胃癌组织中黏附分子整合素CD11c的表达及其与预后的关系

目的 探讨黏附分子家族的成员CD11c在胃癌组织中的表达及其与预后的关系.方法 采用免疫组织化学染色法检测125例胃癌患者癌组织CD11c的表达.结果　CD11c在胃癌中的表达高于胃炎和胃息肉组织.CD11c与患者性别、年龄、病理分级、胃癌部位无关(P＞0.05);与肿瘤大小、浸润深度、淋巴结转移、组织类型、复发和TNM分期明显相关.肿瘤直径＜5 cm组CD11c的表达水平(12.4±6.8)高于肿瘤直径≥5 cm组(7.7±4.6),差异有统计学意义(t=3.98,P＜0.01).浸润深度与CD11c表达水平显著关联,未侵入肌层组胃癌的CD11c表达水平(15.4±8.5)显著高于侵入肌层组(9.5±5.6),差异有统计学意义(t=3.1,P＜0.05).淋巴结转移与CD11c表达水平显著关联,未有淋巴结转移的CD11c表达水平(15.3±6.6)显著高于淋巴结转移组(8.6±5.1),差异有统计学意义(t=5.44,P＜0.01).未复发组的CD11c表达水平(12.3±7.4)高于复发组(9.0±5.3),差异有统计学意义(t=2.90,P＜0.01).分化型组的CD11c表达水平(9.3±5.3)低于低分化型组(12.6±7.8),差异有统计学意义(t=2.68,P＜0.01).CD11c表达随肿瘤临床分期的递增而降低,各组间差异有统计学意义(P＜0.01).多因素COX模型分析,在调整了性别、年龄、肿瘤大小、是否侵及深肌层、分化程度后,与CD11c细胞低表达组比较,高表达组有降低胃癌死亡风险的趋势(RR=0.51,95%CI=0.22～1.16).结论　CD11c表达与胃癌预后有关,可以作为反映患者免疫状态和预后的指标之一.

Abstract：

Objectiye To investigate the expression of CD11c in gastric carcinoma and the clinical significance. Methods The expression of CD11 c in gastric carcinoma, gastritis tissue and gastric polyp was detected by using immunohistochemical assay. Results The expression level of CD11c in gastric carcinoma was higher than that in gastritis tissue and gastric polyp. There was no significant difference in the CD11 c expression among gender, age, pathological grade, primary area (P ＞ 0. 05 ). There was significant difference in the CD11c expression among tumor size, invasive depth, lymph node metastasis, histological type, recurrence and clinical stage. The expression of CD11c in tumor size ＜5 cm group ( 12.4 ±6. 8 ) was higher than that in tumor size ≥ 5 cm group (7.7 ± 4. 6), ( t = 3.98, P ＜ 0. 01 ). The expression of CD11c in non-invasion group ( 15.4 ± 8.5 ) was obviously higher than that in invasion group (9.5 ±5.6), ( t = 3.1 ,P ＜ 0. 05 ). The expression of CD11c in non-lymph node metastasis group ( 15.3 ± 6. 6 )was obviously higher than that in lymph node metastasis group ( 8. 6 ± 5. 1 ), ( t = 5. 44, P ＜ 0. 01 ). The expression of CD11c in non-recurrence group ( 12.3 ±7.4) was higher than that in recurrence group (9. 0± 5.3), ( t= 2.90, P ＜ 0. 01 ). The expression of CD11 c in differentiation group ( 9. 3 ± 5.3 ) was lower than that in low differentiation group ( 12. 6 ± 7. 8 ), ( t = 2. 68, P ＜ 0. 01 ). There was significant diference among groups of clinical stage (P ＜0. 01 ). The multivariate Cox analysis revealed that the risk of death in high expression group was significantly lower than that in low expression group (RR =0. 51, 95% CI=0. 22-1.16). Conclusion Detection of the CD11c expression in gastric carcinoma is beneficial to the judgment of the prognosis of gastric carcinoma and the choice of individualized treatment.     

