Autophagy plays a protective role in advanced glycation end products-induced apoptosis of chondrocytes via regulation of tumor necrosis factor-α,nuclear factor-κ B and reactive oxygen species

Objective: To study the adverse effects of advanced glycation end products(AGEs) on chondrocytes and the role of autophagy in this process. Methods: Chondrocytes were harvested from the human articular cartilage tissues in surgery. AGEs were administered during chondrocytes culture. The rapamycin was used to induce autophagy. The cell viability was determined by 3-[4,5-dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide(MTT) assay.The expression of tumor necrosis factor-α(TNF-α) and nuclear factor-κ B(NF-κ B) was detected by quantitative real-time polymerase chain reaction. The reactive oxygen species(ROS) production and apoptosis of the chondrocytes were determined by fluorescent probe and flow cytometer, respectively. Results: The chondrocytes viability was significantly reduced after 12 h incubation with AGEs(P<0.01)). In contrast, rapamycin pretreatment increased the chondrocytes viability through autophagy. AGEs increased TNF-α and NF-κ B mRNA expression of chondrocytes and autophagy receded or proceeded the change. AGEs increased intracellular ROS accumulation and autophagy reversed the change. AGEs accelerated chondrocytes apoptosis and autophagy suspended apoptosis. Conclusions: Accumulation of AGEs may have an adverse role for chondrocytes by increasing TNF-α and NF-κB expression, ROS accumulation and apoptosis; meanwhile, autophagy ameliorates the AGEsinduced adverse effects.

EFFECTS OF PHYTOLACCA ACINOSA POLYSACCHARIDES I ON CYTOTOXICITY OF MACROPHAGES AND ITS PRODUCTION OF TUMOR NECROSIS FACTOR AND INTERLEUKIN 1

The in vivo effects of Phytolacca acinosa poly-saccharides I (PEP-I) on immunologic cytotoxicity of mouse peritoneal macrophages and its production of tumor necrosis factor (TNF) and interleukin 1 (IL-1) were studied. PEP-I 80 or 160 mg kg was given ip twice every 4 day. Both doses were found to have significant enhancing activity on macrophages cytotoxicity against S180 sarcoma cells and malignant transformed fibroblast L929 cells. Peritoneal activated macrophages were incubated with LPS for 2 and 24 hrs to induce TNF and IL-1, respectively. The TNF and IL-1 activities were tested from cytotoxicity against L929 cells in an absorbence assay of enzymatic reaction and proliferation of thymocytes co-stimulated assay separately. The optimal time for TNF production was found on day 8. Significant increases in TNF and IL-1 were observed. In comparison of the effect of PEP-I on TNF with that of known priming agent BCG, there was no difference between them, but PEP-I had a high effect on IL-1. These results suggest that cytotoxicity of macrophages primed by PEP-I is closely related to its TNF and IL-1 production.

ANTITUMOR MECHANISM OF GEM10 BY THE NATURAL KILLER ACTIVITY AND INTERLEUKIN-2 PRODUCTION

Objective To investigate th e anti-tumor effects of GeM10 by the natural killer(NK) cells activities and th e production of Interleukin-2 (IL-2) in peripheral blood mononuclear cells (PB MNCs). Methods Assay of human NK cells activities by dye reject ion assay in vitro and production of IL-2 in PBMNC by IL-2 bioassay with I L-2 dependent cell line CTLL2 and MTT colorometric method. Results GeM10 could significantly stimulate NK activities (60μg·mL -1 G eM10: 17.077±7.665, 120μg·mL -1 GeM10: 24.9±13.04; control: 7.72±4 .64, P< 0.05). GeM10 could up-regulate the production of IL-2 of PBMNCs in tumor patients(60μg·mL -1 GeM10: 2.965± 1.183; 120μg·mL -1 GeM10: 2.28±0.847; control: 1.792±0.823, P<0.05).Conclu si on The GeM10 not only can stimulate the NK activities but also increase the IL-2 production by PBMNCs in tumor patients. These findings indicate that the GeM10 may have promise as an anti-tumor drug and a biological response modi fier in clinic.

