IL-1β和TNF-α诱导RA成纤维样滑膜细胞ICAM-1和VCAM-1表达调控的机制研究

目的探讨酪氨酸激酶在白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α诱导类风湿关节炎(RA)成纤维样滑膜细胞(FLS)细胞间黏附分子(ICAM-1)和血管细胞黏附分子(VCAM-1)表达中的作用。方法原代培养RA成纤维样滑膜细胞，用Western blot方法检测IL-1β和TNF-α短时间内引起RA FLS蛋白质酪氨酸磷酸化状态改变，并应用genistein，蛋白质酪氨酸激酶(PTK)抑制剂观察对ICAM-1和VCAM-1表达影响。结果IL-1β和TNF—α可以瞬时引起RA FLS蛋白质酪氨酸磷酸化程度增加：IL-1β和TNF-α在1—100U／ml之间呈浓度依赖性促进ICAM-1和VCAM-1的表达；genistein显著抑制两种炎性因子刺激下的VCAM-1表达，而对。ICAM-1的表达仅起中度抑制作用。结论IL-1β和TNF-α在RAFLS信号转导中，可以瞬时导致蛋白质酪氨酸磷酸化程度增加．在炎性因子诱导ICAM-1和VCAM-1的表达调控中蛋白酪氨酸激酶作用不同。     

重症急性呼吸综合征患者血浆细胞因子水平的动态改变

目的　探讨重症急性呼吸综合征 (SARS)患者肺部严重炎症反应的发生机制。方法　选择 2 4例本院确诊的SARS患者 ,于发病第 1、第 2、第 3～ 4周及康复出院后 1个月 (发病 8～ 9周 )收集抗凝静脉血 ,以定量ELISA法检测其血浆白细胞介素 (IL) 1β、IL 2、IL 4、IL 8、IL 10、IL 12 p70、干扰素γ(IFNγ)及肿瘤坏死因子α(TNFα)水平在病程中的改变。并选择 12例正常人作为对照。结果　数据以中位数 (四分位数间距 )表示 ,与正常对照组 [IL 8:6 2 8ng/L( 3 4 3ng/L) ;TNFα :3 77ng/L( 3 4 0ng/L) ]相比 ,所有SARS病人在发病第 1周血浆IL 8浓度明显升高 [31 2 3ng/L( 78 5 1ng/L) ],P <0 0 1,75 % ( 18/2 4 )的病人在发病 3～ 4周达到最高峰 14 9 6 5ng/L( 2 4 5 97ng/L) ,P <0 0 1,至出院后 1个月 (发病 8～ 9周 )平均血浆IL 8浓度降至 8 2 3ng/L( 8 0 7ng/L)。血浆TNFα浓度也有异常升高 ,在发病第 2周为 2 3 12ng/L( 2 6 7 33ng/L) ,P <0 0 1,发病 3～ 4周达到高峰136 35ng/L( 4 76 83ng/L) ,P <0 0 5 ,出院后 1个月下降至 94 88ng/L( 2 77 18ng/L) ,仍高于正常水平 (P <0 0 1)。其他 6种细胞因子与对照组比较 ,差异无显著性。结论　SARS病人体内发生着复杂的细胞因子网络性连锁反应 ,由此产     

宁夏枸杞对代谢综合征患者TNF-α、NF-κB和iNOS水平的影响

目的探讨枸杞干预对代谢综合征（metabolic syndrome,MS）患者血清肿瘤坏死因子α（tumor necrosis factor,TNF-α）、核因子-κB（nuclear factor-κB,NF-κB）和诱导型一氧化氮合酶（inducible nitric oxide synthase,iNOS）水平的影响。方法在横断面研究的基础上,采用病例-对照的方法,选取MS组及健康对照组各119人,比较两组间各测定指标的差异;采用随机对照的研究方法将MS组119人分为对照组和干预组,随后对干预组实施枸杞干预,运用酶联免疫吸附法（enzyme-linked immunosorbent assay,ELISA）测定两组干预0 d、干预30 d、干预45 d和干预75 d 4个时间段人群血清TNF-α、NF-κB和iNOS含量,观察枸杞的干预效果。结果代谢综合征患者血清中TNF-α、NF-κB和iNOS含量明显高于正常人（均有P〈0.05）;干预前后两组人群的TNF-α、NF-κB和iNOS变化有统计学差异（均有P〈0.05）。结论枸杞可以抑制代谢综合征患者TNF-α、NF-κB和iNOS的活化与表达。     

