Effect of IFNα-2a on Fas expression and apoptosis rate of peripheral blood cytotoxic T cells in patients with hepatitis B

Interferon(IFN) with antiviral and im-munomodulatory activities is one of the most important therapeutic agents for the treatment of chronic hepatitis. The apoptotic effect of IFN is influenced by cell type and the types of IFN, which suppresses proliferation and induces apoptosis in some cell types while inhibiting apoptosis in others. The aim of this study was to explore the effect of IFNα-2a on Fas expression and the apoptosis rate of peripheral blood cytotoxic T cells (CTLs) in patients with hepatitis B. METHODS:Peripheral blood mononuclear cells were isolated from 26 patients with hepatitis B including 16 patients with chronic hepatitis B and 10 patients with chronic severe hepatitis B. Fas expression and apoptosis rate of CTLs were analyzed with flow cytometry before and after IFNα-2a treatment. RESULTS:Before IFNα-2a treatment, Fas expression and apoptosis rate of CTLs from patients with chronic hepatitis B were significantly higher than those from patients with chronic severe hepatitis B and healthy controls respectively. No significant difference was observed between Fas expression and apoptosis rate of CTLs from patients with chronic severe hepatitis B and healthy controls. After IFNα-2a treatment,Fas expression and apoptosis rate of CTLs from different groups were compared with those before IFNα-2a treatment, showing no significant difference despite alternation of different degree. CONCLUSIONS:Activation induced cell death (AICD) exists in peripheral blood CTLs from patients with hepatitis B. No effect of IFNα-2a exerts on Fas expression and apoptosis rate of Fas in patients with hepatitis B.

SYNERGIC CYTOTOXICITY TO GASTRIC CANCER CELLS BY COMBINED USE OF TRICHOSANTHIN ANDRECOMBINANT INTERFERON α-2B

Objective To investigate a new approach of the combined use of trichosanthin (TCS) andrecombinant interferon alpha - 2b (rIFN α- 2b) against digestive system cancer cells. Methods Detect separatelythe cytotoxicity of TCS, rIFN α- 2b and their combination against digestive system cancer cell SGC- 7901.Results In the experiment in vitro, TCS, rIFN α- 2b both had direct, dose dependent cytotoxicity againstSGC - 7901. Their combined use demonstrated a toxicity signijicantly higher than that of the two drugs used alone,showing a signilicant synergic effect. This synergic cytotoxicity was confirmed in the animal experiment.Conclusion Combined use of TCS and rIFN α - 2b decreases the therapeutic dose of TCS and its toxic adverseellect, and this synergic effect is favorable to the clinical use of TCS protein against gastric cancer.

Effects of the Cytotoxic T-Cells on the Dynamics of Co-Infection of HIV-1 and Mycobacterium tuberculosis

Enhancement of the Human Immunodeficiency Virus (HIV) specific cytotoxic T-cells mechanisms in an HIV-1 and Mycobacterium tuberculosis (Mtb) co-infected individual seems to improve the clinical picture of an individual by reducing Acquired Immuno Deficiency Syndrome (AIDS) state progression rate. In this paper, we develop a system of deterministic differential equations representing the immune cells involved in an HIV-1 and Mtb co-infected individual. Results show that although the non-lytic arm of the HIV-1 cytotoxic T-cells affects the co-infection dynamics more than the lytic factors, a combination of both factors results in a more positive reduced progression to the AIDS state. This is due to the increased protection of the CD4+ T-cells by the CTL mechanisms by further reducing infections and replications by the HIV. Thus, HIV-1 specific CTLs mechanisms’ involvement is here recommended to be part of a solution to the HIV and Mtb co-infection problems.