抑制记忆性T淋巴细胞共刺激分子OX40延长小鼠胰岛移植物的存活时间

目的 观察抑制记忆性T淋巴细胞共刺激分子OX40延长胰岛移植小鼠存活时间的作用,探讨其作用机制.方法 采用免疫磁珠法制备初始性、类记忆性及记忆性CD8+T淋巴细胞,并采用逆转录聚合酶链反应法检测3种T淋巴细胞上OX40的表达量.将C57BL/6小鼠脾脏T淋巴细胞经尾静脉注射给Rag-/-小鼠.将Rag-/-小鼠分为3组,对照组:给予同型IgG;治疗组:给予抗OX40L单克隆抗体;基因敲除组:输注的T淋巴细胞由OX40基因敲除的C57BL/6小鼠供给.T淋巴细胞输注6周后,使用链脲霉素诱导建立糖尿病模型,然后以DBA/2小鼠为供者,进行胰岛移植,移植后观察和比较3组间胰岛移植物的存活时间.结果 OX40在初始性、类记忆性和记忆性T淋巴细胞上的相对表达量分别为2.87、111.24和146.15,类记忆性和记忆性T淋巴细胞间OX40表达量的差异无统计学意义(P＞0.05),但均显著高于初始性T淋巴细胞(P＜0.01).对照组、治疗组和基因敲除组胰岛移植物的存活时间分别为21、130和125 d,治疗组和基因敲除组间胰岛移植物存活时间的差异无统计学意义(P＞0.05),二者均显著长于对照组(P＜0.05).结论 OX40在记忆性T淋巴细胞表面高表达,且阻断OX40共刺激分子通道可明显延长胰岛移植物的存活时间,抑制OX40/OX40L共刺激通道可能是诱导胰岛移植免疫耐受的关键点之一.

Abstract：

Objective To investigate the role of OX40 in the mechanisms of memory T cells in islet transplant tolerance.Methods The expression of OX40 on native, like memory and memory CD8+T cells was detected by RT-PCR. Splenic T cells from B6 mice were injected into Rag-/- mice via the tail vein, and the Rag-/- mice were divided into three groups (n=8 each): control group, given IgG; treatment group, given anti-OX40L; and OX40 knock-out group, given T cells from OX40 knock-out B6 mice spleen. All recipients were induced into diabetes mellitus model after adoptive transfer. Islet transplantation was performed on all Rag-/- mice as recipients. The mean survival time of islet was observed.Results The expression of OX40 in native T cells, like memory T cells and memory T cells was 2.87, 111.24 and 146.15 respectively. The expression of OX40 in like memory and memory T cells was higher than in native T cells (P<0.05). Comparison with control group , The mean survival time of the DBA/2 islet allografts in treatment group (130 days) and OX40 knock-out group (125 days) was significantly longer than in control group (21 days, P<0.05).Conclusion The OX40 expression is high in memory T cells. The mean survival time of the islet allografts can be prolonged by blocking OX40/OX40L pathway. OX40/OX40L pathway may be the key point of transplant tolerance.     

GW5074对不成熟树突状细胞诱导初始性CD4+T淋巴细胞分化为Treg的影响

目的 建立一种稳定且高效的不成熟树突状细胞(imDC)体外培养方法,探讨细胞外信号调节激酶(ERK)1/2信号传导通路抑制剂GW5074对imDC体外诱导同种初始性CD4+T淋巴细胞分化为调节性T淋巴细胞(Treg)的影响.方法 从健康成人外周血单个核细胞(PBMC)中分离、培养成熟树突状细胞(mDC)和imDC,并对mDC和imDC的免疫表型和功能进行鉴定.取新生儿脐静脉血分离初始性CD4+T淋巴细胞.实验分为5组:(1)空白对照组为单纯培养的初始性CD4+T淋巴细胞,不做任何处理;(2)阳性对照组将imDC与初始性CD4+T淋巴细胞以1∶10的细胞比例混合培养;(3)低浓度GW5074组;(4)中浓度GW5074组;(5)高浓度GW5074组.后3组在阳性对照组基础上,分别加入终浓度为8、24和40μmol/L的GW5074.培养5 d后,用流式细胞仪检测初始性CD4+T淋巴细胞转化为Treg细胞的转化率.结果 imDC呈CD1a高表达,CD80和CD83低表达;mDC呈CD1a低表达,CD80和CD83高表达.imDC和mDC的刺激指数分别为1.12±0.03、2.85±0.07.空白对照组,阳性对照组,低、中及高浓度GW5074组的CD4+CD25+Treg转化率分别为(5.81±1.36)%、(35.73±2.07)%、(22.53±2.11)%、(11.55±1.73)%和(4.97±1.83)%,除空白对照组与高浓度GW5074组间的差异无统计学意义(P＞0.05)外,其余各组之间两两比较,差异均有统计学意义(P＜0.01).结论 通过应用重组人粒细胞-巨噬细胞集落刺激因子和重组人白细胞介素4联合诱导人外周血PBMC可获得高纯度imDC,ERK1/2信号传导通路在诱导免疫耐受中发挥作用,GW5074可抑制初始性CD4+T淋巴细胞向Treg转化.

