Hepatocellular carcinoma-specific immunotherapy with synthesized α1,3-galactosyl epitope-pulsed dendritic cells and cytokine-induced killer cells

AIM: To evaluate the safety and clinical efficacy of a new immunotherapy using both α-Gal epitope-pulsed dendritic cells (DCs) and cytokine-induced killer cells.METHODS: Freshly collected hepatocellular carcinoma(HCC) tumor tissues were incubated with a mixture of neuraminidase and recombinant α1,3-galactosyltransferase (α1,3GT) to synthesize α-Gal epitopes on carbohydrate chains of the glycoproteins of tumor membranes. The subsequent incubation of the processed membranes in the presence of human natural anti-Gal IgG resulted in the effective phagocytosis to the tumor membrane by DCs. Eighteen patients aged 38-78 years with stage Ⅲ primary HCC were randomLy chosen for the study; 9 patients served as controls, and 9 patients were enrolled in the study group.RESULTS: The evaluation demonstrated that the procedure was safe; no serious side effects or autoimmune diseases were observed. The therapy significantly prolonged the survival of treated patients as compared with the controls (17.1 ± 2.01 mo vs 10.1 ± 4.5 mo,P = 0.00121). After treatment, all patients in the study group had positive delayed hyper sensitivity and robust systemic cytotoxicity in response to tumor lysate as measured by interferon-γ-expression in peripheral blood mononuclear cells using enzyme-linked immunosorbent spot assay. They also displayed increased numbers of CD8-, CD45RO-and CD56-positive cells in the peripheral blood and decreased α-fetoprotein level in the serum.CONCLUSION: This new tumor-specific immunotherapy is safe, effective and has a great potential for the treatment of tumors.

Role of Low Dosage Arsenic Trioxide on Pulmonary Dendritic Cells in Asthmatic Mice

Objective: To investigate the distribution and recruitment of pulm onary dendritic cells (DCs) and the influence of low dosage arsenic trioxide (As 2O 3) on them in the airway of asthmatic mice. Methods: Thirty BALB/c mice were randomly divided into 3 groups: the control group, the asthmat ic group and the As 2O 3 treated group. The mice asthmatic model was induced v ia sensitizing with peritoneal injection of ovalbumin (OVA) for two times and th en provocated with aerosol inhalation of OVA for a week. The treated group was p eritoneally injected with 0.2 ml solution of As 2O 3 (4mg/kg) 0.5h after each provocation. The immunohistochemistry and computerised image analysis were appli ed to detect quantitatively the DCs in the lung and airway of mice. Resul ts: All intraepithelial nonlymphoid dendritic cells 145 (NLDC 145) throughout the respiratory tree in the mice of the control group formed a netwo rk with the density of DCs varying from (575±54) cells/mm 2 epithelial surface in the large airway, to (68±12) cells/mm 2 epithelial surface in the small ai rway. The distribution of airway NLDC 145 + in the asthmatic group was simil ar to that in the control group, but its density was significantly upregulated ( P <0.01). The distribution of airway NLDC 145 in the treated group was sim ilar to that in the asthmatic group, only its density was significantly downregu lated ( P <0.01). Conclusion: There is an integral network of N LDC 145 + throughout the respiratory tree. To downregulate the density but n ot change the distribution of pulmonary DCs could be an important therapeutic me chanism of low dosage As 2O 3 in treating asthma.

In-situ observations of dendritic fragmentation during directional solidification of Sn-10wt.% Bi alloy

The stray crystals seriously affect the properties of metals which fabricated by directional solidification(DS). In order to observe the formation of the stray crystals, the growth behavior of Sn-10 wt.%Bi alloy was observed by synchrotron X-ray imaging technique using self-developed directional solidification(DS) apparatus. The experimental results show that Sn–10 wt.%Bi alloy is oriented with the form of dendrites to grow into the liquid phase during the DS process, and dendrites are arranged neatly to form columnar crystal microstructures. It is also found that parts of the dendrite arms begin to crack. These cracked dendrites drift upward to grow up in front of the solid-liquid interface and form stray crystals.

