Influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells of asthmatic rats

Objective： To investigate the influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells（CD4^＋CD25^＋ Tr） of asthmatic rats, and elucidate the possible mechanism of Danshen Injection in treatment of asthma. Methods： 30 Wister rats were randomly divided into control group, asthma group and Danshen Injection treated group. Bronchoalveolar lavage fluids （BALF） were collected, and cytology studies were conducted. Lung tissues were obtained and pathologic analyses were done with hematoxylin and eosin stain （HE）. Flow cytometry was used to detect the CD4^＋CD25^＋ Tr ratio in peripheral blood mononuclear cells （PBMCs）. Results： Total cell, the percentage of lymphocytes, neutrophils and eosinophils （Eos） in BALF of Danshen Injection-treated group were lower than that in asthma group （P〈0.05, P〈0.01）. Compared with asthma group, less infiltration of inflammatory cells in lung tissues was observed in Danshen Injection-treated group. CD4^＋CD25^＋ Tr of asthma group was lower than that of control and Danshen Injection treated group （P〈0.05）. Conclusion： Danshen Injection can suppress airway inflammation of asthmatic rats, probably by increasing the number of CD4^＋CD25^＋ Tr.     

Four-dimensional sonographic assessment of fetal facial expression early in the third trimester.

OBJECTIVE: To evaluate the characteristic patterns of facial expression in fetuses aged from 28 to 34 weeks using 4-dimensional (4-D) ultrasonography. METHODS: The faces of 10 healthy fetuses aged from 28 to 34 weeks were recorded continuously for 15 min with a 4-D ultrasonographic machine performing up to 25 frames per second. The occurrence rates of blinking, mouthing, yawning, tongue expulsion, smiling, scowling, and sucking were evaluated. RESULTS: Mouthing was the most frequent facial expression (median, 6.5; range, 2-19) whereas the least frequent were scowling (median, 1; range, 0-9) and sucking (median, 1; range, 0-2). Mouthing was evident in all fetuses and significantly more frequent than any other movement (P<.05). Yawning (median, 3; range, 0-6), smiling (median, 2; range, 0-9), and blinking (median, 1.5; range, 0-6) were observed in most cases. Tongue expulsion (median, 1.5; range, 0-5), scowling, and sucking were each observed in 6 cases. CONCLUSION: 4-D sonography provides a means of evaluating fetal facial expression early in the third trimester. It may be a key to predicting fetal brain function and well-being and an important modality in future fetal neurophysiologic research.     

Expression of CD28 and cytotoxic T lymphocyte antigen 4 at the maternal-fetal interface in women with unexplained pregnancy loss.

OBJECTIVE: To investigate the expression of CD28 and CTLA-4, two costimulatory molecules involved in T-cell activation at the maternal-fetal interface in women with unexplained pregnancy loss. METHODS: A total of 57 women, 39 with unexplained spontaneous abortion (SA) and 18 with unexplained recurrent spontaneous abortion (RSA), were enrolled in the study. A high-resolution spectratyping analysis of fluorescent bands was performed to detect CD28 and CTLA-4 expression in decidual tissues. RESULTS: The mean expression of CTLA-4 mRNA was significantly higher in women with SA than in controls (P<0.05). Moreover, the expression of CTLA-4 was higher in SA patients with genotype AA and phenotype A+ (AA+AG) than in controls (P<0.01). The expression of CTLA-4 was not significantly different in patients with RSA and in controls. The expression of CD28 was significantly decreased in patients with RSA (P<0.01) and SA (P<0.05) compared with controls. The mean ratios of CTLA-4/CD28 were significantly higher in patients with RSA (P<0.01) and SA (P<0.05) than in controls. CONCLUSIONS: This study suggests that an imbalance in CTLA-4/CD28 expression or suppressed T-cell activity at the maternal-fetal interface may confer susceptibility to unexplained pregnancy loss.     

