大鼠胰腺癌微环境诱导未成熟树突状细胞功能的变化

Objective To study the effect of pancreatic cancer microenvironment on maturation of dendritic cells ( DCs), and to investigate the mechanism by which tumors escape from immune recognition. Methods DCs were cultured with rmGM (40μg/L) ,rmIL-4 (40μg/L) ,and the supernatant of cell line AR42J cells was added on the day 6, then a large number of DCs was gained. DCs cultured and in-duced by routine methods were used as controls. The expression of surface molecules CD86, and CD80 in DCs was detected to observe whether this intervention could delay or block maturation of DCs and this effect could be reversed by stimulation with lipopolysaccharide (LPS). T cell stimulatory effects were also determined. Results Compared to normal DCs, CD86 and CD80 expression positive rate was significantly decreased in DCs induced by AR42J supernatant from (70.88±3.60)% to (7.15±0.71 )%. After stimulation with LPS, the expression of CD86, and CD80 in DCs was still low, suggesting the AR42J super-natant could block the maturation of DCs. The allostimulatory capacity of the immature DCs stimulated with the AR42J superoatant was significantly lower than that of the normal immature DCs. Conclusion The supernatant from pancreatic cancer ceils in rats could inhibit the maturation of DCs, which was not easily reversed.     

大鼠胰腺癌微环境诱导未成熟树突状细胞功能的变化

Objective To study the effect of pancreatic cancer microenvironment on maturation of dendritic cells ( DCs), and to investigate the mechanism by which tumors escape from immune recognition. Methods DCs were cultured with rmGM (40μg/L) ,rmIL-4 (40μg/L) ,and the supernatant of cell line AR42J cells was added on the day 6, then a large number of DCs was gained. DCs cultured and in-duced by routine methods were used as controls. The expression of surface molecules CD86, and CD80 in DCs was detected to observe whether this intervention could delay or block maturation of DCs and this effect could be reversed by stimulation with lipopolysaccharide (LPS). T cell stimulatory effects were also determined. Results Compared to normal DCs, CD86 and CD80 expression positive rate was significantly decreased in DCs induced by AR42J supernatant from (70.88±3.60)% to (7.15±0.71 )%. After stimulation with LPS, the expression of CD86, and CD80 in DCs was still low, suggesting the AR42J super-natant could block the maturation of DCs. The allostimulatory capacity of the immature DCs stimulated with the AR42J superoatant was significantly lower than that of the normal immature DCs. Conclusion The supernatant from pancreatic cancer ceils in rats could inhibit the maturation of DCs, which was not easily reversed.     

大鼠胰腺癌微环境诱导未成熟树突状细胞功能的变化

Objective To study the effect of pancreatic cancer microenvironment on maturation of dendritic cells ( DCs), and to investigate the mechanism by which tumors escape from immune recognition. Methods DCs were cultured with rmGM (40μg/L) ,rmIL-4 (40μg/L) ,and the supernatant of cell line AR42J cells was added on the day 6, then a large number of DCs was gained. DCs cultured and in-duced by routine methods were used as controls. The expression of surface molecules CD86, and CD80 in DCs was detected to observe whether this intervention could delay or block maturation of DCs and this effect could be reversed by stimulation with lipopolysaccharide (LPS). T cell stimulatory effects were also determined. Results Compared to normal DCs, CD86 and CD80 expression positive rate was significantly decreased in DCs induced by AR42J supernatant from (70.88±3.60)% to (7.15±0.71 )%. After stimulation with LPS, the expression of CD86, and CD80 in DCs was still low, suggesting the AR42J super-natant could block the maturation of DCs. The allostimulatory capacity of the immature DCs stimulated with the AR42J superoatant was significantly lower than that of the normal immature DCs. Conclusion The supernatant from pancreatic cancer ceils in rats could inhibit the maturation of DCs, which was not easily reversed.     

大鼠胰腺癌微环境诱导未成熟树突状细胞功能的变化

Objective To study the effect of pancreatic cancer microenvironment on maturation of dendritic cells ( DCs), and to investigate the mechanism by which tumors escape from immune recognition. Methods DCs were cultured with rmGM (40μg/L) ,rmIL-4 (40μg/L) ,and the supernatant of cell line AR42J cells was added on the day 6, then a large number of DCs was gained. DCs cultured and in-duced by routine methods were used as controls. The expression of surface molecules CD86, and CD80 in DCs was detected to observe whether this intervention could delay or block maturation of DCs and this effect could be reversed by stimulation with lipopolysaccharide (LPS). T cell stimulatory effects were also determined. Results Compared to normal DCs, CD86 and CD80 expression positive rate was significantly decreased in DCs induced by AR42J supernatant from (70.88±3.60)% to (7.15±0.71 )%. After stimulation with LPS, the expression of CD86, and CD80 in DCs was still low, suggesting the AR42J super-natant could block the maturation of DCs. The allostimulatory capacity of the immature DCs stimulated with the AR42J superoatant was significantly lower than that of the normal immature DCs. Conclusion The supernatant from pancreatic cancer ceils in rats could inhibit the maturation of DCs, which was not easily reversed.     