咳嗽变异性哮喘与典型支气管哮喘患者外周血树突状细胞的变化

目的:计算咳嗽变异性哮喘(CVA)、典型支气管哮喘(CA)患者,其外周血中树突状细胞(DCs)及亚型髓样树突状细胞(m DC)、浆细胞样树突状细胞(p DC)比例和数量的变化,了解咳嗽变异性哮喘与典型支气管哮喘之间是否有类似的免疫学发病机制.方法:选取CA患者20例,CVA患者17例,对照组健康人17例,使用流式细胞术检测和计算CA组与CVA组受试者外周血中DCs及其两个亚群:p DC和m DC所占比例和数量的变化.结果:咳嗽变异性哮喘与典型支气管哮喘患者相比,其外周血树突状细胞及其亚型的数量和比例均无明显差异,咳嗽变异性哮喘、典型支气管哮喘患者较健康对照组,p DC数量及比例的增高明显,m DCs/p DCs的比值降低有统计学差异.结论:咳嗽变异性哮喘与支气管哮喘的发病机制在免疫学上类似,由于p DCs比例和数量的增多引起Th2细胞数量增多,造成Th1/Th2平衡向Th2细胞偏移.

肝脏滤泡树突状细胞肉瘤临床病理特征

目的:探讨肝脏滤泡树突状细胞肉瘤患者的临床病理学特征、诊断、鉴别诊断、治疗方法和预后.方法:以中国解放军南京军区福州总医院收治的3例肝脏滤泡树突状细胞肉瘤患者为对象,分析其临床表现及诊疗经过,并进行随访,同时对送检标本进行病理学检查,结合有关文献探讨该病的临床病理学特点.结果:肿瘤位于肝脏左叶或右叶,呈浸润性生长.组织学上可见梭形细胞肉瘤成分与炎性细胞成分混合存在;局灶见大片凝固性坏死.免疫组织化学染色显示肿瘤细胞呈CD21、CD23、CD35、vimentin强阳性,CD45、EMA呈弱阳性表达,反应性炎症细胞呈CD20、CD79a、CD2、CD3、CD45RO、CD38阳性.例3肿瘤细胞呈EBER、EBV阳性表达.免疫组织化学染色阴性抗体有:CK7、CK8、CK18、CK(H)、CD34、CD117、Dog-1、actin、SMA、caldesmon、desmin、CD10、CD21、CD23、CD35、CD1a、CD2、CD3、CD20、CD45、ALK、CD68、CD163、CD30、CD15、HMB45、S-100.病理诊断为肝脏滤泡树突状细胞肉瘤.结论:肝脏滤泡树突状细胞肉瘤是一种罕见恶性肿瘤,临床与病理上需要与指突状树突细胞肉瘤、肝肉瘤样癌、炎性肌纤维母细胞瘤、胃肠道外间质瘤、组织细胞肉瘤等相鉴别,预后不定.

Osteoclast fusion and regulation by RANKL-dependent and independent factors

Osteoclasts are the bone resorbing cells essential for bone remodeling.Osteoclasts are formed from hematopoietic progenitors in the monocyte/macrophage lineage.Osteoclastogenesis is composed of several steps including progenitor survival,differentiation to mononuclear pre-osteoclasts,fusion to multi-nuclear mature osteoclasts,and activation to bone resorbing osteoclasts.The regulation of osteoclastogenesis has been extensively studied,in which the receptor activator of NF-κB ligand(RANKL)-mediated signaling pathway and downstream transcription factors play essential roles.However,less is known about osteoclast fusion,which is a property of mature osteoclasts and is required for osteoclasts to resorb bone.Several proteins that affect cell fusion have been identified.Among them,dritic cell-specific transmembrane protein(DC-STAMP)is directly associated to osteoclast fusion in vivo.Cytokines and factors influence osteoclast fusion through regula-tion of DC-STAMP.Here we review the recently discovered new factors that regulate osteoclast fusion with specific focus on DC-STAMP.A better understanding of the mechanistic basis of osteoclast fusion will lead to the development of a new therapeutic strategy for bone disorders due to elevated osteoclast bone resorption.Cell-cell fusion is essential for a variety of cellular biological processes.In mammals,there is a limited number of cell types that fuse to form multinucleated cells,such as the fusion of myoblasts for the formation of skeletal muscle and the fusion of cells of the monocyte/macrophage lineage for the formation of multinucleated osteoclasts and giant cells.In most cases,cellcell fusion is beneficial for cells by enhancing function.Myoblast fusion increases myofiber size and diameter and thereby increases contractile strength.Multinucleated osteoclasts have far more bone resorbing activity than their mono-nuclear counterparts.Multinucleated giant cells are much more efficient in the removal of implanted materials and bacteria due to chronic infection than macrophages.Therefore,they are also called foreign-body giant cells.Cell fusion is a complicated process involving cell migration,chemotaxis,cell-cell recognition and attachment,as well as changes into a fusion-competent status.All of these steps are regulated by multiple factors.In this review,we will discuss osteoclast fusion and regulation.

