制混杂因素。结果   育龄期乳腺癌患者对子代健康和遗传风险担忧得分为（11.40±2.99）分；匹配前后不同子女健康亚量表得分分组的患者在癌症诊断前后生育意愿改变率的差异均不具有统计学意义（P＞0.05）。结论   育龄期乳腺癌患者对子代健康和遗传风险的担忧虽然没有显著影响其生育意愿的改变，但其对子代健康和遗传风险担忧处于较高水平，临床医护人员应提高对这一现象的重视，可通过多渠道科普相关知识，开展遗传咨询服务以减轻患者的担忧。

      Objective  To explore the current status of childbearing age breast cancer patients’  concerns  about their children’s health and genetic risk and its influence on the change of fertility intention before and after cancer diagnosis．Methods  From November 2019 to September 2020，the Chinese version of the Reproductive Concern After Cancer Scale（RCAC）- Children’s Health Sub-scale was used to interview 230 patients with breast cancer of childbearing age in a tertiary hospital in Guangzhou，and SPSS 26.0 and PSM plug-in were used to analyze the results statistically．The  propensity  score matching method was used to control confounding factors when comparing the change  rate of fertility intention of patients with low scores and those with medium and high scores in the children health subscale．Results  The score of concern for the health and genetic risk of children in breast cancer patients of childbearing age was（11.40±2.99）．There was no statistically significant difference in the change  rate of fertility intention before and after cancer diagnosis among patients with different subscale scores of children’s health．Conclusions   Despite not significantly impacting fertility intentions，the notably high level of concern regarding children’s health and genetic risks among childbearing-age breast cancer patients necessitates increased clinical awareness．It is recommended that healthcare professionals address this through multi-channel education and genetically focused counseling within a collaborative model to mitigate patient distress．

目的 探讨乳腺癌术后疼痛综合征（PMPS）的发生率及影响因素。方法 选择2021年1月—2023年2月医院收治的82例乳腺癌患者为研究对象,采取手术治疗,统计PMPS发生率,分析PMPS的特征,比较PMPS患者和非PMPS患者的临床资料,采用 Logistic 逐步回归分析。结果 82例中有20例患者术后发生PMPS,发生率为24.39%,其中患侧腋窝45.00%、麻木样疼痛35.00%、中度疼痛60.00%、术后即刻疼痛50.00%、每日发作疼痛50.00%占比较高。PMPS患者和非PMPS患者的体质指数、教育程度、病理分期、肿瘤占位、手术方式、术前使用非甾体抗炎药、术前化学治疗、术后化学治疗比较差异均无统计学意义（P>0.05）。PMPS患者年龄低于非PMPS患者,负性情绪率30.00%高于非PMPS患者8.06%,清扫腋窝淋巴结率95.00%高于非PMPS患者72.58%,术后放射治疗率30.00%高于非PMPS患者6.45%（P<0.05）。年龄、负性情绪、清扫腋窝淋巴结、术后放射疗为PMPS发生的危险因素（P<0.05）。结论 乳腺癌术后较容易发生PMPS,主要表现为术后即刻腋窝疼痛、麻木,发作频率较高,疼痛较重,其中年龄小、术前焦虑、清扫腋窝淋巴结、术后放疗为引发PMPS的危险因素,需加强监测和针对性处理,研究价值较高。

