慢性牙周炎患者龈下菌斑中伴放线放线杆菌基因型的分析

目的：建立龈下菌斑标本中伴放线放线杆菌（Actinobacillus actinomycetemcomitans，Aa）PCR检测方法，了解慢性牙周炎患者不同牙位的龈下菌斑中Aα 的感染率及其优势基因型。方法：61例慢性牙周炎患者每例采取2个不同牙位共122份龈下菌斑标本，采用培养法分离Aα菌株，以PCR或多重PCR检测16SrDNA基因、lktA基因和fap基因，部分扩增产物克隆后测序。结果：在11例患者的11份龈下菌斑标本中分离到Aα菌株。122份龈下菌斑中Aα16SrDNA、lktA、和fap检测阳性率分别为84.4%、75.4%、和50.0%。38.8%的患者（19/49）不同牙位龈下菌斑中检出的Aα基因型不一致。Aα有4种基因型，其优势基因型是16SrDNA^ /lktA^ /fap^ ，其次为16SrDNA^ /lktA^ /fap^-。部分标本上述3种基因的扩增片段与文献报道核苷酸序列的同源性为93.75%-100%。结论：建立的PCR或多重PCR有较高的敏感性和特异性，适用于龈下菌斑标本中Aα的快速检测。慢性牙周炎患者Aα感染率较高，并存在优势基因型，部分患者可被不同基因型的菌株同时感染。     

应用PCR检测慢性牙周炎患者龈下菌斑中牙龈卜啉单胞菌及感染菌株基因型的分析

目的 建立慢性牙周炎（CP）龈下标本中牙龈卟啉单胞菌（Porphyromonas gingivalis,Pg)的PCR检测方法，了解不同患牙龈下菌斑中Pg基因型的差异。方法 采用培养法分离不同患牙龈下菌斑中Pg，同时采用PCR进行Pg 16S rDNA基因、prtC基因和fimA基因的检测。部分扩增产物T－A克隆后测定核苷酸序列。结果 61例患者122个龈下菌斑标本中，Pg 16S rDNA、prtc和fimA分别扩增的阳性率依次为90．6％、81．9％和78．0％，联合扩增的阳性率高达98．4％，培养法阳性率仅为31．1％。30．0％（18／60）患者不同牙位龈下菌斑中的Pg基因型不一致。Pg 16S rDNA、prtC和fimA扩增片段与文献报道的核苷酸序列比较，同源性98．62％－100％。结论 所建Pg PCR检测方法具有较高的敏感性和特异性，可用于慢性牙周炎龈下标本中Pg的快速临床诊断。部分CP患者可被不同基因型的Pg菌株同时感染。     

应用PCR检测慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌及感染菌株基因型的分析

目的建立慢性牙周炎(CP)龈下标本中牙龈卟啉单胞菌(Porphyromonas gingivalis, Pg)的PCR检测方法,了解不同患牙龈下菌斑中Pg基因型的差异. 方法采用培养法分离不同患牙龈下菌斑中Pg,同时采用 PCR进行Pg 16S rDNA基因、prtC基因和fimA基因的检测.部分扩增产物T-A克隆后测定核苷酸序列. 结果 61例患者122个龈下菌斑标本中,Pg 16S rDNA、prtC和fimA分别扩增的阳性率依次为 90.6%、81.9%和78.0%,联合扩增的阳性率高达98.4%,培养法阳性率仅为 31.1%.30.0%(18/60)患者不同牙位龈下菌斑中的Pg基因型不一致.Pg 16S rDNA、prtC和fimA扩增片段与文献报道的核苷酸序列比较,同源性为98.62%～100%. 结论所建Pg PCR检测方法具有较高的敏感性和特异性,可用于慢性牙周炎龈下标本中Pg的快速临床诊断.部分CP患者可被不同基因型的Pg菌株同时感染.     

