促髓细胞分化的锌指基因单向PCR扩增直接测序的研究

锌指基因是一种造血调节基因,编码锌指结构蛋白,主要在髓细胞中表达,促进髓细胞分化,在急性早幼粒白血病维甲酸治疗中,促使病情缓解。本文报道了我们从基因分子上研究锌指基因作用中,探索并建立了单向聚合酶链反应（ＰＣＲ）扩增特定单链ＤＮＡ,直接测序的新方法。它能产生质和量均佳的单链ＤＮＡ,无需纯化即可直接用于测序,使复杂的测序研究简便易行,可在２,３天内完成。这种单向ＰＣＲ扩增特定单链ＤＮＡ直接测序的方法,经对锌指基因的ｃＤＮＡ测序,得到验证。此法不仅适用于疾病研究中的ＤＮＡ测序,还可制各单链ＤＮＡ探针,更利于基因结构组成的研究。     

Localization of juvenile, but not late-infantile, neuronal ceroid lipofuscinosis on chromosome 16.

The neuronal ceroid lipofuscinoses (NCL) are a group of progressive neurodegenerative disorders characterized by the deposition of autofluorescent proteinaceous fingerprint or curvilinear bodies. We have found that CLN3, the gene underlying the juvenile form of NCL, is very tightly linked to the dinucleotide repeat marker D16S285 on chromosome 16. Integration of D16S285 into the genetic map of chromosome 16 by using the Centre d'Etude du Polymorphisme Humain panel of reference pedigrees yielded a favored marker order in the CLN3 region of qtel-D16S150-.08-D16S285-.04-D16S148-.02-D16S 67-ptel. The most likely location of the disease gene, near D16S285 in the D16S150-D16S148 interval, was favored by odds of greater than 10(4):1 over the adjacent D16S148-D16S67 interval, which was recently reported as the minimum candidate region. Analysis of D16S285 in pedigrees with late-infantile NCL virtually excluded the CLN3 region, suggesting that these two forms of NCL are genetically distinct.     

几种药物对HL-60细胞的诱导分化作用及其过程中蛋白激酶C活力分布的变化

本文就HHT、RA、WB_(852)对HL-60细胞的诱导分化作用及此过程中PKC活力在细胞胞浆部分及膜溶脱部分的变化进行研究。结果表明,在适当的用药浓度下,从细胞生长抑制情况、形态学观察及NBT还原能力测定判断,三种药物对HL-60细胞有明显的诱导分化作用。PKC活力分布变化的研究结果表明,用药组细胞胞浆部分酶活力有不同程度的下降,尤在用药早期(约6h以前)下降显著;而膜部分PKC活力则表现上升、或下降,或活力相差不大的结果。暗示在信息传递过程中起核心作用的PKC对不同的胞外刺激可能采取不同的应答方式。PKC的作用可能主要发生在信息传递的早期。     

血清岩藻糖测定方法学研究

本文对血清岩藻糖测定方法进行了系统研究。血清用量由200μL减至50μL,显色反应4h内稳定。吸收峰在396nm。岩藻糖浓度40μg/mL内线性良好,批间CV=2.1％。以Sephadex-G200层析,血清岩藻糖主要存在于分子量为500kD的组分中。以本法测得30例健康人血清岩藻糖浓度为588.1±172.0μmol/L。     

SLE血清中抗-DNA抗体分离和性质的研究

本文用国产高分子树脂(T)接枝小牛胸腺DNA,通过亲合层析从系统性红斑狼疮SLE患者血清中纯化出抗-ds DNA抗体和抗-ss DNA抗体。酶联免疫吸附分析(ELISA)的研究表明:SLE抗-DNA抗体和DNA结合的差异性很大,是高度非均一性的。抗-ss DNA抗体不仅组成成分比抗-ds DNA抗体复杂,ss DNA/抗-ssDNA亲合能力也明显高于ds DNA/抗-ds DNA。纯化的抗-DNA抗体以IgG类抗体占主导,同时也有其它类型抗体存在(例如IgM等)。抗-ds DNA抗体有较抗-ss DNA抗体高的IgG含量(两者的IgG/IgM分别是7.0和4.0),说明IgG抗-DNA抗体更倾向于同dsDNA结合。     

