白藜芦醇对KBv200细胞多药耐药逆转作用的研究

目的 探讨白藜芦醇对人口咽腔上皮癌KBv200耐药细胞株的多药耐药逆转作用及可能的逆转机制.方法 采用四甲基偶氮唑盐比色法检测KBv200耐药细胞中白藜芦醇对长春新碱、多柔比星和紫杉醇的逆转倍数,流式细胞仪检测细胞的凋亡情况,应用反转录聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blot)检测每组KBv200细胞中肿瘤多药耐药相关基因1(multidrug resistance 1,MDR1)和B细胞淋巴瘤基因2(Bcl-2)mRNA和蛋白水平的表达.结果 白藜芦醇对化疗药物具有协同增效作用,显著逆转KBv200耐药性.200 umol/L白藜芦醇对长春新碱、紫杉醇和多柔比星(阿霉素)的逆转倍数达到77.1、61.3和5.9.白藜芦醇能显著降低Bcl-2、MDR1mRNA和蛋白的表达,100 umol/L、200 umol/L白藜芦醇处理组与未加白藜芦醇组的MDR1、Bcl-2mRNA表达差异均有统计学意义(t值分别为2.98、3.51和3.12、4.56,P值均<0.05).结论 白藜芦醇对口咽腔上皮癌耐药细胞具有耐药逆转作用,该逆转作用可能是通过降低耐药基因表达、促进细胞凋亡实现的.     

维甲酸对小鼠T细胞增殖及骨破坏因子RANKL表达的研究

目的 探讨全反式维甲酸对体外培养的小鼠T淋巴细胞的增殖及核因子-КB受体活化因子配体(RANKL)表达的调节作用及相关机制。方法 体外分离Aa感染的BALB/C小鼠颈T淋巴细胞,分别加入浓度为0、1×10-8、1×10-7、1×10-6、1×10-5mol/L的全反式维甲酸,培养3 d后,取100μl上清保存在-70℃冰箱中,以备sRANKL及IL-10等细胞因子的测定;另加入100μl含有3Hthymidine(0.5μCi/孔)RPMI培养液继续培养18 h,进行T细胞3H增殖率测定。结果 维甲酸处理组与非处理组相比,T细胞3H增殖率下降;上清中RANKL的表达水平下降,IL-10的表达水平增强(P<0.05),二者具有负相关性(r=-0.774,P=0.001),且呈剂量依赖性。结论 维甲酸可通过上调IL-10的表达并下调T细胞上的RANKL的表达水平,抑制T细胞的免疫功能,且呈剂量依赖性。     

Neurogenetics and Clinical Evidence for the Putative Activation of the Brain Reward Circuitry by a Neuroadaptagen: Proposing an Addiction Candidate Gene Panel Map

Abstract

This document presents evidence supporting the role of the KB220/KB220Z neuroadaptagens consisting of amino-acid neurotransmitter precursors and enkephalinase-catecholamine–methyl-transferase (COMT) inhibition therapy called Neuroadaptagen Amino Acid Therapy (NAAT) in brain reward function. It is becoming increasingly clear that this novel formulation is the first neuroadaptagen known to activate the brain reward circuitry. Ongoing research repeatedly confirms the numerous clinical effects that ultimately result in significant benefits for victims having genetic antecedents for all addictive, compulsive and impulsive behaviors. These behaviors are correctly classified under the rubric of “Reward Deficiency Syndrome” (RDS). We are proposing a novel addiction candidate gene map. We present preliminary findings in the United States using qEGG and in China using Functional Magnetic Resonance Imaging (fMRI) regarding the effects of oral NAAT on the activation of brain reward circuitry in victims of SUD. In unpublished data utilizing an fMRI 2X2 design at resting state, NAAT in comparison to placebo shows activation of the caudate brain region and potentially a smoothing out of heroin-induced putamen (a site for emotionality) abnormal connectivity. Although awaiting final analysis, if confirmed by ongoing studies in China coupled with published qEEG results in America, showing an increase in alpha and low beta, NAAT may be shown to impact treatment outcomes.     