高通量测序分析牙周基础治疗对65岁以上糖尿病合并慢性牙周炎患者龈下菌群的影响

目的:采用高通量测序技术分析牙周基础治疗对65岁以上糖尿病合并慢性牙周炎(diabetes mellitus with chronic periodontitis,DMCP)患者龈下菌群的影响.方法:选取100例65岁以上DMCP患者作为研究对象,采用随机数据表法将患者分为研究组与对照组,每组50例.对照组给予强化控制...     

巢式PCR检测龈下菌斑中HCMV、EBV、HSV-1与慢性牙周炎活动性的关系

目的　建立临床检测龈下菌斑标本中人巨细胞病毒 (HCMV)、Epstein Barr病毒 (EBV)、单纯疱疹病毒 1型 (HSV 1 )巢式PCR方法 ,研究这 3种病毒与慢性牙周炎活动性的关系。方法　收集6 2例慢性牙周炎患者 (男性 2 7例、女性 35例 ,平均年龄 5 3岁 )的牙周炎活动部位、牙周炎静止部位的龈下菌斑 ,提取DNA后使用巢式PCR检测HCMV、EBV、HSV 1 ,比较分析其在同一病人不同部位的检出率。结果　牙周炎活动部位的HCMV检出率为 38.7%,EBV的检出率为 5 8%,HSV 1的检出率为30 .6 %,两种以上病毒合并感染的检出率为 4 0 .3%;牙周炎静止部位的HCMV检出率为 1 4 .5 %,EBV为 2 2 .6 %,HSV 1为 1 1 .3%,两种以上病毒合并感染的检出率为 1 1 .3%。这 3种病毒及其合并感染在牙周炎活动部位的检出率均高于牙周炎静止部位 ,差异有统计学意义 (P <0 .0 5 )。结论　提示HC MV、EBV、HSV 1与慢性牙周炎的活动性相关。     

牙周根管超声治疗仪应用于慢性牙周炎的临床疗效

目的 探讨牙周根管超声治疗仪治疗慢性牙周炎患者的临床疗效。方法 选择2016年7月~2018年10月东莞市石碣医院口腔门诊收治的慢性牙周病患者168例,随机分为观察组和对照组,每组84例。对照组给予传统的刮治术进行治疗,观察组给予牙周根管超声治疗仪进行治疗,对比两组的临床疗效及牙周状况。结果 观察组的治疗有效率（94.05%）高于对照组（78.57%）,差异具有统计学意义（P＜0.05）;治疗后两组患者的牙周状况指标（PI,BI,PD,CAL）均较治疗前降低,且观察组均较对照组减轻,差异有统计学意义（P＜0.05）。结论 根管超声治疗仪治疗慢性牙周炎患者可有效控制龈下牙石所致的细菌感染,对促进慢性牙周炎牙周组织愈合具有显著疗效。     

慢性牙周炎龈下菌斑中牙龈卟啉单胞菌胶原酶水平与牙周病变程度的关系

目的 检测慢性牙周炎（chronic periodontitis，CP）患者龈下菌斑中牙龈卟啉单胞菌（Porphyromonas gingivalis,Pg）胶原酶基因prtC及其产物PrtC，了解PrtC水平与牙周病变程度的关系，确定PrtC诱导细胞分泌炎性细胞因子的作用。方法 采用SDS-PAGE检测原核表达系统重组PnC（rPrtC）表达和Ni-NTA亲和层析法提取的rPrtC纯度。rPrtC免疫家兔获得抗血清。建立多重PCR检测56例CP患者209牙位的龈下菌斑标本中Pg 16S rDNA和prtC基因。建立ELISA检测上述标本中的PrtC，并分析PrtC水平与牙周病变程度的关系。采用ELISA检测rPrtC诱导人脐静脉内皮细胞EVC-304分泌IL-1α、IL-8和TNF-α的作用。结果 rPrtC表达产量约占细菌总蛋白的50％。提纯的rPrtC SDS-PAGE后仅见单一的蛋白条带。96．1％（201／209）和92．3％（1931209）的龈下菌斑标本分别Pg 16S rDNA和prtC基因PCR阳性，91．4％的龈下菌斑标本（191／209）PrtC ELISA阳性。重度CP龈下菌斑标本中PrtC含量明显高于轻度和中度CP标本（P〈0．05），但轻度和中度、中度和重度四标本之间PrtC含量差异无统计学意义（P〉0．05）。1μg的rPrtC作用EVC-304细胞24h后，5和10μg的rPrtC作用EVC-304细胞12h后均可使EVC-304细胞分泌的IL-1α、IL-8和TNF-α水平明显增高（P〈0．05）。结论 Pg有很高的prtC基因携带率和表达率。CP牙周病变程度与龈下PrtC水平密切相关。rPrtC有直接诱导细胞合成并分泌IL-1α、IL-8和TNF-α的活性。     