Ectopic Hormone Production from Ovarian Tumor: A Review

Although ectopic hormone-production is uncommon complication, certain tumors can produce symptoms due to the secretion of various bioactive substances accompanied by the aberrantly located tumors. Because of the potential for the ovary to act as a source of aberrant hormone secretion, in the literature, ectopic hormone production from ovarian tumor includes granulocyte-colony stimulating factor (G-CSF), parathyroid hormone-related protein (PTHrP), adrenocorticotropic hormone (ACTH), peptide-YY, gastrin and insulin. All patients may present with syndromes of hormone excess. Failure to localize the ovarian tumor preoperatively may be associated with a significantly higher risk of subsequent unnecessary ablative procedures. Better characterization of hormonal forms relatively specific for neoplasia may enhance the clinical value of ectopic hormones as tumor markers, especially in malignancies that are commonly associated with ectopic hormone production. These circumstances may recommend complete preoperative evaluation of the pelvis in female patients presenting with nonlocalizable endocrine tumors.

Role of the advanced glycation end products receptor in Crohn's disease inflammation

AIM:To investigate the level of mucosal expression and the involvement of the receptor for the advanced glycation end products(RAGE)in delayed apoptosis and tumor necrosis factor(TNF)-αproduction in Crohn’s disease(CD).METHODS:Surgical and endoscopic specimens from both inflamed and non-inflamed areas of the ileum and/or colon were collected from 20 and 14 adult CD patients,respectively,and used for the assessment of RAGE expression by means of immunohistochemistry and western blotting analysis.Normal tissues from 21 control subjects were used for comparison.The same polyclonal anti-human RAGE antibody(R and D System)was used in all experimental conditions.RAGE staining was quantized by a score including both the amount of positive cells and intensity of immunoreactivity;cellular pattern was also described.The effects of RAGE blocking on apoptotic rate and TNF-αproduction were investigated on immune cells freshly isolated from CD mucosa and incubated both with and without the muramyl dipeptide used as antigenic stimulus.Statistical analysis was performed via the test for trend,with regression models to account for intra-patient correlations.A 2-sided P<0.05 was considered significant.RESULTS:In inflamed areas,RAGE expression in both the epithelial and lamina propria compartments was higher than control tissues(P=0.001 and 0.021,respectively),and a cluster of positive cells were usually found in proximity of ulcerative lesions.Similar results were obtained in the lamina propria compartment of non-inflamed areas(P=0.025).The pattern of staining was membranous and granular cytosolic at the epithelial level,while in the lamina propria it was diffuse cytosolic.When evaluating the amount of protein expression by immunoblotting,a significant increase of both surface area and band intensity(P<0.0001 for both)was observed in CD inflamed areas compared to control tissue,while in non-inflamed areas a significant increase was found only for band intensity(P<0.005).Moreover,a significantly lower expression in noninflamed areas in comparison with inflamed areas was found for both surface area and band intensity(P<0.0006 for both).Finally,RAGE blocking largely affects both the apoptotic rate of mucosal cells(towards an increase in both non-inflamed and inflamed areas of P<0.001 and<0.0001,respectively)and TNF-αsecretion(towards a decrease in both non-inflamed and inflamed areas of P<0.05 and<0.01,respectively),mainly in the presence of antigenic stimulation.CONCLUSION:RAGE is up-regulated in CD,especially in inflamed areas,and it appears to play a role in the mechanisms involved in chronic inflammation.

Functional Activity of Circulating Phagocytes as Potential Pretreatment Marker of Tumor Drug Resistance

