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国际免疫学杂志
国际免疫学杂志

李殿俊

双月刊

1673-4394

guojimianyi@126.com

0451-86669596

150081

黑龙江省哈尔滨市南岗区保健路157号

国际免疫学杂志/Journal International Journal of Immunology北大核心CSTPCD
查看更多>>1978年创刊，中华人民共和国卫生部主管，中华医学会、哈尔滨医科大学主办。本刊原名《国外医学》免疫学分册，公开发行的国家级学术期刊。期刊适合于从事免疫学以及相关学科、交叉学科的科研、教学和广大的临床医务工作者参考和交流。
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    丹皮酚调控ITGB3/p38MAPK抑制乳腺癌细胞增殖侵袭与上皮-间质转化

    李睿慧董航王辰囡邵长利...
    1-5页
    查看更多>>摘要：目的 探讨丹皮酚(paeonol，Pae)对三阴乳腺癌细胞系MDA-MB-231细胞增殖、侵袭和上皮-间质转化(epithelial mesenchymal transition，EMT)的影响及可能机制。 方法 不同浓度Pae处理MDA-MB-231细胞后，设立对照样本。采用CCK8方法检测细胞增殖活性；Transwell法检测细胞的侵袭能力；流式细胞技术检测细胞周期；Western blot检测整合素β3 (integrin β3，ITGB3)、p38丝裂原活化蛋白激酶(mitogen-activated protein kinases，p38MAPK)、磷酸化-p38MAPK(phosphorylated-p38MAPK，p-p38MAPK)蛋白的表达以及EMT相关标志物纤维连接蛋白(fibronectin，FN)、波形蛋白(vimentin，Vim)、N-钙粘蛋白(N-cadherin，N-cad)的表达变化。 结果 60 mg/L的Pae对于乳腺癌细胞产生了明显的抑制作用，故本实验使用60 mg/L的Pae浓度。Pae分别处理24、48 h后，G0/G1的细胞周期分别从对照组的50.6%增加到53.0%和65.4%。S期处理48 h后，Pae组肿瘤细胞百分比从37.6%降低至26.6%。Transwell结果显示，与对照组相比，Pae处理乳腺癌细胞24、48 h后细胞侵袭数明显减少，差异有统计学意义[个：(378.73±47.62)比(193.64±32.54)或(174.37±27.83)，t＝5.35、5.75，P值均<0.05]。与对照组相比，Pae处理后乳腺癌细胞ITGB3、p38MAPK、p-p38MAPK表达水平明显降低；EMT标记物FN、N-cad、Vim表达均显著下降，差异有统计学意义[(0.97±0.19)比(0.48±0.10)，(1.24±0.24)比(0.56±0.18)，(0.93±0.18)比(0.51±0.13)，t＝4.08、3.93、3.28，P值均<0.05；(0.88±0.16)比(0.46±0.08)，(0.96±0.17)比(0.43±0.12)，(0.87±0.15)比(0.37±0.13)，t＝4.06、4.42、4.36，P值均<0.05]。 结论 Pae显著抑制三阴乳腺癌细胞系MDA-MB-231细胞增殖，侵袭和EMT，该作用可能与ITGB3/p38MAPK信号通路激活有关。 Objective To explore the effect of paeonol(Pae) on the expansion, invasion and epithelial-mesenchymal transition (EMT) of triple negative breast cancer cell line MDA-MB-231 and its possible mechanism. Methods After MDA-MB-231 cells were treated with Pae at different concentrations, control samples were set up. The proliferation activity was detected by CCK8 method. Transwell method was used to detect the invasiveness of cells. Flow cytometry was used to analyze the cell cycle. Western blot was used to detect the expression of integrin β3 (ITGB3), p38 mitogen-activated protein kinases (p38MAPK), phosphorylated p38MAPK(p-p38MAPK) protein, and the expression changes of EMT related markers fibronectin (FN), vimentin (Vim), and N-cadherin (N-cad). Results The 60 mg/L Pae has obvious inhibitory effect on breast cancer cells, so 60 mg/L Pae concentration was used in this experiment. After 24 h and 48 h of treatment with Pae, the cell cycle of G0/G1 increased from 50.6% in the control group to 53.0% and 65.4% respectively. The percentage of tumor cells in the Pae group decreased from 37.6% to 26.6% after 48 h treatment in the S phase. Transwell results showed that compared with the control group, the number of cell invasion of breast cancer cells after Pae treatment for 24 h and 48 h was significantly reduced [pcs: (378.73±47.62) vs (193.64±32.54) or (174.37±27.83), t=5.35, 5.75, both P values <0.05]. Compared with the control group, the expression levels of ITGB3, p38MAPK, p-p38MAPK in breast cancer cells after Pae treatment were significantly reduced, and the expression levels of EMT markers FN, N-cad, Vim were significantly reduced[(0.97±0.19) vs(0.48±0.10), (1.24±0.24) vs(0.56±0.18), (0.93±0.18) vs(0. 51±0.13), t=4.08, 3.93, 3.28, all P values<0.05 (0.88±0.16) vs(0.46±0.08), (0.96±0.17) vs(0.43±0.12), (0.87±0.15) vs(0.37±0.13),t=4.06, 4.42, 4.36, all P values<0.05]. Conclusion Pae can significantly inhibit the proliferation, invasion and EMT in breast cancer cells, which may be related to ITGB3/p38MAPK signal pathway activation.

