牙齿发育早期间充质细胞的差异蛋白分析

目的:应用蛋白质组学方法分析间充质细胞在牙齿发育早期不同阶段的蛋白质差异表达情况,从蛋白水平进一步了解牙齿发育早期的分子机制.方法:制备C57胎鼠起始期(E11.5)、蕾状期(E13.5)冰冻切片,通过激光捕获显微切割技术(LCM)获取两个时期下颌第一磨牙牙胚间充质细胞,采用双向凝胶电泳方法分离蛋白质,比较分析凝胶图像,通过质谱及生物信息学分析鉴定差异蛋白.结果:激光捕获了较纯的目的细胞,得到双向凝胶电泳图谱,比较分析发现E13.5组8个蛋白点明显上调,E11.5组5个蛋白点发生明显上调.质谱分析初步鉴定了蛋白质种类.结论:LCM技术能较好地获取较为均一的目的细胞,并发现牙胚发育起始期与蕾状期间充质细胞蛋白质表达存在差异,为牙胚早期发生的分子机制研究提供一种新的方法.     

人牙髓细胞向成牙本质细胞分化的蛋白质组学研究

目的 采用蛋白质组学方法分析人牙髓细胞向成牙本质细胞分化过程中细胞蛋白表达谱的改变,揭示特征蛋白在分化过程中所起的作用.方法对人牙髓细胞进行矿化诱导,提取诱导前后细胞总蛋白,双向电泳分离蛋白质,DeCyder V6.0软件确定差异蛋白点,质谱鉴定差异蛋白质.结果双向电泳确认46个差异蛋白质斑点,质谱鉴定20个蛋白斑点,差异蛋白质涉及细胞周期调节、能量代谢、信号传导等细胞生物学过程.结论蛋白质组学技术可高通量筛选与人牙髓细胞向成牙本质细胞分化相关的功能蛋白.     

变异链球菌蛋白质提取方法研究

目的建立稳定、有效的提取变异链球菌蛋白质方法。方法以裂解液为提取介质,分别采用反复冻融超声、超声、热裂解和热裂解超声4种方法提取细菌蛋白质,通过分析细菌破碎程度、蛋白质SDS-PAGE电泳及蛋白质质量浓度,比较各提取方法的效果。结果除热裂解法外,其他3种方法均可有效裂解细菌,其中超声起主要作用,同时这3种方法提取蛋白质的SDS-PAGE条带分布基本相同,但反复冻融超声法提取蛋白质条带丰度最高。蛋白质质量浓度测定显示,反复冻融超声法提取蛋白质质量浓度最高。4种方法均具有较好的稳定性和重复性。结论反复冻融超声法可以有效提取变异链球菌蛋白质,有较好的稳定性和重复性,且操作简单。     

利用shotgun蛋白组学策略构建变形链球菌蛋白质表达谱

目的：建立变形链球菌蛋白质表达谱,为变形链球菌蛋白质组学研究奠定基础。方法：提取变形链球菌总蛋白,溶液内酶解,肽段混合物在色谱上分离后shotgun鉴定,并利用生物信息学软件对鉴定出的蛋白质进行分析。结果：建立了变形链球菌蛋白质表达谱,共鉴定出752种蛋白质,其中高可信度298种,并利用Gene Ontology软件对其按分子功能和生物学途径进行归类。结论：变形链球菌蛋白质表达谱的建立,为变形链球菌蛋白质组学研究提供了重要的信息和资料。     

牙胚细胞的分离培养

目的：分离培养牙胚细胞。方法：选择出生后（PN）8天的SD大鼠,从第一磨牙牙胚上撕脱牙囊和成釉器组织,切取牙胚颈部组织,剁离牙乳头组织,分别进行细胞培养,通过差速消化法分离符种牙胚细胞。结果：牙囊、牙乳头、成釉器和上皮根鞘细胞被分离纯化出来。结论：利用差速消化法可同时分离培养4种主要的牙胍细胞。     

改良热酸酚法提取白色念珠菌早期生物膜总RNA的研究

目的 通过白色念珠菌改良热酸性酚法RNA提取方法与玻璃珠法、超声破碎法和液氮研磨法的比较,确定一种有效提取白色念珠菌早期生物膜总RNA的方法.方法 将培养3 h和6 h白色念珠菌生物膜分为2组,每组分别采用热酸性酚法、玻璃珠法、超声破碎法和液氮研磨法,提取酵母相白色念珠菌的总RNA,用分光光度计及甲醛变性凝胶电泳检测RNA的浓度、纯度以及完整性.结果 1～8号样本RNA总量依次为:9.96、12.72、2.88、4.56、2.76、4.92、7.32、10.68 μg.结论 热酸性酚法是提取白色念珠菌早期生物膜总RNA的理想方法.     