1-甲基色氨酸对胰腺癌荷瘤鼠中调节性T细胞数量变化的影响

目的 观察1-甲基色氨酸(1-MT)对胰腺癌荷瘤鼠中调节性T细胞(Treg)数量变化的影响,比较树突状细胞(DC)疫苗与1-MT联合应用前后抗肿瘤作用的强弱.方法 建立小鼠胰腺癌模型;利用流式细胞术检测荷瘤鼠应用1-MT前后肿瘤组织周围引流淋巴结(TDLNs)及脾脏中CD4~+ CD25~+T细胞占CD4~+T比例;荧光定量聚合酶链反应(PCR)测量Foxp3在TDLNs及脾脏mRNA水平;利用肿瘤细胞裂解物冲击DC制备DC疫苗,并根据是否与1-MT联合应用分组(各组均为n=8);观测各组肿瘤体积的差异.结果 应用1-MT后,荷瘤鼠CD4~+ CD25~+ T细胞占CD~+T细胞的比例明显低于未应用组(TDLNs)分别为(16.01±2.21)%和(25.00±2.16)%(P<0.05);脾脏分别为(13.11±1.93)%和(22.14±2.33)%(P<0.05,P<0.01);应用1-MT组Foxp3 mRNA表达水平显著低于未应用组,应用1-MT组相对表达值:TDLNs0.947±0.216、脾细胞1.198±0.347,而未应用组分别为:1.927±0.256、1.798±0.237(P<0.05);1-MT+DC疫苗组肿瘤生长显著受到抑制,第36天肿瘤体积为(789.0±111.0)mm~3;显著小于DC疫苗组、1-MT组及对照组,肿瘤体积分别为:(1768.0±251.3)、(1854.0±192.1)、(1899.0±201.2)mm~3(P<0.01).结论 1-MT可以有效抑制胰腺癌荷瘤鼠癌组织周围引流淋巴结及脾脏CD4~+ CD25~+ Treg细胞的数量增加,从而增强DC疫苗抗肿瘤作用.     

医学院校临床带教人员教师资格认定问题探析

建立教师质量保障体系,完善教师资格制度,全面提高教师素质是保证教育质量的必由之路?本文分析了我国现阶段的教师资格制度,指出我国的教师资格制度未能充分考虑普通高等医学院校学科及专业特点,使医学院校部分临床带教人员教师资格认定困难,结合医学院校临床教学基地建设,就医学院校临床带教人员在申请高校教师资格认定过程中面临的问题进行了分析并提出相应对策?     

门脉高压患者血浆刺激血管内皮细胞eNOS与Hsp90相互作用的实验研究

目的：用门脉高压患者血浆刺激血管内皮细胞研究内皮型一氧化氮合酶（eNOS）与热休克蛋白90（Hsp90）的相互作用。方法：免疫共沉淀技术是研究蛋白质相互作用的一个有力工具。用肝硬化门脉高压患者血浆处理培养的人脐静脉血管内皮细胞（human umbilical vascular endothelial cell，HUVEC）后，在0、15、30、60和120min时收集细胞，利用该技术检测对Hsp90和eNOS相互作用的影响。细胞裂解液分别用抗eNOS抗体和抗Hsp90抗体沉淀蛋白复合体．经SDS～PAGE电泳和转膜后，再分别用抗Hsp90抗体和抗eNOS抗体进行免疫识别。结果：本实验分别检测出了eNOS和Hsp90的表达,说明Hsp90和eNOS存在相互作用的关系。Hsp90和eNOS对刺激产生的反应有时相性，一般在刺激30min后Hsp90的量有明显增加，进一步说明Hsp90对eNOS调节NO的产生起关键的作用。结论：门脉高压患者血浆可以刺激血管内皮细胞Hsp90与eNOS结合，增强eNOS的生物活性，使NO释放增多。     