肿瘤化疗患者感染病原菌分布及危险因素分析

目的研究肿瘤化疗患者感染病原菌分布及其对抗菌药物的耐药率,探讨发生医院感染的危险因素,分析肿瘤化疗后预防感染对策,控制肿瘤化疗后患者的感染。方法选择2011-2013年肿瘤化疗患者768例,利用BACTECTM9000全自动培养系统分离培养病原菌,采用K-B琼脂法进行药敏试验,研究肿瘤化疗患者医院感染的危险因素,数据采用SPSS 13.0统计软件进行分析,计数资料采用χ2检验。结果 768例肿瘤化疗患者中发生医院感染86例,感染率为11.20%;感染患者病原菌检出革兰阳性菌36株占39.13%,革兰阴性菌56株占60.87%;年龄大、住院时间长、进行手术、合并有其他疾病、进行侵入性操作以及未预防性应用抗菌药物等是肿瘤化疗患者医院感染的危险因素。结论肿瘤化疗患者医院感染率高,医护人员应采取措施控制化疗患者医院感染的发生。     

Fluorescent-tilmanocept for tumor margin analysis in the mouse model

Background

Dendritic cells (DC) are localized in close proximity to cancer cells in many well-known tumors, and thus maybe a useful target for tumor margin assessment.

Materials and methods

[^99mTc]- cyanine 7 (Cy7)-tilmanocept was synthesized and in vitro binding assays to bone marrow-derived DC were performed. Fifteen mice, implanted with either 4T1 mouse mammary or K1735 mouse melanoma tumors, were administered 1.0 nmol of [^99mTc]-Cy7-tilmanocept via tail vein injection. After fluorescence imaging 1 or 2 h after injection, the tumor, muscle, and blood were assayed for radioactivity to calculate percent-injected dose. Digital images of the tumors after immunohistochemical staining for DC were analyzed to determine DC density.

Results

In vitro binding demonstrated subnanomolar affinity of [^99mTc]-Cy7-tilmanocept to DC (K_A = 0.31 ± 0.11 nM). After administration of [^99mTc]-Cy7-tilmanocept, fluorescence imaging showed a 5.5-fold increase in tumor signal as compared with preinjection images and a 3.3-fold difference in fluorescence activity when comparing the tumor with the surgical bed after tumor excision. Immunohistochemical staining analysis demonstrated that DC density positively correlated with tumor percent of injected dose per gram (r = 0.672, P = 0.03), and higher DC density was observed at the periphery versus center of the tumor (186 ± 54 K versus 64 ± 16 K arbitrary units, P = 0.001).

Conclusions

[^99mTc]-Cy7-tilmanocept exhibits in vitro and in vivo tumor-specific binding to DC and maybe useful as a tumor margin targeting agent.     

All-trans-retinoic acid counteract the tumor-stimulating effect of hepatectomy and increases survival of rats bearing liver metastases

Background

We previously demonstrated a stimulating effect of hepatectomy on residual tumor cells after resection of liver metastases. The aim of this study was to analyze the effect of all-trans-retinoic acid (ATRA) on the protumor effect of hepatectomy and survival of hepatectomized rats bearing liver metastases. We also explored whether ATRA interfered with the tumor promoting effect of hepatotropic growth factors (GFs).

Methods

The in vitro effect of ATRA on proliferation of S4MH rhabdomyosarcoma tumor cells was assessed when cultured with laparotomized or hepatectomized rat serum (HRS), or in the presence of GFs (hepatocyte growth factor, insulin growth factor 2, Platelet Derived Growth Factor (PDGF)-BB, and vascular endothelial growth factor). For the in vivo studies, rats were partially hepatectomized on day 10 after metastasis induction, one group being treated with ATRA from day 7 to 14, and a second receiving cyclophosphamide (CY; on days 10 and 14) alone or with ATRA. We determined the size and number of liver and lung metastases. Finally, we analyzed the effect of treatments on rat survival.