血清C反应蛋白在大鼠慢性牙周炎合并动脉粥样硬化模型中的表达

目的 检测和分析大鼠慢性牙周炎（CP）合并动脉粥样硬化（As）模型中血清C反应蛋白（CRP）的表达。方法 60只Wistar大鼠随机分为4组,每组15只,A：正常对照组,B：CP组,C：As组,D：CP+As组,各组接受相应的建模处理。观察牙周组织及动脉血管病理改变,酶联免疫吸附法（ELISA）检测血清CRP水平。结果 牙周病理学观察：B、D组实验牙牙周炎症表现明显,附着丧失（AL）水平较A、C组有明显增加（P<0.05）。动脉病理学观察：C、D组主动脉及胸、腹主动脉形成粥样硬化病变。D组血清CRP水平高于其他组（P<0.05）。相关性分析显示：B、D组CRP与AL均呈正相关。结论 CP与As可能相互影响、促进,两疾病的发生发展与血清CRP水平有关。     

白细胞介素17A在慢性牙周炎和类风湿性关节炎相关性中的研究

目的 探讨白细胞介素-17A（IL-17A）在慢性牙周炎（CP）和类风湿性关节炎（RA）两者间的相关性.方法 按纳入标准选择85名受试者分为四组：①CP组20例;②RA组23例;③RA伴CP组26例;④HP组16侧;记录牙周袋探诊深度（PD）,临床附着丧失（CAL）和出血指数（BI）,收集其晨起空腹血清并用滤纸条法收集受试者龈沟液样本,采用酶联免疫吸附试验法（ELISA）测定血清及龈沟液中IL-17A的水平.结果 RA＋CP组中血清IL-17A水平（2216.0±1520.87）pg/mL与RA组[（851.59±975.81） pg/mL]、CP组[（1039.7±1004.83） pg/mL]和HP组[（209.48±50.02） pg/mL]差异有统计学意义,RA组和CP组中血清IL-17A水平分别与HP组的差异有统计学意义（P＜0.05）,而龈沟液中的IL-17A水平在HP组[（356.75±69.07） pg/mL],RA组[（381.08±79.48） pg/mL],CP组[（397.82±108.53） pg/mL]以及RA＋CP组[（414.71±103.09） pg/mL]间呈现升高趋势,但无统计学差异（P＞0.05）.结论 IL-17A可能在慢性牙周炎和类风湿性关节炎相关性的研究中具有一定的意义.     

慢性牙周炎患者血浆穿透素-3的检测及其临床意义

目的:认识慢性牙周炎患者血浆穿透素-3(PTX-3)浓度的变化并分析其临床意义。方法:收集慢性牙周炎患者作为牙周炎组,共80例。收集同期牙周健康者作为对照组,共80例。ELISA检测血浆PTX-3浓度,进行统计分析。结果:牙周炎组血浆PTX-3浓度(3.4±1.5)ng/ml显著高于对照组(1.8±0.6)ng/ml(P<0.001)。经Pearson相关分析,牙周炎组血浆PTX-3浓度与牙周探诊深度(r=0.376,P<0.01)、附着丧失(r=0.426,P<0.01)和出血指数(r=0.401,P<0.01)呈显著正相关。Logistic回归分析显示,血浆PTX-3浓度(OR=1.791,95%CI=1.172～5.814,P<0.01)是慢性牙周炎发生的独立危险因素。ROC曲线分析显示,血浆PTX-3浓度对慢性牙周炎发生有显著预测价值(曲线下面积=0.834,95%CI=0.764～0.905,P<0.001),且判定血浆PTX-3浓度>2.8ng/ml,对预测慢性牙周炎发生有81.3%的灵敏度和75.0%的特异度。结论:慢性牙周炎患者血浆PTX-3水平升高,PTX-3可能参与慢性牙周炎的病理生理过程。     