The aim of this work was a study of the functional activity of neutrophils and peripheral blood monocytes in rats with the transplanted Walker carcinosarcoma as potential predictors of the sensitivity of this tumor to doxorubicin treatment. This study provides an evidence that such indices of the functional activity of circulating phagocytes of the tumor-bearing rats as the quantity and the phagocytosis intensity of monocytes, as well as the intensity of ROS production by monocytes and neutrophils, may reflect the degree of sensitivity of the tumor to doxorubicin. So it was shown that the growth of the resistant tumor caused a significant increase of the number of circulating phagocytic cells and the intensity of phagocytosis by more than 100% (p < 0.001) compared with the corresponding indices of intact rats and rats with the parental variant of the tumor. The ability of blood mono-cytes and neutrophils of rats with a resistant tumor to produce ROS was also significantly different from that in intact rats and animals with the parental carcinosarcoma variant. The predictive value of these indices is especially important in the dynamic monitoring of the development of tumor drug resistance during long-term cancer chemotherapy. Considering the standard 2 - 3 week interval between the courses of cancer therapy and the short lifetime of circulating phagocytes, an assessment of the indices of their functional activity before each subsequent course can be considered as a pretreatment assessment. Meanwhile, further studies are needed to determine the spectrum of malignant neoplasms for which the degree of tumor drug resistance correlates with the functional activity of circulating phagocytes.

Production of radioactive ~9C ion beam and its optimization

To explore the medical use of a radioactive 9C-ion beam in tumor treatment, which is a double radiation source coming from the external beam itself and the delayed particles emitted internally, some physical experiments are performed at the Secondary Beam Line (SBL) of Heavy Ion Medical Accelerator, Chiba (HIMAC) in Japan be- fore radiobiological research for exhibiting therapeutic value of the 9C beam. Intention of the experiments is for pro- ducing a radioactive 9C-ion beam with higher production rate and purity by means of optimizing the beam line pa- rameters. Finally, a produced 9C beam with the production rate of 9.07×10-6 and purity of 82.88% at full momentum acceptance has been obtained under the optimal conditions of 40 mm-thick beryllium target and 10 mm-thick alumi- num degrader. Both momentum distribution and contaminations for the produced 9C beam under the optimal condi- tions are measured. In order to execute further biological experiments of the 9C beam project, a uniform irradiation field is made with the wobbling magnets and its homogeneity is up to 93.8% inside central area of 20mm in diameter.

天然产物调控肿瘤相关巨噬细胞在抗肿瘤免疫中的研究进展

肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)在调节肿瘤免疫方面作用突出,始终影响着肿瘤的发生、发展,是目前抗肿瘤治疗的重要靶点。天然产物被证实具有显著的免疫调节和抗肿瘤作用,已成为抗肿瘤免疫研究的热门领域。大量研究发现,天然产物如多糖类、黄酮类、皂苷类和生物碱类等可通过调控相关信号通路,调节肿瘤相关巨噬细胞免疫因子及标记物的表达,促进M1巨噬细胞极化,抑制M2巨噬细胞极化,改善肿瘤免疫微环境,发挥抗肿瘤免疫作用。该文通过总结近年来天然产物调控肿瘤相关巨噬细胞以发挥抗肿瘤免疫作用的研究成果,从天然产物调控肿瘤相关巨噬细胞以增强抗肿瘤免疫作用方面进行论述,以期为抗肿瘤免疫治疗提供新的研究思路。

中药常山、蜀漆古今文献考证

目的探究常山、蜀漆的基原、产地、性味归经、功效、毒性特点,以期为常山、蜀漆的研究与合理应用提供参考。方法以第5版中华医典数据库、读秀数据库、中医智库在线古籍数据库为检索范围,考证常山、蜀漆的基原、产地、性味归经、功效、毒性,同时结合现代文献研究进行分析。结果发现常山、蜀漆别名较多,分布广泛,喜阴凉湿润环境,以四川产量最大,质量最优。常山的性味与归经在历代古籍中未发现明显变化,但蜀漆性味记载存在分歧,且归经论述较少,未得统一。古籍载常山、蜀漆功效多为截疟、涌吐,现代药理研究发现两者还具有抗球虫、抗肿瘤、解热、降压、抗流感病毒等药理作用。但因两者均存在一定毒性,为保证安全用药,传统多通过炮制及配伍进行减毒存效,现代多从其活性成分本身的结构修饰及致毒机理实现减毒增效。结论古今文献梳理发现常山及蜀漆,除截疟外还具有抗肿瘤等多种药理活性,研发前景良好,但囿于其存在一定的毒性,制约了其临床应用,传统临床多通过炮制、配伍减毒存效,现代学者尝试通过结构修饰和改变给药途径减毒增效,为其减毒增效提供了新的思路和方向。