Results

Hepatotropic GFs increased cell proliferation in a similar manner to HRS. In vitro, ATRA blocked the protumor effect of both HRS and GFs. In vivo, ATRA reduced the size and number of liver and lung metastases, and significantly increased rat survival. Furthermore, adding ATRA to CY significantly increased survival compared with CY alone.

Conclusions

In our model, ATRA minimizes the tumor-stimulating effect of hepatectomy, reducing the number and size of liver metastases and improving survival. The results suggest that the ATRA may be useful for blocking the growth-promoting effect of hepatotropic GFs released after liver metastasis resection.     

Inter-rater reliability of surgical reviews for AREN03B2: A COG renal tumor committee study

Purpose

The Children's Oncology Group (COG) renal tumor study (AREN03B2) requires real-time central review of radiology, pathology, and the surgical procedure to determine appropriate risk-based therapy. The purpose of this study was to determine the inter-rater reliability of the surgical reviews.

Methods

Of the first 3200 enrolled AREN03B2 patients, a sample of 100 enriched for blood vessel involvement, spill, rupture, and lymph node involvement was selected for analysis. The surgical assessment was then performed independently by two blinded surgical reviewers and compared to the original assessment, which had been completed by another of the committee surgeons. Variables assessed included surgeon-determined local tumor stage, overall disease stage, type of renal procedure performed, presence of tumor rupture, occurrence of intraoperative tumor spill, blood vessel involvement, presence of peritoneal implants, and interpretation of residual disease. Inter-rater reliability was measured using the Fleiss' Kappa statistic two-sided hypothesis tests (Kappa, p-value).

Results

Local tumor stage correlated in all 3 reviews except in one case (Kappa = 0.9775, p < 0.001). Similarly, overall disease stage had excellent correlation (0.9422, p < 0.001). There was strong correlation for type of renal procedure (0.8357, p < 0.001), presence of tumor rupture (0.6858, p < 0.001), intraoperative tumor spill (0.6493, p < 0.001), and blood vessel involvement (0.6470, p < 0.001). Variables that had lower correlation were determination of the presence of peritoneal implants (0.2753, p < 0.001) and interpretation of residual disease status (0.5310, p < 0.001).

Conclusion

The inter-rater reliability of the surgical review is high based on the great consistency in the 3 independent review results. This analysis provides validation and establishes precedent for real-time central surgical review to determine treatment assignment in a risk-based stratagem for multimodal cancer therapy.     