肥胖儿童non-HDL-C、remnant cholesterol与亚临床动脉粥样硬化的关系

目的:了解non-HDL-C和remnant cholesterol与肥胖儿童亚临床动脉粥样硬化(AS)的关系。方法:根据甘油三酯(TG)水平将65例肥胖儿童(肥胖组)分为I组(TG≥2.3 mmol/L)32例,II组(TG<2.3 mmol/L)33例,与肥胖组儿童性别年龄相匹配的正常体质量儿童30例为III组(对照组)。比较3组儿童血脂指标,对肥胖组各项血脂指标与血浆致动脉粥样硬化指数(AIP)进行相关性分析,肥胖患儿体质指数、血脂指标与颈动脉内径、内膜-中层厚度(c-IMT)、血流参数进行相关性分析。结果:I组、II组的TG、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、non-HDL-C、remnant cholesterol、AIP均较对照组升高,高密度脂蛋白胆固醇(HDL-C)较对照组降低,且I组的remnant cholesterol、AIP均较II组升高。肥胖组儿童AIP与TG、non-HDL-C、remnant cholesterol呈正相关,与HDL-C呈负相关。37例行颈动脉超声检查的肥胖儿童的c-IMT较正常儿童的增厚。结论:non-HDL-C、remnant cholesterol在肥胖儿童亚临床AS中有重要作用,在肥胖儿童血脂异常的治疗中应关注non-HDL-C、remnant cholesterol的变化。

Shenqi Fuzheng injection combined with GP chemotherapy in the treatment of advanced non-small cell lung cancer： a meta-analysis

Objective： To evaluate the clinical efficacy of Shenqi Fuzheng injection combined with gemcitabine plus cisplatin（GP） in the treatment of advanced non-small cell lung cancer （NSCLC）. Methods： we performed a systematicsearch in the electronic databases such as Cochrane Library, Pubmed, Embase, Chinese Journal Full-text Database,Chinese Biomedical Literature Database, Chinese Science and Technology Periodical Full-text Database andWanfang Database up to 30 January 2017. Randomized controlled trials （RCT） of Shenqi Fuzheng Injectioncombined with GP chemotherapy in the treatment of advanced NSCLC were searched, and all the RCTs wereconducted on methodological quality assessment. Data extraction and data analysis were according to standards ofCochrane systematic review. Results： Eight trials were included including a total of 701 patients. Meta-analysisresults： Shenqi Fuzheng injection combined with GP chemotherapy could significantly improve the functionalstatus of patients with NSCLC （OR = 3.44, 95% CI [2.26, 5.25], P 〈 0.0001） and clinical treatment efficacy （OR =（OR = 0.31, 95%CI [0.20, 0.47], P 〈 0.0001. The rate of leukopenia （OR = .31, 95%CI [0.20,0.47], P 〈 0.0001）,thrombocytopenia （OR = 0.58, 95%CI [0.37, 0.91], P = 0.020）, hemoglobin decline （（OR = 0.31, 95%CI [0.16,0.59], P = 0.0004） and incidence of gastrointestinal reactions （OR = 0.58,P 〈 0.05） could be reduced. Conclusion：Shenqi Fuzheng injection combined with GP chemotherapy in the treatment of advanced NSCLC obtainedsignificantly clinical efficacy. The quality of the literature incorporated is low, the conclusion requires high-qualityresearch to further prove.

ISOLATION AND IDENTIFICATION OF CYTOTOXIC COMPONENTS FROM BRYOPHYLLUM PINNATUM

A potent cytotoxic bufadienolide orthoacetate has been Isolated from Bryophyllum pinnatum. Baaed on spectral study methods, its structure was identified as bersaldegenin 1,3, 5-orthoacetate.

Targeted immunotherapy for non-small cell lung cancer

Targeted therapies that deliver the expected anti-tumor effects while mitigating the adverse effects are taking the cancer world by storm. The need for such therapies in non-small cell lung cancer(NSCLC), where systemic cytotoxic chemotherapies still remain the backbone of management, is felt more than ever before. Runway success of immunotherapies such as Ipilimumab for melanoma has brought excitement among oncologists. Immune-based treatments are in various stages of evaluation for NSCLC as well. Immunotherapies using strategies of antigen based or cell based vaccines, and blocking immune checkpoints are of substantial interest. Meaningful clinical responses are yet to be reaped from these new treatment modalities.