    丹皮酚乳腺癌上皮-间质转化ITGB3/p38MAPK

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    2型糖尿病与HLA-E基因相关性研究

    何柳媚孙丽艳贾黎静徐筠娉...
    6-11页
    查看更多>>摘要：目的 探讨人类白细胞抗原(human leukocyte antigen，HLA)-E基因与2型糖尿病(type 2 diabetes mellitus，T2DM)的相关性。 方法 选择深圳市人民医院确诊的T2DM患者254例作为病例组，同期选取深圳市血液中心参加无偿献血的健康献血者272例作为对照组。采用聚合酶链式反应-直接测序分型法(polymerase chain reaction-sequence based typing，PCR-SBT)对两组人群进行HLA-E基因第3外显子和启动子区进行序列测定，比较分析病例组与对照组的基因频率。 结果 HLA-E*01：01与HLA-E*01：03的差异主要体现在第3外显子756碱基处，当756位碱基只有G峰时，为HLA-E*01：03纯合子，当756位碱基只有A峰时，为HLA-E*01：01纯合子，当756位碱基是G＋A峰时，为HLA-E*01：01/E*01：03杂合子。HLA-E*01：03基因频率比较结果显示，病例组中的HLA-E*01：03阳性基因频率显著高于健康对照组，差异有统计学意义[例(%)：189(87.10)比209(76.84)，χ2＝8.39，P<0.05]。当NT-26位只有G时，为HLA-E*01：01：01：01纯合子，当NT-26位碱基只有T峰时，为HLA-E*01：01：01：06纯合子，当NT-26位碱基是G＋T峰时，为HLA-E*01：01：01：01/E*01：01：01：06杂合子。NT-26T基因频率比较结果显示，病例组与对照组NT-26T基因频率的组间差异无统计学意义(P>0.05)。 结论 HLA-E*01：03基因是T2DM的易感基因，其可能成为预测、诊断或治疗T2DM的一项指标。 Objective To explore the correlation between human leukocyte antigen (HLA)-E gene and type 2 diabetes mellitus (T2DM). Methods A total of 254 T2DM patients from Shenzhen People’s Hospital were selected as the patient group, while 272 healthy blood donors participating in the blood donation from Shenzhen Blood Center were selected as the normal control group. Polymerase chain reaction-sequence based typing (PCR-SBT) was used to sequence the third exon and promoter region of HLA-E gene in both group, and the gene frequencies of the two groups were compared and analyzed. Results The difference between HLA-E*01: 01 and HLA-E*01: 03 is mainly reflected at the 756 base of exon 3. When the 756 base has only a G peak, it is a homozygous HLA-E*01: 03, the 756 base has only an A peak, it is a homozygous HLA-E*01: 01, and the 756 base has a G+ A peak, it is a heterozygous HLA-E E*01: 01/E*01: 03. The comparison of HLA-E*01: 03 gene frequency showed that the frequency of HLA-E*01: 03 positive genotype in the patient group was significantly higher than that in the healthy control group[case(%): 189 (87.10) vs 209 (76.84), χ2=8.39, P<0.05]. When the NT-26 base has only G, it is HLA-E*01: 01: 01: 01 homozygous, the NT-26 base has only T peak, it is HLA-E*01: 01: 01: 06 homozygous, and the NT-26 base has G+ T peak, it is HLA-E*01: 01: 01: 01/E*01: 01: 01: 06 heterozygous. The comparison of NT-26T gene frequency showed no statistically significant difference between the patient group and the healthy control group (P>0.05). Conclusions HLA-E*01: 03 gene is a susceptibility gene for T2DM, and it may be a marker for the prediction, diagnosis or treatment of T2DM.