Abstract：

Objective To establish a stable and efficient method of culturing imDCs in vitro,and to explore the effect of GW5074, which blocks ERK1/2 signal pathway in the process of imnature dentritic cells (imDCs) on inducing differentiation of the na(i)ve allogeneic CD4+ T cells into Treg cells in vitro. Methods The imDCs and mature DCs (mDCs) were isolated and cultured from the peripheral blood mononuclear cells (PBMC) derived from a healthy adult male volunteer, and they were identified by cell morphology, cell surface marker and cell functions respectively. Na(i)ve CD4+ T cells were isolated from newborn umbilical vein blood and were divided into 5 groups to be cultured: (1) Blank control group: Na(i)ve CD4+ T cells were cultured alone;(2) Positive control group: The irrDCs were Middle-concentration GW5074 group;(5) High-concentration GW5074 group. In the last three groups, imDCs and na(i)ve CD4+ T cells were co-cultured, the same as the positive control group, but these groups were added by GW5074 dilution at the concentrations of 8, 24, and 40μmol/Lrespectively. After co-culture for 5 days, the transformation ratio from naive CD4+T cells to Treg T cells was detected by flow cytometry. Results On the surface of imDCs, there was stronger pression of CD1a, but weaker expression of CD80 and CD83. On the contrary, on the surface of mDCs, there was weaker expression of CD1a, but stronger expression of CD80 and CD83. The stimulation index in imDCs group and mDCs group was 1.12±0.03 and 2.85±0. 07 respectively. The transformation ratio of Treg T cells in blank control group, positive control group, low-concentration GW5074 group, middle-concentration GW5074 group and high-concentration GW5074 group was (5. 81±1.36)%, (35.73±2.07)%, (22.53±2.11)%, (11.55±1.73)%, and (4.97±1.83)%respectively. One-way ANOVA analysis revealed that there was no significant difference between high-concentration GW5074 group and blank control group, P＞0. 05, but significant difference between the remaining groups, P＜0.01. Conclusion High purity of imDCs can be obtained from PBMC by induction with rhGM-CSF and rhIL-4. ERK1/2 signal pathway plays a role in inducing the immune tolerance. GW5074 can inhibit differentiation of na(i)ve CD4+ T cells into Treg T cells.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

胃癌外周血淋巴细胞亚群表达水平对患者生存率的影响

目的探讨胃癌患者外周血淋巴细胞亚群表达与生存率的关系。方法用流式细胞仪检测833例胃癌首诊患者外周血的淋巴细胞亚群CD3^＋、CD4^＋、CD8^＋、CD4^＋／CD8^＋、CD19^＋、CD25^＋、CD44^＋及NK^＋细胞．并根据96名健康对照者的平均检测值分为高表达组和低表达组．比较各亚群高表达组与低表达组患者的生存率。结果与健康对照者相比．胃癌患者CD3^＋、CD8^＋低表达。而CD4^＋,CDl9^＋,CD25^＋、CD4^＋／CD8^＋、CD44^＋,NK^＋高表达（P〈0．05）。CD19^＋高表达与低表达者分别为444例和389例,3年生存率为36．4％和18．5％,差异有统计学意义（P〈0．05）：而其他7种淋巴细胞亚群表达水平则与患者生存率无关（均P〉0．05）。结论与健康人群相比,胃癌患者外周血淋巴细胞亚群发生显著变化．其中CDl9^＋高表达患者具有明显的生存优势。     

