胃癌术后免疫化疗的远期疗效评价

目的：为探讨术后免疫化疗在胃癌综合治疗中的远期疗效，本文对428例胃癌进行回顾性分析。方法：全部资料来自我院、西安交通大学第一附属医院1989／2001年收治的428例胃癌患者，并随机分为单纯手术组144例，术后化疗组138例，术后化疗免疫组146例。化疗组均给于术后化疗1～3a，观察其5a生仔率。结果：（1）单纯手术、术后化疗及术后免疫化疗组5a生存率分别为24．3％、38．4％和53．4％。术后免疫化疗组与单纯手术组间差异有显著性（P〈0.01）；5a生存率术后免疫化疗组较术后化疗组提高15．0％。（2）无淋巴结转移组中，单纯手术组，术后化疗组及术后免疫化疗组5a生存牢分别为29．3％、51．9％、66．7％。术后免疫化疗组与单纯手术组差异有显著性（P〈0．01），较术后化疗组5a生存率提高14．8％；有淋巴结转移组中，单纯下术组，术后化疗组及术后免疫化疗组5a生存率分别为14．8％、29．8％、44．2％。术后免疫化疗组与单纯手术组差异有显著性（P〈0.01），较术后化疗组5a生存率提高14．4％。结论：胃癌术后免疫化疗呵提高患者的生存期，其远期疗效明显优于术后化疗及单纯手术。对有／无淋巴结转移者，术后免疫化疗均有明显的远期疗效。

肠外营养联合精氨酸对晚期大肠癌化疗患者抗肿瘤免疫应答及癌细胞活力的影响

目的探讨肠外营养联合精氨酸对晚期大肠癌化疗患者抗肿瘤免疫应答及癌细胞活力的影响。方法收集2014年5月至2016年5月间在本院接受化疗的晚期大肠癌患者78例，按照双盲随机对照法分为接受常规肠外营养干预的对照组39例、接受肠外营养联合精氨酸干预的观察组39例。干预前、干预1个月后，采用流式细胞仪测定其中细胞免疫指标水平，采用酶联免疫吸附法（ELISA）测定血清Th1／Th2免疫指标含量，采用发光免疫分析仪检测血清肿瘤标志物含量。结果干预前，两组患者的细胞免疫指标水平、Th1／Th2免疫指标含量、肿瘤标志物含量差异无统计学意义（P〉0．05）。干预1个月后，观察组外周血中细胞免疫指标CD4^＋、CD4^＋／CD8^＋水平高于对照组，CD8^＋水平低于对照组（P〈0．05）；观察组血清中Th1免疫指标IFN-γ、IL-12含量高于对照组，Th2免疫指标IL-4、IL－10含量低于对照组（P〈0．05）；观察组血清中肿瘤标志物糖类抗原724（CA724）、糖类抗原242（CA242）、糖类抗原50（CA50）、癌胚抗原（CEA）含量低于对照组（P〈0．05）。结论肠外营养联合精氨酸可提升晚期大肠癌化疗患者的抗肿瘤免疫应答能力，降低癌细胞活力。

Expression of HMGB1 Protein in Human Cervical Squamous Epithelium Carcinoma

OBJECTIVE To investigate the expression of the high mobility group boxl（HMGB1） in human cervical squamous epithelial carcinoma （CSEC） and to explore the relationship of HMGB1 expression to the differentiation degree, size, invasion and metastasis of CSEC. METHODS Immunohistochemical staining of tissue microarrays and Western blot analysis were conducted to detect the expression of HMGB1 in the following tissue samples： 30 carcinoma in situ, 90 invasive CSEC without metastasis, 30 invasive CSEC with metastasis, 30 cases of normal cervical squamous epithelia. RESULTS The positive-expression rate of HMGB1 was 58.7% （88/150） in CSEC, showing a significant difference compared to normal cervical squamous epithelia. The expression of HMGB1 was correlated with tumor size, invasion and metastasis of CSEC （respectively, P〈0.01）, but had no relationship with the degree of differentiation （P〉0.05）. CONCLUSION The over-expression of HMGB1 in CSEC might be a useful parameter as an indication of tumor invasion, metastasis, prognosis and overall biological behavior of human CSEC, as well as a noval target site for gene therapy.