纳米二氧化锆超细颗粒物对KB细胞活性的影响

目的 评估纳米二氧化锆(nano-ZrO2)对人类口腔细胞活性的影响.方法 采用不同浓度的纳米ZrO2颗粒(0、10、30、60、100、150、250 μg·ml-1)对Human epidermoid carcinoma(KB)细胞分别进行24、48、72 h的染毒,应用MTT法检测细胞活性.结果 ①24、48 h低浓度纳米ZrO2染毒时KB细胞活性较高;②24、48 h高浓度nano-ZrO2染毒时没有对KB细胞的活性产生明显影响;③染毒72 h之后KB细胞活性没有发生显著变化,nano-ZrO2对KB细胞也不具有细胞毒性.结论 在适当浓度下,nano-ZrO2对KB细胞活性无明显毒作用.     

Overcoming qEEG Abnormalities and Reward Gene Deficits during Protracted Abstinence in Male Psychostimulant and Polydrug Abusers Utilizing Putative Dopamine D2 Agonist Therapy: Part 2

Abstract

Background: It is well established that in both food- and drug-addicted individuals there is “dopamine resistance” associated with the DRD2 gene A1 allele. Based on earlier studies, evidence is emerging wherein the potential of utilizing a natural, nonaddicting, safe, putative D2 agonist may play a significant role in the recovery of individuals with reward deficiency syndrome, including those addicted to psychoactive chemicals. Findings: Positive outcomes demonstrated by quantitative electroencephalographic (qEEG) imaging in a randomized, triple-blind, placebo-controlled, crossover study involving oral Synaptose Complex KB220Z? showed an increase of alpha waves and low beta wave activity in the parietal brain region. Using t statistics, significant differences observed between placebo and Synaptose Complex KB220Z? consistently occurred in the frontal regions after week 1 and then again after week 2 of analyses (P = 0.03). This is the first report to demonstrate involvement of the prefrontal cortex in the qEEG response to a natural putative D2 agonist (Synaptose Complex KB220Z?), especially evident in dopamine D2 A1 allele subjects. Independently, we have further supported this finding with an additional study of 3 serious polydrug abusers undergoing protracted abstinence who carried the DRD2 A1 allele. Significant qEEG differences were found between those who received 1 dose of placebo compared with those who were administered Synaptose Complex KB220Z?. Synaptose Complex KB220Z? induced positive regulation of the dysregulated electrical activity of the brain in these addicts. The results are indicative of a phase change from low amplitude or low power in the brain to a more regulated state by increasing an average of 6.169 mV² across the prefrontal cortical region. In the first experiment we found that while 50% of the subjects carried the DRD2 A1 allele, 100% carried ≥ 1 risk allele. Specifically, based on the proposed addiction risk score for these 14 subjects, 72% had moderate-to-severe addiction risk. Similar findings were obtained by repeating the experiment in 3 additional currently abstinent polydrug abusers carrying the DRD2 A1 allele. Conclusion: This seminal work will provide important information that may ultimately lead to significant improvement in the recovery of individuals with psychostimulant and polydrug abuse problems, specifically those with genetically induced dopamine deficiency. Based on this small sample size, we are proposing that with necessary large populations supporting these initial results, and possibly even additional candidate genes and single nucleotide polymorphisms, we may eventually have the clinical ability to classify severity according to genotype and possession of risk alleles, along with offering a safe, nonaddicting, natural dopaminergic receptor agonist that potentially upregulates instead of downregulates dopaminergic receptors, preferably the D2 subtype.     