高效液相色谱-示差折光法测定NMM抗肿瘤DNA疫苗中甘露醇含量

目的 建立测定NMM抗肿瘤DNA疫苗中甘露醇含量的高效液相色谱-示差折光法。方法 检测器为示差折光检测器,色谱柱为SUGAR SC1011阳离子钙型交换柱(300 mm×8 mm,6μm),流动相为纯化水,流速为1.0 mL/min,柱温为80℃,检测池温度为37℃,进样量为10μL。结果 甘露醇的质量浓度在0.02~5.00 mg/mL(R2=1.000 0,n=6)范围内与峰面积线性关系良好;定量限为13.37μg/mL;精密度、重复性、稳定性试验结果的RSD均小于2.0%;加样回收率为97.76%,RSD为0.97%(n=9)。3批小试样品和中试样品中甘露醇的含量分别为20.02,19.95 mg/mL。结论 该方法操作简便、重复性好、结果准确,可用于NMM抗肿瘤DNA疫苗中甘露醇的含量测定。

急性缺血性脑卒中溶栓后脑出血转化患者血清AGEs、IBIL、CTRP-3水平观察

目的观察急性缺血性脑卒中(AIS)溶栓后脑出血转化患者血清晚期糖基化终末产物(AGE)、间接胆红素(IBIL)、补体C1q肿瘤坏死因子相关蛋白-3(CTRP-3)水平。方法回顾性分析2022年1月至2023年1月在临汾市中心医院接受治疗的92例AIS患者的临床资料,所有患者均进行溶栓治疗,经磁共振成像检查,依据溶栓治疗后24 h是否发生脑出血转化将患者分为脑出血转化组(n=22)和非脑出血转化组(n=70)。比较两组基础资料信息[性别、年龄、体重指数、糖尿病、高血压、高血脂、吸烟史、发病至溶栓时间、收缩压、舒张压、溶栓前美国国立卫生研究院卒中量表(NIHSS)评分]和AGE、IBIL、CTRP-3水平,采用多因素Logistic回归分析对影响AIS患者溶栓后脑出血转化的危险因素进行分析,绘制受试者工作特征(ROC)曲线评价血清AGE、IBIL、CTRP-3水平预测AIS溶栓后脑出血转化的效能。结果两组性别构成比、年龄、体重指数、糖尿病、高血压、高血脂、吸烟史、发病至溶栓时间、收缩压、舒张压比较,差异均无统计学意义(P>0.05);脑出血转化组溶栓前NIHSS评分、AGE、IBIL水平分别为(14.68±2.41)分、(510.91±51.16)ng/L、(13.69±2.27)μmol/L,均显著高于非脑出血转化组[(11.64±1.91)分、(447.69±49.46)ng/L、(10.17±1.79)μmol/L],而CTRP-3水平(250.13±25.69)ng/mL,显著低于非脑出血转化组[(284.57±28.92)ng/mL],差异均有统计学意义(P<0.05)。经多因素Logistic回归分析证实,溶栓前NIHSS评分、AGE、IBIL水平及CTRP-3水平是影响AIS患者溶栓后脑出血转化的危险因素(P<0.05);经ROC曲线分析证实,血清AGE、IBIL、CTRP-3水平均可用于AIS溶栓后脑出血转化的预测,曲线下面积分别为0.829、0.927、0.836(P<0.05)。结论溶栓前NIHSS评分及AGE、IBIL、CTRP-3水平的异常表达会增大AIS患者溶栓后脑出血转化的风险,可将以上指标作为评估患者病情和溶栓后脑出血转化情况的标志物,为临床病情评估和治疗提供参考。