Analysis of Numerical Results in High Temperature Congealment and Chemistry Non-equilibrium Flow Field

Using the air plasma ignition technique, physicochemical process of burning can be accelerated; concentration limit ofretrofires both can be extended; reliability of retrofires and stability of burning can be improved. In this paper, using internalequivalent heat area in place of electric are that created Ohm heat, the flow fields of thermodynamic equilibrium chemistry con-gealment and chemistry non-equilibrium in the plasma generator were simulated. The influences of the inlet prerotation angleof air, the inlet total pressure of air and the airflow compression angle of spray nozzle on the temperature on the surface of elec-

Turbulent Flow Action of Pulp in Wet - Laid Non - Woven Processes

The paper deals with the fluid field of web forming in wet-laid non-woven production.The influence of the turbulent flow on blending fiber and occluded fluid produced in pulp flow has been discussed in theory and practice.The suitable use of the imported velocity of pulp is very important in producing wet-laid products of good quality.

CYFRA 21-1 as an early predictor of first line chemotherapy response in advanced non small cell lung cancer

Objective： In an era of ever evolving, promising new therapies for advanced non small cell lung cancer （NSCLC）, early predictors of response to therapy, are needed. We evaluated early variations in CYFRA 21-1 serum levels of patients with advanced NSCLC receiving first line chemotherapy and correlated the results with objective tumor response. Methods： 29 consecutive, previously untreated, patients of advanced non small cell lung cancer, with measurable disease on CT scan were evaluated. All patients were treated with conventional systemic chemotherapy, although the choice of chemotherapy was left to the discretion of the treating physicians. Serum samples were obtained immediately before the start of 1st and 2nd cycles of chemotherapy. CYFRA 21-1 was measured with an electrochemiluminescense immunoassay on an automatic analyzer （Elecsys 2000; Roche Diagnostics）. Response was evaluated using Response evaluation criteria in solid tumors （RECIST） criteria. Results： 10 patients had partial response, 9 patients had stable disease and 9 had progressive disease. None of the patients had complete response. 21/29 （72%） patients had an elevated baseline value of CYFRA 21-1.62% patients （18/29） had a decrease in CYFRA 21-1 after 1 cycle of chemotherapy. The average reduction in the 2nd reading was irrespective of whether baseline value was normal or not. The average reduction was statistically significant （P = 0.002; 95% CI, from 0.8369 to 3.49464; t test）. 8 out of 10 （80%） patients with partial response had a reduction in their 2nd reading of. CYFRA （P = 0.019; 95% CI, from 0.81965 to 7.20035; t test） which was significant. We also observed that 6/9 （66%） patients whose disease remains stable also had a decrease in their subsequent reading （P = 0.0106; 95% CI, from -0.44942 to 3.82720; t test）, though it was not significant statistically. Although 5 out of 9 （55%） patients, who had an increase in their CYFRA 21-1 level, had progressive disease, but it was not statistically significant （P = 0.537; 95% CI, from -1.20021 to 2.13354; ttest）. 14 out of 19 （73%） who either had partial response or had stable disease, had a reduction in their 2nd value of CYFRA 21-1 and was significant statistically （P = 0.004; 95% CI, from 0.74792 to 3.50208; t test）. We also observed that except for 1 patient, all patients who had a decrease of 42% or more in their subsequent CYFRA 21-1 level, were those who had either responded to chemotherapy or had stable disease （P = 0.001）, which was statistically significant. Conclusion： We can conclude that monitoring of serum marker CYFRA 21-1, early dudng first-line chemotherapy may be a useful prognostic tool for evaluation of early tumor response in patients with advanced NSCLC.

Isolation and structural elucidation of cytotoxic compounds from the root bark of Diospyros quercina (Baill.) endemic to Madagascar

Objective:To isolate and characterize the cytotoxic compounds from Diospyros quercina(Baill.)G.E.Schatz&Lowry(Ebenaceae).Methods:An ethno-botanical survey was conducted in the south of Madagascar from July to August 2010.Bio-guided fractionation assay was carried out on the root bark of Diospyros quercina,using cytotoxicity bioassay on murine P388 leukemia cell lines as model.The structures of the cytotoxic compounds were elucidated by 1D and 2D NMR spectroscopy and mass spectrometry.Results:Biological experiments resulted in the isolation of three bioactive pure compounds(named TR-21,TR-22,and TR-23)which exhibited very good in vitro cytotoxic activities with the IC_(50)values of(0.017 5±0.0060)μg/mL,(0.089±0.005)μg/mL and(1.027±0.070)μg/mL respectively.Thus,they support the claims of traditional healers and suggest the possible correlation between the chemical composition of this plant and its wide use in Malagasy folk medicine to treat cancer.Conclusions:The ability of isolated compounds in this study to inhibit cell growth may represent a rational explanation for the use of Diospyros quercina root bark in treating cancer by Malagasy traditional healers.Further studies are,therefore,necessary to evaluate the in vivo antineoplastic activity of these cytotoxic compounds as effective anticancer drugs.

