健康青少年口腔正常菌群的初步研究

本文采用非选择性培养基对22名健康青少年的唾液、沟裂菌斑、龈上菌斑及龈下菌斑中的需氧菌、兼性厌氧菌及专性厌氧菌进行了分离培养,并计算其在不同标本中占可培养菌的百分比及检出率。结果共分离到包括18个菌属的35种细菌。其中,链球菌、放线菌、奈瑟氏球菌、二氧化碳噬纤维菌、类杆菌、梭杆菌,奴卡氏菌及棒状杆菌在口腔4个部位的检出率及所占比例均较高,是健康青少年口腔中的优势菌群.通过比较还发现,其中一些菌在口腔4个部位的分布存在一定差异.本文还采用刚果红负性染色涂片法,镜下观察龈上、龈下菌斑中的螺旋体,并计算其相对比例.结果龈下菌斑中螺旋体的相对比例明显高于龈上菌斑.     

镍铬合金烤瓷冠修复牙与对照牙牙周微生态及龈沟液蛋白水平的比较

目的观察比较镍铬合金烤瓷冠（Ni-Cr based porcelain-fused-metal crown）修复患牙与牙周健康对照牙龈下菌斑微生物分布、龈沟液总蛋白水平及牙周临床指标的差异。方法选择临床病例13例,前牙区咬合关系基本正常。采用镍铬合金烤瓷冠修复上颌切牙17颗,对侧同名健康天然牙作为对照。烤瓷冠戴入6～8个月后复诊,进行牙周健康状况的临床检查,分别取患牙、对照牙龈沟液及龈下菌斑盐水涂片。龈沟液定量并检测总蛋白水平。龈下菌斑涂片革兰染色,分类计数革兰阴性球菌、杆菌、弯曲菌、螺旋体及革兰阳性球菌、杆菌,共计数200个细菌。结果修复患牙与对照牙比较探诊深度、龈沟出血指数、龈沟液量、龈沟液总蛋白水平、龈下菌斑中革兰阴性杆菌增加,革兰阳性球菌减少,差异均有显著性（P〈0．05）。而菌斑指数及其他形态的细菌数差异无显著性。结论临床合格的镍铬合金烤瓷冠修复亦使患牙龈下菌群分布发生变化,可能是导致患牙牙龈炎症的原因之一;龈沟液量及总蛋白水平增加;牙周临床指标显示不良改变。     

正常口腔中的可疑牙周致病菌

本实验对49例正常口腔的龈上和龈下菌斑内的可疑牙周致病菌(SPB)的分布情况进行了观察分析。 49例研究对象男21例,女28例;6～25岁,无龋,无牙周病的健康人。三月内未服用抗生素,未接受任何牙周治疗。定对在右上颌第一磨牙(No.3)的龈上及龈下菌斑。用BHI培养基厌氧培养37℃,4天以后进行细菌菌落计数,转种分离和鉴定。龈上菌斑内SPB的检出率按其高低顺序依次为:二氧化碳噬纤维菌、梭杆菌、消化链球菌、唾液弯曲杆菌、产黑色素类杆菌、优杆菌,溶蚀艾肯氏菌、生疾月形单胞菌。二氧化碳噬纤维菌中依次为牙龈二氧化碳噬纤维菌、黄褐二氧化碳噬纤维菌及生疾二氧化碳噬纤     

牙周炎病人龈下韦荣球菌的实验观察

目的 探讨牙周炎病人龈下韦荣球菌的变化及牙周炎的细菌因素。方法 采用厌氧培养及活菌计数（CFU）的方法检测莲球菌（Streptococcus）、韦荣球菌（Veillonella）、乳杆菌(Lactobacillus）及类杆菌（Bacteroids)的含量。结果牙周炎病人龈下莲球菌含量显著低于正常对照组（P＜0.001）,韦荣球菌、乳杆菌及类杆菌的含量显著高于正常对照组（P＜0.001）。结论 牙周炎病人龈下菌群处于严重失调状态,韦荣球菌对牙周炎发病的作用应引起高度重视。     