牙本质磷蛋白和成釉蛋白在组织工程牙胚的表达模式研究

目的：采用大鼠牙胚细胞与胶原蛋白构建组织工程牙齿,通过免疫荧光组织化学分析牙齿特异性蛋白在组织工程牙齿的表达模式。方法：采用出生后4dSD仔鼠磨牙牙胚细胞,培养后将牙胚细胞与胶原蛋白溶液混合构建组织工程牙胚,2W后取材,HE染色观察移植物中的成牙能力,免疫荧光组织化学染色观察牙本质磷蛋白DSP和成釉蛋白AM的表达模式。结果：组织工程牙胚移植后2W可在肾被膜下形成乳白色矿化组织,HE染色可见典型的牙髓牙本质复合体样组织和釉质样结构,免疫荧光组织化学染色观察可见成釉蛋白AM阳性的釉质样结构和牙本质磷蛋白DSP阳性的牙乳头样组织,与正常牙齿组织表达模式一致。结论：采用牙胚细胞可与胶原蛋白进行重新排列形成组织工程牙胚,本实验从组织和蛋白分析上证实了其具有较强的成牙能力,为以后的临床应用提供了理论基础。     

唾液蛋白质组学唾液样本制备的方法学初探

目的：通过对唾液淀粉酶单克隆抗体的制备和鉴定,寻找蛋白质组学研究中样本制备关键环节的可行性方案,为日后唾液蛋白质组学研究奠定基础。方法：将腮腺液经超滤法浓缩蛋白后,进行SDS-PAGE分析,切取分子量为50～65kDa的高丰度蛋白条带,研磨后注射BALB/c小鼠,诱导产生免疫应答并制备人唾液淀粉酶单克隆抗体,应用酶联免疫吸附试验和蛋白免疫印记等方法鉴定。结果：成功制备和鉴定了人唾液淀粉酶单克隆抗体。结论：成功制备和鉴定了唾液淀粉酶单克隆抗体,为唾液蛋白质组学研究中样本制备提供解决方案。     

小鼠釉丛蛋白基因不同mRNA剪接体的克隆

目的：克隆小鼠釉丛蛋白基因不同mRNA剪接体，分析小鼠釉丛蛋白的多态性。方法：采用异硫酸氰胍一步法从新生小鼠磨牙牙胚组织中提取总RNA，反转录成cDNA第一链，经PCR扩增出小鼠釉丛蛋白基因特异片段，播入pBS质粒，筛选阳性克隆，并经酶切和核苷酸序列分析鉴定阳性克隆。结果：克隆T1序列与已发表的完整釉从蛋白mRNA序列一致。克隆T2序列与完整釉丛蛋白mRNA序列比较，缺失序列的84-158的75个碱基片段。结论：小鼠釉从蛋白基因在转录mRNA时，可产生不同的剪接体。     

维甲酸诱导鼠磨牙牙胚midkine基因表达的研究

探讨ＲＡ与ｍｉｄｋｉｎｅ（ＭＫ）基因表达在牙胚发育过程中的相互关系。方法：将１６ｄ胎龄的鼠胎下颌第一磨牙牙胚分别置于含不同浓度的外源性ＲＡ（分别５＊１０＾－８,５＊１０＾－７和５＊１０＾－６ｍｏｌ／Ｌ）的ＲＰＭＩ１６４０半固态培养基表面。培养ｄ后分别提取总ＲＮＡ,经ＲＴ－ＰＣＲ扩增２５个循环,采用Ｓｏｕｔｈｅｒｍ印迹法检测ＭＫｍＲＮＡ在牙胚组织中的表达变化。结果体外培养ｄ后,对照组牙胚ＭＫｍＲＡＮ     

Oral cancer: molecular technologies for risk assessment and diagnosis

Purpose: The effective biomarkers related to diagnosis, metastasis, drug resistance and irradiation sensitivity of oral cancers will help the pathologist and oncologist to determine the molecular taxonomy diagnosis and design the individualization treatment for the patients with oral cancers.     

Stimulated whole saliva components in children with Down syndrome

The authors report on the components of stimulated whole saliva from children with Down syndrome—including pH, flow rate, sialic acid and protein concentrations, and amylase and peroxidase activity. Saliva samples were collected from 35 children aged 6–10 years. Of the participants, 17 had Down syndrome and 18 did not. To stimulate saliva production, the children chewed a piece of parafilm for 10 minutes before the sample was collected. Soon after collecting the saliva sample, the authors measured pH using a portable pH-meter. Sialic acid levels were determined with a thiobarbituric acid assay. Protein content was determined with Folin's phenol reagent. Amylase was assayed and the authors measured the maltose produced by the breakdown of starch and peroxidase using ortho-dianisidine.
No statistically significant difference was observed in levels of sialic acid (free and total) between the two groups. Protein concentration was about 36% higher in the group with Down syndrome. However, the salivary flow rate, pH, and amylase and peroxidase activities were lower among the children with Down syndrome.     