复方硫酸铝注射液局部注射的药效学和全身吸收实验研究

目的 考察复方硫酸铝注射液体内对小鼠移植肿瘤生长的抑制作用 ,以及局部注射后的全身吸收情况 ,并评价用药的安全性。方法 采用瘤内一次注射给药法考察药物体内抑制肿瘤生长的作用 ;采用兔背部皮下注射后测定血铝浓度的方法考察药物的全身吸收 ;血铝浓度的测定采用等离子体质谱法。结果 复方硫酸铝注射液对小鼠移植肿瘤的生长有显著的抑制作用 ,硫酸铝是抑制肿瘤的有效成分 ,其抑制作用与剂量呈明显的正相关 ,浓度为 0 .117,0 .175 ,0 .2 34 ,0 .2 92mol·L-1时平均抑制率分别为 2 3.0 % ,32 .7% ,45 .4%和 5 3.3% ,处方中其它成分不影响肿瘤的生长。复方硫酸铝注射液兔背部皮下注射0 .46 8mmol·kg-1(以无水硫酸铝计 ) ,血铝浓度没有明显的峰值 ,最高浓度为 30 0ng·mL-1(n =5 )。结论 复方硫酸铝注射液局部注射后显著抑制肿瘤生长 ,并且硫酸铝的全身吸收量很小 ,是一种有效。

β-胡萝卜素氧化降解产物及其抑制癌细胞活性的研究

研究了β-胡萝卜素氧化降解产物的生理活性。对利用OsO4为催化剂,将β-胡萝卜素在OsO4-H2O2-Ether体系中进行控制型氧化降解,对氧化降解产物进行薄层分离后,采用体外细胞培养技术观察β-胡萝卜素氧化降解产物对Hela细胞以及bel-7402细胞增殖的影响。结果发现,β-胡萝卜素氧化降解产物在高浓度时对Hela以及bel-7402的抑制作用强于β-胡萝卜素的,且对bel-7402的作用强于对Hela的,显示β-胡萝卜素氧化降解产物对癌细胞的增殖有抑制作用。

风湿免疫病生物制剂治疗进展

生物制剂是一种选择性针对参与免疫反应或炎症过程的分子或以受体为靶目标的单克隆抗体或天然抑制分子的重组产物。生物制剂种类繁多,主要针对细胞因子,B细胞和共刺激信号分子,包括:肿瘤坏死因子抑制剂、抗白细胞介素6(IL-6)受体单克隆抗体和抗CD20单克隆抗体利妥昔单抗等多种药物,主要用于治疗强直性脊柱炎、类风湿关节炎和系统性红斑狼疮等自身免疫性疾病。生物制剂的使用应关注其安全性,用药前进行结核筛查,除外活动性感染和肿瘤。生物制剂的减停药问题有待探讨。

柴胡皂苷a药理学研究进展

柴胡是我国传统中药,其正品为伞形科植物柴胡或狭叶柴胡的干燥根。由于柴胡广泛的药理活性及临床应用范围,使得关于柴胡及其有效成分的研究受到广泛重视。柴胡皂苷a是从柴胡中分离得到的具有多种药理活性的天然化合物,在抗肿瘤、抗炎、抗癫痫、保肝护肝等方面均展现出良好的应用前景。基于CNKI与ISI Web of Knowledge数据库查找及追溯了近年来关于柴胡皂苷a的最新研究文献报道,归纳总结了其药理学研究进展,以期为相关研究人员提供参考。

科学表征细胞体内过程,推动细胞治疗产品高质量发展

细胞治疗技术的突飞猛进引领了复杂难治性疾病治疗范式的重要转变,以CAR-T为代表的细胞治疗产品陆续获批上市,标志着药物治疗史上一个新时代的到来。与临床治疗新技术不同,用于上市申请的细胞治疗产品应遵循药品注册相关法规要求,开展必要的临床药理学研究,科学表征其体内过程。为推动我国细胞治疗产品高质量发展,国家药品监督管理局药品审评中心组织起草了《细胞治疗产品临床药理学研究技术指导原则(试行)》,针对细胞治疗产品体内过程特点,重点阐述开展必要的细胞动力学、药效学及影响因素研究,探索细胞治疗产品安全有效性可控关键要素的相关考虑。