Objective To investigate the incidence and influencing factors of post-mastectomy pain syndrome（PMPS）in breast cancer patients．Methods A total of 82 patients with breast cancer admitted to our hospital from January 2021 to February 2023 were selected as the research objects,and underwent surgical treatment．The incidence of PMPS was counted,the characteristics of PMPS were analyzed,and the clinical data of PMPS patients and non-PMPS patients were compared．Results Among the 82 patients,20 patients had PMPS after surgery,with an incidence of 24.39%．Among them,the affected axilla accounted for 45.00%,numbness pain 35.00%,moderate pain 60.00%,immediate postoperative pain 50.00%,and daily pain 50.00%．There were no significant differences in body mass index,education level,pathological stage of disease,tumor location,surgical method,preoperative use of non-steroid anti-inflammatory drugs,preoperative chemotherapy and postoperative chemotherapy between PMPS patients and non-PMPS patients（P>0.05）．The age of PMPS patients was significantly higher than that of non-PMPS patients,the rate of negative emotion was 30.00%,the rate of axillary lymph node dissection was 95.00%,and the rate of postoperative radiotherapy was 30.00%,which was significantly higher than that of non-PMPS patients（P<0.05）．Age,negative emotion,axillary lymph node dissection and postoperative radiotherapy were independent risk factors for PMPS（P<0.05）．Conclusions PMPS is prone to occur after breast cancer surgery,mainly characterized by immediate postoperative axillary pain and numbness,with a high frequency and severe pain．Young age,preoperative anxiety,axillary lymph node dissection,and postoperative radiotherapy are independent risk factors for PMPS,which need to be strengthened monitoring and targeted treatment．

目的 探讨动态对比增强磁共振成像（DCE-MRI）多参数定量特征对乳腺癌腋窝淋巴结转移（ALNM）风险的预测价值。方法 回顾性收集2020年3月—2022年11月在佛山市高明区人民医院经手术病理确诊的155例乳腺癌患者临床资料,根据患者是否发生ALNM分为ALNM 组（n=39）和无ALNM 组（n=116）。采用单因素分析和多因素Logistic回归分析乳腺癌发生ALNM的影响因素。结果 ALNM组和无ALNM 组患者的肿块质地、肿块直径、肿块部位、肿块形状、肿块内部强化特征等指标比较差异无统计学意义（t/χ²=2.249、0.977、1.369、0.524、2.158,P>0.05）。两组患者肿块表观扩散系数（ADC）值、腋窝淋巴结（ALN）短径、肿块边缘、动态增强时间-信号强度曲线（TIC）曲线等指标比较,差异有统计学意义（t/χ²=6.573、9.873、29.441、2.031,P<0.05）。二元Logistic回归模型结果显示,肿块ADC值、ALN 短径（≥5 mm）、TIC曲线（流出型）为乳腺癌ALNM发生的危险因素（OR=0.251、0.106、0.002,P<0.05）。结论 DCE-MRI多参数定量特征中,乳腺癌患者的肿块ADC值低、ALN 短径（≥5 mm）、TIC曲线（流出型）为乳腺癌ALNM发生的危险因素。

Objective To investigate the predictive value of multi-parameter quantitative features of dynamic contrast-enhanced magnetic resonance imaging（DCE-MRI）in the risk of axillary lymph node metastasis（ALNM）in breast cancer．Methods Clinical data of 155 patients with breast cancer diagnosed by surgery and pathology in Foshan Gaoming District People's hospital from March 2020 to November 2022 were retrospectively collected．According to whether the patients had ALNM,they were divided into ALNM group（n=39）and non-ALNM group（n=116）．Univariate analysis and multiple Logistic regression models were used to explore the influencing factors of ALNM in breast cancer．Results There was no significant difference in mass texture,mass diameter,mass location,mass shape and internal enhancement between the ALNM group and the non-ALNM group（t/χ²=2.249,0.977,1.369,0.524,2.158,P>0.05）．There were significant differences in ADC value,ALN short diameter,tumor margin and TIC curve between the two groups（t/χ²=6.573,9.873,29.441,2.031,P<0.05）．Binary Logistic regression model showed that ADC value,ALN short diameter（≥5 mm）and tumor margin（blur）were risk factors for the occurrence of breast cancer ALNM（OR=0.251,0.106,0.002,P<0.05）．Conclusions Among the multi-parameter quantitative features of DCE-MRI,the ADC value of breast cancer,the short diameter of ALN（≥5 mm）,and the edge of the tumor（blur）are the risk factors for the occurrence of ALNM in breast cancer．