柠檬精油对牙周致病菌抑制作用及其安全性初步研究

目的:探讨柠檬精油对牙周致病菌的体外抗菌活性及对细胞增殖的影响。方法:采用微量液体稀释法测定柠檬精油对Pg、Fn、Aa、Pi的最小抑菌浓度(minimal inhibitory concentration,MIC)及最小杀菌浓度(minimum bactericidal concentration,MBC);以较低浓度的MIC为标准稀释LEO作为实验组,采用MTT法测定柠檬精油对HUVECs的毒性作用,明确抑菌浓度下LEO的安全性。结果:柠檬精油对牙周主要致病菌均有抑菌作用,Pg、Fn、Aa、Pi的MIC分别是9.0 g/L、4.5 g/L、4.5 g/L、9.0 g/L,Aa、Fn的 MBC是9.0 g/L,Pg、Pi的MBC未测得。1/2MIC、1/20MIC浓度的LEO能够抑制人脐静脉内皮细胞的生长,而低于1/200MIC浓度的LEO则对人脐静脉内皮细胞的生长没有影响,其中1/200MIC浓度的LEO作用明显优于0.02%的CHX。结论:体外环境中,柠檬精油对牙周致病菌Pg、Fn、Aa、Pi具有抗菌活性,低浓度应用对机体相对安全。     

Clinical and pathological aspects on some odontogenic tumors

Odontogenic tumors constitute a very diverse group of lesions that reflects the complex processes of odontogenesis. Controversies over their classification/subtyping, terminology and diagnosis have been persisted, which has direct bearings on therapeutic and/or prognostic implications.     

牙髓再生的研究现状

目前,根管治疗术在临床上已得到了广泛的应用并成功的保存了患牙,但其仍有许多术后并发症。随着组织工程技术在口腔领域的应用,构建一种组织工程化牙髓进行牙髓-牙本质复合体的再生治疗成为可能。近年来国内外已有很多关于牙髓再生的研究,并且已经取得了一定成果。本文将对目前国内外牙髓再生的研究进展现状作一综述。     

Computer-assisted planning for maxillofacial prosthodontic rehabilitation

Computerized tomography (CT) planning and the use of CT derived surgical templates for implant placement have shown promise for restoring function within months after surgical reconstruction of acquired post-oncologic defects.     

Effect of Pingyangmycin on human venous malformation endothelial cells

Purpose: Venous malformations are common vascular anomalies with a propensity of the head and neck. Intralesional injection of Pingyangmycin (PYM, bleomycin A5 hydrochloride) is a widely used sclerotherapy method for the treatment of venous malformation.     

Can you be too old for oral implants? An update on ageing and plasticity in the oro‐facial sensorimotor system

This review focuses on the capacity of the brain for plasticity and the utility and efficacy of oral implants in helping to restore oro‐facial sensorimotor functions, especially in elderly patients. The review first outlines the components of the oro‐facial sensorimotor system which encompasses both oro‐facial tissues and a number of brain regions. One such region is the sensorimotor cortex that controls the activity of the numerous oro‐facial skeletal muscles. These muscles are involved in a number of functions including reflexes and the more complex sensorimotor functions of mastication, swallowing and speech. The review outlines the use by the brain of sensory inputs from oro‐facial receptors in order to provide for exquisite sensorimotor control of the activity of the oro‐facial muscles. It highlights the role in this sensorimotor control played by periodontal mechanoreceptors and their sensory inputs to the brain, and how oral implants in concert with the plastic capacity of the brain may, at least in part, compensate for reduced sensorimotor functioning when teeth are lost. It outlines findings of ageing‐related decrements in oro‐facial sensorimotor functions and control. The changes in oro‐facial tissues and the brain that underlie these ageing‐related functional alterations are also considered, along with adaptive and compensatory processes that utilise the brain's capacity for plasticity. The review also notes the evidence t hat rehabilitation that incorporates adjunctive approaches such as sensorimotor training paradigms in addition to oral prostheses such as implants may enhance these processes and help maintain or facilitate recovery of sensorimotor functioning in the elderly.     

Tumor microenvironment in oral cancer: novel approaches for immunotherapy

Squamous cell carcinoma of the head and neck (SCCHN), which arises from the squamous mucosal epithelium of the oral cavity, pharynx and larynx, is a major health problem in the US and other parts of the world, especially in developing countries.