抵抗素及其结合多肽对大鼠骨骼肌细胞糖摄取功能的影响

目的探讨抵抗素及其结合多肽(RBP)对大鼠骨骼肌细胞糖摄取功能的影响。方法体外培养的大鼠骨骼肌细胞(L6)随机分为对照组,抵抗素(10 nmol/L)组,RBP(1 nmol/L)组、抵抗素(10 nmol/L) RBP(1 nmol/L)组,培养0.5 h后,液闪计数法(LSC)观察L6细胞糖摄取功能的变化,RT-PCR、Western blot法检测葡萄糖转运载体(GLUT)4基因的表达及转位变化。结果与对照组相比,抵抗素组糖摄取功能下降,RBP组及抵抗素 RBP组糖摄取功能无明显变化(P<0.05);与抵抗素组相比,抵抗素 RBP组糖摄取功能明显上调(P<0.05)。4组L6细胞的GLUT4基因的蛋白表达及转位均无明显变化(P>0.05)。结论抵抗素能抑制骨骼肌细胞的糖摄取;但对正常骨骼肌细胞糖摄取功能无明显影响;RBP但能有效拮抗抵抗素抑制骨骼肌细胞糖摄取的作用。     

哮喘患儿血清白细胞介素-4、12与细胞免疫的相关性

目的探讨不同病期哮喘患儿血清IL-12、IL-4水平、细胞免疫功能及其相互关系。方法随机选择哮喘患儿50例。婴幼儿哮喘13例,儿童哮喘37例。发作期30例,经吸入激素治疗后缓解期患儿20例。健康对照组22例。IL-12、IL-4测定采用酶联免疫双抗体夹心法(ELISA)。采用碱性磷酸酶抗碱性磷酸酶桥联酶标试验(APAAP)检测T细胞亚群,采用郭氏法检测红细胞免疫功能。结果1.哮喘患儿发作期、缓解期和对照组IL-12水平分别为(24.44±13.26)、(42.30±12.65)、(44.68±28.28)ng/L,3组比较有显著性差异(F=8.92P<0.01),发作期明显低于其他两组,缓解期与对照组无显著性差异。2.IL-4在发作期、缓解期、对照组阳性率分别为70%、35%、9%,发作期明显高于缓解期(χ2=5.96P<0.05),缓解期仍高于对照组(χ2=4.17P<0.05)。3.哮喘患儿与健康儿童相比,CD3(t=3.18P<0.01)、CD4(t=5.51P<0.01)、CD8(t=10.38P<0.01)均降低,CD4/CD8升高(t=4.28P<0.01),RBC-C3bR降低(t=4.65P<0.01),RBC-IC增高(t=10.22P<0.01)。4.IL-4与IL-12呈显著负相关(r=-0.43P<0.05),与RBC-IC呈显著正相关(r=0.49P<0.05)。结论哮喘患儿存在Th1/Th2类细胞因子失衡和细胞免疫功能紊乱状态,哮喘患儿缓解期IL-12水平恢复至接近健康儿童,IL-4水平仍高于健康儿童。应坚持长期抗感染治疗。     

黄芪对哮喘大鼠模型支气管肺泡灌洗液白细胞介素-4、-12表达的影响

目的研究哮喘大鼠模型支气管肺泡灌洗液（BALF）白细胞介素-4（IL-4）和IL—12水平变化及黄芪对其影响。方法雄性SD大鼠30只随机分为正常对照组、哮喘组和黄芪组（AM组）。以卵蛋白（OVA）致敏激发法制备大鼠哮喘模型．对PALF进行嗜酸性粒细胞（EOS）计数和分类计数；应用双抗体夹心酶联免疫吸附试验法测定BALF中IL-4、-12水平。结果1．哮喘组BALF细胞总数、EOS绝对值和EOS占细胞总数百分比（EOS％）均高于对照组（P均〈0．01），AM组上述指标均低于哮喘组（P均〈0．01）；2．BALF中IL-4水平哮喘组显著高于对照组（P〈0．01），AM组显著低于哮喘组（P〈0．01）；3．IL-12水平哮喘组显著低于对照组（P〈0．01），AM组最著高于哮喘组（P〈0．01）。结论IL-4高表达、IL-12低表达参与哮喘的病理生理过程；黄芪下调IL-4、上调IL-12表达水平，可能为其抑制哮喘呼吸道炎症的重要作用机制之一。     

