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国家自然科学基金(30430690)

作品数:9 被引量:29H指数:3
相关作者:王松灵夏登胜张春梅杨圣辉孙启鸿更多>>
相关机构:首都医科大学附属北京口腔医院首都医科大学美国国立卫生院更多>>
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小型猪与小鼠腮腺、泪腺、垂体超微结构的观察与比较
2009年
目的研究小型猪及小鼠腮腺的超微结构特点,并将腮腺与泪腺、垂体的超微结构对比观察,旨为涎腺基因治疗转基因表达的生物学特性提供腺体形态学的依据。方法选用成年小型猪、昆明种小鼠各5只,分别取腮腺、泪腺、垂体进行超微结构观察。结果小型猪较小鼠的腮腺腺泡细胞内分泌颗粒密度大,质地均匀,细胞器少见,血管内皮细胞内有较多分泌颗粒存在。小型猪与小鼠泪腺超微结构均与各自腮腺相似,小型猪与小鼠垂体结构特点为细胞排列散在,细胞闻充满血窦及毛细血管。结论小型猪腮腺腺泡细胞分泌颗粒多,其毛细血管内皮细胞中也可见较多分泌颗粒,提示这些分泌颗粒可能进入血液发挥内分泌功能。
颜兴海波孙异临张春梅王松灵
关键词:腮腺泪器垂体
唾液硝酸盐、亚硝酸盐代谢及其对人体影响的研究被引量:8
2005年
外源性硝酸盐经口腔进入胃内,经过胃肠循环进入血液,被唾液腺主动吸收,然后分泌到口腔。唾液硝 酸盐在口腔内经硝酸盐还原菌还原成亚硝酸盐,亚硝酸盐对机体的影响具有两重性。
夏登胜王松灵
关键词:唾液腺亚硝酸盐口腔内胃内胃肠
Antimicrobial effect of acidified nitrate and nitrite on six common oral pathogens in vitro被引量:7
2006年
Background Salivary nitrate is positively correlated with plasma nitrate and its level is 9 times the plasma level after nitrate loading. Nitrate in saliva is known to be reduced to nitrite by oral bacteria. Nitrate and nitrite levels in saliva are 3-5 times those in serum in physiological conditions respectively in our previous study. The biological functions of high salivary nitrate and nitrite are still not well understood. The aim of this in vitro study was to investigate the antimicrobial effects of nitrate and nitrite on main oral pathogens under acidic conditions. Methods Six common oral pathogens including Streptococcus mutans NCTC 10449, Lactobacillus acidophilus ATCC 4646, Porphyromonas gingivalis ATCC 33277, Capnocytophaga gingivalis ATCC 33624, Fusobacterium nucleatum ATCC 10953, and Candida albicans ATCC 10231 were cultured in liquid medium. Sodium nitrate or sodium nitrite was added to the medium to final concentrations of 0, 0.5, 1, 2, and 10 mmol/L. All of the microorganisms were incubated for 24 to 48 hours. The optical densities (OD) of cell suspensions were determined and the cultures were transferred to solid nutrient broth medium to observe the minimum inhibitory concentration and minimum bactericidal/fungicidal concentration for the six tested pathogens. Results Nitrite at concentrations of 0.5 to 10 mmol/L had an inhibitory effect on all tested organisms at low pH values. The antimicrobial effect of nitrite increased with the acidity of the medium. Streptococcus mutans NCTC 10449 was highly sensitive to nitrite at low pH values. Lactobacillus acidophilus