MiR-16-5p plays an inhibitory role in human non-small cell lung cancer through Fermitin family member 2

Increasing evidence indicates that aberrant expressions of some microRNAs are associated with cancer progression.However,the roles and biological mechanisms of miRNA-16-5p in human non-small cell lung cancer(NSCLC)are not to be well studied.Here,we validated that the expression of miR-16-5p was decreased significantly in NSCLC samples and cell lines.The correlation between the clinicopathological features of NSCLC and the miR-16-5p expression showed that the expression of miR-16-5p in non-small cell lung cancer was linked with the advanced TNM stage,positive lymph node metastasis,with short overall survival(OS).Also,a negative correlation between miR-16-5p and Fermitin family member 2(FERMT2)was observed,implying there may be a potential link about their regulation.The hypothesis was further confirmed by in-silico analysis and dual-luciferase reporter assay.Moreover,we demonstrated that the transfections of miR-16-5p mimics could alter some biological characteristics of NSCLC cells remarkably accomplished by the expression variance of FERMT2 in vitro and in vivo assays.Summarily,this study demonstrated that miR-16-5p,as a tumor suppression factor in NSCLC by targeting FERMT2,could serve as one promising biomarker in the prediction for NSCLC patients.

外周血sCTLA-4作为晚期非小细胞肺癌患者的预后因子

目的：通过检测晚期非小细胞肺癌（non-small-cell lung cancer，NSCLC）患者外周血可溶性细胞毒性T淋巴细胞相关抗原4（soluble cytotoxic T lymphocyte associated antigen- 4，sCTLA-4）的表达，探讨其与晚期NSCLC患者生存期的关系。方法：采集2010年8月至2013年6月上海中医药大学附属龙华医院肿瘤科经病理证实的58例晚期NSCLC患者和30例正常人的外周血，运用ELISA法检测血中sCTLA-4含量，通过电话随访或上海市疾控中心获得患者生存期数据，分析sCTLA-4表达与NSCLC生存期的关系。结果：NSCLC患者外周血sCTLA-4表达率高于正常人（70.7% vs 6.7%， χ2=32.44，P〈0.01）,外周血sCTLA-4增高的患者中位生存期（MST）较短（41.63个月vs 31.57个月， χ2=7.765，P〈0.01），死亡风险高于其他患者（RR=305， χ2=8.01, P〈0.01）。结论：NSCLC患者外周血中sCTLA-4高表达，sCTLA-4可能成为晚期NSCLC患者的预后因子之一。

非小细胞肺癌T细胞CTLA-4免疫靶向治疗基础与转化性研究现状的思考

细胞毒T淋巴细胞相关抗原4(CTLA-4)是一种白细胞分化抗原,参与免疫反应的负调节。随着非小细胞肺癌(NSCLC)的治疗模式由驱动基因靶向治疗向免疫靶向治疗的转变,CDLA-4已成为一个新的研究热点。本文梳理近年来CTLA-4在NSCLC领域的研究情况,并结合相关基础及临床转化性研究进展作一综述。

肝细胞靶向性Gal-Bu对大鼠肝细胞的转染效率检测

目的检测肝细胞靶向性半乳糖基化聚乙烯亚胺衍生物Gal-Bu对大鼠肝细胞(BRL-3A)的转染效率。方法对PEI-Bu化学修饰以半乳糖基团制备Gal-Bu,琼脂糖凝胶电泳检测其复合质粒DNA的能力;MTT法检测Gal-Bu对BRL-3A的细胞毒性;以荧光素酶质粒作为报告基因,测定Gal-Bu在BRL-3A细胞的转染效率;半乳糖竞争抑制实验观察Gal-Bu对肝细胞的靶向性。结果琼脂糖凝胶电泳表明在质量比>15时,Gal-Bu具有复合DNA的能力。在5～100μg/mL浓度范围内,Gal-Bu和PEI 25 000的细胞毒性随浓度升高而增大;在同一浓度下,Gal-Bu的细胞毒性小于PEI 25 000(P<0.01)。Gal-Bu/DNA在质量比为50时达到最高转染活性,其数值为PEI 25 000的5.6倍(P<0.01),且接近Lipofectamine 2000。半乳糖竞争抑制实验表明,100 mmol/L的半乳糖可显著降低Gal-Bu的转染效率(P<0.01),但是并不影响PEI-Bu的转染效率(P>0.05)。结论 Gal-Bu是一种低细胞毒性、高转染活性的非病毒肝细胞靶向基因载体,在肝脏疾病的基因治疗领域中具有潜在的应用前景。