Prevalence of Aggregatibacter actinomycetemcomitans,Porphyromonas gingivalis and Tannerella forsythia in Japanese patients with generalized chronic and aggressive periodontitis

This study aimed to investigate the prevalence and levels of major periodontal pathogens, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia in subgingival plaque samples of a group of Japanese patients with aggressive periodontitis (AgP) and chronic periodontitis (CP). A total of 40 patients with clinical diagnosis of AgP or CP and 10 periodontally healthy volunteers were subjected to clinical and microbiological analysis. Subgingival plaque samples were analyzed for A. actinomycetemcomitans, P. gingivalis and T. forsythia with a real-time polymerase chain reaction (PCR) technique. The prevalence of P. gingivalis and T. forsythia was relatively high in patients with periodontitis: over 60% of AgP or CP patients harbored these pathogens whereas they were not detected in the subgingival plaque samples from periodontally healthy individuals. P. gingivalis and T. forsythia were relatively frequently detected together in AgP and CP patients. No significant differences in the prevalence or level of the 3 pathogens were found between periodontitis groups. The proportion of T. forsythia was approximately 4-fold higher in CP group than in AgP group (P = 0.02). In periodontitis patients, a significant positive correlation was found between periodontal parameters (probing depth and clinical attachment level) and the numbers of total bacteria, P. gingivalis and T. forsythia. No distinct pattern of the subgingival profile of these pathogens was discerned between the two disease entities, except for the difference in the proportion of T. forsythia. The red complex bacteria, P. gingivalis and T. forsythia were highly prevalent in this population of Japanese AgP and CP patients, collaborating their roles in periodontitis.     

Microbiological characteristics of subgingival microbiota in adult periodontitis, localized juvenile periodontitis and rapidly progressive periodontitis subjects

Objective   To describe the prevalence of the cultivable subgingival microbiota in periodontal diseases and to draw attention to the polymicrobial nature of periodontic infections.
Methods   The study population consisted of 95 patients, 51 females and 44 males, aged 14–62 years. Twenty-nine patients exhibited adult periodontitis (AP), six localized juvenile periodontitis (LJP), and 60 rapidly progressive periodontitis (RPP). Two to four pooled bacterial samples were obtained from each patient. Samples were collected with sterile paper points from the deepest periodontal pockets. The samples were cultured under anaerobic and microaerophilic conditions using selective and non-selective media. Isolates were characterized to species level by conventional biochemical tests and by a commercial rapid test system.
Results   Prevotella intermedia and Capnocytophaga spp. were the most frequently detected microorganisms in all diagnostic groups. Porphyromonas gingivalis and Peptostreptococcus micros were found more frequently in AP and RPP patients, while Actinobacillus actinomycetemcomitans and Eikenella corrodens were associated with AP, LJP and RPP patients. The other bacterial species, including Actinomyces spp., Streptococcus spp. and Eubacterium spp., were detected at different levels in the three disease groups.
Conclusions   The data show the complexity of the subgingival microbiota associated with different periodontal disease groups, indicating that the detection frequency and levels of recovery of some periodontal pathogens are different in teeth affected by different forms of periodontal disease.     

