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目的探讨多重PCR技术用于检测牙龈卟啉单胞菌(Porphyromonasgingivalis,P.gingivalis)致病岛rag基因的可行性,并研究致病岛rag基因在毒力株和非毒力株中的分布情况。方法本研究于2011年1—6月在山东省口腔生物医学重点实验室进行,对P.gingivalis毒力株和非毒力株进行厌氧培养,采用普通PCR和多重PCR分别对他们的致病岛rag基因进行检测,对比检测结果,并对临床标本进行预实验研究。结果普通PCR和多重PCR均能对P.gingivalis致病岛rag基因进行检测,并且结果一致。多重PCR可用于临床标本的检测。致病岛rag基因不同基因型在毒力株和非毒力株中的分布不同,rag-1型存在于高毒力株P.gingivalisW83中,而rag-4型存在于低毒力株P.gingivalisATCC33277中。结论致病岛rag基因与细菌致病性密切相关;多重PCR技术省时省力、简单快速,为后续临床标本中rag基因的快速检测提供实验依据。     

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目的评价Flexitime硅橡胶印模材料的临床操作性能。方法对2011年11月至2012年1月于中国医科大学附属口腔医院就诊的60例上颌磨牙单冠修复患者,分别采用Flexitime硅橡胶印模材料（Flexitime组）、Express^TM硅橡胶印模材料(Express^TM组)和Impregum聚醚橡胶印模材料（Impregum组）单步取模法制取上颌印模,根据操作时间、口内凝固时间和从口内脱模难易程度对3种印模材料的临床操作性能进行综合分析比较。结果 Flexitime组、Express^TM组及Impregum组操作时间和口内凝固时间两两比较差异均有统计学意义（P<0.01）。Flexitime组口内凝固时间与操作时间无相关关系（P>0.05）;Express^TM组及Impregum组均呈负相关（P<0.01）。Flexitime组及Express^TM组与Impregum组口腔内脱模难易程度比较差异有统计学意义（P<0.01）。结论 Flexitime硅橡胶印模材料操作时间短,口内凝固快,且口内凝固时间不受操作时间影响,从口内脱模容易,临床操作性能优越。     

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全颌曲面断层片现在已经广泛应用于口腔正畸临床工作中,它是根据口腔颌面部左右对称、颌骨牙弓呈弓形等解剖特点而设计的三轴固定连续转换的弧面断层摄影,正畸医生可以通过曲面断层片观察全口牙齿发育情况以及上下颌骨情况。以往对于运用全颌曲面断层片进行定量分析研究学术界上存在一定争议。本综述就是根据各学者的研究,总结探讨全颌曲面断层片在下颌骨对称性分析的可行性。     

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变色牙的美容修复是临床中经常遇到的问题。对变色牙的治疗主要包括漂白、釉质微打磨技术、贴面和全冠修复等方式。牙漂白术作为一种最保守的治疗方法常成为变色牙治疗的首选。有研究表明,高浓度的漂白药物对牙体组织的表面形态、化学组成、微硬度及釉质抗折性等都有影响;漂白药物甚至还可透过牙本质对牙髓造成刺激。本文着重以漂白药物对牙体硬组织的影响做一综述,以探讨漂白药物对牙体结构的影响。     

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烤瓷熔附金属全冠(porcelain-fused-to-metal,PFM)也称金属烤瓷全冠,是一种由低熔烤瓷真空条件下熔附到铸造金属基底冠上的金一瓷复合结构的修复体.烤瓷熔附金属全冠兼具有金属全冠的机械强度好和全瓷冠美观的优点[1].     