α-干扰素对慢性髓细胞白血病来源的树突状细胞的免疫调节作用(Ⅱ)

目的 于慢性髓细胞白血病 (CML)来源的树突状细胞 (DCs)的无血清培养基中 ,加入α 干扰素 (IFN α) ,观察IFN α对CML DCs表达Fas/FasL的影响 ,以及体外IFN α合并DCs瘤苗对CML祖细胞的抑制作用。方法 CML DCs的培养基中除加入SCF、GM CSF、TNF α及IL 4外 ,还加入IFN α。培养 10～ 14d ,鉴定细胞免疫表型、Ph1染色体比例和混合淋巴细胞反应 ,流式细胞仪检测细胞表达Fas/FasL比例 ,碘化丙锭 (PI)染色分析细胞凋亡 ,ELISA法测上清液sFas含量 ,甲基纤维素半固体集落培养法鉴定IFN α加DCs瘤苗对CML的CFU GEMM的体外抑制作用。结果 加入IFN α后 ,CML DC免疫表型及刺激T细胞增殖能力均显著改善 ;CML DCsFas的表达上调 ,sFas含量却下降 ,FasL表达阴性 ;PI染色显示亚二倍体峰的数值升高 ,即凋亡比增加。IFN α合并DCs瘤苗组对CML的CFU GEMM体外抑制作用优于IFN α组或DCs瘤苗组。结论 IFN α在改善CML DCs免疫表型及功能的同时 ,亦可通过Fas途径促进Ph1阳性细胞凋亡。IFN α与DCs瘤苗有协同作用 ,体外二者联合应用对CML祖细胞有明显的抑制作用。

Computer aided analysis system for drilling

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氯化胆碱-尿素低共熔溶剂中电解制备超细铜粉

利用1ChCl∶2Urea低共熔溶剂作为电解质,以单质铜作为阳极,钛片作为阴极,草酸(H2C2O4·2H2O)作为添加剂,在恒电流密度下电解制备出树枝状超细铜粉。用电化学工作站对[1ChCl∶2Urea]/0.1 mol·L-1 H2C2O4·2H2O低共熔溶剂体系进行了相关的电化学测试。研究了电解温度、电流密度及草酸浓度对电解所得铜粉的粒度、电流效率和直流电耗的影响;利用扫描电镜和激光粒度仪分别对铜粉的形貌和粒度进行了测试表征。结果表明:优化工艺条件为控制温度在80℃,电流密度为8.6mA·cm-2以及草酸浓度0.10mol·L-1的条件下,可电解得到粒度小于15μm的树枝状超细铜粉,电解过程的电流效率高达95.6%,电耗为1746kW·h·t-1。

THE VARIATIONAL PRINCIPLE OF THIN SHELL IN STABILITY

this paper gives the resultant forces and moments strain energy and work of external forces on the basis of the deformation theory of flexible body therefore in aqcfordance with the principle of virtual displacement the energy criterion of critical lad is obtained and the equilibrium equation and boundary conditions of stability problem are derived

真核表达载体pEGFP-GPC3的构建及表达

目的:构建磷脂酰肌醇蛋白聚糖3(GPC3)真核表达载体pEGFP-GPC3,并在小鼠树突状细胞(DC)表达。方法:将质粒pCMV-SPORT6-GPC3和载体pEGFP-N1分别进行双酶切获得目的基因GPC3片段和空载体,回收纯化后用T4连接酶将两者连接并转化E.coli DH5α菌株,用EcoR I、BamH I双酶切鉴定后通过脂质体法转染到DC中,然后进行荧光检测和Western blot分析。结果:构建的真核表达载体pEGFP-GPC3,转染DC后,荧光显微镜下可见转染的DC中有EGFP-GPC3融合蛋白的表达,Western blot分析发现有相对分子质量(Mr)大小约为67 000的蛋白条带。结论:成功地构建了真核表达载体pEGFP-GPC3并在转染DC后能在其中表达,为进一步研究GPC3基因的功能提供实验基础。

ICU重症肺炎的中医证候分布特征研究

目的总结ICU重症肺炎患者的中医证型及证候要素分布特点,对ICU重症肺炎患者开展中医药方面的治疗提供依据。方法收集2015年1月~2019年9月安徽省中医院ICU收治的重症肺炎患者病例,采用描述性统计学方法计算证候及症状的频数和构成比,总结分析其中医辨证分型及证候要素的分布特征。结果纳入研究患者158例,中医证型分布包括痰热壅肺证、肺胃热盛证、气阴两虚证、风热犯肺证、热闭心包证。其中实证120例,占总体的75.95%;虚证38例,占总体的24.05%。证候要素分布中,实性证素包括痰、热、瘀、湿、风,虚性证素包括气虚、阴虚、阳虚、血虚。预后情况方面,痰热壅肺证死亡10例,肺胃热盛证死亡6例,气阴两虚证死亡4例,热闭心包证死亡4例,风热犯肺证死亡0例。结论重症肺炎的中医证型以实证为主,其中痰热壅肺证是主要证型,气阴两虚证也不可忽视;在治疗中应主要采用清热化痰药物,还要注意益气养阴药物的使用。在预后方面,热闭心包证死亡率最高,在治疗中应未病先防、既病防变,防止毒邪入里。

