瘤苗

摘要： 目的 探讨mRNA高通量测序的树突状细胞(DC)原位胰腺癌细胞(BxPC-3)瘤苗(DC-BxPC-3)抗胰腺癌的机制.方法 2018年12月至2019年12月取新鲜外周静脉血(红十字会),胰腺癌BxPC-3细胞购自上海生命科学研究院.以淋巴细胞分离液分离50 ml外周静脉血,贴壁培养获得DCs和去DCs的系统免疫效应细胞(SIECs),并诱导增殖.DCs致敏时,BxPC-3∶ DCs=1∶1;抑制与凋亡实验时,DCs∶ SIECs∶靶细胞=1∶20∶2.以mRNA高通量测序,细胞计数试剂盒(CCK-8)、膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)、碘化丙锭(PI)进行检测.组间比较采用LSD或Dunnett T3检验.结果 CCK-8实验显示,致敏DCs组、DCs-BxPC-3瘤苗组的BxPC-3生存率分别为56.7％、23.2％,两组比较差异均有统计学意义(t=11.160,P＜0.05).凋亡实验显示,致敏DCs组、DCs-BxPC3瘤苗组对BxPC-3的凋亡率分别为(23.12±1.05)％、(80.86±5.09)％,组间比较差异有统计学意义(t=19.240,P＜0.05).差异mRNA的基因本体论(GO)分析显示融合瘤苗组的催化活性、分子信号传导活性、免疫系统、代谢过程、生物调节等功能富集;京都基因与基因组百科全书(KEGG)结果显示差异基因主要富集于干扰肿瘤细胞氨基酸代谢与脂质转运.结论 DC瘤苗诱导免疫效应细胞抗胰腺癌效能优于肿瘤裂解物致敏DC.

摘要： 目的 体外观察诱导扩增的健康人或正常人(统称为健康人)外周静脉血树突状细胞(DCs)杂交瘤苗诱导系统免疫效应细胞(SIECs)对原位胰腺癌细胞株(BxPC-3)的抑制与凋亡作用.方法 以淋巴细胞分离液梯度离心获得健康人外周静脉血单核细胞(PBMCs),贴壁法获取DCs和去DCs的单核细胞(即SIECs),并分别用人粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素4(IL-4)、肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)、BxPC-3肿瘤特异性抗原(BxPC-3TSA)培养DCs和SIECs.5d后收集DCs并计数,用PEG+ 10％二甲基亚砜(DMSO)做为融合剂,按10∶1、5∶1、1∶5将BxPC-3TSA致敏的DCs(简称敏DCs)、DCs分别与BxPC-3进行融合,获得敏DCs-BxPC-3、DCs-BxPC-3融合瘤苗.第7天,SIECs分别和敏DCs-BxPC-3、DCs-BxPC-3、敏DCs、DCs以40∶1混合培养1d,分别收集混合培养细胞,并以混合细胞中的SIECs∶ BxPC-3=10∶1的效靶比对BxPC-3进行膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)/碘化丙锭(PI)法凋亡实验和细胞计数试剂盒(CCK-8)法毒性实验.结果 敏DCs、DCs与BxPC-3融合,以1∶1的比例融合时融合效果最好,融合率为40.0％.CCK-8法实验显示:敏DCs-BxPC-3、DCs-BxPC-3、敏DCs诱导SIECs对BxPC-3抑制率分别为53.1％、45.3％、78.8％,吸光度(A)值Dunnett T3检验显示组之间差异均有统计学意义(P＜0.05).凋亡实验显示:敏DCs瘤苗、DCs瘤苗、敏DCs、DCs诱导SIECs对BxPC-3的凋亡率分别为24.17％,11.43％,85.87％,29.94％,各组间差异有统计学意义(P＜0.05).结论 肿瘤抗原序贯法致敏DCs使其获取抗原时出现叠加效应,从而发挥更好的抗肿瘤结果;早期DCs融合瘤苗在一定程度上可诱导SIECs抑制肿瘤细胞效果.%Objective To observe the inhibition and apoptosis effects of systematic immune effector cells (SIECs) induced by increased expansion of normal human peripheral venous blood dendritic cells (DCs) hybridoma vaccine on orthotopic pancreatic cancer cell line (BxPC-3) in vitro.Methods The peripheral blood mononuclear cells (PBMCs) of healthy human were obtained by gradient centrifugation of lymphocyte separation fluid,and DCs and DCs derived monocytes cells (SIECs) were obtained by adherent method.DCs and SIECs were respectively cultured with granulocyte macrophage-colony stimulating factor (GM-CSF),interleukin (IL)-4,tumor necrosis factor-α (TNF-α),IL-2,BxPC-3 tumor specific antigen (pc-3tsa).After five days,DCs were collected and counted.BxPC-3TAA-sensitized DCs (referred to as sensitive DCs),and DCs were fused with BxPC-3 by PEG + 10％ dimethylsurfoxide (DMSO) with 10∶ 1,5∶1,1∶5 to obtain sensitive DCs-BxPC-3 and DCs-BxPC-3 fusion tumor vaccines.On the 7th day,SIECs were respectively mixed with sensitive DCs-BxPC-3,DCs-BxPC-3,sensitive DCs,and DCs for 1 days in a 40∶1 mixture.The mixed cultured cells were collected,and the Annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay and cell counting kit-8 (CCK-8) toxicity assay were performed on BxPC-3 in the mixed target SIECs∶ BxPC3 =10∶1.Results The sensitives DCs,DCs and BxPC-3 were fused,and the fusion effect was the best when the ratio was 1 ∶ 1,and the fusion rate was 40.0％.The CCK-8 assay showed that the inhibition rates of BxPC-3 induced by SIECs and SIECs by sensitive DCs-BxPC-3,DCs-BxPC-3,sensitive DCs and DCs were 53.1％,45.3％ and 78.8％,respectively.The Annexin V-FITC/PI assay showed that apoptosis rates of sensitive DCs-BxPC-3,DCs-BxPC-3,sensitive DCs,DCs and SIECs were 24.17％,11.43％,85.87％ and 29.94％.Conclusion Tumor antigen sequential sensitization DCs have a superimposed effect on antigen acquisition,which can play a better anti-tumor effect.Early DCs fusion tumor vaccine can induce SIECs to inhibit tumor cells to a certain extent.

