白细胞介素2基因转染细胞因子诱导的杀伤细胞对恶性黑素瘤细胞的杀伤作用

目的 探讨白细胞介素2(IL-2)基因转染细胞因子诱导的杀伤细胞(CIK)对恶性黑素瘤的杀伤能力.方法 提取小鼠脾细胞,分离淋巴细胞,培养CIK细胞,用携带IL-2的质粒PEGF-N1-IL-2转染CIK细胞,荧光显微镜观察质粒转染情况,反转录-聚合酶链反应(RT-PCR)鉴定IL-2基因的表达.将效应细胞(CIK细胞或IL-2转染CIK细胞)和靶细胞(B16黑素瘤细胞)分别按照效靶比10∶1、20∶1和40∶1混合培养,采用4h乳酸脱氢酶释放测定法,检测两种CIK细胞对B16细胞的细胞毒活性.按照效靶比40∶1混合,采用酶联免疫吸附测定法(ELISA)检测两种CIK细胞IL-2、干扰素γ(IFN-γ)和肿瘤坏死因子α(TNF-α)水平.建立小鼠黑素瘤模型,将28只模型小鼠平均分为4组:对照组(瘤旁注射0.2 ml生理氯化钠溶液)、IL-2组(瘤旁注射100 IU IL-2)、CIK组(瘤旁注射细胞数约1×106 CIK细胞悬液)、IL-2转染CIK组(瘤旁注射细胞数约1×106 IL-2转染CIK细胞悬液),通过肿瘤形态学和抑瘤率、细胞凋亡率来评价荷瘤小鼠肿瘤生长情况.两组正态分布计量资料的比较行t检验,多组计量资料的比较采用方差分析,两两间多重比较采用LSD-t检验.结果 荧光显微镜及RT-PCR均显示IL-2转染CIK细胞成功.效靶比实验显示,40∶1时IL-2转染CIK细胞对B16细胞的毒性最强,IL-2转染CIK细胞组分泌IL-2 (1107.26±6.49 pg/ml)、IFN-γ(50.01±3.35 pg/ml)和TNF-α(39.86±3.25 pg/ml)的能力明显高于CIK细胞组(分别为51.09±3.85、32.71±2.43、30.11±3.08 pg/ml),两组比较,t值分别为442.60、14.93和6.89,差异均有统计学意义(P＜0.01).动物实验显示,与干预前相比,干预后对照组小鼠肿瘤体积明显增大(P＜0.05),而IL-2组、CIK组和IL-2转染CIK组小鼠肿瘤体积明显减小(P＜ 0.001),且IL-2转染CIK组肿瘤体积显著小于其他3组(均P＜0.01),但IL-2组和CIK组差异无统计学意义(P＞0.05).CIK组、IL-2组和IL-2转染CIK组的细胞凋亡率均显著大于对照组(P＜0.01),IL-2转染CIK组的细胞凋亡率及抑瘤率均显著大于IL-2组和CIK组(P＜ 0.01),而IL-2组和CIK组细胞凋亡率及抑瘤率差异无统计学意义(P＞0.05).结论 IL-2转染CIK细胞对恶性黑素瘤有更强的杀伤作用.     

Evaluation of D-dimer serum levels among patients with chronic urticaria,psoriasis and urticarial vasculitis

BACKGROUND

It has been demonstrated that neutrophils, eosinophils and monocytes, under appropriated stimulus, may express tissue factor and therefore, activate the extrinsic pathway of coagulation. We performed a transversal and case-control study of patients with chronic urticaria and patients with psoriasis, in our outpatient clinic to evaluate the production of D-dimer.

OBJECTIVE

To evaluate D-dimer serum levels in patients with chronic urticaria and its possible correlation with disease activity.

PATIENTS AND METHODS

The study was conducted from October 2010 until March 2011. We selected 37 consecutive patients from our Allergy Unit and Psoriasis Unit, and divided them into three groups for statistical analysis: (i) 12 patients with active chronic urticaria (CU); (ii) 10 patients with chronic urticaria under remission and (iii) 15 patients with psoriasis (a disease with skin inflammatory infiltrate constituted by neutrophils, lymphocytes and monocytes). Another five patients with urticarial vasculitis were allocated in our study, but not included in statistical analysis. The serum levels of D-dimer were measured by Enzyme Linked Fluorescent Assay (ELFA), and the result units were given in ng/ml FEU.