透平机的稳定性运行技术探讨

通过剖析在生产运行中遇到的一些问题及采取的一些解决措施,对提高透平机的运行可靠性展开一系列的探讨。

子宫颈癌的免疫治疗

近几年有研究以免疫治疗运用于子宫颈恶性肿瘤并取得一定的成效,其中包括检查点抑制剂(immune checkpoint inhibitors)、基于树突状细胞的治疗(dendritic cell-based therapy)、免疫T细胞移植(adoptive T-cell transfer)、肿瘤浸润淋巴细胞免疫细胞移植(tumor infiltrating lymphocytesadoptive cell transfer)、基于细胞毒性T淋巴细胞的免疫疗法(cytotoxic T lymphocytes-based immunotherapy)、使用DC-CIK治疗因子诱导的杀伤细胞的免疫疗法(cytokine-induced killer cell-based immunotherapy)、基于DNA的免疫疗法(DNAbased immunotherapy)。免疫治疗的方法包括结合不同免疫治疗方法,或与放化疗结合使用。这些方法为子宫颈癌的治疗提供了新的希望。然而,免疫治疗的机制和制备过程复杂,且个体化治疗要求高,制备效率低、价格昂贵、周期长等问题亟待解决,需要进一步研究和临床共同努力。

树突状细胞疫苗治疗肝细胞癌的研究进展

肝细胞癌（HCC）为肝脏最常见的恶性肿瘤，早期诊断困难，进展迅速，患者疾病负担较重，预后较差。目前，HCC的主要治疗手段包括手术切除、肝移植、介入治疗、化疗、消融治疗、靶向治疗等，但其改善预后的效果不佳。近年来，树突状细胞在肿瘤免疫治疗中的作用逐渐被认识，并将成为有前景的改善HCC患者预后的治疗方法。本文就树突状细胞疫苗治疗HCC的研究现状进行综述。

无关第三方耐受型树突细胞对小鼠异基因骨髓移植后急性移植物抗宿主病的预防作用

目的探讨无关第三方耐受型树突细胞（tDC）的生物学特性及其对急性移植物抗宿主病（aGVHD）的预防作用。方法用GM—CSF、IL-10和TGF—B1诱导培养D1小鼠骨髓源tDC，用流式细胞术检测细胞表型，用RT—PCR方法检测细胞因子基因表达情况。采用混合淋巴细胞反应法检测其体外对异基因小鼠CD4＋T细胞增殖的影响；将D1小鼠骨髓源tDC按不同剂量分组输注给aGVHD小鼠模型（B6--D2），观察受鼠一般状况、生存率、临床GVHD评分、血清中Thl／Th2细胞因子水平的变化，并以单纯aGVHD模型为对照组进行比较。结果流式细胞术检测结果表明tDC表达低水平的MHCⅡ分子（I—A／I—E）和共刺激分子（CD80、CD86、CD40）。RT—PCR结果表明，tDC低水平表达炎性细胞因子IL-12p40，但高水平表达免疫抑制性因子IL-10、TGF—β、FasL、IDO和精氨酸酶。无关第三方tDC在体外能够有效抑制异基因CD4＋T细胞的增殖，并且这种抑制效应与tDC的剂量有关；小鼠体内实验显示，aGVHD对照组小鼠在移植后18d内全部死亡；而接受无关第三方tDC输注组小鼠aGVHD症状均有所改善，其中输注10。tDC的受鼠有60％能存活至60d以上，无明显的GVHD症状。而输注10^3 tDC的小鼠仅有20％能存活至移植后第60天，输注10^5 tDC的小鼠则在移植后37d内全部死亡。移植21d时，输注10^3、10^4、10^5无关第三方tDC的小鼠血清内IL-10水平分别为（114．23±7．78）、（646．18±212．02）、（121．97±10．47）ng／L，10^4 tDC组显著高于其他组（P〈0．05）。结论经IL—10和TGF-β1诱导的无关第三方tDC在体外能够抑制CD4＋T细胞的增殖，对aGVHD有显著的预防作用，这种作用与tDC剂量和IL-10的高表达有关。过继性输注一定量的无关第三方tDC能显著延长aGVHD小鼠的生存期，有效预防异基因骨髓移植小鼠aGVHD的发生。