乳腺癌是女性最常见的原发恶性肿瘤之一。目前,通过采用综合治疗手段,包括手术、新辅助治疗、辅助放化疗等多种手段,乳腺癌已成为疗效最佳的实体肿瘤之一。其中,新辅助治疗（NAT）包括新辅助化疗、靶向治疗和内分泌治疗,目的是使肿瘤降期、保乳、保腋窝,并可观察药物敏感性,是当前乳腺癌综合治疗中非常重要的组成部分,其治疗疗效对患者手术方式的选择和预后至关重要。尽管病理学评价在评估NAT疗效方面被公认为金标准,但其局限性在于只能通过有创手段在治疗后进行,无法在治疗前对患者做出准确预测。磁共振成像（MRI）作为一项广泛使用的乳腺成像技术,在评估NAT疗效中扮演着关键角色。近年来,人工智能技术,尤其是影像组学（Radiomics）和深度学习（Deep Learning）,在医学影像分析领域取得了显著进展。这些技术能够从医学图像中提取大量肉眼难以识别的特征,揭示病变内部的微观结构和生物学行为,全面反映肿瘤的异质性,这不仅有助于临床医生更准确地区分良、恶性肿瘤,还能对恶性肿瘤的预后进行更为精确的评估。本文系统综述了近年来基于MRI的人工智能技术在乳腺癌新辅助治疗疗效评估中的应用研究,旨在促进人工智能在NAT临床实践中的应用和发展,为乳腺癌NAT治疗策略的优化和个性化医疗的实现提供科学依据。

Breast cancer is one of the most common primary malignant tumors in women．Currently,breast cancer has become one of the most effective solid tumors by using comprehensive treatment methods,including surgery,neoadjuvant therapy,adjuvant radiotherapy and chemotherapy．Among them,neoadjuvant therapy（NAT）,including neoadjuvant chemotherapy,targeted therapy and endocrine therapy,is a very important part of the current comprehensive treatment of breast cancer．It aims to reduce the tumor stage,preserve the breast,preserve the armpit,and observe the drug sensitivity．Its therapeutic effect is crucial to the choice of surgical methods and prognosis of patients．Although pathological evaluation is recognized as the gold standard in evaluating the response to NAT,its limitation is that it can only be performed after treatment by invasive means,and cannot accurately predict response before treatment．As a widely used breast imaging technology,magnetic resonance imaging（MRI）plays a key role in evaluating the response to NAT．However,traditional MRI evaluation methods are limited by the individual differences of interobserver and the low repeatability of evaluation results,which affects the accuracy of efficacy evaluation to a certain extent．In recent years,artificial intelligence technology,especially radiomics and deep learning,has made significant progress in the field of medical image analysis．These techniques can extract a large number of features that are difficult to be recognized by the naked eye from medical images,reveal the internal microstructure and biological behavior of the lesion,and fully reflect the heterogeneity of the tumor．This not only helps clinicians to distinguish benign and malignant tumors more accurately,but also makes a more accurate assessment of the prognosis of malignant tumors．This article reviews the application and progress of MRI-based artificial intelligence technology in evaluating the response to neoadjuvant therapy for breast cancer in the past five years,aiming to promote the application and development of artificial intelligence in NAT clinical practice,and provide a scientific basis for the optimization of NAT treatment strategy and the realization of personalized medicine for breast cancer．

目的 分析TIMELESS、鼠肉瘤病毒家族相关蛋白2A（RAB2A）、异常纺锤体样小头畸形相关基因（ASPM）在乳腺癌组织中的表达及与临床特征相关性。方法 选取2019年2月—2021年2月我院乳腺癌组织标本84例作为研究组、正常乳腺组织标本53例作为对照组,采用荧光定量聚合酶测定TIMELESS、ASPM,采用Western blot检测RAB2A蛋白表达情况,分析上述三个指标在乳腺癌中表达及与临床特征相关性。结果 对比对照组,研究组TIMELESS、ASPM表达较高,RAB2A较低（P<0.05）。TIMELESS、RAB2A、ASPM与乳腺癌淋巴结浸润、TNM分期、分化程度相关（P<0.05）。TIMELESS、RAB2A、ASPM为影响乳腺癌发生的危险因素（P<0.05）。TIMELESS、RAB2A负相关（r=-0.383、P=0.001）;TIMELESS、ASPM正相关（r=0.397、P=0.001）;RAB2A、ASPM负相关（r=-0.257、P=0.018）。对比TIMELESS、RAB2A、ASPM单一检测,三者联合检测对乳腺癌的诊断价值较高（P<0.05）。结论 乳腺癌患者TIMELESS、ASPM呈高表达,RAB2A呈低表达,上述三个指标与乳腺癌高度相关,可作为乳腺癌发生的检测指标。