ATCC 4646 and Candida albicans ATCC 10231 were relatively resistant to acidified nitrite. Nitrate at the given concentrations and under acidic conditions had no inhibitory effect on the growth of any of the tested pathogens. Conclusion Nitrite, at a concentration equal to that in human saliva, is both cytocidal and cytostatic to six principal oral pathogens in vitro, whereas nitrate at a similar concentration has no antimicrobial effect
XIA Deng-shengLIU YinZHANG Chun-meiYANG Sheng-huiWANG Song-lin
关键词:NITRITENITRATEACIDIFICATION
小型猪、大鼠及人口腔常见菌群的比较研究被引量:11
2006年
目的观察小型猪口腔不同部位常见菌群的分布情况,并与大鼠及人进行比较。方法选取小型猪、大鼠及人前牙、后牙、舌背和唾液的细菌标本进行培养,对培养结果进行比较分析。结果选取的口腔链球菌、放线菌、核梭菌、二氧化碳嗜纤维菌及乳杆菌等5种人口腔中的常见菌群在小型猪口腔中均可发现。其中小型猪与人5种菌群的构成比在前牙区无显著性差异(P>0.05);小型猪后牙区放线菌的构成比明显低于人(P<0.05);唾液中的变链菌占口腔链球菌的百分比低于人(P<0.05);小型猪舌背二氧化碳嗜纤维菌的构成比高于人(P<0·05);前后牙菌斑及唾液中产黑色素菌占厌氧菌的比例高于人(P<0.05)。结论大鼠的菌群组成与人差异较大,小型猪是目前口腔疾病研究中较为理想的一种大型实验动物模型。
李岩涛李金陆杨圣辉夏登胜张春梅王松灵
关键词:小型猪口腔微生态口腔菌群
Effect of Same-dose Single or Dual Field Irradiation on Damage to Miniature Pig Parotid Glands
2009年
Aim To evaluate the effect of single or dual field irra- diation (IR) with the same dose on damage to miniature pig parotid glands. Methodology Sixteen miniature pigs were divided into two IR groups (n=6) and a control group (n=4). The irradi- ation groups were subjected to 20 Gy X-radiation to one parotid gland using single-field or dual-field modality by linear accelerator. The dose-volume distributions between two IR groups were compared. Saliva from parotid glands and blood were collected at 0, 4, 8 and 16 weeks after irradiation. Parotid glands were removed at 16 weeks to evaluate tissue morphology.Results The irradiation dose volume distributions were significantly different between single and dual field irradi- ation groups (t----4.177, P=0.002), although dose volume histogramin (DVH) indicated the equal maximal dose in parotid glands. Saliva flow rates from IR side decreased dramatically at all time points in IR groups, especially in dual field irradiation group. The radiation caused changes of white blood cell count in blood, lactate dehydrogenase and amylase in serum, calcium, potassium and amylase in saliva. Morphologically, more severe radiation damage was found in irradiated parotid glands from dual field irradi- ation group than that from single field irradiation group. Conclusion Data from this large animal model demons- trated that the radiation damage from the dual field irradiation was more severe than that of the single field irradiation at the same dose, suggesting that dose-volume distribution is an important factor in evaluation of the radiobiology of parotid glands.