新型非病毒纳米基因载体PEI-β-CyD对软骨细胞和骨髓间充质干细胞转染有效性评估

目的评估新型非病毒纳米基因载体聚乙烯亚胺-β-环糊精(PEI-β-CyD)对软骨细胞系C5.18和骨髓间充质干细胞系C3H10T1/2转染的有效性。方法体外培养软骨细胞系C5.18和骨髓间充质干细胞系C3H10T1/2,MTT法观察并比较PEI-β-CyD和相对分子质量为25 000的聚乙烯亚胺(PEI25KDa)对C5.18细胞和C3H10T1/2细胞的细胞毒性。在不同的N/P(载体有效氮含量/外源基因有效磷含量)复合条件下,利用PEI-β-CyD和PEI25KDa分别转染C5.18细胞和C3H10T1/2细胞,倒置荧光显微镜结合流式细胞仪分析转染效率。结果 PEI-β-CyD对C5.18细胞和C3H10T1/2细胞的细胞毒性均小于PEI25KDa。PEI-β-CyD与PEI25KDa对C5.18细胞的转染效率比较,差异无统计学意义(P>0.05);PEI-β-CyD对C3H10T1/2细胞的转染效率显著高于PEI25KDa(P<0.05)。结论 PEI-β-CyD对于软骨细胞系C5.18及骨髓间充质干细胞系C3H10T1/2是一种低毒有效的非病毒纳米基因载体,在软骨组织工程和细胞治疗研究及应用领域具有良好的开发前景。

丙型肝炎病毒NS3区C末端HLA-11限制性CTL表位的作用

目的 研究丙型肝炎病毒 (hepatitis C virus,HCV)非结构 3区 (NS3) C末端 HL A- 11限制的细胞毒 T细胞(CTL )表位在 HCV感染中的作用 .方法 血清学方法检测 3例患者的 HL A分型 ,他们均存在 HL A- A11表型 ,以 HL A- 11结合基序为依据 ,预测了 HL A- 11限制的 CTL 表位 . 2例慢性患者 5 a内 3个时间点和 3a内 2个时间点及转阴患者的血清样本 ,用反转录 PCR扩增出 HCV基因片段 ,Sanger法测定核苷酸序列 ,并翻译成蛋白质 .结果 慢性 HCV感染者 2例预测的 HL A- A11限制的 CTL 表位都未发生明显改变 ,而在1例 HCV转阴者存在 1个改变的 HL A- 11限制性 CTL表位(序列为 IIL THPVTK) ,然而经 T细胞增殖实验表明此表位不是 T细胞表位 .结论 在 HCV NS3蛋白 C末端可能不存在与 HCV转归有关的 HL A- A11限制的 CTL