摘要： 目的：探讨从胃癌干细胞中提取伴侣分子⁃抗原肽的方法及其免疫功能。方法低温超声裂解筛选胃癌干细胞和胃癌细胞，经盐析粗提蛋白及透析后，采用十二烷基硫酸钠⁃聚丙烯酰胺凝胶电泳（ SDS⁃PAGE）分析伴侣分子⁃抗原肽表达量，采用溴化氰活化琼脂糖凝胶4B亲和层析分离、纯化伴侣分子⁃抗原肽，采用反向高压液相色谱、SDS⁃PAGE和Western blot法进行蛋白纯度分析，采用淋巴细胞增殖实验和细胞毒活性实验检测伴侣分子⁃抗原肽的免疫活性。结果成功制备并鉴定胃癌干细胞的伴侣分子⁃抗原肽，主要组分为HSP60⁃抗原肽、HSP70⁃抗原肽、HSP90⁃抗原肽和HSP110⁃抗原肽。0．75μg HSP70⁃抗原肽、1．00μg HSP70⁃抗原肽和1．00μg HSP90⁃抗原肽对淋巴细胞有明显的增殖作用，其A490值分别为0．26±0．03、0．45±0．05和0．32±0．04，而相应剂量的 HSP60⁃抗原肽和HSP110⁃抗原肽不能激活淋巴细胞。1．00μg HSP70⁃抗原肽组和1．00μg HSP70组的细胞杀伤率分别为（45．0±2．0）％和（16．0±2．0）％，差异有统计学意义（P＝0．012）。1．00μg HSP90⁃抗原肽组和1．00μg HSP90组的细胞杀伤率分别为（36．0±5．0）％和（13．0±4．0）％，差异有统计学意义（P＝0．048）。结论胃癌细胞中伴侣分子⁃抗原肽含量极低，而胃癌干细胞中伴侣分子⁃抗原肽含量明显增多，经提纯后可制备纯度较高的伴侣分子⁃抗原肽。提取的伴侣分子⁃抗原肽具有较强的免疫原性，可在体外制作瘤苗，可能在胃癌靶向治疗上有较好的应用价值。%Objective To explore the method of extracting chaperone antigen peptide complexes from gastric cancer stem cells and its immune function. Methods Gastric cancer stem cells and gastric cancer cells were screened by low temperature ultrasonic lysis. After salting out and dialysis, the lysate supernatant was processed with SDS⁃PAGE to analyze the expression of chaperone antigen peptide complexes, and then was separated and purified with CNBr⁃activated SepharoseTM 4B. Reverse high pressure liquid chromatography ( HPLC) , SDS⁃PAGE and Western blotting were used to analyze the purity and nature of the acquired albumen. Lymphocyte proliferation assay and lymphocytotoxicity assay were used to ditermine the immunological activity of the chaperone⁃antigen peptide complexes. Results The chaperone antigen peptide complexes of gastric cancer stem cells were prepared and identified successfully, of which the main components were the antigen peptides of HSP60, HSP70, HSP90 and HSP110. 0.75 μg and 1.00 μg HSP70⁃antigen peptide and 1. 00 μg HSP90⁃antigen peptide activated lymphocytes significantly. Their A490 values were 0.26±0.03, 0.45±0.05 and 0.32±0.04, respectively, while the corresponding doses of HSP60⁃antigen peptide and HSP110⁃antigen peptide did not activate lymphocytes. The killing rates of 1. 00 μg HSP70⁃antigen peptide and 1.00μg HSP70 were (45.0±2.0)% and (16.0±2.0)%, respectively, showing a significant difference (P=0.012). Similarly, the killing rates of 1.00 μg HSP90⁃antigen peptide and 1.00 μg HSP90 were (36.0±5.0)% and (13.0±4.0)%, respectively, also showing a significant difference (P=0.048 ) . Conclusions The amount of chaperone antigen peptide complexes in gastric cancer cells is extremely low, but it is obviously increased in gastric cancer stem cells. After purification, the chaperone antigen peptide complexes with high purity can be prepared. The extracted chaperone antigen peptide complexes have stronger immunogenicity, and can be used to make tumor vaccine in vitro, which may have a good application value in the targeted therapy of gastric cancer.