紧密连接蛋白1与重症急性胰腺炎大鼠胰腺微血管损伤关系的实验研究

目的研究重症急性胰腺炎（severe acute pancreatitis,SAP）大鼠紧密连接蛋白1（ZO-1）随时间的变化及其与SAP微血管损伤和胰腺组织病理学评分的关系。方法将48只Wistar大鼠随机均分为假手术组（SO组）和SAP组。SO组大鼠开腹后仅翻动胰腺;SAP组大鼠开腹后,以逆行胰胆管微泵注射5%牛磺胆酸钠法制备SAP模型。2组大鼠分别于手术后6、12及24 h各处死8只大鼠,取腹主动脉血测定外周血中淀粉酶、胰蛋白酶、白介素-8（IL-8）、肿瘤坏死因子-α（TNF-α）及ZO-1蛋白水平;取胰腺组织行HE染色,观察其病理学变化并评分;同时行免疫组化染色检测ZO-1蛋白的表达情况。结果同时点与SO组比较,各时点SAP组的血清淀粉酶、胰蛋白酶、IL-8、TNF-α及ZO-1蛋白水平均较高（P〈0.05）。SAP-6 h组和12 h组的血清淀粉酶水平均低于24 h组（P〈0.05）;SAP组内大鼠的胰蛋白酶、IL-8和ZO-1蛋白水平均随时点延长逐渐升高,各时点组间两两比较差异均有统计学意义（P〈0.05）;SAP组内3个时点组间TNF-α水平的差异无统计学意义（P〉0.05）。多重线性回归模型结果显示,SAP组血清ZO-1蛋白水平与胰腺组织病理学评分（b=0.96,P〈0.05）、血清淀粉酶水平（b=0.87,P〈0.05）、胰蛋白酶水平（b=0.72,P〈0.05）及IL-8水平（b=0.69,P〈0.05）均呈正相关,而与TNF-α水平的关系无统计学意义（P〉0.05）。HE染色结果显示,各时点SAP组大鼠的胰腺组织损伤程度重于SO组,且随时间延长,SAP大鼠胰腺组织的病理学损伤加重;SAP-12 h组和24 h组的胰腺组织病理学评分均高于6 h组（P〈0.05）。SP免疫组化染色结果显示,随时间延长,SAP组大鼠胰腺腺泡细胞间及毛细血管壁的ZO-1蛋白颗粒数量减少,且在毛细血管中的表达不连续。结论在SAP的进程中,血清中ZO-1蛋白的水平上升,同时其在胰腺组织中的表达呈下调趋势,表明其与SAP时的胰腺微血管损伤有关。     

Changes of serum Tau, GFAP, TNF-α and malonaldehyde after blast-related traumatic brain injury

Objective： To determine the changes of serum Tau protein, glial fibrillary acidic protein （GFAP）, tumor necrosis factor alpha （TNF-α）, and malonaldehyde （MDA） in rats after blast-related traumatic brain injury （BTBI） and to provide relative information for further studies on BTBI mechanism and seek specific biomarkers for BTBI. Methods： Ninety male Sprague-Dawley rats were randomly assigned into three groups： control group, moderate blast injury group, and severe blast injury group （n=30 for each）. Rats in the moderate and severe blast injury groups were respectively exposed to corresponding levels of BTBI. After explosion, serum levels of Tau, GFAP, TNF-α, and MDA in each group were determined by Elisa assay at different time points after injury （8 h, 24 h, 3 d, and 6 d）. The extent of brain damage was detected by Nissl staining and TUNEL assay. Results： Serum levels of Tau and GFAP rapidly increased and reached the peak at 24 h after either moderate or severe blast injury. All the values were significantly higher than control group at all time points （P〈0.05）. Serum TNF-α level of both injury groups peaked at 8 h after BTBI and stayed significantly higher than control group at all time points （P〈0.05）. Serum MDA of two injury groups began to significantly increase at 3 d and the level stayed significantly higher than control group until 6 d （P〈0.05）. Moreover, unlike the other biomarkers, serum MDA of severe blast injury group was significantly higher than moderate blast injury group at 6 d （P〈0.05）. Conclusion： The changes of serum Tau, GFAP, and TNF-α showed a good sensitivity at the acute phase after BTBI （within 24 h）. However, their specificity and correlation with the extent of injury were limited in this experiment. Moreover, although the change of serum MDA showed a poor sensitivity and specificity to the diagnosis of BTBI during the first few days, it can reflect the injury degree at 6 d after injury. Therefore, further studies are nee     

下咽癌中差异表达的蛋白激酶及抑制剂的研究进展

在头颈恶性肿瘤中,下咽癌恶性程度高,患者生存质量较差且通常预后不良。下咽癌的治疗采用手术为主的方式,并辅助以放化疗。术后患者喉功能受损严重,寻找能够尽量保护喉功能的治疗方法是目前研究的重点。靶向治疗对下咽癌患者保护喉功能、提高生存质量意义重大。研究发现,下咽癌发生过程中涉及多种蛋白激酶介导的信号通路及基因,对可能在这一过程中有重要意义的蛋白激酶PKC-β、CDK4/6、Cdc42、Rac1、STK33、CK2和PHLPP作一综述,以期为蛋白激酶抑制剂作为下咽癌治疗的靶点提供新的方向。