Bcl-2基因在口腔上皮癌多药耐药细胞的表达

[目的]通过检测Bcl-2基因在口腔上皮癌敏感细胞株（KB）与多药耐药细胞株（KBV）的表达差异,探讨Bcl-2基因在口腔上皮癌细胞多药耐药性发生中的作用。[方法]体外培养口腔上皮癌多药耐药株及敏感株,通过RT-PCR法及免疫荧光法检测Bcl-2基因在KB及KBV细胞中转录及翻译水平的差异,通过MTT法及流式细胞检测技术检测KB及KBV细胞对长春新碱（VCR）敏感度的差异。[结果]免疫荧光图像显示Bcl-2蛋白在KB细胞中弱表达,而在KBV细胞中强表达;RT-PCR检测结果显示,KB细胞Bcl-2/β-actin比值是25.0%,KBV组Bcl-2/β-actin比值为36.1%,两组细胞间差异具有统计学意义（P〈0.01）;MTT法检测结果显示KBV组细胞对长春新碱的耐药性为KB组的16.5倍,流式细胞仪检测结果显示KB细胞凋亡率达（98.8±1.2）%;KBV细胞凋亡率达（23.5±2.1）%,两组间差异具有统计学意义（P〈0.01）。[结论]Bcl-2基因与口腔上皮癌耐药性的发生密切相关。     

Antitumor Agents 126. Novel 4β-Substituted Anilino Derivatives of 3′,4′ -O,O-Didemethylpodophyllotoxin as Potent Inhibitors of Human DNA Topoisomerase II

A series of derivatives of 3,4 0,0-didemethylpodophyllotoxin have been synthesized and evaluated for their inhibitor activity against neoplastic cell growth (KB) and against human DNA topoisomerase II as well as for their activity in causing cellular protein-linked DNA breakage. The results show that the compounds possessing a 4-anilino moiety either unsubstituted or substituted at the para (F, COOCH₃, COCH₃, CN, CH₂CN, NO₂) or meta (OH) positions or with an ethylenedioxy moiety showed the same or greater activity than etoposide in causing cellular protein-linked DNA breakage and in inhibiting DNA topoisomerase II. However, compared to the corresponding 4-0-demethyl analogues, the 3,4-O,O-didemethyl compounds have a similar potency in inhibition of DNA topoisomerase II but are less active in causing cellular protein-linked DNA breakage. Complete correlation between the three biological activities–cytotoxicity, inhibition of DNA topoisomerase II, and induction of protein-linked DNA breakage–was also not observed. This supports the possibility that the biological determinants of action among these compounds may be different.     

40种香豆素类化合物对人鼻咽癌细胞株KB和人白血病细胞株HL-60细胞生长抑制活性的筛选

目的:从中药中筛选有抗肿瘤活性的化合物。方法:应用人鼻咽癌细胞株KB和人白血病细胞株HL-60筛选40种香豆素类化合物对其生长的抑制作用。结果:马栗树皮苷、去甲基呋喃皮纳灵、前胡素和二氢山芹醇当归酸酯对人鼻咽癌细胞株KB细胞的生长有一定程度的抑制作用,且呈浓度-效应关系;伞形花内酯刘寄奴酸酯、牛防风素和考迈斯托醇对人白血病细胞株HL-60细胞的生长有一定程度的抑制作用。结论:马栗树皮苷、去甲基呋喃皮纳灵、前胡素和二氢山芹醇当归酸酯对人鼻咽癌细胞株KB细胞的生长有抑制作用;伞形花内酯刘寄奴酸酯、牛防风素和考迈斯托醇对人白血病细胞株HL-60细胞的生长有抑制作用。     

β-连环蛋白在KB细胞及其肿瘤球细胞中的表达

目的：研究信号通路β-连环蛋白在人口腔上皮样癌KB细胞及其肿瘤球细胞中的表达。方法：培养KB细胞及其肿瘤球,分别提取KB细胞和肿瘤球细胞的总RNA,设计引物,使用RT-PCR方法检测β-连环蛋白在KB细胞和肿瘤球细胞中的转录,使用Western blot方法检测β-连环蛋白在两种细胞中的翻译。结果：RT-PCR方法检测显示两种细胞的β-连环蛋白的结果均为阳性,KB细胞中β-连环蛋白mRNA含量多于肿瘤球细胞。Western bolt方法检测显示结果均为阳性,KB细胞中β-连环蛋白含量少于肿瘤球细胞。结论：在KB细胞及其肿瘤球细胞中β-连环蛋白转录并积累。