Expression and clinical significance of PTEN protein in osteosarcoma

Objective： To investigate the expression and significance of cancer inhibitory gene PTEN protein in osteosar-coma. To analyze the level of its expression in different histological classification of osteosarcoma. To determine the possibility of taking PTEN protein as a marker gene for diagnosing osteosarcoma. To observe the clinical value of PTEN expression levels as a reference index for osteosarcoma classification. Methods： 43 specimens collected from osteosarcoma excision were studied. 30 specimens collected during the same period from benign lesion of bone （osteochondroma） operation were taken as the control group. Immunohistochemistry staining （ElivisonTM two steps method） was used to detect the expression of PTEN protein in 43 cases of osteosarcoma. SPSS 10.0 was used in statistical analysis. Results： Immunohistochemistry staining showed that the positive reaction of PTEN protein was all oriented to cytoplasm, which were brown or yellowish- brown granules. By way of X^2 test, the significant difference of the positive expressions of PTEN protein between bone benign lesion and osteosarcoma （X^2 = 7.976, P 〈 0.01） was observed. Osteosarcoma with different degrees of histodifferentiation showed different level expression of PTEN protein. There was significant difference between well-differentiated osteosarcoma （grades Ⅰ-Ⅱ） and poorly-differentiated osteosarcoma （grade Ⅲ） statistically （P 〈 0.01）. The level of expression of PTEN was negatively correlated to the histological grade of osteosarcoma. There was great significance statistically （rs=-0.4922, P 〈 0.01）. Conclusion： PTEN protein may be used as candidate gene of cancer inhibitory gene： PTEN protein is a cancer suppressor gene protein which has expression in bone tumors. It might not only be used in the study of pulmonary carcinoma and neurogliocytoma, but also in the study of bone tumor; the expression of PTEN is related to benignancy or malignancy of bone tumor and their degree of differentiation. The expression of PTEN is positively correlated with degree of differentiation.

Resection of the uncinate process of the pancreas due to a ganglioneuroma

A 33-year-old woman who presented with epigastric discomfort and diarrhea underwent an abdominal ultrasound(US).This investigation and subsequent contrastenhanced computed tomography,magnetic resonance imaging and endoscopic US with fine needle aspiration (FNA)revealed a 40 mm well-circumscribed mass in the uncinate process of the pancreas.Findings were suggestive of a mucinous or solid-cystic pseudopapillary tumor of the pancreas,although other lesions such as a nonfunctioning neuroendocrine tumor could not be ruled out.FNA samples were negative for malignant cells,but of limited value due to poor cellularity.It was decided to surgically remove the tumor because malignancy could not be discounted.Multiple intraoperative biopsies were suggestive of mesenchymal tumor and consequently a conservative resection(uncinatectomy)was performed. The postoperative course was uneventful.The definitive diagnosis was ganglioneuroma.Immunocytochemistry showed positive staining with vimentin,S-100 protein, neurofilament and neuron-specific enolase.Ganglioneuroma is a rare benign tumor that can also present as a pancreatic tumor.Uncinatectomy is feasible,safe and a good surgical technique for the treatment of nonmalignant tumors located in the uncinate process of the pancreas.

KIT-negative gastrointestinal stromal tumors with a long term follow-up:A new subgroup does exist

AIM： To investigate the incidence of KIT immunohostochemical staining in （GI） stromal tumors （GISTs）, and to analyze the clinical manifestations of the tumors and prognostic indicators. METHODS： We retrospectively analyzed 50 cases of previously diagnosed GISTs. Tissue samples were assessed with KIT （CDl17 antigen）, CD34, SMA, desmin, S-100, NSE, PCNA, Ki-67, and BCL-2 for immunohistochemical study and pathological characteristics were analyzed for prognostic factors. RESULTS： Fifteen tumors （30%） were negative in KIT staining. A significant association was observed between gender （male patients： 14/15） and KIT-negative staining （P = 0.003）.The patients＇s mean age was 56.6 years. Tumors developed in stomach （n = 8）, small intestine （n = 5）, large intestine （n = 1） and oesophagus （n = 1）. The mean tumor size was 5.72 cm. The mitotic count ranged from 0-29/50 HPF （mean： 3.4） and 73% of tumors showed no necrosis. The majority of the tumors （67%） had dual or epithelioid differentiation. Tumors were classified as very low or low risk （n = 7）, intermediate risk （n = 5）, and high risk （n = 3） groups. Twelve （80%） patients were alive without evidence of residual tumor for an average period of 40.25 mo （12-82 too）; three patients developed metastatic disease to the liver and eventually died within 2-12 mo （median survival： 8.6 too）.CONCLUSION： A small subgroup of GISTs fulfils the clinical and morphological criteria of these tumors, and lacks KIT expression. These tumors predominantly developed in the stomach, being dual or epithelioid in morphology, which are classified as low risk tumors and presented a better survival status than KIT-positive tumors. The ability to diagnose GISTs still depends on immunohistochemical staining but the research should extend in gene mutations.