摘要： Objective To explore the effect of tumor vaccine combined with CpG ODN on change of regulatory T(Treg) cells in mice with leukemia.Methods Fifty mice were randomly divided into control group,PBS group,tumor vaccine group,CpG 1826 group and tumor vaccine with CpG group(combination group).The animal model of mouse with leukemia was established by subcutaneous injection with erythroleukemia FBL-3 cells.Tumor vaccine was prepared by mitomycin C inactivating FBL-3 cells.The mice in PBS group,tumor vaccine group,CpG 1826 group and combination group were treated with PBS,tumor vaccine,CpG 1826 and tumor vaccine + CpG,respectively.The mice in control group received no treatment.The percentages of Treg cells were detected by flow cytometry and the expression levels of Foxp3 mRNA were detected by reverse transcription polymerase chain reaction(RT-PCR).Serum IL-10 levels were measured by enzyme-linked immunosorbent assay(ELISA).Results Compared with normal control group,the percentages of CD4 + FoxP3 + Treg cells,the mRNA expression of Foxp3 and the serum IL-10 levels in PBS group and tumor vaccine group were statistically increased(P ＜ 0.05),but there was no significant difference between PBS group and tumor vaccine group(P ＞ 0.05).The percentage of CD4 + FoxP3 + Treg cells,the mRNA expression of Foxp3 and the serum IL-10 levels in CpG 1826 group and combination group were statistically lower than those in PBS group and tumor vaccine group (P ＜ 0.05).Conclusion The tumor vaccine combined with CpG ODN may reduce Treg cells in mice with leukemia,and Treg cells may play roles in lukemia and immunotherapy.%目的 探讨红白血病FBL-3细胞瘤苗联合CpG 1826治疗白血病小鼠时调节性T细胞(regulatory T cells,Treg)的变化及其意义. 方法 50只小鼠随机分为对照组、PBS组、瘤苗组、CpG 1826组和瘤苗加CpG 1826组,皮下注射红白血病FBL-3细胞建立小鼠白血病模型,丝裂霉素C灭活FBL-3细胞制备瘤苗.PBS组、瘤苗组、CpG 1826组和瘤苗加CpG 1826组分别利用PBS、瘤苗、CpG 1826和瘤苗加CpG 1826进行治疗,而对照组不进行任何处理.流式细胞术检测小鼠脾细胞中Treg细胞百分率,逆转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)测定Foxp3 mRNA的表达,ELISA法检测血清中IL-10水平. 结果 与对照组小鼠相比,PBS组、瘤苗组小鼠CD4+ FoxP3+ Treg细胞百分率、Foxp3 mRNA的表达水平、血清中IL-10水平明显增高,差异具有统计学意义(P＜0.05),而PBS组与瘤苗组小鼠之间差异无统计学意义(P＞0.05).CpG 1826组、瘤苗联合CpG 1826组小鼠CD4+ FoxP3+ Treg细胞百分率、Foxp3 mRNA的表达水平、血清中IL-10水平比PBS组及瘤苗组明显下降,差异具有统计学意义(P＜0.05). 结论 瘤苗联合CpG 1826治疗小鼠白血病时Treg细胞减少,Treg细胞在白血病及免疫治疗中可能发挥了重要作用.