RESULTS

Patients with active chronic urticaria had the highest serum levels of D-dimer (p<0.01), when compared to patients with CU under remission and the control group (patients with psoriasis).

CONCLUSIONS

Patients with active chronic urticaria have higher serum levels of D-dimer, when compared to patients with chronic urticaria under remission and patients with psoriasis. We found elevated serum levels of D-dimer among patients with urticarial vasculitis.     

荨麻疹性血管炎临床及发病机理探讨

目的探讨荨麻疹性血管炎临床特征及发病机理。方法对１６例荨麻疹性血管炎临床及实验室检查资料进行分析。结果本病符合Ⅲ型变态反应引起的免疫复合物疾病,ＩｇＧ和补体Ｃ３在发病和皮损的形成中起了重要的作用,局部组织低垂或受压至组织液体静压增大与皮损的形成亦有密切的关系。结论本病好发于成年女性,皮损以风团和环状红斑样损害为特征,四肢关节游走性疼痛和眼结膜充血为常见的表现。     

低氧诱导肾细胞癌786-O细胞HIF-1α的表达变化及意义

目的探讨缺氧条件下肾透明细胞癌786-O细胞缺氧诱导因子1α（HIF-1α）的表达变化及意义。方法786-O细胞在缺氧条件下培养不同时间（0、8、16、24h）后,RT-PCR法检测HIF-1α mRNA的转录情况;Western blot法检测HIF-1α蛋白表达变化。结果常氧状态下HIF-1α mRNA和蛋白均有一定表达。缺氧处理一定时间（8、16、24h）后786-O细胞HIF-1α mRNA表达没有明显改变（P〉0．05）,而HIF-1α蛋白表达水平显著升高（P〈0．01）。结论缺氧条件下786-O细胞HIF-1α表达变化主要在翻译水平而非转录水平。在蛋白水平对HIF-1α表达进行调控可作为肾透明细胞癌治疗的新靶点。     

以精神障碍为首发症状的抗mGluR5脑炎1例

本文目的是通过报道一例罕见的以精神障碍为首发症状的抗代谢型谷氨酸受体5(mGluR5)脑炎患者的临床诊疗过程,为临床识别和诊断提供参考。一例38岁的男性患者,以咽痛为前驱症状,记忆力快速下降,伴焦虑抑郁、易激惹及冲动伤人精神运动性兴奋,病程进行性加重,最严重时意识呈浅昏迷状态,认知功能进一步下降,伴幻觉妄想,自知力缺乏,脑脊液和血清抗mGluR5抗体均强阳性（1∶100）。先后两次使用激素冲击治疗后,患者意识、认知功能、情绪、精神行为等均有改善。     

基于matK序列的积雪草与其易混品的分子鉴定方法分析

目的:积雪草容易与多种混淆品相互混淆,本研究拟基于mat K序列探讨积雪草与其混淆品鉴定的新方法。方法:从Gen Bank核酸数据库下载积雪草、过路黄、连钱草和乌蔹莓的mat K序列,应用Clustal X 2.1软件进行SNP鉴别位点分析,应用MEGA5.0软件计算种内种间(K2P)遗传距离和构建邻接(NJ)系统聚类树。结果:获得积雪草与过路黄各5个mat K序列及连钱草和乌蔹莓各1个mat K序列,分析显示积雪草与其混伪品的mat K序列存在较大差异,具有近百个SNP位点,且其中多是积雪草特有的,可用于分子鉴定。且积雪草最大种内K2P遗传距离0.001,远远小于其与混伪品的种间K2P遗传距离0.242~0.293,构建的系统发育树显示积雪草与其混淆品可明显分开。结论:综合以上分析,mat K序列为鉴定积雪草及其混淆品提供了新的分子鉴定方法。     