Objective To analyze the expression of TIMELESS,murine sarcoma virus family related protein 2A（RAB2A）and abnormal spindle like microcephaly related gene（ASPM）in breast cancer tissues and their correlation with clinical features．Methods A total of 84 breast cancer tissue samples from our hospital from February 2019 to February 2021 were selected as the study group and 53 normal breast tissue samples were selected as the control group．Time,ASPM and RAB2A protein expression were determined by fluorescent quantitative polymerase,and RAB2A protein expression was detected by Western blot．The expression of the above three indicators in breast cancer and their correlation with clinical characteristics were analyzed．Results Compared with the control group,the study group had higher TIMELESS and ASPM expression levels and lower RAB2A level（P<0.05）．TIMELESS,RAB2A and ASPM expressions were correlated with lymph node infiltration,TNM stage and differentiation of breast cancer（P<0.05）．TIMELESS,RAB2A and ASPM were the risk factors of breast cancer（P<0.05）．TIMELESS and RAB2A were negatively correlated（r=-0.383,P=0.001）;TIMELESS and ASPM were positive correlated（r=0.397、P=0.001）;RAB2A and ASPM were negatively correlated（r=-0.257,P=0.018）．Compared with the single detection of TIMELESS,RAB2A and ASPM,the combined detection had higher diagnostic value for breast cancer（P<0.05）．Conclusions Patients with breast cancer had high expression of TIMELESS and ASPM,and low expression of RAB2A．The above three indicators were highly correlated with breast cancer and can be detection indicators for breast cancer．

目的 构建尺寸可变纳米递送系统PAMAM/DOX-pep并进行表征,检测其理化性质并评价其体外抗肿瘤效果与靶向性。方法 将阿霉素（DOX）物理包埋在阳离子聚合物PAMAM的疏水空腔内,以4-（N-马来酰亚胺基甲基）环己烷羧酸N-羟基琥珀酰亚胺酯（SMCC）作为交联剂,采用金属基质蛋白酶（MMP-2）敏感的多肽pep（CPLGVRGC）串联小粒径纳米颗粒形成大尺寸纳米递送系统（PAMAM/DOX-pep）,对各纳米颗粒的粒径、电位、理化性质以及对小鼠乳腺癌细胞（4T1）的抑制作用、细胞摄取效果和核靶向作用进行检测。结果 PAMAM/DOX粒径约为10 nm,载药率为23%,多肽pep交联后形成的PAMAM/DOX-pep粒径约为200 nm,可在低pH下缓释DOX,7天内体外保持稳定且溶血率低、安全无毒,其与MMP-2共孵育后细胞摄取量与核靶向性显著增加。结论 尺寸可变纳米颗粒有助于克服尺寸所引发的递送障碍,将药物靶向递送至乳腺癌细胞核内并发挥作用,为纳米递送系统的设计提供了新策略。