    2型糖尿病人类白细胞抗原-E基因

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    肠道菌群紊乱对小鼠SLE发生发展的影响及机制

    李亚彤赵珈华桂金秋刘洋...
    12-18页
    查看更多>>摘要：目的 探讨肠道菌群紊乱对小鼠系统性红斑狼疮(systemic lupus erythematosus，SLE)发生发展的影响及机制。 方法 40只雌性美国癌症研究所(Institute of Cancer Researc，ICR)小鼠随机分为正常对照组(normal control，NC)、菌群紊乱模型组(disorder flora model，DFM)、SLE模型对照组(SLE model control group，SM)和菌群紊乱SLE组(disorder flora SLE model，DS)，每组10只。实验结束后，进行小鼠器官指数检测；HE染色观察肾脏病理改变；粪便活菌选择性培养计数分析小鼠肠道菌群的变化；酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)检测血清中抗核抗体和抗组蛋白抗体；实时荧光定量PCR(real-time quantitative PCR，RT-qPCR)检测脾脏组织中白细胞介素(interleukin，IL)-2 mRNA的表达水平。 结果 成功建立肠道菌群紊乱小鼠模型，与DS组相比，NC组、DFM组、SM组大肠杆菌、粪肠球菌数量明显降低，双歧杆菌和乳酸杆菌数量明显升高，差异均具有统计学意义(F＝289.60、143.90、504.40、1687.00，P值均<0.05)；与DFM组/SM组相比，DS组大肠杆菌和粪肠球菌数量明显增加，双歧杆菌和乳酸杆菌数量明显减少，差异均具有统计学意义[logCFU/g：(6.94±0.01)或(6.87±0.01)比(6.96±0.01)，(5.85±0.01)或(5.77±0.01)比(5.89±0.01)，(8.91±0.01)或(8.98±0.01)比(8.73±0.02)，(8.77±0.01)或(8.92±0.00)比(8.60±0.01)，P值均<0.05]。与DS组比较，NC组、DFM组、SM组小鼠体重显著升高，脾指数、肾指数和胸腺指数显著降低，差异均有统计学意义(F＝92.02、26.99、29.43、17.06，P值均<0.05)。与NC组相比，DFM组、SM组和DS组小鼠的抗核抗体、抗双链DNA抗体水平显著增加，差异具有统计学意义(F＝332.10、1151.00，P值均<0.05)；与DFM组相比，SM组和DS组小鼠的抗双链DNA均显著增加，差异有统计学意义[pg/mL：(1.86±0.23)比(6.49±0.56)或(8.58±0.42)；ng/ mL: (7.76±0.70)比(17.66±0.14)或(18.51±0.10)，P值均<0.05]。HE染色结果显示，与NC组和DFM组相比，SM和DS组小鼠肾脏可见肾小球内细胞数量明显增多，肾间质及血管周围可见炎性细胞浸润。与NC组比较，DFM组、SM组和DS组小鼠脾脏组织IL-2 mRNA表达均显著下降，差异有统计学意义[(2.05±0.14)比(1.69±0.20)比(1.52±0.11)比(1.01±0.13)，F＝16.83，P<0.05 ]。 结论 肠道菌群紊乱能够影响相关细胞因子表达，促进SLE的炎症反应，进而加重SLE的发生发展。 Objective To investigate the effect and mechanism of intestinal flora disturbance on the development of systemic lupus erythematosus (SLE) in mice. Methods Forty female Institute of Cancer Research (ICR) mice were randomly divided into normal control group (NC), disorder flora model group (DFM), SLE model control group (SM), and disorder flora SLE model(DS), with 10 animals in each group. After the experiment, the mouse organ index was detected HE staining was used to observe renal pathological changes. Selective culture counting of fecal viable bacteria was used to analyze the changes of intestinal flora in mice. Enzyme-linked immunosorbent assay (ELISA) was used to detect antinuclear and antihistone antibodies in serum, and real-time quantitative PCR (RT-qPCR) was used to detect the expression level of interleukin (IL) -2 mRNA in spleen tissue. Results A mouse model of intestinal microbiota disorder was successfully established. Compared with the DS group, the NC group, DFM group, and SM group showed a significant decrease in the number of Escherichia coli and Enterococcus faecalis, while the number of Bifidobacterium and Lactobacillus increased significantly (F=289.60, 143.90, 504.40, 1687.00, all P values<0.05). Compared with the DFM group/SM group, the DS group showed a significant increase in the number of Escherichia coli andEnterococcus faecalis, while the number of Bifidobacterium and Lactobacillus decreased significantly[logCFU/g: (6.94±0.01) or (6.87±0.01) vs(6.96±0.01), (5.85±0.01) or (5.77±0.01) vs(5.89±0.01), (8.91±0.01) or (8.98±0.01) vs(8.73±0.02), (8.77±0.01) or (8.92±0.00) vs(8.60±0.01), all P values<0.05]. Compared with the DS group, the NC group, DFM group, and SM group showed a significant increase in body weight, while the spleen index, kidney index, and thymus index decreased significantly (F=92.02, 26.99, 29.43, 17.06, all P values<0.05). Compared with the NC group, the levels of anti-nuclear antibodies and anti-double-stranded DNA antibodies in the DFM, SM, and DS groups were significantly increased(F=332.10, 1151.00, both P values<0.05). Compared with the DFM group, the anti-double-stranded DNA levels in both SM and DS groups were significantly increased[pg/mL: (1.86±0.23) vs(6.49±0.56) or (8.58±0.42) ng/mL: (7.76±0.70) vs(17.66±0.14) or (18.51±0.10), bothP values<0.05]. The HE staining results showed that compared with the NC and DFM groups, the SM and DS groups showed a significant increase in the number of glomerular cells in the kidneys, and inflammatory cell infiltration was observed in the renal interstitium and surrounding blood vessels. Compared with the NC group, the expression of IL-2 mRNA in the spleen tissue in the DFM group, SM group, and DS group decreased significantly[(2.05±0.14) vs (1.69±0.20) vs (1.52±0.11) vs (1.01±0.13),F=16.83, P<0.05]. Conclusion The disturbance of gut microbiota can affect the expression of related cytokines, promote the inflammatory response of SLE, and further exacerbate the occurrence and development of SLE.

    系统性红斑狼疮肠道菌群菌群紊乱菌群培养

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    CD47胞外区蛋白多克隆抗体的制备

    刘海静苏雨萌王芊艺
    19-24页
    查看更多>>摘要：目的 制备和鉴定抗CD47胞外区蛋白多克隆抗体。 方法 通过逆转录PCR(reverse transcription-PCR，RT-PCR)扩增CD47分子胞外区基因序列，分别重组到原核表达载体pET32a(＋)和pET31b(＋)中，利用大肠杆菌表达CD47蛋白，并优化异丙基硫代半乳糖苷(isopropyl-β-D-thiogalactopyranoside，IPTG)诱导蛋白表达工艺，表达出CD47蛋白。用纯化的CD47蛋白免疫Balb/c小鼠，获得小鼠多克隆抗体，使用亲和层析法纯化CD47多抗，并对多抗进行酶联免疫吸附试验(enzyme-linked immunosorbent assay，ELISA )效价检测和Western blot特异性鉴定。 结果 成功构建pET32a(＋)-CD47和pET31b(＋)-CD47重组质粒。按照实验设置不同条件诱导表达蛋白后，选出最佳表达载体pET32a(＋)-CD47。在大肠杆菌中获得表达最佳蛋白表达工艺。表达制备CD47蛋白，经过五次免疫后，得到CD47多抗。用ELISA法检测制备多抗的效价能达到1∶128 000。Western blot检测结果显示，自制的CD47多抗能准确检测出小鼠心脏组织样品。 结论 成功制备了CD47胞外区蛋白多抗，为进一步研究CD47生物学功能奠定了基础。 Objective To prepare and identify antibody of CD47 extracellular domain protein. Methods The extracellular domain gene fragment of CD47 was amplified by reverse transcription PCR (RT-PCR) and recombined into prokaryotic expression vector pET32a(+ ) and pET31b(+ ) respectively. The CD47 protein was expressed in Escherichia coli(E.coli) and the isopropyl-β-D-thiogalactopyranoside (IPTG) induced protein expression process was optimized to express the CD47 protein. Balb/c mice were immunized with purified CD47 protein to obtain mouse polyclonal antibodies. Anti-CD47 polyclonal antibodies were purified by affinity chromatography. Enzyme-linked immunosorbent assay (ELISA) and Western blot were used for potency detection and specificity identification respectively. Results Recombinant plasmids pET32a (+ )-CD47 and pET31b(+ )-CD47 were successfully constructed. After inducing the expression of proteins in different experimental conditions, the optimal expression vector PET32a(+ )-CD47 was selected. The optimal expression technology of PET32a(+ )-CD47 was obtained in E. coli. Express and prepare CD47 protein, after five immunizations, obtain CD47 polyclonal antibody. The titer of CD47-polyclonal antibody can reach 1∶128 000 using ELISA. Western blot results showed that the self-made CD47 can identify mouse heart tissue samples accurately. Conclusion The polyclonal antibody of CD47 extracellular domain protein was successfully prepared, laying the foundation for further study on the biological function of CD47.