晚期糖化终产物受体在小鼠急性肝损伤中的表达及意义

目的探讨晚期糖化终产物受体(RAGE)在刀豆蛋白A(ConA)诱导的小鼠急性肝损伤中的表达及意义。方法将72只昆明种小鼠随机分为对照(NS)组、ConA组,分别于注射后2、6、12、18、24、48 h眼球取血并留取肝脏标本,用赖氏法检测血清中谷氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)活力;HE染色法检查肝组织病理学改变;RT-PCR技术检测肝组织高迁移率族蛋白1(HMGB1)、RAGE、肿瘤坏死因子α(TNF-α)、白介素-1β(IL-lβ)mRNA的表达;免疫组化法检测肝组织HMGB1蛋白的表达;Westernblot法检测肝组织RAGE蛋白的表达;ELISA法检测血清中TNF-α、IL-lβ含量。结果成功构建ConA诱导小鼠肝损伤模型;注射ConA 2 h肝组织中HMGB1 mRNA表达即增加,至18 h达峰值(P<0.01);HMGB1蛋白表达明显聚集于坏死区域;肝组织RAGE mRNA于12 h表达上调;RAGE蛋白的表达于24 h达到峰值,与NS组相比差异有统计学意义(P<0.01);肝组织TNF-αmRNA表达分别在6、24 h出现2个峰值,血清TNF-α含量在注射ConA 2 h达峰值(455.25 ng/L),随后逐渐减低,至48 h与NS组相比无明显差异;注射ConA各时间点小鼠肝组织与血清中IL-1β水平均显著高于NS组(P<0.01)。结论在ConA诱导小鼠急性肝损伤模型中,损伤肝组织RAGE的表达显著增加,这可能与诱导肝组织损伤中炎症因子的进一步产生有关。

晚期氧化蛋白产物通过活性氧诱导单核细胞分泌肿瘤坏死因子

目的探讨晚期氧化蛋白产物(AOPP)对单核细胞分泌肿瘤坏死因子(TNFα)的影响及可能机制。方法以THP-1和人外周血单核细胞为单核细胞模型,与用次氯酸氧化牛血清白蛋白(BSA)制备的AOPP-BSA共同培养,用ELISA法检测培养上清TNFα的释放量,用VICTORWallac1420多标记分析系统检测细胞氧化2,7-二氢二氯荧光素产生的荧光量反映活性氧的产生量;用抗氧化剂吡咯烷二硫代氨基甲酸盐(PDTC),NADPH氧化酶抑制剂apocynin和P38磷酸化抑制剂SB203580预处理细胞,探讨AOPP诱导单核细胞分泌TNFα的可能机制。结果AOPP-BSA能诱导单核细胞TNFα的分泌和细胞内活性氧(ROS)的产生。用PDTC预处理细胞,可以清除AOPP-BSA诱导产生的ROS,同时抑制TNFα的分泌。用apocynin抑制NADPH氧化酶及用SB203580抑制P38磷酸化均能有效地抑制AOPP-BSA诱导的TNFα分泌。结论AOPP可能通过激活单核细胞NADPH氧化酶,释放ROS,使P38磷酸化,导致TNFα的分泌。

天然来源萘醌类化合物抗肿瘤活性研究进展

天然来源萘醌类化合物是一类具有明显生物活性的物质,多具有抗肿瘤作用。通过查阅近期国内外文献,综述了天然来源萘醌类抗肿瘤化合物的自然界来源、化合物结构、抗肿瘤作用及其机制,以期为进一步研究与开发提供有价值的参考。

盐生海芦笋来源真菌Salcoli6发酵产物抗肿瘤活性成分研究

采用活性追踪的方法,对一株盐生海芦笋来源真菌Salcoli6发酵产物中的抗肿瘤活性成分进行分离和鉴定。利用溶剂萃取、硅胶柱色谱及制备HPLC等分离手段从发酵产物中分离得到7个化合物,通过理化性质分析及波谱学方法鉴定其化学结构分别为枝孢菌素(1)、异枝孢菌素(2)、2-乙酰基喹唑啉(3)、萘满酮(4)、3-羟基-2-甲级色原酮(5)、3-甲级异苯并呋喃(6)、对-羟基苯乙酸(7);以SRB法评价化合物的抗肿瘤活性,化合物3、4和7对p388细胞具有强的细胞毒活性,IC50值分别为12、24μg/mL和6μg/mL,其他化合物没有活性。