单纯型多数牙缺失的PAX9基因新突变一例

苏州大学附属第二医院口腔中心收治单纯型多数牙缺失1例,患者先天缺失恒牙16颗,其他系统无异常,其母与患者有相似表型。收集患者、患者父母及外祖父母外周血,提取DNA,采用PCR结合直接测序进行PAX9、MSX1、WNT10A、WNT10B、AXIN2和EDA等候选基因突变检测,PCR-Sanger测序发现PAX9基因的起始密码子存在1个杂合突变,即NM006194.3(PAX9):c.2T>C(p.Met1Thr),使起始密码子突变为苏氨酸,患者母亲有相同突变,可能是该患者先天单纯型多数牙缺失的遗传致病原因。患者拔除口内滞留乳牙后进行正畸-修复序列治疗,疗效满意。     

社区常见恶性肿瘤筛查研究

目的探寻一种简便的恶性肿瘤初筛自查方法。方法将人体肿瘤分为三类:体表肿瘤约占15％;空腔脏器肿瘤占65％;深层实体脏器肿瘤20％。通过科普宣传教育,让群众学会自查体表部位肿瘤,每年用自查盒助查各空腔脏器有无微量出血,隐血阳性时,去医院进一步精查。北京部分大学选40～70岁居民1万人,分成试验组、对照组各5千人。结果试验四年后:试验组共查出79例癌,年平均癌检出率482.5人/10万,癌死亡率为12.2/10万;对照组癌死亡率206.2人/10万,两组有显著性差异。在癌症高发区,同时作扩大普查试验:广东四会市肿瘤所,在门诊普查1669人,检出25例癌,鼻咽癌占24例。1999～2000年,江苏省食管胃癌高发区,普查近8万人,检出480例癌。加上“九五”以前的普查统计,用秦氏自查盒已筛查431075人,检出1272例癌,癌前病变1万多例。结论试验证明该方案设计合理,在当前是一种最简便的恶性肿瘤初筛自查方法。     

Differential expression profiling of lncRNAs related to Epstein-Barr virus infection in the epithelial cells

Epstein-Barr virus (EBV) is a prevalent γ-herpesvirus associated with a variety of cancers, including epithelial nasopharyngeal carcinoma (NPC) and gastric carcinoma (GC). Long noncoding RNAs (lncRNAs) are emerging as powerful regulators that have demonstrated to play crucial roles in cancer development. In this approach, based on genome-wide RNA sequencing, we examined the lncRNA expression profiles in four EBV genome-infected and EBV-negative 293 cells. A series of lncRNAs were found to be dysregulated in a comparative analysis. Then, eight typical lncRNAs were selected for validation of expression levels by real-time quantitative polymerase chain reaction and were detected in EBV-positive or EBV-negative GC and NPC cells. The differential expression patterns of the eight lncRNAs for validation were fundamentally identical to those revealed in RNA sequencing. Particularly, the differential expression of these lncRNAs in GC and NPC cells indicated their possible roles in EBV infection and tumorigenesis. In addition, a predicted lncRNA-microRNA-messenger RNA network suggested their potential interactions. This study reveals the first lncRNAome related to EBV infection in the epithelial cells and provides novel clues for the study of viral role in epithelial cancers through the interaction between EBV and host lncRNAs.     

Nuclear factor kappaB p65 phosphorylation in orthodontic tooth movement

Osteoclasts play a vital role in orthodontic tooth movement. Transactivation of nuclear factor kappaB (NFkappaB) by phosphorylation of the p65 component of NFkappaB at amino acid 536 (p65*(536)) plays a role in osteoclast differentiation stimulated by receptor activator of nuclear factor kappaB-ligand (RANK-L). We hypothesized that this transactivation pathway might be involved in the responses of alveolar bone cells during orthodontic tooth movement. We detected sharp increases in the levels of p65*(536) 3 and 12 hrs after the application of orthodontic stimuli in rats. In cell culture, osteoclast-like cells displayed no changes in p65*(536) in response to RANK-L, but levels rapidly increased after the cells were mechanically scraped. We conclude that p65*(536) is produced rapidly in response to orthodontic stimuli and mechanical insults, and may be important in bone remodeling associated with orthodontic tooth movement.     