Dominance of CD4+ lymphocytic infiltrates with disturbed effector cell characteristics in the tumor microenvironment of prostate carcinoma

BACKGROUND: Prostate cancer is the most common cancer of men in the Western world. Despite the over-expression of tumor-associated antigens, like PSA or PSMA, immune activation is inefficient. The goal of this investigation was to assess in situ characteristics of prostate cancer-infiltrating lymphocytes and to determine their activation status and effector function. METHODS: We compared 17 carcinoma containing tissues, four benign prostatic hyperplasia tissues and eight healthy prostate tissues regarding lymphocyte subset composition, locoregional distribution, and functional status using immunohistological staining of cryopreserved tissues. For determination of lymphocyte subsets, serial sections were stained with CD3, CD4, and CD8 antibodies. Activation status and effector function were studied using CD69, interferon-gamma (IFN gamma), perforin, and CD3 zeta chain antibodies. T-cell-receptor repertoire (TCR) analysis was made to determine the complexity of infiltrating lymphocytes. RESULTS: CD3+, CD4+, and CD69+ T lymphocytes were prominent in tissues derived from patients with prostate carcinoma. CD8+ lymphocytes were significantly less than CD4+ lymphocytes. IFN gamma and perforin were downregulated on infiltrating lymphocytes compared to cells of healthy prostate tissue. Very few lymphocytes were detected within cancerous lesions whereas surrounding tissues showed extensive lymphocyte cluster formation. The TCR repertoire of infiltrating lymphocytes was broad and similar to that of healthy prostate tissue, giving no evidence for specific lymphocyte recruitment. CONCLUSIONS: In the prostate cancer microenvironment, CD4+ T lymphocytes dominated while CD8+ T cells were sparse. The lymphocytes exhibited signs of disturbed effector function. Consequently, the immune response against autologous tumor cells is likely to be inefficient in controlling tumor growth.     

吲哚胺2,3双加氧酶对小鼠CD4+T细胞增殖的抑制作用

目的 探讨吲哚胺2,3双加氧酶(IDO)抑制T细胞增殖的机制。方法 通过低色氨酸(10μmol/L)培养基(LTM)和/或添加不同浓度(50、100、200和400μmol/L)色氨酸代谢产物(TC),观察小鼠树突细胞(DC)与异系CD4+T细胞培养体系中后者的增殖。Annexin-V及碘化丙锭(PI)双染法测定CD4+T细胞凋亡。结果 LTM中CD4+T细胞增殖指数(2.718 ±0.010)及抑制CD4+T细胞增殖的TC浓度(200μmol/L KYN或50 μmol/L 3-HAA)较正常色氨酸培养基(NTM)(3.385 ±0.013,400 μmol/L KYN或100 μmol/L 3-HAA)显著降低,差异有统计学意义(P＜0.01)。LTM中CD4+T细胞早期凋亡率(33.163±0.556)％显著高于NTM(8.867 ±0.565)％ (P ＜0.01)。NTM中CD4+T细胞凋亡率随3-HAA浓度升高而增加[3-HAA浓度50、100、200和400 μmol/L组中分别为(14.433 ±0.640)％、(22.273±0.629)％、(37.363±0.953)％和(46.643±0.633)％]。结论 LTM及TC均可通过诱导凋亡来抑制CD4+T细胞增殖,两者具有叠加效应。     