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目的研究能够敏感反映患者侧貌美观程度的头影测量指标及其标准。方法中国医科大学口腔医学院正畸科2002—2005年117例矫治后患者侧貌剪影图,经16名正畸医师、63名大学生进行美观程度评分,按评分分为高分组和低分组,测量两组患者10项头影测量指标,分析组间各项指标的差异,通过Logistic回归分析,筛选与颜面侧貌美观程度密切相关的指标。结果高分组U1-AP(6.80±1.87)mm,L1-NB(3.29±2.30)mm,FMIA(56.56±6.70)°;低分组U1-AP(8.7±2.17)mm,L1-NB(4.20±3.88)mm,FMIA(49.50±10.28)°,3项指标在两组间均有统计学差异(P均<0.05),其余7项指标组间均无统计学差异(P均>0.05)。Logistic回归分析结果显示,U1-AP值与侧貌美观程度有显著相关性。结论上牙突度是影响侧貌美观的关键因素,治疗目标应以U1-AP突距(6.80±1.87)mm为参考标准。     

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??Objective    To investigate the relationship between the depth of curve of Spee??COS?? and temporomandibular joint disorders??TMD?? in elderly patients. Methods    Eighty-nine elderly patients with TMD visiting the Hospital of Stomatology of China Medical University from January 2013 to June 2013 were recruited in the study by random number table. Muscle pain and temporomandibular joint??TMJ??sounds were examined according to Research Diagnostic Criteria for TMD??and the depth of COS were measured on the dental casts. The mean depth of COS of patients without muscle pain and patients with pain at all levels were compared??and the unilateral depth of COS of patients with and without TMJ sounds were compared. Related data were analyzed statistically with one-way analysis of variance and t-test. Results           The depth of ipsilateral COS of patients with TMJ sounds was significantly smaller than those of patients without TMJ sounds??left??P < 0.001??right??P = 0.008????while the depth of contralateral COS of patients with TMJ sounds had no significant differences with those of patients without TMJ sounds??left??P = 0.481??right??P = 0.905??. In addition??there were no significant differences between the mean depth of COS of patients without muscle pain and patients with pain at all levels??P = 0.327??. Conclusion    TMJ sounds are closely associated with COS??and the depth of ipsilateral COS of patients with TMJ sounds is smaller??and the COS is flatter. There is no significant association between the mean depth of COS and muscle pain.     

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??Objective    To evaluate the effect of HMME-mediated adjunctive sonodynamic therapy??SDT??on periodontitis in rats??in order to provide theoretical and experimental evidence for the clinical application of SDT. Methods    Ligatures were placed on the subgingival regions of the first maxillary molar in 54 rats to induce the periodontitis models. Then the rats were randomly divided into three groups with 18 rats in each group.??1??The control group was treated by normal saline??NS?? to wash the periodontal pocket????2??the scaling and root planing??SRP??group received SRP and was washed with normal saline????3??the SRP plus SDT group was treated by SRP and washed with normal saline?? and then was treated through subcutaneous injection of HMME 1 mL??40 μg/mL??. After 90 min of incubation in the dark??the ultrasonic treatment at the frequency of 1 MHz and the intensity of 3W/cm2 was given for 10 minutes. SDT was performed every other day. Six rats from each group were sacrificed at 6??10??or 14 days postoperatively and periodontal tissue samples were taken for histological examination and immunohistochemical analysis. Results    SRP plus SDT group had reduced number of inflammatory cells. The expression of receptor activator of nuclear factor kappa-B ligand ??RANKL??was weakly positive??and the number of positive Tartrate-resistant acid phosphatase??TRAP?? was significantly reduced ??P < 0.05????compared to the SRP and the control group??P < 0.05??. Conclusion    SDT is an effective adjunctive therapy for periodontitis in rats.     

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??Objective    To explore the relationship between curve of Spee and craniofacial morphology.  Methods    Lateral cephalograms and dental models of 88 patients between the age of 14 and 32 were evaluated. Correlation analysis and a multiple linear regression analysis were performed to determine the relationship between the curve of Spee and 13 selected cephalometric variables. Results    The curve of Spee was not affected by sex. The depth of the curve of Spee significantly differed among different Angle classifications in which class ?? was obviously higher than class ??. SNA?? ANB?? AO-BO?? post. facial height/ant. facial height?? ODI and upper lip length/lower lip length had statistically significant positive correlation with the depth of the curve of Spee. APDI ?? IMPA and mentolabial angle had negative correlation with the depth of the curve of Spee. Conclusion    The depth of the curve of Spee is correlated with craniofacial morphology?? which should be taken into consideration during orthodontic diagnosis and treatment.     