口源性口臭患者主要厌氧菌的分布

目的分析口臭患者龈下菌斑和舌苔上主要相关厌氧菌的分布情况。方法选择口腔门诊中口臭患者29例,鼻闻法来确定产臭部位和非产臭部位;分别采集龈下菌斑和舌苔标本接种在非选择性培养基和核梭杆菌选择性培养基,厌氧培养5d后记录非选择性培养基上生长的细菌总数、产黑色素细菌总数及核梭杆菌选择性培养基上生长的目的菌总数。结果29例患者中,15例患者的口臭主要来源于龈缘菌斑,10例主要来源于舌苔,4例患者的口臭由龈缘菌斑和舌苔共同产生;产臭部位和非产臭部位相比,细菌总数、产黑色素菌和具核梭杆菌数都明显上升（P〈0．01）。结论口源性口臭患者口气变化与产黑色素细菌、核梭杆菌相关。     

成年牙周炎病人龈下菌斑优势菌群的探讨

本文用2种非选择性培养基和12种选择性培养基对17名北京市成年牙周炎病人和9名健康对照者龈下及龈沟菌斑菌群进行了培养、分离和鉴定。结果,牙周炎患者龈下菌斑的菌群主要由专性厌氧的革兰氏阴性杆菌组成(60.8％),优势菌为牙龈拟杆菌等;而对照组的菌群主要由兼性厌氧的革兰氏阳性球菌组成(69.2％),优势菌为链球菌属等。这种菌群构成,两组样本存在显著性差异,对于我们认识牙周炎的病原及有关微生态学问题可能有重要意义。     

牙周炎治疗过程中龈下菌斑的动态观察

目的　通过对牙周炎治疗过程中龈下菌斑内螺旋体和球菌的百分比组成的改变进行动态观察研究,为牙周炎的诊断和疗效评价提供依据。方法　选择１８ 例经门诊确诊为成人牙周炎的患者,随机分为Ａ、Ｂ、Ｃ 三组,均施以龈上洁治、龈下刮治和根面平整术。其中Ａ 组和Ｂ 组分别给予艾利克及洗必太含漱液含漱。所有患者每周复诊一次,采集龈下菌斑,进行刚果红染色,镜检计数螺旋体及球菌的百分比,同时检测患处牙龈指数（ＧＩ） 。结果　Ａ、Ｂ 组患者经治疗１ 周后,ＧＩ值变化极为显著（Ｐ＜ ０．０１） ,螺旋体的减少及球菌的增加也极具显著性（Ｐ＜ ０．０１） ,至３ 周时趋于稳定;Ｃ 组则在１ 周时ＧＩ值及菌斑组成的变化有显著性（Ｐ＜ ０．０５） ,至２ 周时变化才最为明显（Ｐ＜ ０．０１） ,４ 周时趋于稳定。结论　牙周基础治疗能明显改善临床症状,使螺旋体百分比显著降低,球菌百分比显著增加。口腔含漱液作为一种牙周炎的辅助用药,可明显改善菌斑的组成,促进正常口腔卫生环境的建立。     

牙周炎龈下菌群的初步探讨

牙周炎为口腔常见病,其发展结果为牙周膜破坏、牙槽骨吸收而造成牙列缺失。对牙周炎的病因进行了一个多世纪的研究,现普遍认为微生物及其产物是引起牙周炎的最主要原因。为进一步探讨牙周炎龈下菌群的组成、种类及变化,我们于1987年2月～7月对临床检查为牙周炎的患者12名(患牙71颗、健康牙52颗)进行普通光学显微镜直接涂片检查并作刚果红负性染色及革兰氏染色检查以比较分析;另设健康人9名共50颗牙作对照组。按微生物形态分五类计数:球菌、杆菌、螺旋体、活动菌及其它(包括丝状菌、梭状菌     