黄芪多糖对分泌白细胞介素12树突细胞亚群的免疫调控效应与机制

目的 观察黄芪多糖(APS)对分泌IL-12树突细胞(DC)亚群CD11chighCD45RBlowDC功能的影响.方法磁珠分选技术获得BALB/c小鼠脾脏CD11chighCD45RBlowDC和CD4+T淋巴细胞.在CD11 chighCD45RBlowDC中加入不同浓度APS(50、100、200μg/mL)处理,以不加APS的细胞作为对照,应用ELISA法检测细胞培养上清液中IL-12水平,流式细胞仪检测细胞表面分子CD40、CD80、CD86、I-A/E及Toll样受体4(TLR4)的表达.将CD4+T淋巴细胞分为正常对照组(未行任何处理)、未刺激组(加入未经APS处理的CD11chighCD45RBlowDC与CD4+T淋巴细胞混合培养)、高浓度APS刺激组(加入经200μg/mL APS处理后的CD11chighCD45RBlowDC与CD4+T淋巴细胞混合培养)、高浓度APS刺激+抗体1组(加入经200μg/mL APS处理后的CD11chighCD45RBlowDC、IL-12抗体与CD4+T淋巴细胞混合培养)和高浓度APS刺激+抗体2组(加入经200 μg/mL APS处理后的CD11chighCD45RBlowDC、IL-12同型对照抗体与CD4+T淋巴细胞混合培养).采用噻唑蓝法测定CD4+T淋巴细胞增殖能力,流式细胞仪检测细胞培养液中IL-4和γ干扰素水平.对数据行多组间单因素方差分析.结果与未加APS刺激相比,3种浓度APS均显著增强CD11chighCD45RBlowDC表面分子CD40、CD80、I-A/E及TLR4表达及IL-12分泌,其中IL-12分泌呈APS浓度依赖性;CD86表达无明显变化.高浓度APS刺激组CD4+T淋巴细胞增殖能力高于未刺激组(F=13.438,P＜0.05);高浓度APS刺激组细胞γ干扰素水平为(2784±137)pg/mL,高于未刺激组[(1952±101)pg/mL,F=12.177,P＜0.05];高浓度APS刺激组细胞IL-4水平为(172±20)pg/mL,明显低于未刺激组[(193±19)pg/mL,F=11.963,P＜0.05].高浓度APS刺激+抗体1组前述3项指标表达水平较未刺激组明显改善,高浓度APS刺激+抗体2组前述3项指标表达水平与高浓度APS刺激组接近.结论 APS能够通过促进CD11chighCD45RBlowDC中IL-12的表达,诱导CD4+T淋巴细胞向Th1型反应分化,通过激活CD11chighCD45RBlowDC增强免疫活性.

Abstract：

Objective To investigate immunomodulatory effect of Astragalus polysaccharides (APS) on IL-12-secreting dendritic cell (DC) subset CD11chigh CD45RBlow DC. Methods Spleen CD11chighCD45RBlow DC and CD4 +T lymphocytes in BALB/c mice were purified by magnetic beads sorting,and were treated with 0 (as control), 50, 100, 200 μg/mL APS. Immunofluorescence staining and flow cytometry were used to determine expressions of CD11chighCD45RBlow DC surface molecules, including CD40,CD80, CD86, I-A/E, and Toll-like receptor (TLR) 4. IL-12 level in CD11chighCD45RBlow DC culture supernatant was determined by ELISA. The CD4+ T lymphocytes were divided into: normal control group,non-stimulation group ( CD4 + T lymphocytes cocultured with APS-unstimulated CD11 chigh CD45RBlow DC ) ,high-dose APS stimulation group (CD4+T lymphocytes cocultured with 200 μg/mL APS-stimulated CD11ch'ghCD45RBlow DC) , high-dose APS stimulation + antibody 1 group ( CD4 + T lymphocytes cocultured with 200 μg/mL APS-stimulated CD11chighCD45RBlow DC and IL-12 antibody), high-dose APS stimulation +antibody 2 group (CD4 +T lymphocytes cocultured with 200 μg/mL APS-stimulated CD11chigh CD45RBlow DC and IL-12 antibody isotype). Proliferation ability of CD4 + T lymphocytes was determined with MTT method.IL-4 level as well as IFN-γ level in CD4 + T lymphocyte culture supernatant was determined by flow cytometry. Data were processed with one-way analysis of variance. Results Compared with those in control, the expressions of CD 11 chigh CD45 RBlow DC surface molecules ( except for CD86 ) on CD 11 chigh CD45RBlow DC surface, as well as IL-12-secreting level with dose-dependence were increased in cells stimulated with 50,100, 200 μg/mL APS. Proliferation ability of CD4 +T lymphocytes in high-dose APS stimulation group was higher as compared with that in non-stimulation group ( F = 13. 438, P ＜0.05). IFN-γlevel in high-dose APS stimulation group [(2784 ± 137 ) pg/mL] was higher than that in non-stimulation group [(1952 ±101 ) pg/mL, F = 12. 177, P ＜0.05]. IL-4 level in high-dose APS stimulation group was (172 t 20) pg/mL,which was lower than that in non-stimulation group [( 193 ± 19) pg/mL, F = 11.963, P ＜0.05]. Proliferation ability of CD4+ T lymphocytes, IFN-γ level, and IL-4 level in high-dose APS stimulation + antibody 1 group were all ameliorated when compared with those in non-stimulation group; while levels of the 3 indexes in high-dose APS stimulation + antibody 2 group were similar to those in high-dose APS stimulation group.Conclusions APS can activate IL-12-producing CD11 chighCD45RBlowDC, and further induce the activation of immune function of T lymphocyte with shifting of Th2 to Th1 in vitro. APS can enhance the immune response via promoting the phenotypic and functional maturation of CD11 chighCD45RBlow DC.     