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目的研究前方牵引联合唇挡矫治替牙期骨性Ⅲ类错的临床疗效。方法选择2010—2011年大庆油田总医院集团五官医院口腔正畸科门诊就诊的替牙期上颌发育不足的骨性Ⅲ类错患者30例。随机分为试验组和对照组,各15例。试验组使用前方牵引联合唇挡进行矫治;对照组则仅使用前方牵引矫治。在治疗前后对所有患者进行X线头影测量分析并比较组间差异。结果试验组治疗前后变化差值与对照组比较,X线头影测量分析结果显示,SNA、ANB、U1-SN、U1-NA角、U1-NA距、Ptm-U6差异均有统计学意义(P<0.01);模型测量结果显示,TAL、AD、拥挤度差异亦均有统计学意义(P<0.01)。结论采用前方牵引和唇挡联合矫治替牙期骨性Ⅲ类错,可有效消除单一采用前方牵引矫治带来的负面影响,是一种切实可行的办法。     

正畸治疗中牙龈蛋白K与牙龈炎性反应的关系

目的分析牙龈蛋白K（gingipain K,Kgp）在正畸治疗中对牙龈炎性反应的影响。方法选择青少年牙龈健康者45例,分别取矫治器戴入前与矫治器戴入后3个月的龈沟液,应用16S rDNA PCR技术检测各样本中的牙龈卟啉单胞菌（P.g）及Kgp,采用SPSS17.0软件包对数据进行统计学分析。结果矫治器戴入前Kgp检出率为35.71%,矫治器戴入后Kgp检出率为67.86%,两者之间差异显著（P<0.05）。结论正畸治疗患者在矫治器戴入后Kgp检出率增加,出现牙龈炎性反应。     

Spontaneous gingival antibody production to Fusobacterium nucleatum outer membrane in patients with adult periodontitis

The local antibody response to Fusobacterium nucleatum outer membrane (FnOM) was analyzed in patients with adult periodontitis (AP) at the single cell level. Furthermore, we analyzed whether periodontal hygienic treatment could alter the antibody response. The number of IgG- and IgM-producing cells were investigated in gingival samples collected from 20 patients with AP. The patients were divided into 2 groups, before (BT, n =9) and after (AT, n =11) periodontal hygienic treatment. Four healthy gingival samples were used as controls. The results obtained showed that local antibody production against FnOM occurred in gingiva of patients with AP, but not in healthy gingiva. The IgG anti-FnOM was the predominant isotype observed. However, there was no statistically significant difference between the BT and AT groups. These results indicate that periodontal hygienic treatment was not sufficient to alter significantly the number of IgG- arid IgM-secreting cells present in gingival tissue of AP patients, but it promoted a reduction of IgG anti-FnOM secreting cells. The presence of anti-FnOM antibodies in AP but not in control patients indicates that this bacteria may play a role in the pathogenesis of periodontal disease.     

具核梭杆菌生物学特性及检测手段的研究进展

具核梭杆菌是牙周炎主要致病菌之一,在口腔乃至全身感染性疾病中检出率极高,与临床厌氧菌感染的关系十分密切。具核梭杆菌具有明显的毒力或致病性,可通过多种机制干扰宿主防御能力,引发牙周组织破坏。本文描述了具核梭杆菌的一些重要特性,包括其生物学特征、分类和毒性特征以及主要的生物学检测手段,着重描述了聚合酶链反应技术在具核梭形杆菌检测中的应用。     