Effects of Nidus Vespae extract and chemical fractions on glucosyltransferases, adherence and biofilm formation of Streptococcus mutans

Nidus Vespae (the honeycomb of Polistes olivaceous, P. japonicus Saussure and Parapolybiavaria fabricius) have been extensively used in traditional Chinese medicine, given their multiple pharmacological activities, including antimicrobial, anti-inflammatory, anti-virus, anti-tumor and anesthetic properties. The present study evaluated the anti-glucosyltransferases (GTFs) activity, anti-adherence and anti-biofilm properties of 95% ethanol/water extract, cyclohexane/ethyl acetate, petroleum ether/ethyl acetate and chloroform/methanol fractions of Nidus Vespae. Chloroform/methanol fraction showed a remarkable capacity for inhibiting the adherence of Streptococcus mutans ATCC 25175 to saliva-coated hydroxyapatite disc (S-HA) at sub-MC concentrations. In addition, the Nidus Vespae extract and chemical fractions significantly inhibited the activity of cell-associated and extracellular GTFs at sub-MIC concentrations, and the chloroform/methanol fraction was the most effective one. For the anti-biofilm activity assays, minimum biofilm inhibition concentrations (MBIC50) and minimum biofilm reduction concentrations (MBRC50) were determined using the microdilution method. The chloroform/methanol fraction showed the highest anti-biofilm activities with a MBIC50 of 8mg/ml and a MBRC(50) of 16mg/ml against Streptococcus mutans ATCC 25175. The significant inhibition of GTFs activity and biofilm formation demonstrated by Nidus Vespae shows it to be a promising natural product for the prevention of dental caries.     

布鲁顿酪氨酸激酶对破骨细胞增殖及分化作用的实验研究

目的 观察布鲁顿酪氨酸激酶（BTK）对破骨细胞增殖及分化的影响,探讨BTK对根尖周炎骨破坏的作用。方法 破骨前体细胞RAW264.7经100 ng·L ^-1核因子κB受体活化因子配体（RANKL）诱导5 d后,通过观察细胞形态、抗酒石酸酸性磷酸酶（TRAP）染色及实时荧光定量PCR（RT-qPCR）的方法,验证破骨细胞是否诱导成功。破骨细胞诱导成功后,转染BTK-小干扰RNA（siRNA）24 h,利用RT-qPCR检测TRAP mRNA的表达,采用CCK-8和TRAP酶活性检测法检测破骨细胞的增殖及分化情况。结果 RAW264.7细胞经RANKL诱导5 d后,可见大量体积较大,外观呈圆形、椭圆形,周围有不规则突起及TRAP染色阳性的多核破骨细胞。经BTK-siRNA转染24 h后,破骨细胞TRAP mRNA的表达水平显著降低（P<0.05）,其增殖及分化能力也明显受到抑制（P<0.05）。结论 抑制BTK表达,可以抑制破骨细胞的增殖及分化;BTK可作为抑制破骨细胞的新靶点。     

妇科千金胶囊毒理试验研究

妇科千金胶囊小鼠ig给药最大耐受量为384g生药/kg,相当于50公斤成人临床一日用量的960倍,未见明显的急性毒性反应.经大鼠ig给药(40g生药/kg)连续13周,大鼠的行为活动、食欲、二便、体重、血液和生化学指标及主要脏器系数和病理组织学检查均未见异常.     

原花青素对放射性心脏损伤大鼠血清cTn-Ⅰ、SOD、MDA、GSH-px的影响

目的:观察原花青素(proanthocyanidins)对放射性心脏损伤(radiation-induced heart damage,RIHD)大鼠血清肌钙蛋白Ⅰ(cardiac troponin Ⅰ,cTn-Ⅰ)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-px)表达水平的影响,以及对RIHD大鼠的可能作用及作用机制.方法:60只雄性远交群(sprague dawley,SD)大鼠,随机数字表法分为6组,每组10只,分别为正常对照组、单纯照射组、OPC小剂量(50 mg/kg)组、OPC中剂量(100 mg/kg)组、OPC大剂量(150 mg/kg)组、蓝莓(200 mg/kg)组.除正常对照组外,其余各组大鼠给予6MVX线单次大剂量照射心脏,总剂量(total dose,Dt)20 Gy,OPC组和蓝莓组大鼠给予相应药物灌胃.放疗结束后4周,HE染色、Masson染色观察大鼠心肌组织病理改变,ELISA检测血清cTn-Ⅰ、SOD、MDA、GSH-px表达水平.结果:与正常对照组比较,单纯照射组、OPC组、蓝莓组病理评分和心肌胶原容积分数(collagen volume fraction,CVF)评分升高(P＜0.05);与单纯照射组比较,OPC组、蓝莓组病理评分和CVF评分升高(P＜0.05),差异均具有统计学意义.与正常对照组相比,单纯照射组、OPC组、蓝莓组cTn-Ⅰ、MDA表达水平升高,单纯照射组、OPC组、蓝莓组SOD、GSH-px表达水平下降;与单纯照射组相比,OPC组、蓝莓组cTn-Ⅰ、MDA表达水平升高,OPC组、蓝莓组SOD、GSH-px表达水平下降,差异均具有统计学意义(P＜0.05).结论:OPC可通过抑制氧化应激,下调RIHD大鼠血清cTn-Ⅰ、MDA表达水平,上调血清SOD、GSH-px表达水平,抑制RIHD大鼠心肌损伤,疗效优于蓝莓.