Objective To construct and characterize the size-variable nano-delivery system PAMAM/DOX-pep,examine its physicochemical properties and evaluate its antitumor and targeting effects in vitro. Methods Small particle size PAMAM/DOX was obtained by physically encapsulating DOX within the hydrophobic cavity of the cationic polymer PAMAM. The large size nano-delivery system(PAMAM/DOX-pep)was formed by tandem linking small size nanoparticles by MMP-2 sensitive peptide pep(CPLGVRGC)using SMCC as a cross-linker. The particle size,potential,physical and chemical properties,inhibitory effect,cell uptake and nuclear targeting effect of each nanoparticle on mouse breast cancer cells(4T1)were detected. Results The particle size of PAMAM/DOX was about 10 nm,and the drug loading rate was 23%. PAMAMAM/DOX-pep,formed after cross-linking of peptide,had a particle size of about 200 nm,which could release DOX slowly at low pH,and remained stable,safe and non-toxic in vitro for 7 days with low hemolysis rate,and its cellular uptake amount and nuclear targeting rate increased significantly after co-incubation with MMP. Conclusions Size-variable nanoparticles overcome size-induced delivery barriers to target and deliver drugs to the 4T1 nucleus,providing a new strategy for the design of nano delivery systems.

目的 观察程序性死亡受体1（PD-1）联合细胞毒性T淋巴细胞相关蛋白4（CTLA-4）双免疫疗法对改善晚期乳腺癌近期疗效及远期预后的影响。方法 选择2020年5月—2022年5月商丘市第一人民医院收治的124例晚期乳腺癌患者为研究对象,经随机数字表法将其分为对照组（60例）和观察组（64例）,对照组予以常规PD-1单抗免疫疗法治疗,观察组采用PD-1联合CTLA-4双免疫疗法治疗,比较2组患者治疗前后肿瘤标志物水平、治疗后病灶缓解情况,对所有患者开展为期1年随访,统计并对比2组的不良反应发生情况及远期生存情况。结果 治疗前,2组患者的肿瘤标志物水平比较差异均无统计学意义（均P>0.05）;治疗后,观察组的癌胚抗原为（3.36±0.17）ng/mL,糖类抗原15-3为（25.33±5.28）U/mL,糖类抗原19-9为（38.77±5.62）U/mL,均低于对照组[（5.27±1.36）ng/mL、（28.44±5.18）U/mL、（41.25±5.46）U/mL,均P<0.05]。治疗后,观察组的完全缓解率为21.88%（14/64）,部分缓解率为31.25%（20/64）,病情稳定率为37.50%（24/64）,均高于对照组[8.33%（5/60）、13.33%（8/60）、23.33%（14/60）],肿瘤生长率为（30.27±5.18）%,肿瘤超进展率为6.25%（4/64）,均低于对照组[（33.49±5.32）%、18.33%（11/60）,均P<0.05]。治疗后,观察组的不良反应发生率为34.38%（22/64）,略高于对照组33.33%（20/60）,组间比较差异无统计学意义（P>0.05）;观察组的中位无进展生存期为（9.33±2.25）月,中位总生存期为（10.76±3.32）月,均高于对照组[（7.25±2.31）月、（7.41±1.62）月,均P<0.05]。结论 PD-1联合CTLA-4双免疫疗法能有效改善晚期乳腺癌的近期疗效及远期预后,此疗法未明显增加不良反应发生风险,安全性高。