Understanding the function and dysfunction of the immune system in lung cancer: the role of immune checkpoints

Survival rates for metastatic lung cancer, including non-small cell lung cancer （NSCLC） and small cell lung cancer （SCLC）, are poor with S-year survivals of less than 5%. The immune system has an intricate and complex relationship with tumorigenesis; a groundswell of research on the immune system is leading to greater understanding of how cancer progresses and presenting new ways to halt disease progress. Due to the extraordinary power of the immune system-- with its capacity for memory, exquisite specificity and central and universal role in human biology--immunotherapy has the potential to achieve complete, long-lasting remissions and cures, with few side effects for any cancer patient, regardless of cancer type. As a result, a range of cancer therapies are under development that work by turning our own immune cells against tumors. However deeper understanding of the complexity of immunomodulation by tumors is key to the development of effective immunotherapies, especially in lung cancer.

Immuno-oncology combinations： raising the tail of the survival curve

There have been exponential gains in immuno-oncology in recent times through the development of immune checkpoint inhibitors. Already approved by the U.S. Food and Drug Administration for advanced melanoma and non-small cell lung cancer,immune checkpoint inhibitors also appears to have significant antitumor activity in multiple other tumor types. An exciting component of immunotherapy is the durability of antitumor responses observed, with some patients achieving disease control for many years. Nevertheless, not all patients benefit, and efforts should thus now focus on improving the efficacy of immunotherapy through the use of combination approaches and predictive biomarkers of response and resistance. There are multiple potential rational combinations using an immunotherapy backbone, including existing treatments such as radiotherapy, chemotherapy or molecularly targeted agents, as well as other immunotherapeutics. The aim of such antitumor strategies will be to raise the tail on the survival curve by increasing the number of long term survivors, while managing any additive or synergistic toxicities that may arise with immunotherapy combinations. Rational trial designs based on a clear understanding of tumor biology and drug pharmacology remain paramount. This article reviews the biology underpinning immuno-oncology, discusses existing and novel immunotherapeutic combinations currently in development, the challenges of predictive biomarkers of response and resistance and the impact of immuno-oncology on early phase clinical trial design.

Effects and possible anti-tumor immunity of electrochemotherapy with bleomycin on human colon cancer xenografts in nude mice

AIM: To evaluate the anti-tumor effects and possible involvement of anti-tumor immunity of electrochemotherapy (ECT) employing electroporation and bleomycin in human colon cancer xenografts in nude mice, and to establish the experimental basis for clinical application of ECT. METHODS: Forty nude mice, inoculated subcutaneously human colon cancer cell line LoVo for 3 wk, were allocated randomly into four groups: B+E+ (ECT), B+E- (administration of bleomycin alone), B-E+ (administration of electric pulses alone), and B-E- (no treatment). Tumor volumes were measured daily. The animals were killed on the 7* d, the weights of xenografts were measured, and histologies of tumors were evaluated. Cytotoxicity of spleen natural killer (NK) and lymphokine-activated killer (LAK) cells was then assessed by lactic dehydrogenase release assay. RESULTS: The mean tumor volume of group B+E+ was statistically different from the other three groups after the treatment (F= 36.80, P<0.01). There was one case of complete response, seven cases of partial response (PR) in group B+E+, one case of PR in group B+E- and group B-E+ respectively, and no response was observed in group B-E-. The difference of response between group B+E+ and the other three groups was statistically significant (X2 = 25.67,P<0.01). Histologically, extensive necrosis of tumor cells with considerable vascular damage and inflammatory cells infiltration were observed in group B+E+. There was no statistical difference between the cytotoxicity of NK and LAK cells in the four treatment groups. CONCLUSION: ECT significantly enhances the chemosensitivity and effects of chemotherapy in human colon cancer xenografts in nude mice, and could be a kind of novel treatment modality for human colon cancer. The generation of T-cell-dependent, tumor-specific immunity might be involved in the process of ECT.

实体肿瘤进展小析

1 免疫治疗 近年来，免疫治疗在肿瘤治疗中由默默无闻到大放异彩，经历了漫长的过程。免疫治疗抗体，抗肿瘤细胞免疫治疗足其蕈要的部分。抗CTLA-4、PD-1／PD-L1抗体作为首当其中的先驱，在黑色素瘤、肺癌、肾痛的临床研究及基础研究中给出了不错的答案，且二者联用取得了更优的结果。

热化疗栓塞影响中晚期肝癌患者机体免疫功能的研究

目的观察经肝动脉热化疗栓塞对中晚期肝癌患者的机体免疫功能的影响。方法原发性肝癌患者40例完全随机分为2组，均采用经股动脉途径插管至肝动脉，A组（20例）ADM40mg＋MMC 10mg溶于37℃的生理盐水灌注；B组（20例）ADM40mg＋MMC 10mg溶于60℃的生理盐水灌注；A或B组灌注后再用适量碘化油栓塞。采用流式细胞仪及双抗体夹心法，检测治疗前、后外周血T细胞亚群（CD3^＋、CD4^＋、CD8^＋）、NK细胞的百分率及sIL-2R的变化。采用LDH释放法检测NK细胞的杀伤活性。结果与术前及普通化疗栓塞组比较，热化疗栓塞治疗后CD4^＋、NK细胞的百分率明显升高（P〈0．05），NK细胞杀伤活性也升高（P〈0．05），而CD8^＋细胞的百分率及sIL-2R的水平明显降低（P〈0．05）。结论热化疗栓塞具有免疫激活作用，可使机体的免疫功能得到一定程度的改善。