Clonality of Fusobacterium nucleatum in root canal infections

Fusobacterium nucleatum is a gram-negative non-spore-forming, non-motile, obligate anaerobic rod that is normally isolated from the oral cavity. Several studies have reported a significant heterogeneity within the F. nucleatum species. The aim of the present study was to analyze the clonal diversity of F. nucleatum strains isolated from intracanal infections and to evaluate the presence of Enterobacterial Repetitive Intergenic Consensus (ERIC)-like sequences in the genome of F. nucleatum . Samples were collected from 13 single-root teeth from adult patients, all having carious lesions, necrotic pulps and radiographic evidence of periradicular bone loss. F. nucleatum was isolated from two different patients (subjects 5 and 7) by culture. Amplification of 19 colonies from subject 5 and 15 colonies from subject 7 using ERIC primers resulted in four clonal types, two per subject. An intense amplicon of approximately 700 bp was generated by ERIC-PCR for all F. nucleatum isolates and F. nucleatum ssp. polymorphum ATCC 10953. The amplification reaction using primer 1254 confirmed the results obtained with the ERIC primer. Our findings indicate that DNA fingerprints provided by ERIC- and Arbitrarily Primed (AP)-PCR may constitute a powerful tool for investigating F. nucleatum clonal diversity.     

Identification of components in Fusobacterium nucleatum chemostat-culture supernatants that are potent inhibitors of human gingival fibroblast proliferation

The present investigation concerned the effect of chemostat-culture cell-free supernatants of Fusobacterium nucleatum on the growth and synthetic activity of human gingival fibroblasts in vitro. Human gingival fibroblasts were cultured in fetal calf serum supplemented Dulbecco-Vogt medium containing various dilutions of conditioned or unconditioned bacterial culture medium. Cell proliferation was monitored by assessing cell growth over 5 days or incorporation of [3H]-thymidine into DNA. Protein and proteoglycan synthesis were monitored by the incorporation of [3H]-proline and [35S]-sulfate, respectively, into macromolecules. While the conditioned culture medium caused a complete inhibition of cell growth and incorporation of [3H]-thymidine DNA, there was no discernible effect on protein or proteoglycan synthesis. This indicated that the cells remained viable yet unable to divide. Such a view was supported by the observation that the inhibitory effect was reversible upon removal of the conditioned medium. This activity had a molecular size less than 30,000, was heat-stable and nonvolatile. Chemical analysis of the conditioned bacterial culture supernatants indicated that high proportions of butyrate, ammonium, and acetate were present. When these components were added to unconditioned medium and tested, most of the inhibitory activity could be attributed to ammonium and butyrate. Since many bacteria which constitute the subgingival microflora release ammonium and butyrate, a very high concentration of these metabolites may well accumulate. Clearly, the potential for inhibition of fibroblast proliferation has ramifications related to diminished tissue repair following bacterially-induced periodontal destruction.     

The distribution and frequency of Fusobacterium nucleatum subspecies in the human oral cavity

Three reference strains of Fusobacterium nucleatum and 32 human oral isolates were compared by a variety of physiological tests, enzyme electrophoretic profiles, SDS-PAGE patterns, DNA base composition and hybridization to test their possible site specificity and frequency of the recently described subspecies of F. nucleatum. Nine of the 11 isolates assigned to F. nucleatum subspecies nucleatum were from diseased sites, whereas isolates from healthy sites were all identified as F. nucleatum subspecies polymorphum or F. nucleatum subspecies fusiforme. Strains of the latter subspecies were the least frequently isolated (2 of 32). These results, although still inconclusive because of the relatively small sample size, nevertheless confirmed the heterogeneity of F. nucleatum and indicate that most human oral isolates from subgingival sites probably belong to F. nucleatum subspecies nucleatum.     

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目的检测慢性阻塞性肺疾病急性加重期（AE—COPD）患者呼吸道内具核梭杆菌（h．）的定植,探讨n．与AE—COPD间的关系。方法选择2008年10月至2009年4月中国医科大学附属第一医院、盛京医院确诊为AE—COPD的患者53例,采集呼吸道分泌物,提取细菌DNA,应用SYBR。GreenIPremixExTaq＇…模式的实时PCR技术,对n．进行定量检测,并计算其所占总菌的比例。同时记录患者FEVl占预计值比例（％）及口腔牙周指标。结果53例样本中有32例检出Fn．,Fn．检出率为60．38％,占总菌的比例为（44．58±16．47）％。Fn．相对含量与FEVl占预计值比例呈负相关关系（P〈0．05）。F凡．相对含量与简化口腔卫生指数及附着丧失水平呈正相关关系（P〈0．05）;与牙龈出血指数、探诊深度、缺失牙数无相关性（P〉0．05）。结论AE—COPD患者呼吸道内存在Fn,Fn相对含量随着肺功能的减弱而增加;Fn．与AE—COPD密切相关。     