    CD47胞外区蛋白多克隆抗体

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    白花蛇舌草多糖通过调控STAT3/β-catenin通路抑制喉鳞状细胞癌细胞生长和转移的机制

    韩建存郝淑娟
    25-32页
    查看更多>>摘要：目的 探讨白花蛇舌草多糖(hedyotis diffusa polysaccharide，HDP)调控STAT3/β-catenin通路对喉鳞状细胞癌(laryngeal squamous cell carcinoma，LSCC)细胞生长和转移的影响。 方法 依据HDP干预剂量将LSCC TU177细胞分为NC组、HDP-L组、HDP-M组、HDP-H组和HDP-H＋Colivelin组。依据HDP注射剂量将Balb/c裸鼠分为M-NC组、M-HDP-L组、M-HDP-M组、M-HDP-H组、M-HDP-H＋Colivelin组，每组5只。CCK-8、平板克隆实验，流式细胞术，划痕实验，Transwell分别检测TU177细胞增殖、凋亡、迁移及侵袭；Western blot检测TU177增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)、Bcl-2相关X蛋白(Bcl-2-associated X protein，Bax)、基质金属蛋白酶(matrix matalloproteinases，MMP)-2、MMP-9、磷酸化信号传导子及转录激活子3(solubility signal transducer and activator of transcription 3，p-STAT3)、β-连环素(β-catenin)蛋白表达；体内裸鼠移植瘤实验检测肿瘤生长情况。 结果 与NC组比较，HDP-L组、HDP-M组、HDP-H组、HDP-H＋Colivelin组TU177细胞OD450值、克隆形成率显著降低，细胞凋亡率显著升高，细胞划痕愈合率及细胞侵袭数目明显降低，且呈剂量依赖性，差异均有统计学意义(F＝83.31、339.30、448.13、251.27、451.10，P值均<0.001)；与HDP-H组比较，HDP-H＋Colivelin组TU177细胞OD450值、克隆形成率显著升高，细胞凋亡率显著降低，细胞划痕愈合率及细胞侵袭数目明显升高，差异有统计学意义[(0.38±0.04)比(0.75±0.06)，(24.42±1.23)%比(52.29±2.57)%，(42.69±2.50)%比(24.43±1.18)%，(19.23±1.05)%比(33.78±1.62)%，(21.23±1.12)个比(48.86±2.33)个，P值均<0.001]。与NC组比较，HDP-L组、HDP-M组、HDP-H组、HDP-H＋Colivelin组TU177细胞中PCNA、MMP-2、MMP-9、p-STAT3、β-catenin表达显著降低，Bax表达显著升高，且呈剂量依赖性，差异有统计学意义(F＝202.57、70.14、57.48、253.92、90.97、181.72，P值均<0.001)；与HDP-H组比较，HDP-H＋Colivelin组TU177细胞中PCNA、MMP-2、MMP-9、p-STAT3、β-catenin表达显著升高，Bax表达显著降低，差异有统计学意义[(0.31±0.03)比(0.84±0.06)，(0.52±0.04)比(0.91±0.08)，(0.45±0.04)比(0.79±0.06)，(0.16±0.01)比(0.52±0.04)，(0.26±0.02)比(0.63±0.06)，(1.36±0.14)比(0.69±0.05)，P值均<0.001]。与M-NC组比较，M-HDP-L组、M-HDP-M组、M-HDP-H组、M-HDP-H＋Colivelin组LSCC肿瘤质量明显降低，且与M-HDP-H组比较，M-HDP-H＋Colivelin组肿瘤质量明显升高，差异有统计学意义[g：(0.68±0.05)比(0.54±0.05)比(0.42±0.03)比(0.28±0.03)比(0.45±0.04)，F＝65.42，P<0.001]。 结论 HDP可能通过抑制STAT3/β-catenin通路抑制TU177细胞增殖、迁移、侵袭，促进细胞凋亡。 Objective To investigate the influences of hedyotis diffusa polysaccharide (HDP) on the growth and metastasis of laryngeal squamous cell carcinoma (LSCC) cells by STAT3/β-catenin pathway. Methods LSCC TU177 cells were divided into NC group, HDP-L group, HDP-M group, HDP-H group, and HDP-H+ Colivelin group according to the intervention dose of HDP. According to the injection dose of HDP, Balb/c nude mice were divided into M-NC group, M-HDP-L group, M-HDP-M group, M-HDP-H group and M-HDP-H+ Colivelin group, with 5 mice in each group. The proliferation, apoptosis, migration and invasion of TU177 cells were detected by CCK-8, plate cloning, flow cytometry, scratch assay and Transwell respectively. Proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (Bax), matrix matalloproteinases (MMP)-2, MMP-9, phosphorylation signal transducer and activator of transcription 3 (p-STAT3), and β-catenin protein expression in TU177 cells were detected by Western blot. Tumor growth was detected by transplant tumor experiment of nude mice in vivo. Results Compared with NC group, the OD450 value, clone formation rate were significantly decreased, the cell apoptosis rate was significantly increased, the cell scratch healing rate and the number of cell invasions were significantly decreased in TU177 cells of HDP-L, HDP-M, HDP-H and HDP-H+ Colivelin groups with a dose-dependent manner (F= 83.31, 339.30, 448.13, 251.27, 451.10, all P values<0.001). Compared with the HDP-H group, the OD450 value and clone formation rate were significantly increased, the apoptosis rate was significantly reduced, and the scratch healing rate and number of cell invasions were significantly increased in TU177 cells of the HDP-H+ Colivelin group [(0.38±0.04) vs (0.75±0.06), (24.42±1.23)% vs (52.29±2.57)%, (42.69±2.50)% vs (24.43±1.18)%, (19.23±1.05)% vs (33.78±1.62)%, (21.23±1.12) pcs vs (48.86±2.33) pcs, all P values<0.001]. Compared with NC group, the expression levels of PCNA, MMP-2, MMP-9, p-STAT3 and β-catenin in TU177 cells of HDP-L, HDP-M, HDP-H and HDP-H+ Colivelin groups were significantly decreased, while the expression of Bax was significantly increased with a dose-dependent manner(F=202.57, 70.14, 57.48, 253.92, 90.97, 181.72, all P values<0.001). Compared with the HDP-H group, the expression levels of PCNA, MMP-2, MMP-9, p-STAT3 and β-catenin were significantly increased, while the expression of Bax was significantly decreased in TU177 cells of the HDP-H+ Colivelin group [(0.31±0.03) vs (0.84±0.06), (0.52±0.04) vs (0.91±0.08), (0.45±0.04) vs (0.79±0.06), (0.16±0.01) vs (0.52±0.04), (0.26±0.02) vs (0.63±0.06), (1.36±0.14) vs (0.69±0.05), allP values<0.001]. Compared with M-NC group, LSCC tumor mass in M-HDP-L group, M-HDP-M group, M-HDP-H group and M-HDP-H+ Colivelin group was significantly reduced, and compared with the M-HDP-H group, the tumor quality in the M-HDP-H+ Colivelin group was significantly increased[g: (0.68±0.05) vs (0.54±0.05) vs (0.42±0.03) vs (0.28±0.03)vs (0.45±0.04),F=65.42, P<0.001]. Conclusion HDP may inhibit the proliferation, migration and invasion of TU177 cells and promote cell apoptosis by inhibiting the STAT3/β-catenin pathway.