长链非编码RNA MIR22HG通过微小RNA-9-3p调节CTLA4抑制前列腺癌

目的探讨长链非编码RNA(LncRNA)MIR22HG通过微小RNA-9-3p(miR-9-3p)调节细胞毒性T淋巴细胞相关蛋白4(CTLA4)抑制前列腺癌的机制。方法收集26例前列腺癌组织样本和26例前列腺增生组织样本,培养VCAP、PC3、LNCap和DU145细胞。采用实时荧光定量聚合酶链反应(qRT-PCR)检测前列腺癌组织、前列腺增生组织及4种前列腺癌细胞的MIR22HG和miR-9-3p mRNA水平。筛选MIR22HG和miR-9-3p mRNA表达较高的细胞。将筛选出的细胞按不同干预方法分为OE-CTLA4(CTLA4过表达组)、OE-MIR22HG(MIR22HG过表达组)、miR-9-3p mimic(miR-9-3p过表达组)、OE-MIR22HG+miR-9-3p mimic(MIR22HG及miR-9-3p联合过表达组)、OE-NC+NC mimic(转染对照组)、NC mimic(miR-9-3p过表达对照组)。采用TargetScan数据库及荧光素酶报告基因实验验证MIR22HG与miR-9-3p及miR-9-3p与CTLA4是否存在靶向抑制关系。采用qRT-PCR及Western blot检测各组细胞MIR22HG和miR-9-3p mRNA及CTLA4蛋白表达。取裸鼠96只,按干预方法不同分为OE-NC+NC mimic组(转染对照)、OE-MIR22HG组(MIR22HG过表达)及OE-MIR22HG+miR-9-3p mimic组(MIR22HG及miR-9-3p联合过表达),每组32只。接种后4周期间,每周检测小鼠原位移植瘤并统计瘤体的质量和体积变化。结果前列腺癌细胞VCAP、PC3、LNCap和DU145细胞中,PC3细胞MIR22HG mRNA相对较低,miR-9-3p mRNA相对较高,因此选择PC3细胞进行研究。在PC3细胞中成功过表达MIR22HG,荧光素酶报告基因实验验证了MIR22HG和miR-9-3p的靶向抑制关系,过表达MIR22HG,miR-9-3p表达下调,差异有统计学意义(P<0.05)。TargetScan数据库预测免疫基因CTLA4和miR-9-3p的靶向关系,荧光素酶报告基因实验验证miR-9-3p和CTLA4的靶向抑制关系,过表达miR-9-3p,CTLA4表达上调,差异有统计学意义(P<0.05)。过表达MIR22HG的裸鼠皮下肿瘤的质量降低,差异有统计学意义(P<0.05)。OE-MIR22HG+miR-9-3p mimic组裸鼠皮下肿瘤的质量、体积与OE-NC+NC mimic组比较,差异无统计学意义(P>0.05)。结论MIR22HG与miR-9-3p存在靶向负调节关系,miR-9-3p与CTLA-4存在靶向调节关系,MIR22HG/miR-9-3p可能通过抑制CTLA4达到免疫治疗前列腺癌的目的。

化疗联合DC-CTL治疗不同分期非小细胞肺癌的疗效分析

目的:分析化疗联合树突状细胞诱导的细胞毒性T淋巴细胞(dendritic cells and cytotoxic lymphocytes,DC-CTL)对非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的疗效及安全性。方法:收集我院2013年5月至2015年12月期间住院病理确诊为Ⅰb(具有高危因素)、Ⅱa、Ⅱb、Ⅲa、Ⅲb、Ⅳ期的NSCLC患者114例,其中接受肺癌根治术的患者分为术后化疗联合DC-CTL组20例和术后化疗组32例,未行手术患者分为化疗联合DC-CTL组23例和化疗组39例。回顾分析了114例患者的临床特点、疾病控制率(disease control rate,DCR)、中位无疾病生存期(median-disease free survival,M-DFS)、中位无进展生存期(medianprogression free survival,M-PFS)、副作用等资料。随访截止至2016年11月。结果:1年DCR:术后化疗联合DC-CTL组(75.0%)vs术后化疗组(43.8%)(P=0.044)。M-DFS:术后化疗联合DC-CTL组(19.5个月)与术后化疗组(11.5个月)相比,差异具有明显统计学意义(P=0.025 5)。化疗联合DC-CTL治疗与化疗组相比,DCR及M-PFS不具有统计学差异。DC-CTL治疗的副作用显著低于化疗。结论:术后化疗联合DC-CTL治疗可显著提高DCR及延缓NSCLC患者的疾病进展,且安全性高。

口腔非贵金属材料对L-929细胞毒性的研究

目的:评价5种口腔修复非贵金属材料的细胞毒性。方法:采用细胞培养法、MTY法和流式细胞仪检测细胞周期的方法,研究5种合金的细胞毒性。结果:MTT比色法结果揭示:纯钛、钛合金具有良好的生物相容性,而钴铬合金、镍铬合金、铜基合金抑制了细胞的生长;细胞周期所测各组细胞G2期比例与MTI结果相符。结论:不同金属合金的细胞毒性不同。