Objective To observe the effect of programmed cell death protein-1（PD-1）combined with cytotoxic T lymphocyte-associated antigen-4（CTLA-4）dual immunotherapy on the short-term efficacy and long-term prognosis of advanced breast cancer．Methods A total of 124 patients with advanced breast cancer who were admitted to the First People's Hospital of Shangqiu City from May 2020 to May 2022 were selected as the research objects．They were randomly divided into the control group（60 cases）and the observation group（64 cases）by the method of random number table．The control group was treated with conventional PD-1 monoclonal antibody immunotherapy,and the observation group was treated with PD-1 combined with CTLA-4 double immunotherapy．The levels of tumor markers before and after treatment and the focal remission after treatment were compared between the two groups．All patients were followed up for one year,the incidence of adverse reactions and long-term survival between the two groups were compared．Results Before treatment,there was no statistically significant difference in the levels of tumor markers between two groups（all P>0.05）．After treatment,the carcino-embryonic antigen content of the observation group was（3.36±0.17）ng/mL,CA153 was（25.33±5.28）U/mL,and CA199 was（38.77±5.62）U/mL,which were lower than those of the control group [（5.27±1.36）ng/mL,（28.44±5.18）U/mL,（41.25±5.46）U/mL,all P<0.05]．After treatment,the complete remission rate of the observation group was 21.88%（14/64）,partial remission rate was 31.25%（20/64）,and stable disease rate was 37.50%（24/64）,all higher than those of the control group [8.33%（5/60）,13.33%（8/60）,23.33%（14/60）];tumor growth rate of the observation group was（30.27±5.18）%,hyper progressive disease rate was 6.25%（4/64）,both lower than those of the control group [（33.49±5.32）%,18.33%（11/60）,both P<0.05]．After treatment,the incidence of adverse reactions in the observation group was 34.38%（22/64）,slightly higher than that in the control group 33.33%（20/60）（P>0.05）．The median progression free survival of the observation group was（9.33±2.25）months,and the median overall survival was（10.76±3.32）months,both higher than those of the control group [（7.25±2.31）months and（7.41±1.62）months]（P<0.05）．Conclusions PD-1 combined with CTLA-4 dual immunotherapy can effectively improve the short-term efficacy and long-term prognosis of advanced breast cancer．This therapy does not significantly increase the risk of side effects,which is safe．

目的 探讨核结合蛋白2(NUCB2)介导的下游信号分子和通路,为阐明NUCB2在乳腺癌中的功能提供依据。方法 构建NUCB2-RNAi慢病毒载体,感染MDA-MB-231细胞株。然后将MDA-MB-231分为阴性对照病毒感染细胞组(NC组)、感染NUCB2基因shRNA病毒细胞组(KD组),用Affymetrix基因表达谱芯片对NUCB2下游基因进行筛选,并对所有数据进行独创性通路分析(IPA)分析。用qPCR测定mRNA水平。统计采用SPSS 20.0软件。结果 Path-Array研究筛选了KD组与NC组的差异基因,其中上调基因186个,下调基因356个,部分差异表达基因的检测表明,这些基因的mRNA水平与Path-Array筛选结果一致。IPA分析显示,经典途径中差异表达基因的显著富集表明胆固醇生物合成的超途径被显著抑制。上游调节因子分析显示了所有不同表达基因的上游调节因子,包括转录因子、细胞因子、小RNA、受体、激酶、化学分子和药物。疾病和功能差异表达基因的显著丰富表明,与NUCB2相关的差异表达基因与41种疾病和功能显著相关,更多与癌症、组织损伤和异常相关。结论 NUCB2的功能涉及多种基因和多种信号通路。

Objective In order to further explore the downstream signal molecules and pathways mediated by nucleobindin-2 (NUCB2), to provide a basis for elucidating the significance of NUCB2 in breast cancer. Method NUCB2-RNAi lentivirus vector was constructed and infecting MDA-MB-231 cell line.Then MDA-MB-231 cells were divived into two group, cells with negative control virus infection (NC group) and cells infected with NUCB2 gene shRNA virus (KD group). NUCB2 downstream gene screening was conducted by Affymetrix gene expression profiling Path-Array chip and all data were analyzed by ingenuity pathway analysis (IPA). The mRNA level was detected by qPCR. SPSS 20.0 software was used for statistics. Results Path-Array study screened out differential genes between KD and NC group which the number of up-regulated genes was 186, the number of down-regulated genes was 356.Detection of some differentially expressed genes showed that the mRNA levels of these genes were consistent with the results of Path-Array screening.IPA analysis revealed that significant enrichment of differentially expressed genes in the classical pathway showed superpathway of cholesterol biosynthesis was significantly inhibited.The upstream regulatory factor analysis showed the upstream regulatory factors of all the differentially expressed genes, including transcription factors, cytokine, small RNA, receptors, kinases, chemical molecules and drugs.The significant enrichment of differentially expressed genes in disease and function showed that NUCB2 associated differentially expressed genes were significantly related with 41 diseases and functions, which were more related with cancer, organismal injury and abnormities. Conclusion The function of NUCB2 involved multiple genes and multiple signaling pathways.