    白花蛇舌草多糖喉鳞状细胞癌STAT3/β-catenin转移

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    呼吸道合胞病毒滴度双抗体夹心ELISA检测方法的建立

    王婉唐悦赵忆宁陈玥如...
    33-38页
    查看更多>>摘要：目的 建立双抗体夹心酶联免疫吸附试验(double antibody sandwich enzyme-linked immunosorbent assay，DAS-ELISA)检测呼吸道合胞病毒(respiratory syncytial virus，RSV)滴度。 方法 以RSV融合前F蛋白的两种抗体AM14与D25作为捕获抗体和检测抗体，通过对其抗体浓度的优化来确定DAS-ELISA较适反应条件，并对该方法的特异性、重复性、灵敏度及适用性进行验证。 结果 选择用AM14抗体作为捕获抗体，D25作为检测抗体。捕获抗体AM14较适工作浓度为20 μg/mL，检测抗体D25的稀释比例为1∶3 000。建立的DAS-ELISA与轮状病毒(rotavirus，RV)和人3型副流感病毒(human parainfluenza virus type 3，HPIV-3)无交叉反应，且不同稀释倍数的RSV病毒进行批内与批间重复试验的变异系数(coefficient of variation，CV)均<10%；不同亚型的RSV及蛋白进行试验，均能检测到病毒滴度；检测结果与CCID50比较，可检测的稀释倍数略高。 结论 建立了检测RSV滴度的DAS-ELISA，该方法可为实验室RSV病毒滴度检测提供依据。 Objective In order to develop a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for detecting respiratory syncytial virus(RSV) titer. Methods Two antibodies AM14 and D25 of RSV pre-fusion protein were used as capture antibody and detection antibody separately. The optimal reaction condition of DAS-ELISA was determined through antibody concentration optimization, and the specificity, repeatability, sensitivity and applicability were validated. Results Choosing AM14 antibody as capture antibody and D25 as detection antibody, the working concentration of AM14 was determined to be 20 μg/mL, and the working concentration of D25 was determined to be 1∶3 000 dilution. The established DAS-ELISA had no cross-reaction with rotavirus(RV) and human parainfluenza virus type 3(HPIV-3). The coefficient variation ( CV) of RSV viruses with different dilution ratios in both intra and inter batch repeated experiments were less than 10%. The virus titer of different subtype RSV and protein were detected by this method. Compared with CCID50, the detectable dilution ratio is slightly higher. Conclusion DAS-ELISA in detection of RSV titer was successfully developed, which provided a basis for RSV titer detection in laboratory.