Brn-4在偏侧帕金森病大鼠模型纹状体中的表达变化

目的:探讨Brn-4在偏侧帕金森病大鼠模型纹状体中的表达变化。方法:用6-OHDA注入80只SD大鼠右侧前脑内侧束(medialforebrainbundle,MFB),行为学检测得成功PD模型48只,随机分成7d、14d、21d、28d4组,每组各12只。每组中的6只动物和6只正常对照大鼠的纹状体进行Brn-4Westernblot检测;另6只动物的纹状体进行Brn-4免疫组化染色。结果:Westernblot检测发现Brn-4的相对表达量(Brn-4/β-actin)正常对照组为0.123±0.02,7d组为0.538±0.04,14d组为0.192±0.05,21d组为0.122±0.02,28d组为0.084±0.02;而模型侧/正常侧纹状体Brn-4阳性细胞的比值7d、14d、21d和28d组分别为1.560±0.14,0.855±0.15,0.638±0.11和0.369±0.10。结论:PD大鼠模型纹状体中Brn-4蛋白于前脑内侧束注射6-OHDA后7d表达量急剧增高,而14d后则急剧下降,到28d时已低于正常水平。     

磁酶免系统测定血清FT3、FT4及其临床应用

血清游离3，5，3’-三碘甲状腺氨酸（FT3）、游离四碘甲状腺氨酸（FT4）是甲状腺疾病诊断与疗效评定的重要指标。在国内，一般采用放射免疫法（RIA）检测，其优点是敏感可靠，缺点为斌剂受同位素半衰期的限制，有效期短，放射性辐射危害；磁分离酶联免疫测定系统（SEROZYME）检测血清FT3、FT4，是一种超微量的非放射性物质标记的免疫化学定量分析技术。本文用磁酶免法检测血清FT3、FT4，并与放射免疫法对比研究，现将结果报告如下。     

咪喹莫特治疗哮喘作用机制的实验研究

目的：观察咪喹莫特对小鼠哮喘模型气道炎症和肺组织n2型胸腺和活化调节趋化因子（thymus and activation regulated chemokine，TARC）和其受体CCR4表达的影响。方法：建立哮喘模型．自第14天雾化吸入OVA前0．5h．干预组分别雾化吸入咪喹莫特30min及腹腔注射地塞米松。OVA雾化结束后24h，每组小鼠眼球摘除采血收集血清，采用酶联免疫吸附法测定血清中TARC水平；收集肺泡灌洗液进行细胞计数和分类；HE染色观察肺组织炎症改变；用免疫组化方法检测肺组织CCR4和TARC的表达；用逆转录．聚合酶链反应半定量检测肺组织IL-4、IFN-γ、CCR4和TARC mRNA表达水平。结果：①哮喘小鼠HE染色可见气道上皮不完整，黏膜下有大量的炎细胞浸润，以淋巴细胞为主。地塞米松组和咪喹莫特组炎症程度减轻。②哮喘组小鼠支气管肺泡灌洗液（BALF）中细胞总数及各种炎症细胞较正常组显著增加。眯喹莫特治疗组的嗜酸性粒细胞、单核细胞租淋巴细胞比哮喘组明显减少。③哮喘组小鼠血清TARC水平较正常组升高，咪喹莫特治疗组小鼠血TARC水平较哮喘组降低，与正常组比较差异无显著性。④CR4和TARC均在气道上皮细胞表达．咪喹莫特和地塞米松治疗组能减少CCR4和TARC的表达。⑤哮喘组小鼠肺组织IL-4、CCR4和TARC mRNA表达较正常组增强。咪喹莫特和地塞米松治疗组小鼠肺组织IL-4、CCR4和TARCmRNA表达较哮喘组降低，比正常组表达增加。结论：雾化吸入咪喹莫特可在一定程度上减轻哮喘小鼠的气道炎症。抑制肺组织CCR4和TARC蛋白和mRNA的过度表达。