摘要： 目的 观察胰腺癌细胞PC3-树突状细胞(DC)杂交瘤苗诱导免疫效应细胞对PC3的抑制作用.方法 将DC与胰腺癌PC3细胞融合制备PC3-DC融合细胞(简称DC瘤苗),磁式分选器分选DC瘤苗；酶法检测DC瘤苗诱导免疫效应细胞对PC3的毒性作用；分别用18只裸鼠进行DC瘤苗体内致瘤实验及DC瘤苗诱导免疫效应细胞对PC3移植瘤生长抑制效果实验.结果 PC3-DC杂交瘤苗诱导免疫效应细胞对PC3的杀伤效率最高,其杀伤效率高达76.6％.裸鼠实验发现阴性对照组(Ⅰ)、细胞治疗组(Ⅲ)均于第12天发生移植瘤；预防治疗组(Ⅱ)观察30 d时,6例只有1例发生移植瘤；于移植瘤产生后的第45天处死所有裸鼠并称瘤体的重量,各组比较差异有统计学意义(P＜0.05),组Ⅰ分别与组Ⅱ、Ⅲ比较差异有统计学意义(P＜0.001),组Ⅱ与组Ⅲ比较差异有统计学意义(P＜0.05).结论 DC瘤苗能有效诱导免疫效应细胞抑制恶性肿瘤生长.%Objective To observe the inhibition of immunoeffector cells on the.growth PC3 cells induced by dendritic cell fusion hybrids.Methods DCs were fused with PC3 cells and fusion cells (DC fusion hybrids) were marked with CD11c MicroBeads.In vitro,cytotoxin study,the immunoeffector cells were divided into A1 group (without DC stimulated),A2 group (DC stimulated),A3 group (DC fusion hybrids stimulated).During the in vivo study,18 nude mice were allocated to 3 groups:preventive group (Ⅱ),1-2 days before inoculating PC3,giving DC fusion hybrids activated immunoeffector cells; treated group (Ⅲ),giving DC fusion hybrids activated immunoeffector cells after emergence of implanted tumor in all nude mice; control group (Ⅰ),giving equivalent amount of 1640 cultured liquid.Results In vitro,the maximal killing rate was A3 group (76.6％).In vivo,12 mice developed implanted tumor on the day 12 in group Ⅰ and Ⅲ,separately; In group Ⅱ,on the 30th day,1 of the 6 mice developed implanted tumor.On the 45th day after development of implanted tumor,all the nude mice were.sacrificed and the tumor body weighed.There was significant difference among group Ⅰ,Ⅱ,Ⅲ (P ＜ 0.05).Conclusion DC fusion hybrids may play an important role in anti-tumor therapy.

摘要： 目的 观察Hepal-6 RNA 脉冲BMDCs瘤苗能否激发有效的机体抗肿瘤免疫反应.方法 用Hepal-6 RNA脉冲骨髓分离培养5d的BMDCs,瘤苗的抗肿瘤效应通过C57BL/6J 小鼠肝细胞癌(HCC)模型验证,即C57BL/6J 小鼠皮下接种Hepal-6细胞8d后,成瘤小鼠分为2组:对照组(注射无血清RPMI-1640)和实验组(足垫部注射Hepal-6 RNA脉冲BMDCs瘤苗),30d后处死小鼠并取淋巴结、脾和瘤体冰冻切片,进行CD4、CD8、CD25和Foxp3免疫组织化学染色.结果 免疫组织化学检测显示,淋巴结、脾、肿瘤内有大量CD4+T细胞、CD8+T细胞、CD25+T细胞和Foxp3+Tregs细胞浸润.与对照组小鼠比较,足垫部注射Hepal-6 RNA脉冲BMDCs瘤苗的实验组小鼠淋巴结、脾、瘤内CD4+T细胞和CD8+T细胞浸润显著增加,CD25+T细胞和Foxp3+Tregs浸润明显减少.结论 Hepal-6 RNA 脉冲BMDCs瘤苗能下调小鼠HCC瘤内CD25+T细胞和Foxp3+Tregs浸润,促进CD4+T细胞和CD8+T细胞的增殖和活化,增强D4+T细胞、CD8+T细胞归巢至淋巴结和脾,具有抗瘤免疫效应.