    呼吸道合胞病毒融合前F蛋白双抗体夹心ELISA病毒滴度

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    白塞病患者外周血T淋巴细胞亚群及细胞因子与其临床特征关系分析

    毛玉景焦占峰程盼盼宋芹...
    39-45页
    查看更多>>摘要：目的 检测白塞病(Behcet's disease，BD)患者外周血T细胞亚群及细胞因子水平，并分析BD患者不同临床特征对外周血T细胞亚群及细胞因子水平的影响。 方法 选取2018年1月至2021年12月在济宁医学院附属医院诊治的BD患者80例作为病例组，另选取本院健康体检中心40例健康志愿者作为对照组。采用流式细胞术检测外周血辅助性T细胞(helper T cell，Th)1、Th17、调节性T细胞(regulatory T cell，Treg)亚群表达。酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)法测定血清细胞因子干扰素-γ(interferon-γ，IFN-γ)、白细胞介素(interleukin，IL)-4、IL-10、IL-17、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)水平。 结果 与对照组相比，BD患者总T淋巴亚群、CD8＋T及Th1细胞表达水平均显著升高，差异有统计学意义{%：[68.75(54.23，80.32)]比[72.10(62.23，86.45)]，[23.44(12.89，33.76)]比[31.46(20.13，45.26)]，[10.67(8.23，12.35)]比[17.96(10.26，25.39)]，Z＝－3.13、－4.54、－3.97，P值均<0.05}，而Th、Th17及Treg细胞表达的组间差异无统计学意义(P>0.05)。与对照组相比，BD患者IL-17、IL-6、TNF-α、IL-4、IFN-γ血清水平均显著升高，血清IL-10水平显著降低，差异均有统计学意义(Z＝－4.32、－5.01、－8.18、－8.70、－3.48、－8.30，P值均<0.05)。与无相关表现患者相比，有关节炎表现的BD患者和活动性BD患者IL-6表达显著升高，差异有统计学意义{pg/mL：[3.23(2.98，5.35)]比[10.51(6.72，23.21)]，[6.32(4.79，8.93)]比[9.68(6.97，18.73)]，Z＝5.47、8.76，P值均<0.05}。与无相关临床表现患者相比有关节炎表现、眼部表现、消化道表现和活动性BD患者TNF-α水平显著升高，同时有消化系统病变的BD患者IFN-γ水平显著升高，差异均有统计学意义{ pg/mL：[7.74(6.89，10.19)]比[39.84(30.37，50.61)]，[6.12(5.36，9.89)]比[31.35(15.71，30.46)]，[6.49(4.78，10.21)]比[19.89(14.36，36.21)] ，[5.89(4.61，8.96)]比[27.91(15.32，37.81)]，[6.89(5.43，14.86)]比[26.79(15.41，31.56)]，Z＝7.70、6.84、6.94、9.47、5.70，P值均<0.05}。而无相关临床表现和有相关临床表现的BD患者IL-4和IL-17表达水平的组间差异无统计学意义(P>0.05)。 结论 BD患者存在着T淋巴细胞亚群及细胞因子的失衡，且病情活动和临床分型可能参与了T细胞亚群及细胞因子失衡。 Objective To detect peripheral blood T lymphocyte subsets and cytokines in Behcet's disease (BD) patients, and analyze the impact of different clinical characteristics on peripheral blood T cell subpopulations and cytokine levels in BD patients. Methods Eighty patients with BD who were diagnosed and treated at the Affiliated Hospital of Jining Medical University from January 2018 to December 2021 were selected as the case group. Another forty healthy volunteers from the health examination center of our hospital were selected as control group. Helper T cell (Th) 1, Th17, and regulatory T cell (Treg) lymphocyte subsets were detected by flow cytometry. Enzyme linked immunosorbent assay (ELISA) was used to detect the serum levels of interferon-γ(IFN-γ), interleukin(IL)-4, IL-10, IL-17, IL-6, tumor necrosis factor-α(TNF-α). Results Compared with the control group, the expression levels of total T lymphocyte subpopulations, CD8+ T and Th1 cell in BD patients were significantly increased{%: [68.75 (54.23, 80.32)] vs [72.10 (62.23, 86.45)], [23.44 (12.89, 33.76)] vs [31.46 (20.13, 45.26)], [10.67 (8.23, 12.35)] vs [17.96 (10.26, 25.39)],Z=-3.13, -4.54, -3.97, all P values<0.05}, while there was no statistically significant difference in the expression of Th, Th17 and Treg cells between the groups (P>0.05). Compared with the control group, the serum levels of IFN-γ、IL-4、IL-17、IL-6 and TNF-α were increased and IL-10 was decreased significantly in the BD group(Z = -4.32, -5.01, -8.18, -8.70, -3.48, -8.30, all P values <0.05). Compared to patients without related manifestations, the expression levels of IL-6 in patients with osteoarthritis and active BD were significantly increased{pg/mL: [3.23 (2.98, 5.35)] vs [10.51 (6.72, 23.21)], [6.32 (4.79, 8.93)] vs [9.68 (6.97, 18.73)], Z=5.473, 8.763, both P values<0.05}. Compared to patients without relevant clinical manifestations, the expression levels of TNF-α were significantly increased in patients with arthritis, ocular manifestations, gastrointestinal manifestations and active BD, and the IFN-γ was significantly increased in BD patients with digestive system disorders {pg/mL: [7.74 (6.89, 10.19)] vs [39.84 (30.37, 50.61)], [6.12 (5.36, 9.89)] vs [31.35 (15.71, 30.46)], [6.49 (4.78, 10.21)] vs[19.89 (14.36, 36.21)], [5.89 (4.61, 8.96)] vs [27.91 (15.32, 37.81)], [6.89 (5.43, 14.86)] vs [26.79 (15.41, 31.56)],Z=7.70, 6.84, 6.94, 9.47, 5.70, all P values<0.05}. However, there was no statistically significant difference in the expression levels of IL-4 and IL-17 between BD patients without or with related clinical manifestations(P>0.05). Conclusion BD patients have imbalances in T lymphocyte subpopulations and cytokines, and disease activity and clinical classification may be involved in the imbalance of T lymphocyte subpopulations and cytokines.