Xing YanBo HaiZhao-chen ShanChang-yu ZhengChun-mei ZhangSong-lin Wang
兔抗人唾液酸转运蛋白抗体的制备及特性鉴定
2006年
目的:制备兔抗人唾液酸转运蛋白(sialin)的抗体,并进行特性鉴定。方法:应用RT—PCR从培养的人颌下腺细胞系HSG中扩增编码sialin N端1~38氨基酸的DNA序列,构建重组表达质粒pGEX-5X-1-sialin,测序鉴定后,转化大肠杆菌JM109,在0.1 mmol/L IPTG的诱导下表达GST—sialin融合蛋白。经SDS-PAGE鉴定后,利用GSTrap FF^TM柱纯化融合蛋白,并以其免疫家兔制备抗人sialin抗体。用ELISA法测定兔抗sialin血清的效价;用Western blot及免疫细胞化学等方法鉴定抗血清的特异性。结果:构建了重组表达质粒pGEX-5X-1-sialin;以其转化大肠杆菌在IPTG诱导下,获得以可溶性形式表达的GST-sialin融合蛋白;表达的GST—sialin经GSTrap FF^TM柱纯化后,免疫家兔制备出抗sialin抗血清,ELISA法测定抗血清的效价为1:32000。Western blot鉴定表明,制备的抗sialin抗体可特异地识别HSG细胞中相对分子质量(Mr)约55000的sialin蛋白。免疫细胞化学检测表明,sialin抗血清识别的抗原定位于HSG细胞的细胞质和胞核。结论:成功地制备出兔抗人sialin抗血清,为进一步研究sialin在涎腺组织的功能奠定了基础。
孙其飞高媛刘细保孙启鸿王松灵
关键词:融合蛋白原核表达抗体制备
小型猪双侧腮腺萎缩后唾液流率及口腔细菌变化的观察
2007年
目的观察小型猪腮腺萎缩后混合唾液流率的变化以及对口腔主要致病菌的影响。方法10只小型猪分成两组,每组5只,其中一组双侧腮腺注入甲紫致腺体萎缩,另5只作为健康对照组。分别在萎缩后第12、24个月收集混合唾液并观察唾液流率的变化;同时检测小型猪龈下菌斑和混合唾液中口腔主要致病菌数量并与健康对照组比较。结果腮腺萎缩后第12个月混合唾液流率自(1.36±0.74)ml/min下降到(1.32±0.65)ml/min,第24个月唾液流率自(0.72±0.34)ml/min上升到(0.86±0.57)ml/min;龈下菌斑中的核梭杆菌、后牙龈下菌斑中厌氧菌总数、产黑菌数量均明显高于健康对照组;第12个月前牙和第24个月后牙的龈下菌斑中变形链球菌有短暂升高,而需氧菌总数、乳酸菌和放线菌数量无明显变化。唾液中除变形链球菌和产黑菌在腮腺萎缩早期有轻度上升外,其余细菌数量均无明显变化。结论双侧腮腺萎缩后明显降低口腔中唾液总量,导致牙菌斑中的致龋菌和牙周致病菌数量上升。
夏登胜刘颖张春梅杨圣辉王松灵
关键词:腮腺唾液分泌动物
唾液硝酸盐、亚硝酸盐检测方法的探讨被引量:3
2007年
目的通过比较几种不同测定唾液硝酸盐、亚硝酸盐含量的方法来寻求最佳测定唾液微量硝酸盐、亚硝酸盐的标准方法。方法比较经典的Griess法、镉柱法和高压液相色谱法(HPLC)对含不同浓度的硝酸盐、亚硝酸盐标准品以及10位健康志愿者混合唾液样本中硝酸盐、亚硝酸盐含量的最低检出限和硝酸盐、亚硝酸盐回收率。结果Griess法和镉柱法测定唾液硝酸盐、亚硝酸盐的最低检出限NO3-为10ng/μl,NO2-为1ng/μl,回收率为91.2%;HPLC法测定唾液硝酸盐、亚硝酸盐的最低检出限为NO3-为0.8ng/μl,NO2-为0.1ng/μl,回收率达到97.6%。结论HPLC在微量硝酸盐、亚硝酸盐检测中的灵敏度明显高于经典的Griess法。
夏登胜崔秀玉邓大君王松灵
关键词:硝酸盐亚硝酸盐混合唾液高压液相色谱
Sialin(SLC17A5)在腮腺、颌下腺及颌下腺细胞系HSG表达的初步分析被引量:1
2006年
目的研究唾液酸转运蛋白Sialin在正常涎腺组织和颌下腺细胞系HSG的表达。方法应用基因芯片检测正常人腮腺、颌下腺组织Sialin编码基因SLC17A5的表达;提取腮腺、颌下腺组织及颌下腺细胞系(HSG)总RNA,通过RT-PCR在基因水平上验证SLC17A5基因的表达;提取HSG细胞总蛋白,用免疫印迹法检测Sialin在蛋白水平表达。结果基因芯片检测到Sialin编码基因SLC17A5基因在正常人腮腺、颌下腺均有表达,表达比值分别为226、206·9,不存在表达差异;RT-PCR验证芯片结果并在其他涎腺样本及颌下腺细胞系HSG中能扩增出其全部翻译区1485bp的片段,应用商业化的抗体能够在HSG细胞检测到Sialin相应的约54KDa蛋白。结论正常腮腺、颌下腺组织和颌下腺细胞系HSG均表达SLC17A5基因,但是唾液酸转运蛋白Sialin在组织、细胞内定位和功能需要进一步研究。
孙其飞刘细保孙启鸿王松灵
关键词:RT-PCR免疫印迹
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