摘要： Objective To explore the efficacy ofβ-GC combined with HBsAg gene-modified DC vaccine on animal model with HCC using different immunization routes. Methods Recombinant adenovirus vector-mediated HBsAg gene-modified DC was used to construct HBsAg-DC which was infused into C57BL/6J mice bearing HBsAg-related HCC by subcutaneous injection. Mice in group A, B and C were vaccinated with PBS, group D, E and F were vaccinated with pAd-HBsAg-DC twice before HepG2 22.1.5 inoculation as described above. Starting at the day of inoculation, mice were treated with daily intra-peritoneal. (ip, 1.5μg per mouse)β-GC injection in group B and E or daily oralβ-GC doses(po, 15μg per mouse) in group C and F. Group A and D receiving vehicle (po) were set as control forβ-GC treatment.The effectiveness of the immune therapy on tumor growth was compared among different groups. Results Tumor size measured 36 days after inoculation was (364.2±3.06)mm3 in group A, (236.5±8.96)mm3 in group B, (251.0±5.76)mm3 in group C, (75.0±5.9)mm3 in group D, (35.3±4.46)mm3 in group E, and (38.5±5.47)mm3 in group F. The application ofβ-GC demonstrated anti-tumor activity by itself (group B and C), and also enhanced the tumor preventive effect of pAd-HBsAg-DC (group E and F). No difference in tumor growth was observed between ip and po application ofβ-GC. Conclusion β-GC may serve as an immune enhancer to augment the anti-tumor immunity triggered by HBsAg gene-modified DC vaccine. It's antitumor effect may be related to activation of NKT cells.%目的探讨β-葡萄糖神经酰胺（β-GC）联合乙型肝炎表面抗原（HBsAg）基因修饰树突状细胞（dendritic cell, DC）瘤苗治疗肝癌的作用。方法以重组腺病毒为载体构建HBsAg-DC瘤苗。将C57BL/6J小鼠随机分为6组（A-F，6只/组），其中A、B、C组接种PBS液2次，D、E、F组接种HBsAg-DC瘤苗2次。皮下注入HepG222.1.5肝癌细胞当天始， B、E组接受β-GC（1.5μg）腹腔注射，C、F组接受β-GC（15μg）灌胃给药，A、D组为安慰剂对照，比较不同组间移植瘤生长情况。结果 B、C组移植瘤生长较之A组明显受抑（P＜0.05），E、F组移植瘤生长较之D组明显受抑（P＜0.05）。A-F组移植瘤大小（mm3）分别为364.2±3.06，236.5±8.96，251.0±5.76，75.0±5.9，35.3±4.46，38.5±5.47。经腹腔注射给药与灌胃给药相比在抗瘤作用方面差异无统计学意义。结论β-GC经腹腔或灌胃给药均具有提高HBsAg-DC瘤苗的免疫治疗作用，其协同抗肿瘤效应的机制可能通过激活NKT细胞。