    白塞病临床特征T细胞亚群细胞因子

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    抑郁症患者外周血T细胞亚群、Tregs及细胞因子水平变化及意义

    杨娟许华斌张光满孙斌...
    46-51页
    查看更多>>摘要：目的 研究抑郁症患者外周血中T细胞亚群、调节性T细胞(regulatory T cells，Tregs)和白细胞介素(interleukin，IL)-6、IL-10、干扰素-γ(interferon-γ，INF-γ)以及肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)的表达水平，探讨其在郁抑症发病机制中的作用。 方法 选择2020年1月至2022年6月于皖西卫生职业学院附属医院就诊的初诊抑郁症患者82例作为病例组，同期选取我院体检中心体检健康的40例志愿者作为对照组。根据汉密尔顿抑郁量表(Hamilton depression scale，HAMD)评分，将病例组患者分为重度抑郁组(23例)和轻中度抑郁组(59例)。采用流式细胞仪检测两组研究对象外周血中T细胞和Tregs水平，采用酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)检测血浆中IL-6、IL-10、INF-γ以及TNF-α水平，对比分析T细胞亚群、Tregs及细胞因子水平的变化及意义。 结果 与对照组相比，初诊抑郁症患者外周血中CD8＋T、Tregs细胞比例显著升高，CD4＋/CD8＋比值显著降低，差异具有统计学意义[(23.13±9.24)%比(26.72±8.12)%，(5.31±1.65)%比(12.41±2.52)% ，(1.71±0.32)比(1.28±0.51) ，t＝3.22、5.94、4.20，P值均<0.05]，CD3＋T、CD4＋T细胞比例的组间差异无统计学意义(P>0.05)。抑郁症患者外周血中IL-6和INF-γ表达水平较对照组明显升高，差异具有统计学意义[(2.89±2.11) pg/mL比(1.81±1.17) pg/mL，(28.12±11.02) pg/mL比(13.18±6.41) pg/mL，t＝5.10、7.59，P值均<0.05]，IL-10、TNF-α的组间差异无统计学意义(P>0.05)。重度组抑郁症患者Tregs和IL-6水平较轻中度抑郁症患者显著升高，差异具有统计学意义[(13.11±2.92)%比(8.31±2.05)%，(3.21±2.37) pg/mL比(2.59±1.91) pg/mL，t＝7.43、5.93，P值均<0.05]，而CD8＋T、INF-γ的组间差异无统计学意义(P>0.05)。与治疗前相比，治疗后抑郁症患者CD8＋T、Tregs细胞比例，IL-6、INF-γ水平显著下降，差异有统计学意义[(26.72±8.12)%比(22.18±8.14)%，(12.41±2.52)%比(8.11±1.95)%，(2.89±2.11) pg/mL比(2.21±1.87) pg/mL，( 28.12 ±11.02) pg/mL比(17.21±9.46) pg/mL，t＝21.04、51.33、35.38、7.11，P值均<0.05]。 结论 CD8＋T、Tregs和IL-6参与了郁抑症患者免疫耐受异常的形成，抑郁症的发生与免疫系统的异常改变密切相关。 Objective To investigate the expression levels of T-cell subsets, regulatory T cells (Tregs), interleukin(IL)-6, IL-10, interferon-γ (INF-γ), and tumor necrosis factor-α (TNF-α) in peripheral blood of patients with depression, in order to explore their roles in the pathogenesis of depressive disorders. Methods A total of 82 patients who visited the Affiliated Hospital of Health Vocational College of West Anhui University from January 2020 to June 2022 were selected as the case group. 40 healthy volunteers from the hospital's physical examination center were selected as the control group during the same period. According to the Hamilton depression scale (HAMD) scores, the patients in the case group were divided into a severe depression group (23 cases) and a mild-to-moderate depression group (59 cases). Flow cytometry was used to detect the levels of T cells and Tregs in the peripheral blood. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of IL-6, IL-10, INF-γ, and TNF-α in the plasma. Changes and significance in the T-cell subsets, Tregs, and cytokine levels were compared and analyzed. Results Compared with the control group, the proportion of CD8+ T and Tregs cells in the peripheral blood of patients with newly diagnosed depression was significantly increased, and the CD4+ /CD8+ ratio was significantly reduced [(23.13±9.24)% vs (26.72±8.12)%, (5.31±1.65)% vs(12.41±2.52)%, (1.71±0.32)vs(1.28±0.51), t=3.22, 5.94, 4.20, all P values<0.05], but there was no statistically significant difference in the proportions of CD3+ T and CD4+ T cells between groups (P>0.05). The expression levels of IL-6 and INF-γ in the peripheral blood of patients with depression were significantly higher than those in the control group[ (2.89±2.11) pg/mL vs (1.81±1.17) pg/mL, (28.12±11.02) pg/mL vs (13.18±6.41) pg/mL,t=5.10, 7.59, both P values <0.05], there was no statistical significance in the expression of IL-10 and TNF-α between groups ( P >0.05). The levels of Tregs and IL-6 in patients with severe depression were significantly higher than those in patients with mild-to-moderate depression [(13.11±2.92)% vs (8.31±2.05)%, (3.21±2.37) pg/mL vs (2.59±1.91) pg/mL, t=7.43, 5.93, both P values <0.05], while no differences have been found in the levels of CD8 + T and INF-γ between groups (P>0.05). Compared with before treatment, the proportion of CD8+ T and Tregs cells, and the levels of IL-6 and INF-γ in patients with depression decreased significantly after treatment[(26.72±8.12)% vs (22.18±8.14)%, (12.41±2.52)% vs (8.11±1.95)%, (2.89±2.11) pg/mL vs (2.21±1.87) pg/mL, ( 28.12±11.02 ) pg/mL vs (17.21±9.46) pg/mL,t=21.04, 51.33, 35.38, 7.11, all P values<0.05]. Conclusion CD8+ T cells, Tregs and IL-6 are involved in the development of immune tolerance abnormalities in patients with depression. The occurrence of depression is closely associated with abnormal changes in the immune system.