尼美舒利对慢性牙周炎治疗效果的评价

目的:评价尼美舒利对慢性牙周炎的治疗效果。方法:选择136名慢性牙周炎患者，所有患者均进行牙周基础治疗，然后分别应用尼美舒利和甲硝唑治疗4周。在牙周治疗前及治疗2和4周后常规临床检查，记录探诊出血、菌斑指数和探诊深度，分别比较两种药物对慢性牙周炎的临床治疗效果。 结果:在临床治疗中观察到尼美舒利和甲硝唑均能有效减轻牙周炎症程度，但尼美舒利能显著降低探诊深度（P<0.05）;尼美舒利治疗慢性牙周炎的显效率明显高于甲硝唑治疗组（P<0.05）。 结论:与甲硝唑相比较，尼美舒利对慢性牙周炎的临床治疗效果更显著。     

Single nucleotide polymorphisms in interleukin-1gene cluster and subgingival colonization with Aggregatibacter actinomycetemcomitans in patients with aggressive periodontitis

Periodontitis is initiated by the subgingival occurrence of periodontopathogens. It is triggered by a specific host-dependent immune response that is influenced by genetic predisposition. Polymorphisms in the interleukin-1 (IL-1) gene cluster have been suggested to influence the pathogenesis of periodontitis. A total of 159 periodontitis patients (chronic disease: n = 73, aggressive disease: n = 86) and 89 periodontitis-free controls were included in the study. Polymorphisms IL-1α (rs1800587), IL-1β (rs16944, rs1143634), IL-1 receptor (rs2234650), and IL-1 receptor antagonist (rs315952) were determined by polymerase chain reaction with sequence-specific primers (PCR-SSP). Subgingival bacterial colonization was assessed using a polymerase chain reaction/DNA probe test (micro-Ident). Haplotype block structure was determined using Haploview 4.2. Statistical analyses were performed applying SPSS 17.0 considering dominant, recessive, and codominant genetic models. In this case-control study, no association between genomic variants of the IL-1 gene cluster and the incidence of severe periodontitis could be shown. Carriers of the rare genotypes of rs1800587 (p(corr) = 0.009), rs1143634 (p(corr) = 0.009) and composite genotype (rs1800587+rs1143634) (p(corr) = 0.031) had a twofold higher risk for subgingival occurrence of Aggregatibacter actinomycetemcomitans. In forward stepwise binary logistic regression analyses considering age, gender, smoking, and approximal plaque index as potential confounders these significant associations were demonstrated. Despite the genetic background of IL-1 gene cluster could be shown to be associated with subgingival colonization of A actinomycetemcomitans, there is no evidence that it is an independent risk indicator for periodontitis.     

Treatment of chronic periodontitis decreases serum prohepcidin levels in patients with chronic kidney disease

OBJECTIVE:

To determine the impact of periodontal treatment on serum levels of prohepcidin (the prohormone of hepcidin) and systemic inflammation markers, as well as correlations among these markers, in patients with chronic periodontitis and chronic kidney disease who were not undergoing dialysis.

METHODS:

We included 56 chronic periodontitis patients, 36 with chronic kidney disease and 20 without systemic diseases and with normal renal function (control group). Chronic kidney disease was defined as suggested by the clinical practice guidelines in the National Kidney Foundation. Chronic periodontitis was defined through clinical attachment level and by probing pocket depth, according to the American Association of Periodontology. The inflammatory markers ultrasensitive C-reactive protein, interleukin-6, and prohepcidin were evaluated before and 3 months after periodontal treatment.

RESULTS:

The efficacy of periodontal treatment was confirmed by the improvement in clinical parameters of chronic periodontitis in the control and chronic kidney disease groups. Periodontal treatment resulted in significant reductions in ultrasensitive C-reactive protein, interleukin-6 and serum prohepcidin levels in both groups. Moreover, in multivariate linear regression, the reduction in prohepcidin after periodontal treatment was significantly and independently associated with interleukin-6 levels in the control group.

CONCLUSIONS:

By inducing a decline in the systemic inflammatory response and a decrease in serum prohepcidin, successful periodontal treatment may represent an important means of ameliorating the inflammatory burden seen in patients with chronic kidney disease. Trial registration: ISRCTN59866656.     

Subgingival microbiota in individuals with severe chronic periodontitis

Background/Purpose

Subgingival microorganisms are potentially associated with periodontal diseases. However, the correlation between the variance in the periodontal microbiome and the prevalence and severity of periodontitis remains unclear. The aim of this study was to determine the subgingival microbiota in Taiwanese individuals with severe chronic periodontitis (SP).