摘要： The study was to enrich MCF 7 breast cancer stem cells by mammosphere culture using serum free mediumand prepare mRNA nucleic acid antigen for preparation of dendritic cell vaccine targeting on breast cancer stem cell. MCF 7 breast cancer stem cell was cultured and enriched by using serum free medium, and confirmed by identification of cellular sur face marks by using flow cytometer, clone forming capability test by agar assay, and tumorigenic ability test in vivo by using NOD SCID mouse. The mRNA was amplified by using T7 mMESSAGE mMACHINE kit with template of total RNA extracted from MCF 7 breast cancer stem cells. The result showed that MCF 7 breast cancer cell grouped and formed mammosphere in cultural suspension by using serum free media. About 90. 16% cells within mammosphere possessed CD+44/CD-24 phenotype. The ability of clone forming and capability of tumor initiating in NOD/SCID mouse with low dose were stronger in suspension cells than that in adhesive cells. The mRNA molecule amplified in vitro could be as specific nucleic acid antigen of breast cancer stem cells. We proposed that the high percent of MCF 7 breast cancer stem cells could be obtained by mammosphere culture u sing serum free medium, and mRNA amplified in vitro could be as specific nucleic acid antigen of breast cancer stem cells. It lay a foundation for preparation of dendritic cell vaccine targeting on breast cancer stem cell.%目的 应用无血清培养基细胞球培养法富集MCF-7乳腺癌千细胞,制备乳腺癌干细胞mRNA核酸抗原,为制备靶向乳腺癌干细胞树突细胞瘤苗奠定基础.方法 应用无血清培养基悬浮培养法富集MCF-7乳腺癌细胞,经单克隆形成、表面标志检测、NOD-SCID小鼠成瘤等实验对其肿瘤干细胞特性进行鉴定后,利用T7 mMESSAGE mMACHINE试剂盒进行mRNA体外扩增.结果 乳腺癌细胞MCF-7在无血清培养基中可形成悬浮状细胞球,其中具有CD44+CD24-表面标志的细胞约占90.16%.该细胞亚群具有较贴壁细胞更强的克隆形成能力和低剂量体内成瘤能力.细胞总RNA经体外扩增获得的mRNA,可作为核酸抗原.结论应用无血清悬浮细胞球培养法可以获得高含量乳腺癌干细胞.通过体外扩增方法获得该细胞亚群的mRNA,可作为肿瘤核酸抗原,为下一步制备靶向乳腺癌干细胞的树突细胞瘤苗奠定了基础.

摘要： 目的制备膜表达聚苯丙赖氨酸蛋白瘤苗并研究其抗肿瘤治疗作用.方法人工合成表达聚苯丙赖氨酸的DNA片断,扩增成带酶切位点的目的基因.选择载体pDisplay,将目的基因插入载体多克隆酶位点BglⅡ、PstⅠ之间.DNA测序证实后,以脂质体转染的方法转染到鼠CT26细胞.采用G418抗性筛选并扩增阳性克隆,化疗药处理后采用化学偶连两步法使跨膜型多聚苯丙赖氨酸与肠癌细胞膜共价偶连,构建成新型瘤苗,观察瘤苗体外免疫效应.结果酶切电泳和DNA测序证实载体构建正确,目的基因的上游带信号肽序列,下游带跨膜序列.RT-PCR产物电泳、流式细胞术、激光共聚焦显微镜证实聚苯丙赖氨酸表达在转染的鼠CT26细胞膜上,体外免疫学实验显示瘤苗细胞体外对淋巴细胞增殖刺激效应显示有较强的淋巴增殖效应,产生的多克隆抗体对CT26肠癌细胞有特异性杀伤作用.结论聚苯丙赖氨酸跨膜修饰的瘤苗能够产生特异、较强的免疫效应.

摘要： @@ 随着现代医学发展和分子生物学技术的提高,人们逐渐从分子、基因水平对肿瘤的发病机制有了,肿瘤的靶向治疗技术应运而生,成为近年肿瘤治疗领域研究的热点.肿瘤靶向治疗理论机制不断完善,多种治疗手段应用于肿瘤的治疗,并在临床实践中取得了的成就.就肿瘤靶向治疗技术的现状简明分类并阐述如下.

摘要： 随着肿瘤基因治疗研究的进展,肿瘤基因治疗的安全问题日益成为人们关心的焦点问题.本文对以逆转录病毒和腺病毒为载体的转基因细胞的DNA及培养上清液进行了体内和体外的致突变性研究,目的是为转基因肿瘤细胞作为瘤苗进行临床应用的安全性检测提供依据.

摘要： 本发明公开了苗药廖氏化风丹在制备防治黑色素瘤药物中的应用，通过在体外运用黑色素瘤细胞WM35、WM239检测发现，廖氏化风丹具有显著的抑制黑色素瘤作用，并通过流式细胞检测其在细胞周期方面的作用，结果表明廖氏化风丹对黑色素瘤细胞周期影响显著，并且药物浓度越高，其阻滞越明显，可作为防治黑色素瘤制剂，制备成药剂学上允许的口服液、注射剂、片剂、胶囊剂、滴丸剂等。