牙周炎患者菌斑细菌的药敏分析

目的:了解牙周炎患者龈下菌斑细菌对于口腔常用抗菌药物的敏感性.方法:从慢性牙周炎患者龈下菌斑中分离培养牙周可疑致病菌,测定6种口腔常用抗菌药物对其的最低抑菌浓度,分析以上致病菌对这几类抗菌药物的敏感性.结果:共从50名慢性牙周炎患者牙周袋中分离培养72株革兰阴性厌氧菌进行药物敏感试验.全部实验菌株都对阿莫西林/克拉维酸敏感.对甲硝唑、替硝唑、奥硝唑未见耐药株产生.结论:对于慢性牙周炎患者龈下菌斑中分离的厌氧菌,阿莫西林/克拉维酸和硝基咪唑类药物甲硝唑、替硝唑和奥硝唑的抑菌作用强.     

642株自胆道分离病原菌的病原学分析

目的调查引起胆道感染的主要致病细菌群分布及耐药性状况。方法用WHONET统计软件回顾性分析2000年1月至2004年12月5年问引起胆道感染的致病细菌的特点。结果5年问共分离出642株细菌,阳性率为20．1％;其中革兰阴性杆菌354株,革兰阳性球菌288株。分离率最高的革兰阴、阳性细菌分别为：肺炎克雷伯菌、铜绿假单胞菌、粪肠球菌、凝固酶阴性葡萄球菌和屎肠球菌。药敏统计结果显示各主要致病革兰阴性杆菌对亚胺培南、美罗培南、头孢哌酮／舒巴坦和头孢吡肟总的敏感率最高。各主要革兰阳性球菌对万古霉素、替考拉宁的总的敏感率最高。结论引起胆道感染的致病细菌发生明显的变迁．革兰阳性球菌的构成比有逐渐升高的趋势,胆道手术应合理进行预防性用药,以有效控制胆道感染。     

临床分离的表皮葡萄球菌产膜株检出方法及耐药性比较

目的对临床分离表皮葡萄球菌产膜株的检出方法及耐药性进行比较。方法采用刚果红培养基检出PIA阳性株;结晶紫染色进行生物膜定量测定;WalKAway-40全自动微生物鉴定仪检测表皮葡萄球菌临床株对17种抗菌药物耐药性。结果定量测定法及刚果红培养基I、II、III检出产膜株分别为16、10、8和12株;产膜与不产膜的表皮葡萄球菌对17种抗菌药物的耐药性差异无统计学意义(P0.05)。结论定量测定法及添加少量葡萄糖、氯化钠的刚果红培养基均能较好检出产膜菌;在浮游状态下产膜和不产膜表皮葡萄球菌对抗菌药物的耐药性差异无统计学意义。     

Methods for staining amyloid in tissues: a review

The traditional way of identifying amyloid in tissue sections has been staining with Congo red and demonstration of green birefringence under crossed polarizers. The original method of Congo red staining, described by Bennhold in 1922, has undergone several modifications to improve its sensitivity, specificity, and reliability. The most common modification is the alkaline Congo red method described by Puchtler and co-workers in 1962. Specificity is improved by using freshly prepared stain and a staining solution fully saturated with sodium chloride. Amyloid proteins can be further distinguished by autoclaving or by treating the tissue with potassium permanganate or alkaline guanidine. Autoclaving the tissues at 120 C for 30 min causes protein AA to lose its affinity for Congo red. Prolongation of autoclaving to 120 min abolishes the Congophilia of protein AL, but prealbumin-related amyloid shows little or no change. Treatment of the tissue with potassium permanganate causes protein AA and B2-microglobulin amyloid to lose their affinity to Congo red. Protein AA fails to stain with Congo red after treatment with alkaline guanidine for 1 min and protein AL and systemic senile amyloid protein (SSA) after 2 hr. Familial amyloid protein (FAP), prealbumin type, can stand 2 hr of alkaline guanidine treatment without losing its ability to stain with Congo red. Other methods of detection of amyloid include fluorescent stains, e.g., thioflavin T or S, and metachromatic stains such as crystal violet. Immunofluorescence and immunoperoxidase methods are used to identify and classify amyloid proteins in tissues. Antibodies against the P component, proteins AA and AL and FAP have been used with great precision. Due to cross-reactivity, these methods do not differentiate between some types of familial and senile systemic amyloidosis.     