止咳清肺口服液对25例慢性阻塞性肺疾病患者痰液炎症细胞白细胞介素8、肿瘤坏死因子α的影响

目的:观察中药止咳清肺口服液治疗慢性阻塞性肺疾病(COPD)气道炎症的效果.方法:50例稳定期COPD患者随机分成治疗组25例,对照组25例.两组氧疗、强心、利尿、止咳平喘等常规治疗相同.治疗组加止咳清肺口服液20ml,每日3次,连服6周.治疗前后记录临床症状计分,检测肺功能、痰液炎症细胞计数、分类和白细胞介素8(IL-8)、肿瘤坏死因子α(TNF-α)的变化.结果:治疗组临床症状计分、1秒钟用力呼吸容积(FEV1)和FEV1/用力肺活量(FVC)均有明显改善(P＜0.05或P＜0.01),而对照组治疗前后变化不明显.痰液中的细胞总数、中性粒细胞和IL-8、TNF-α水平比治疗前均有显著下降(P＜0.05或P＜0.01),而对照组下降不明显.结论:止咳清肺口服液治疗COPD气道炎症疗效明显.     

莪芪抗瘤方剂对白血病HL-60细胞凋亡的作用及机理研究

目的：研究中药复方莪芪抗瘤方剂在体外对白血病HL-60细胞凋亡影响，并探讨凋亡发生的可能机制。方法：通过血清药理学方法，选择10％浓度的含药血清与白血病HL-60细胞共同培育24、48、72、96h后，应用荧光显微镜观察细胞形态，流式细胞仪检测凋亡峰（亚二倍体峰），用Fura-2荧光负载方法测定细胞内Ca^2＋浓度的变化；比色法检测凋亡相关基因Caspase-3活性变化。结果：10％浓度的含药血清处理组细胞体积缩小，细胞核固缩，膜起泡，核断裂及形成凋亡小体等，以72h最为明显；可见DNA含量低于G1期的凋亡细胞群，这群细胞的凋亡百分率随时间的递增而增加。10％浓度的含药血清处理组的细胞内Ca^2＋浓度也明显高于对照组（P〈0．01），Caspase-3基因活性明显高于对照组相（P〈0．01．P〈0．05）。结论：莪芪抗瘤方剂含药血清诱导白血病HL-60细胞发生凋亡，这种变化可能与细胞内Ca^2＋水平上调，Caspase-3活性增强有关。     

Effect of blending sewage sludge with coal on combustion and ash slagging behavior

Blending sewage sludge (SS) with Zhundong coal (ZDC) for combustion in coal-fired power plants is a recent approach that can alleviate the shortage of high-quality coal resources and achieve the harmless treatment of SS, while also having a significant influence on combustion and ash slagging. Due to the high content of alkali and alkaline earth metals (AAEMs) in ZDC, its combustion ash has a strong likelihood of slagging. This study aims to investigate the effect of blending SS with ZDC on combustion and ash slagging. Thermogravimetry (TG) results indicate that blending with SS could lower the ignition and burnout temperatures of ZDC. With an increase in the ratio of sludge, the comprehensive combustion index (S) first increases and then decreases, showing that blending SS with ZDC in an appropriate proportion could improve the overall combustion. Through the analysis of the interaction, it is confirmed that SS and ZDC could complement each other during co-combustion due to their different components. X-ray fluorescence (XRF) was used to test the ash components of different blending ratios (10–30%) and combustion temperatures (800–1100 °C). Slagging indices including alkali acid ratio (B/A), silicon ratio (G), and silica–alumina ratio (SiO₂/Al₂O₃) were also calculated. The results suggest that the slagging behavior of ZDC is greatly reduced even if the blending ratio is only 10%. However, with an increase in the blending ratio, the effect on slagging gradually weakens. Considering the dual influence of SS blending on combustion and slagging, this study assumes the optimal blending ratio of 20%. Influenced by the components of the combustion ash, B/A and SiO₂/Al₂O₃ are more suitable for evaluating the slagging tendency of ash; however, there is great deviation in the results for G. This research is beneficial to coal-fired power plants for the selection of operation parameters during co-combustion with SS.

Blending sewage sludge (SS) with Zhundong coal (ZDC) for combustion in coal-fired power plants is a recent approach that can alleviate the shortage of high-quality coal resources and achieve the harmless treatment of SS, while also having a significant influence on combustion and ash slagging.