Purification of arginine-sensitive hemagglutinin from Fusobacterium nucleatum and its role in coaggregation

Hemagglutinin of Fusobacterium nucleatum was extracted from Triton X-100-pronase P-treated cell envelopes, and was purified by affinity chromatography on L-arginine agarose. The hemagglutinin was inactivated by heating at 70°C for 1 min. The activity was inhibited by L-arginine but was not affected by any sugars or by EDTA. The hemagglutinin aggreggated 14 out of 17 strains of oral streptococci tested, and the bacterial aggregating activity was also inhibited by L-arginine. The results indicate the dominant role of this hemagglutinin in the adherence of this bacterium both to host cells and to other bacteria.     

Aspects of the growth and metabolism of Fusobacterium nucleatum ATCC 10953 in continuous culture

Fusobacterium nucleatum ATCC 10953, a type strain of one of the newly proposed subspecies of this group of organisms, was grown anaerobically in continuous culture in a chemically defined medium. Its response to conditions of varying pH, nutritional environment, and imposed growth rate were then examined. The organism failed to grow at pH 7.8 but grew at pH 5.8, although the cell yield was greatly reduced. At pH 6.8 the cell yield was halved and less than 50% of available glucose was consumed. The optimum growth pH was around 7.4 when the culture appeared to be limited for both glucose and the amino acids glutamate, histidine and serine. Some intracellular polyglucose (IP) was produced and acetate, butyrate and ammonia were the major fermentation end-products, as they were under all growth conditions tested. Increasing the available glucose or amino acids did not alter cell numbers but the amount of IP was greatly increased. When glucose was omitted from the medium, the cell yield was halved and the culture then became limited for lysine as well as for glutamate, histidine and serine. Growth rate had little overall effect on the organism's physiology and the maximum growth rate at pH 7.4 was 0.20 h-1, a doubling time of 3.5 h. Glucose was thus channelled into stable IP synthesis only when the growth limitation imposed by lack of fermentable amino acids was relieved. The metabolism of IP and the ability to obtain carbon and energy from a variety of substrates may explain why F. nucleatum is one of the most commonly detected organisms in subgingival dental plaque.     

Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates

F. nucleatum is the most commonly isolated microorganism from subgingival plaque, but the role of this microorganism in periodontal diseases remains undefined. Arbitrarily primed polymerase chain reaction (AP-PCR) was evaluated as a method for fingerprinting F. nucleatum isolates and for use in clonal analysis. Pulsed field gel electrophoresis was used to further differentiate F. nucleatum isolates, with identical AP-PCR patterns. Extremely heterogeneous AP-PCR fingerprints were observed among the 98 F. nucleatum isolates, with 36 different genotypes observed with primer CI and 30 different genotypes detected with primer C2. Combining the results of the AP-PCR genotype analysis from C1 and C2 primer amplifications revealed that up to 7 different genotypes could be distinguished from isolates from the same oral cavity and that up to 4 different genotypes were observed within a single site. An intense amplicon at approximately 450 bp generated in AP-PCR amplification with primer C2 was associated with F. nucleatum subsp. nucleatum (ATCC 25586) and with 15 F. nucleatum isolates from diseased sites and 2 isolates from healthy sites. Pulsed field gel electrophoresis confirmed the AP-PCR genotypes and demonstrated increased discriminatory power over AP-PCR. The results indicated that AP-PCR and pulsed field gel electrophoresis provide a simple and sensitive means for differentiating oral F. nucleatum isolates and further demonstrate the heterogeneity of this species. These techniques may serve as useful tools in the clonal and epidemiological analysis of F. nucleatum isolates, which may help define the role of these microorganisms in periodontal diseases.