不同浓度γ干扰素诱导大鼠脾脏来源树突状细胞表达IDO的变化及对T淋巴细胞增殖的影响

背景：DC可通过产生吲哚胺-2 ,3-双加氧酶（IDO）,抑制T淋巴细胞增殖诱导免疫耐受。因此,上调DC细胞的IDO表达可能成为诱导移植后免疫耐受的新策略。本研究的目的在于探讨不同浓度γ干扰素（IFN-γ）作用下大鼠脾脏来源的树突状细胞（DC）中吲哚胺2,3-双加氧酶（IDO）mRNA和蛋白表达的变化及IDO对同种异体T淋巴细胞增殖的影响。方法：应用细胞因子体外诱导培养大鼠脾脏来源DC,应用流式细胞仪测定大鼠DC特异性分子OX62和表面分子CD80、CD86的表达。分别用不同浓度（0、100、300、500U/ml）的IFN-γ诱导作用DC后,Real-time PCR测定DC中IDO mRNA的相对表达水平,Western Blot检测DC中IDO蛋白在DC中的表达水平。同种异体混合淋巴细胞反应（MLR）检测不同浓度IFN-γ诱导后的DC对同种异体T淋巴细胞增殖的影响。结果：体外诱导培养的DC光学显微镜下观察,具有典型的树枝状突起。OX62表达率达到80%以上,CD80、CD86的阳性表达也在80%左右;DC的IDO mRNA和蛋白的相对表达量随着IFN-γ作用浓度的增大逐渐增大,不同浓度组间差异有统计学意义（P<0.05）;各组IFN-γ作用后DC与对照组比较,T淋巴细胞的增殖率显著降低（P<0.05）;且随着IFN-γ作用浓度的增大T淋巴细胞的增殖率逐渐降低,各组间差异有统计学意义（P<0.05）。结论：采用改良的培养黏附法体外成功地获得了纯度较高的大鼠脾脏来源的DC;IFN-γ可以诱导大鼠脾脏来源DC表面活性IDO的表达增加,减弱脾脏DC对同种T细胞增殖的刺激能力。     

癌相关成纤维细胞在乳腺癌转移及耐药作用的研究进展

目的 总结癌相关成纤维细胞（cancer-associated fibroblasts,CAFs）在乳腺癌侵袭转移及耐药方面的最新研究进展及相关机制,以寻求乳腺癌转移及耐药患者的最佳治疗策略。方法 检索近年来有关乳腺癌中CAFs研究的文献并进行综述。结果 CAF是肿瘤微环境（tumor microenvironment,TME）中的主要基质细胞,通过改变TME从而改变CAFs的生物学特性,诱导乳腺癌细胞的生长和侵袭;乳腺癌中的CAFs通过与炎性因子的相互作用,以及促进转移前生态位的形成来促进乳腺癌细胞的侵袭及转移,并且CAFs通过分泌各种细胞因子介导乳腺癌的化疗耐药、靶向耐药、内分泌耐药以及放射抗性。结论 目前,在乳腺癌中CAFs的相关研究已取得一些进展,但离临床应用还有一定差距。CAFs具有多种功能表型,因此在研究新的抗CAFs治疗策略时,需确定和表征特定的CAFs亚型。且已有研究证明将CAFs作为乳腺癌治疗的特异性靶标具有巨大潜力,但CAFs仍缺乏特异性的生物学标志物。因此深入了解CAFs的生物学特性及其异质性,可为开发靶向CAFs的药物提供可靠的理论依据。     

大鼠胰腺癌微环境诱导未成熟树突状细胞功能的变化

Objective To study the effect of pancreatic cancer microenvironment on maturation of dendritic cells ( DCs), and to investigate the mechanism by which tumors escape from immune recognition. Methods DCs were cultured with rmGM (40μg/L) ,rmIL-4 (40μg/L) ,and the supernatant of cell line AR42J cells was added on the day 6, then a large number of DCs was gained. DCs cultured and in-duced by routine methods were used as controls. The expression of surface molecules CD86, and CD80 in DCs was detected to observe whether this intervention could delay or block maturation of DCs and this effect could be reversed by stimulation with lipopolysaccharide (LPS). T cell stimulatory effects were also determined. Results Compared to normal DCs, CD86 and CD80 expression positive rate was significantly decreased in DCs induced by AR42J supernatant from (70.88±3.60)% to (7.15±0.71 )%. After stimulation with LPS, the expression of CD86, and CD80 in DCs was still low, suggesting the AR42J super-natant could block the maturation of DCs. The allostimulatory capacity of the immature DCs stimulated with the AR42J superoatant was significantly lower than that of the normal immature DCs. Conclusion The supernatant from pancreatic cancer ceils in rats could inhibit the maturation of DCs, which was not easily reversed.