PEP3-KLH负载小鼠树突状细胞疫苗抗前列腺癌实验研究

目的 探讨PEP3-KLH负载树突状细胞（DCs）疫苗诱导的特异性抗前列腺癌（PCa）免疫作用.方法 用PEP3-KLH或KLH体外冲击小鼠骨髓来源DCs,构建DCs瘤苗.PEP3-KLH-DC皮下免疫C57BL/6J小鼠3次,每次间隔2周.末次免疫后1周,ELISA法检测小鼠血清IL-2、IL-12、及IFN-γ含量;乳酸脱氢酶（LDH）法检测小鼠脾T细胞、肿瘤浸润淋巴细胞（TILs）特异性细胞毒性T淋巴细胞（CTL）活性;观察不同抗原负载DC疫苗对肿瘤攻击小鼠免疫保护作用,计算存活期肿瘤平均体积和小鼠生存率;流式细胞术（FCM）检测肿瘤局部TILs的CD4＋、CD8＋T细胞阳性表达百分率.结果 PEP3-KLH-DC免疫可诱导分泌较高水平IFN-γ和IL-12;CTL杀伤肿瘤细胞活性增强,其中TIL细胞毒活性明显增强;FCM检测该组肿瘤局部TILs的CD4＋、CD8＋T细胞阳性表达率明显高于DC-KLH组和DC＋ PBS组（P＜0.01）,而对照组之间比较差异无统计学意义（P＞0.05）.通过比较肿瘤生长曲线、生存曲线可见PEP3-KLH-DC组小鼠肿瘤体积明显较其它各组小,生存率明显提高（P＜0.01）.结论 PEP3-KLH-DC疫苗能有效抑制肿瘤生长,诱导和增强机体抗PCa特异性细胞免疫功能.

Hurdles of CAR-T cell-based cancer immunotherapy directed against solid tumors

Recent reports on the impressive efficacy of chimeric antigen receptor(CAR)-modified T cells against hematologic malignancies have inspired oncologists to extend these efforts for the treatment of solid tumors. Clinical trials of CAR-T-based cancer immunotherapy for solid tumors showed that the efficacies are not as remarkable as in the case of hematologic malignancies. There are several challenges that researchers must face when treating solid cancers with CAR-T cells, these include choosing an ideal target, promoting efficient trafficking and infiltration, overcoming the immunosuppressive microenvironment, and avoiding associated toxicity. In this review, we discuss the obstacles imposed by solid tumors on CAR-T cell-based immunotherapy and strategies adopted to improve the therapeutic potential of this approach. Continued investigations are necessary to improve therapeutic outcomes and decrease the adverse effects of CAR-T cell therapy in patients with solid malignancies in the future.

DNA methylation in the tumor microenvironment

The tumor microenvironment （TME） plays an important role in supporting cancer progression. The TME is composed of tumor cells, the surrounding tumor-associated stromal cells, and the extracellular matrix （ECM）. Crosstalk between the TME components contributes to tumorigenesis. Recently, one of our studies showed that pancreatic ductal adenocarcinoma （PDAC） cells can induce DNA methylation in cancer-associated flbroblasts （CAFs）, thereby modifying tumor-stromal interactions in the TME, and subsequently creating a TME that supports tumor growth Here we summarize recent studies about how DNA methylation affects tumorigenesis through regulating tumor- associated stromal components including fibroblasts and immune cells. We also discuss the potential for targeting DNA methylation for the treatment of cancers.

Dynamics on tumor immunotherapy model with periodic impulsive infusion

A periodic pulse differential equation model of tumor immunotherapy is established by considering the periodic and transient behavior of infusing immune cells. Using comparison theorem and Floquet multiplier theory of the impulsive differential equation, the boundedness of the model solution, the existence and stability of the free-tumor periodic solution are given. Furthermore, the persistence of the system is analyzed. Numerical simulations are carried to confirm the main theorems.

Tumor immunology and immunotherapy: a journey I started from Hangzhou

This short article is dedicated to the 90th Anniversary of the School of Life Sciences at Zhejiang University,China.Immunotherapy of cancer is currently a hot topic in the biomedical field,and a research focus of my laboratory is on developing new and effective combinatorial immunotherapeutic strategies for liver cancer.Of note,my interest in immunotherapy of cancer stems from the training as an undergraduate student at Hangzhou University,China,almost 40 years ago.