    抑郁症T细胞亚群调节性T细胞细胞因子

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    非霍奇金淋巴瘤PD-1抗体治疗患者免疫功能变化及不良反应预防研究

    史玉李天一王亚秋肖建波...
    52-56页
    查看更多>>摘要：目的 探讨程序性细胞死亡受体-1(programmed cell death-1，PD-1)抗体对非霍奇金淋巴瘤(non-Hodgkin lymphoma，NHL)患者免疫功能表达的影响，并分析不良反应的预防。 方法 选取2013年4月至2016年8月于河北省唐山市人民医院接受治疗的NHL确诊患者共计100例，并将其分为研究组和对照组，每组各50例。研究组接受PD-1抗体治疗，对照组患者依据病情给予NHL化疗治疗。免疫组化法检测治疗前后两组患者免疫检查点OX40、糖皮质激素诱导的肿瘤坏死因子受体(glucocorticoid-induced tumor necrosis factor receptor，GITR)、CD28、PD-1、血清T细胞免疫球蛋白黏液素3(T cell immnoglobulin and mucin 3，TIM-3)、淋巴细胞活化因子-3(lymphocyte activation gene-3，LAG-3)表达情况，并统计分析两组患者治疗后不良反应情况。 结果 治疗前，研究组与对照组患者共刺激性免疫检查点OX40、GITR、CD28表达水平，共抑制性免疫检查点PD-1、TIM-3、LAG-3表达水平及美国东部肿瘤合作组(Eastern Cooperative Group，ECOG)评分比较差异均无统计学意义(P>0.05)。治疗后，研究组患者OX40、GITR、CD28、TIM-3水平明显高于对照组，PD-1水平明显低于对照组，差异均有统计学意义[%：(9.75±3.02)比(6.35±2.85)，(7.48±1.62)比(5.37±1.56)，(16.37±3.42)比(13.46±3.56)，(15.48±3.45)比(13.24±3.83)，(0.87±0.22)比(1.32±0.36)，t＝5.79、6.63、4.17、3.07、7.54 ，P值均<0.05]，两组患者LAG-3水平差异无统计学意义(P>0.05)。治疗后，研究组ECOG评分明显高于对照组，不良反应发生率明显低于对照组，差异有统计学意义[(4.61±1.46)分比( 2.74±0.66)分，10.00%比26.00%，t ＝8.25、χ2＝4.34，P值均<0.05]。 结论 PD-1抗体治疗可有效提高NHL患者免疫共刺激分子的表达，同时抑制PD-1表达，下调患者免疫共抑制，以此改善NHL患者免疫系统状态。 Objective To explore the effect of programmed cell death-1 (PD-1) antibody on the immune function expression of patients with non-Hodgkin lymphoma (NHL), and to analyze the prevention of adverse reactions. Methods A total of 100 patients with NHL who received treatment at Tangshan People's Hospital in Hebei Province from April 2013 to August 2016 were selected, and divided into a study group and a control group, with 50 patients in each group. The research group received PD-1 antibody treatment, while the control group received NHL chemotherapy based on their condition. Immunohistochemical method was used to detect the expression of immune checkpoint OX40, glucocorticoid induced tumor necrosis factor receptor (GITR), CD28, PD-1, serum T cell immunoglobulin and mucin 3 (TIM-3), and lymphocyte activation gene-3 (LAG-3) in two groups of patients before and after treatment, and statistically analyze the adverse reactions. Result Before treatment, there were no statistically significant differences between the study group and the control group in the expression levels of OX40, GITR, CD28 at the co-stimulating immune checkpoint, and the expression levels of PD-1, TIM-3, and LAG-3 at the co-inhibitory immune checkpoint, as well as the Eastern Cooperative Group (ECOG) scores. After treatment, the levels of OX40, GITR, CD28, and TIM-3 in the study group were significantly higher than those in the control group, while the levels of PD-1 were significantly lower than those in the control group[%: (9.75±3.02) vs (6.35±2.85), (7.48±1.62) vs (5.37±1.56), (16.37±3.42) vs (13.46±3.56), (15.48±3.45) vs (13.24±3.83), (0.87±0.22) vs (1.32±0.36), t=5.79, 6.63, 4.17, 3.07, 7.54, all P values<0.05]. There was no statistically significant difference in LAG-3 levels between the two groups of patients (P>0.05). After treatment, the ECOG score of the study group was significantly higher than that of the control group, and the incidence of adverse reactions was significantly lower than that of the control group[(4.61±1.46) points vs (2.74±0.66) points, 10.00% vs 26.00%,t =8.25, χ2=4.34, both P values<0.05]. Conclusion PD-1 antibody therapy can effectively improve the immunocostimulation of NHL patients, inhibit the expression of PD-1, and down-regulate the immunocosuppression of patients, so as to improve the immune system status of NHL patients.

    非霍奇金淋巴瘤程序性细胞死亡受体-1抗体抑制性免疫检查点不良反应

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    光子照射对免疫炎症双向调控的研究进展

    王杏容黄彩凤程喜平
    57-61页
    查看更多>>摘要：光子是传递电磁相互作用的基本粒子，光子照射包括紫外线(ultraviolet，UV)、红外线(infrared，IR)、可见光(visible light，VL)等，在医学领域应用极为广泛。研究表明光子照射对机体免疫炎症具有双向调控的作用，双向影响免疫炎症性疾病的发生发展，文章就光子照射对免疫炎症双向调控的研究进展进行综述。 Photons are elementary particles that mediate electromagnetic interactions. Photon irradiation encompasses ultraviolet(UV), infrared(IR), and visible light(VL), which find extensive applications in the medical domain. Research demonstrates the bidirectional regulatory effects of photon radiation on immune inflammation, influencing the development of immune-inflammatory diseases. This review summarizes the bidirectional regulation of immune inflammation by photons.

    光子照射紫外线免疫炎症淋巴细胞

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