硫酸铝钾对小鼠肝损害的实验研究

本文主要观察硫酸铝钾——饮用水净化剂引致小鼠肝酶组化,超微结构和组织结构的变化。动物30只,分为正常组、硫酸铝钾大、小剂量组。实验结果:硫酸铝钾10mg/kg/日(大剂量组)与5mg/kg/日(小剂量组)动物用药10天、80天均可使肝SDH酶活性降低;肝细胞线粒体肿胀、嵴断裂和溶解。在光学显微镜下用药80天后大部分肝细胞肿胀,胞浆疏松淡染、肝小叶内可见呈小灶状分布的炎性细胞浸润,主要为淋巴细胞及浆细胞,门管区偶见少许炎性细胞,但肝小叶和门管区未见结缔组织增生。为此,硫酸铝钾作为净水剂,不宜用量过大。     

慈竹无性系种群能值特点及其影响能值计测因素的研究

 本文引用Harper(1977)的构件结构理论,从构件结构单位、无性系分株和无性系三个层次,对四川南充市郊慈竹无性系种群的能值特点及其影响能值的计测因素进行了定量研究。研究结果表明：慈竹无性系种群中,各构件单位的去灰分热值(AFCV)分别为：根15349.42J／g、根茎16372.92J／g、秆17106.06J／g、枝18111.99J／g和叶19451.90J／g;慈竹无性系分株的AFCV(J／g)随龄级增大而递增;慈竹无性系水平上的AFCV为：Ⅰ龄占16.47%、Ⅱ龄占25.76%、Ⅲ龄占36.32%、Ⅳ龄为13.08%及Ⅴ龄为8.37%。用恒容燃烧法测定热值时,其能值变化与氧分压密切相关。用经验公式计算的能值较作图法高;用AFCV表示能值较总干重热值(GCV)准确。     

枯草杆菌BS224菌在防治Ⅲ°烧伤创面感染中痂下组织细菌数量的动态观察

对48例Ⅲ°烧伤病人的创面,定量植入枯草杆菌BS224菌后,分别在24h、48h、72h及96h做痂下组织细菌定量检测。结果显示:枯草杆菌对痂下组织的致病菌有明显的拮抗作用。感染创面的BS224菌体数量24—48小时显著增加,72—96小时而下降。与清洁创面的BS224菌动态变化上相同,呈常态曲线的规律变化。     

Blockage of MDM2-mediated p53 ubiquitination by yuanhuacine restrains the carcinogenesis of prostate carcinoma cells by suppressing LncRNA LINC00665

Prostate cancer (PCa) is a challenging issue for men's health worldwide due to its uncontrolled proliferation and high metastatic potential. Increasing evidence has supported plant extracts and natural plant derivatives as promising antitumor therapy with less toxic side effects. Yuanhuacine is an active component isolated from Daphne genkwa and can effectively suppress the tumorigenesis of several cancers. However, its role in PCa remains unclear. In this study, yuanhuacine dose-dependently inhibited the proliferation and induced apoptosis of PCa cells. Moreover, yuanhuacine also restrained the invasion and migration of PCa cells. Mechanically, yuanhuacine decreased the ubiquitination and degradation of p53 protein, and ultimately increased p53 levels, which was regulated by inhibiting the phosphorylation and total protein levels of mouse double minute 2 (MDM2). Moreover, elevation of MDM2 reversed the suppressive efficacy of yuanhuacine in PCa cell viability, invasion, and migration. The network pharmacologic and bioinformatics analysis confirmed that MDM2 might be a common target of D. genkwa and LINC00665. Furthermore, yuanhuacine inhibited LINC00665 expression. Upregulation of LINC00665 reversed yuanhuacine-mediated inhibition in MDM2 protein expression and suppressed p53 levels by enhancing its ubiquitination in yuanhuacine-treated cells. Importantly, the inhibitory effects of yuanhuacine on cell viability and metastatic potential were offset after LINC00665 elevation. Together, the current findings highlight that yuanhuacine may possess tumor-suppressive efficacy by inhibiting LINC00665-mediated MDM2/p53 ubiquitination signaling. Therefore, this study indicates that yuanhuacine may be a promising candidate for the treatment of PCa.