固定矫治器戴入前后牙周可疑病原菌和牙周临床指数的变化

目的:固定矫治器戴入前后的不同时间进行龈缘菌斑的微生物学检查和牙周状况的临床检查,以探讨固定矫治器戴入前后牙周可疑病原菌和牙周状况的动态变化过程.方法:选择18名固定正畸患者,于矫治器戴入前和戴入后1、3、6月分别检查16、41牙位的菌斑指数、牙龈指数、探诊深度、探诊出血,并在近中颊侧颊轴角处采集龈缘菌斑样本,采用细菌培养鉴定方法测定牙周可疑病原菌的检出量(CFU/g)和检出率.结果:与基线相比,观察期内龈缘菌斑的G￣产黑色素厌氧杆菌检出量和检出率在两个牙位均升高(P＜0.05),41的优杆菌、弯曲杆菌、拟杆菌、普氏菌亦有升高(P＜0.05).临床指标中16的牙龈指数(颊侧)和探诊深度(颊、舌侧)升高;41的菌斑指数(舌侧)降低(P＜0.05).结论:固定矫治器戴入后虽然可通过严格的口腔卫生指导有效控制牙面菌斑,但仍可引起牙周可疑病原菌增加.     

Reproducibility of morphometric measurements of amyloid after various staining methods

Four histologic staining methods used for detecting amyloid (Congo red, viewed in both normal and polarized light, Sirius red, Crystal violet and Thioflavine T) were applied to heart muscle autopsy samples from 19 patients who suffered from amyloidosis. The amount of amyloid present was evaluated with morphometry (point counting) by five pathologists, and the interobserver reproducibility and variation of point counting in these staining methods were analyzed. The Sirius red method showed the least variation and was the most suitable stain for demonstrating amyloid with respect to reproducibility. Thioflavine T showed the greatest variation and was the least suitable stain with respect to reproducibility. The range of variation was considerable in all staining methods. The results show that stains differ in their specificity and sensitivity in staining amyloid, observers differ in their interpretation of staining results and certain stains result in more uniform interpretations than do others.     

痛风的临床病理特征分析并文献复习

目的:总结痛风临床病理特点。方法:回顾性分析1例痛风患者的生化机制、临床病理特征、刚果红染色、PAS染色特点、鉴别诊断要点,并复习相关文献。结果:患者主要临床表现为间断性多关节肿痛3年,加重伴发热3个月。体格检查发现患者有多发性皮下结节、多关节肿胀压痛。左腕、左肘皮下结节活检,经HE染色后光镜查见大量肉芽肿性病变,有的多核巨细胞内查见被吞噬的异物,有的病灶尚查见呈均质状物(尿酸盐结晶),其周围有较多异物巨细胞及纤维结缔组织包绕呈结节状,在结节的周边纤维血管周围可查见残留分化成熟的淋巴细胞及少数嗜酸性粒细胞。刚果红、PAS染色均为阴性。偏光显微镜下,刚果红未查见绿色强折光晶体,但见多量略呈淡黄色具有强折光性的晶体呈棒状或梭形。结论:痛风在刚果红染色偏光显微镜下观察呈淡黄色梭形或针状结晶,具有强折光性晶体,但这是否是痛风在刚果红染色的特征尚有待于进一步研究。     

赖夫生鞭毛染色法的改进

赖夫生染色法是细菌鞭毛染色的一种常用方法,最近发现了延长赖失生染剂使用寿命及改进染色效果的方法,并在此基础上进一步论证了乙醇在染色过程中所起的作用。     

Activity staining of endoglucanases in polyacrylamide gels.