Sema6D与受体PlexinA1在胃癌中的表达及其临床病理意义

目的 探讨Sema6D及其受体PlexinA1在胃癌中的表达及它们与肿瘤细胞增殖和血管生成的关系.方法 应用逆转录.聚合酶链反应(RT-PCR)和Western blot方法检测20例胃癌患者的癌组织及相应胃切缘正常胃黏膜Sema6D及其受体PlexinA1的mRNA和蛋白表达;免疫组织化学方法检测50例胃癌组织和20例胃正常黏膜中Sema6D、PlexinA1、肿瘤细胞增殖指数(Ki-67)和第Ⅷ因子(微血管密度MVD)的表达.结果 PT-PCR和Western blot示胃癌组织中的Sema6D mRNA和蛋白的表达明显高于胃正常黏膜[(0.24±0.06)比(0.19±0.07),P＜0.05,和(0.45±0.16)比(0.29±0.08),P＜0.01];同时PlexinA1 mRNA和蛋白的表达亦明显高于胃正常黏膜[(0.71±0.37)比(0.60±0.25),P＜0.05,和(0.47±0.16)比(0.21±0.08),P＜0.01].肿瘤细胞增殖指数(Ki-67)随着Sema6D和PlexinA1表达的增高而增高(r=0.5996,P＜0.01和r=0.5024,P＜0.05);胃癌组织中MVD与Sema6D和PlexinA1存在明显正相关(r=0.5759,P＜0.01和r=0.7286,P＜0.01).结论 Sema6D及其受体PlexinA1在胃癌发生发展中发挥重要作用,与促进肿瘤细胞增殖和调节血管生成有关.     

K252a通过下调脊髓背角钾氯共转运体-2的表达缓解大鼠术后持续性痛

目的 观察鞘内注射酪氨酸激酶受体B(tyrosine kinase receptor B,TrkB)抑制剂K252a对皮肤/肌肉切口牵拉术(skin/muscle incision and retraction,SMIR)诱发的术后持续性痛大鼠脊髓背角钾氯共转运体-2(K+-Cl-cotransporter 2,KCC2)蛋白表达的影响. 方法 采用随机数字表法将52只成年雄性SD大鼠随机分成假手术组(对照组)、SMIR组、SMIR+二甲基亚砜(dimethyl sulfoxide,DMSO)组和SMIR+K252a组,每组13只.于术前1d及术后3、7、12、22、32 d测定大鼠机械缩足反射阈值(mechanical withdrawal threshold,MWT),Western blot测定术后7d大鼠脊髓背角KCC2蛋白的表达. 结果 与对照组比较,术后7、12、22 d时MWT在SMIR组[(22.5±2.3)、(24.9±1.4)、(29.5±2.4)g]和SMIR+DMSO组[(24.0±1.9)、(24.8±2.3)、(26.7±2.1)g]明显降低(P＜0.05);与SMIR组比较,术后7、12、22 d时MWT在SMIR+K252a组[(31.6±1.7)、(36.1±2.0)、(38.1±2.1)g]明显上调(P＜0.05).与对照组比较,术后7d时SMIR组和SMIR+DMSO组的脊髓背角KCC2表达明显下调(P＜0.05),SMIR+K252a组未检测到明显变化(P＞0.05).与SMIR组比较,SMIR+K252a组的脊髓背角KCC2表达明显上调(P＜0.05).结论 脊髓背角KCC2的表达变化可能参与了大鼠术后持续性痛的形成.     