摘要： 本发明公开了一种治疗脑纤维瘤的苗药组方，本发明所述药物由艾叶、嘎炯豆丢劳独、见血清制备而成。本发明具有祛风除湿，温经散寒的功效，适用于患有脑纤维瘤的人群，制成的药物疗效显著，无毒副作用。

摘要： 本发明公开了苗药廖氏化风丹在制备防治黑色素瘤药物中的应用，通过在体外运用黑色素瘤细胞WM35、WM239检测发现，廖氏化风丹具有显著的抑制黑色素瘤作用，并通过流式细胞检测其在细胞周期方面的作用，结果表明廖氏化风丹对黑色素瘤细胞周期影响显著，并且药物浓度越高，其阻滞越明显，可作为防治黑色素瘤制剂，制备成药剂学上允许的口服液、注射剂、片剂、胶囊剂、滴丸剂等。

摘要： 本发明涉及一种DC瘤苗制备方法及其在肿瘤治疗中的应用。本发明针对肝癌细胞特异性的制备得到了通过白藜芦醇提高了DC成熟度的DC瘤苗，所述瘤苗除了本身具有较好的抑制肝癌细胞增殖的效果之外，通过与PD‑1单克隆抗体的组合使用，能够显著的抑制肿瘤细胞的生长以及提高相关细胞因子的分泌，具有较好的应用价值。

摘要： 本发明涉及DC瘤苗联合单克隆抗体在制备用于癌症治疗的药物组合物中的用途。本发明特异性的制备针对胃癌的DC瘤苗，该瘤苗具有较好的刺激免疫反应，与CLDN18.2单克隆抗体联合应用或者单独使用均可以有效的行使杀伤肿瘤细胞的功能，具有较好的应用价值。

摘要： 本发明涉及一种DC瘤苗及其制备的药物组合物。本发明特异性的制备针对胃癌的DC瘤苗，该瘤苗具有较好的刺激免疫反应，与CLDN18.2单克隆抗体联合应用或者单独使用均可以有效的行使杀伤肿瘤细胞的功能，具有较好的应用价值。

摘要： 本发明涉及一种DC瘤苗制备方法及其在肿瘤治疗中的应用。本发明针对肝癌细胞特异性的制备得到了通过白藜芦醇提高了DC成熟度的DC瘤苗，所述瘤苗除了本身具有较好的抑制肝癌细胞增殖的效果之外，通过与PD‑1单克隆抗体的组合使用，能够显著的抑制肿瘤细胞的生长以及提高相关细胞因子的分泌，具有较好的应用价值。

摘要： 本发明涉及含有DC瘤苗的药物组合物及其在治疗癌症中的应用。本发明针对肝癌细胞特异性的制备得到了通过白藜芦醇提高了DC成熟度的DC瘤苗，所述瘤苗除了本身具有较好的抑制肝癌细胞增殖的效果之外，通过与PD‑1单克隆抗体以及顺铂的组合使用，能够显著的抑制肿瘤细胞的生长以及提高相关细胞因子的分泌，具有较好的应用价值。

摘要： 本发明公开了一种非小细胞肺癌全细胞抗原树突状细胞瘤苗的制备方法，具体步骤为：全细胞肿瘤抗原制备：得到细胞悬液；将细胞悬液置于恒温水浴锅中；水浴锅中取出细胞悬液，放到‑75‑‑85°C超低温冰箱中，1h之后，转入37°C水浴锅中，轻轻摇晃，至完全融化，再次放入超低温冰箱中，如此反复3次；在12000rpm的条件下，离心10min；得到热休克肿瘤细胞冻融抗原；DC细胞制备；DC瘤苗制备。本发明具有免疫效果好、无副作用等优点。

摘要： 本发明公开了一种制备自噬小体型瘤苗的培养基添加剂，冻干粉制剂包含一组组合物；组合物包含蛋白酶体抑制剂、自噬诱导剂、溶酶体腔碱化剂、赋形剂和抗氧化剂；本发明具有如下突出的优点：本发明所述的添加剂组合物中含有蛋白酶体抑制剂、溶酶体腔碱化剂、自噬诱导剂，三者协同，一方面，可同时抑制肿瘤细胞的蛋白酶体降解途径和自噬溶酶体降解途径，使肿瘤抗原得以保存；另一方面，诱导自噬小体形成，使大量肿瘤抗原保存在自噬小体中，通过提取这些自噬小体，即可制备自噬小体型亚细胞肿瘤疫苗。