The endoglucanases of Penicillium funiculosum were analyzed for the presence of multiple forms using a modified version of the Congo red method. Postelectrophoretic slab gels were directly incubated in a solution of carboxymethylcellulose for a period as short as 15 min and then the activities were visualized by staining with Congo red. Ten distinct bands of clearances were obtained indicating the presence of at least as many multiple forms.     

Conversion of Congo red and 2-azoxyfluorene to mutagens following in vitro reduction by whole-cell rat cecal bacteria

Congo red, an azo dye derived from benzidine, and 2-azoxyfluorene, a derivative of 2-aminofluorene, were reduced during overnight incubation with a suspension of rat intestinal bacteria. High performance liquid chromatography and ultraviolet spectral analysis verified the presence of benzidine in extracts of the Congo red incubations and 2-aminofluorene in extracts of the 2-azoxyfluorene incubations. Extracts of the Congo red incubations were mutagenic toward Salmonella typhimurium TA1538 in the presence of a post-mitochondrial activating system, but Congo red was not mutagenic without this reductive pretreatment. Thus, the utility of the Ames test in screening for potential mutagens may be expanded by a reductive pretreatment utilizing cecal bacteria.     

Red light kills bacteria via photodynamic action.

With the increase in the number of antibiotic resistant strains of microorganism, the search for alternative treatments of microbial infections becomes all the more important. We report a novel method for bacterial inactivation based on the optical excitation of the naturally occurring (endogenous) photosensitzing porphyrins by red light. In particular, the pathogenic Gram-positive porphyrin producing ATCC strains Propionibacterium acnes, Actinomyces odontolyticus and Porphyromonas gingivalis were investigated. Sensitive autofluorescence spectroscopy revealed that these bacteria naturally synthezise the fluorescent photosensitizer protoporphyrin IX. In addition, bacterial plaque samples of periodontitis patients were studied. Non-labeled fluorescent bacterial colonies were exposed to red light at 632.8 nm, 100 mW/cm2 light intensity and 360 J/cm2 energy density using a helium-neon laser. The survival rate after a single phototreatment with red light was found to be 0.58 +/- 0.09 in the case of Propionibacterium acnes, 0.30 +/- 0.04 in Actinomyces odontolyticus and 0.59 +/- 0.10 in Porphyrormonas gingivalis compared to non-exposed bacteria suspensions. No photoeffect was found for the bacterium Streptococcus mutans which exhibited no detectable porphyrin autofluorescence. Red-light exposed plaque samples of patients showed significant reduction of colony forming units by 50% as well as a pronounced photoeffect on the pigmented species Prevotella intermedia. Taken together, these results suggest the treatment with red light can be potentially employed as an therapeutic method to inactivate certain pathogenic strains of porphyrin producing bacteria without the use of external photosensitizers.     

Detection of cellulolytic fungi by using Congo red as an indicator: a comparative study with the dinitrosalicyclic acid reagent method

Cellulolytic fungi can easily be screened within 2 d for the production of cellulolytic enzymes by staining with Congo red, or by measuring the amount of reducing sugar (glucose) produced with the dinitrosalicyclic acid reagent method. Endoglucanase activity is visible in carboxymethyl cellulose agar plates after staining with Congo red, and fixing with HCL or NaOH. This method is essentially based on the interaction of Congo red with intact β-(1-4)-D-glucans in carboxymethyl cellulose. Endoglucanase and exoglucanase activities of cellulases are quantitatively measured with the dinitrosalicyclic acid reagent method. The enzymatic activity detected with Congo red is compared with that obtained by the dinitrosalicyclic acid reagent method.