羟苯磺酸钙对慢性马兜铃酸肾病大鼠肾组织CD34和血管性血友病因子表达的影响

目的 观察羟苯磺酸钙对慢性马兜铃酸肾病(CAAN)大鼠肾小管周毛细血管内皮细胞特异抗原CD34和血管性血友病因子(vWF)表达的影响,探讨羟苯磺酸钙改善CAAN大鼠模型肾脏微循环障碍的作用和机制.方法 关木通水煎剂灌胃12周制作马兜铃酸肾病大鼠模型,随机分成未治疗组和治疗组.未治疗组(n=8)予饮用水灌胃4周;治疗组(n=8)予羟苯磺酸钙灌胃4周.另设健康对照组(n=8).实验第16周处死所有大鼠,留取血、尿、肾组织标本行生化、病理及免疫组织化学检查.结果 与未治疗组比较,治疗组肾功能明显改善,肾间质纤维化程度减轻[(38.22±5.17)×103比(69.97±17.69)×103,P＜0.01],CD34阳性表达显著增加[(16.72±4.17)×103比(3.19±1.40)X103,P＜0.01];vWF阳性表达显著减少[(10.16±1.67)×103比(18.66±4.65)×103,P＜0.01)].结论 羟苯磺酸钙能增加CAAN大鼠肾组织CD34的阳性表达,增加肾小管周围毛细血管密度;减少CAAN大鼠肾组织vWF的阳性表达,减少微血栓形成.     

Nod样受体蛋白3炎性体抑制剂格列苯脲对机械通气致小鼠急性肺损伤的保护作用

目的 探究Nod样受体蛋白3(Nod-like receptor protein-3,NLRP3)炎性体抑制剂格列苯脲对机械通气导致的小鼠急性肺损伤是否具有保护作用. 方法 28只7～9周的清洁级ICR雄性小鼠,按完全随机分组法分为4组:对照组(CON组,6只)、格列苯脲组(GLY组,6只)、机械通气组(VEN组,8只)和格列苯脲+机械通气组(GLY+VEN组,8只).VEN组和GLY+VEN组机械通气4h后与CON组及GLY组麻醉插管后4h测定肺泡灌洗液中蛋白含量及炎性细胞数量,测量肺组织湿/干重比(wet/dry,W/D),观察肺组织病理学改变,ELISA法检测肺组织IL-1β、IL-6、TNF-α的含量. 结果 VEN组肺泡灌洗液中蛋白浓度和细胞数量[(0.534±0.104) g/L和(3.4±0.7)×105/ml]比CON组[(0.167±0.021) g/L和(1.9±0.5) ×105/ml]升高(P＜0.01);GLY+VEN组肺泡灌洗液中蛋白浓度和细胞数量[(0.425±0.083) g/L和(2.4±0.6) ×105/ml]比VEN组下降(P＜0.05).VEN组肺组织W/D(5.1±0.5)与CON组(4.4±0.4)比较,差异有统计学意义(P＜0.01),GLY +VEN组肺组织W/D(4.7±0.4)与VEN组比较,差异有统计学意义(P＜0.05).VEN组和GLY+VEN组肺组织中IL-1β、IL-6和TNF-α蛋白表达与CON组比较,明显升高(P＜0.05),GLY+VEN组IL-1β和IL-6表达与VEN组比较,表达明显降低(P＜0.05).结论 机械通气前给予格列苯脲可有效减少小鼠肺组织炎症细胞聚集,减轻肺水肿,机制可能与其抑制炎性体的激活有关.