DGGE法分析慢性化脓性中耳炎耳道菌群变化及药敏试验研究

目的解析正常成人及慢性化脓性中耳炎（CSOM）患者耳道内菌群结构的差别,寻找有效抑制致病菌同时对皮肤主导菌群影响不大的抗生素,指导临床合理用药。方法取20例正常成人及20例CSOM患者耳道分泌物行血琼脂平板划线法分离鉴定细菌,选择典型病例进行DGGE法分析耳道菌群多样性,对培养的常见细菌进行药物敏感试验,分析药敏结果。结果（1）20例正常成人耳道分泌物中18例培养分离出表皮葡萄球菌（90％）,2例培养阴性（10％）;20例CSOM患者耳道分泌物中离出金黄色葡萄球菌8例（40％）,铜绿假单胞菌5例（25％）,溶血性葡萄球菌、表皮葡萄球菌各2例（各10％）,阴沟肠杆菌、洋葱伯克霍尔德菌各1例（各5％）,无菌生长1例（5％）。（2）DGGE分析显示,正常成人耳道菌群种类比CSOM明显增多。（3）金黄色葡萄球菌、表皮葡萄球菌、铜绿假单胞菌对环丙沙星均敏感。表皮葡萄球菌对阿莫西林、头孢唑林、左旋氧氟沙星耐药而金黄色葡萄球菌对其敏感。结论（1）正常人耳道正常菌群多样性高于CSOM。（2）目前培养出的正常成人主要耳道菌群为表皮葡萄球菌,CSOM主要菌群为金黄色葡萄球菌、铜绿假单胞菌。（3）环丙沙星同时抑制正常菌群和致病菌的生长。阿莫西林、头孢唑林、左旋氧氟沙星在抑制金黄色葡萄球菌等致病菌的同时,可以在一定程度上保护正常菌群成员表皮葡萄球菌。     

高盐稀醪酱油发酵原油中微生物区系研究

对传统的高盐稀醪酱油发酵过程中的正常原油和异常原油中的微生物区系进行了分析,并对主要的细菌和酵母菌进行了菌种鉴定。同时,初步探讨了温度对原油中微生物区系的影响。实验结果表明,正常原油和异常原油中细菌总数、芽孢菌数、肠道杆菌、乳酸菌数和厌氧菌数没有明显差异,而酵母菌数和耐盐菌数有明显的差异。正常原油中优势酵母为球拟酵母属(Torulopsis)和酵母属(Saccharomyces),分别占酵母总数的55.9%和35.3%。异常原油中优势酵母为毕赤酵母属(Pichia)、假丝酵母属(candida)和酵母属(Saccharomyces),分别占酵母总数的62.8%、17.9%和9.0%。     

Numerical simulation of wave propagation in a realistic model of the human external ear

In this study, a numerical investigation is performed to evaluate the effects of high-pressure sinusoidal and blast wave's propagation around and inside of a human external ear. A series of computed tomography images are used to reconstruct a realistic three-dimensional (3D) model of a human ear canal and the auricle. The airflow field is then computed by solving the governing differential equations in the time domain using a computational fluid dynamics software. An unsteady algorithm is used to obtain the high-pressure wave propagation throughout the ear canal which is validated against the available analytical and numerical data in literature. The effects of frequency, wave shape, and the auricle on pressure distribution are then evaluated and discussed. The results clearly indicate that the frequency plays a key role on pressure distribution within the ear canal. At 4 kHz frequency, the pressure magnitude is much more amplified within the ear canal than the frequencies of 2 and 6 kHz, for the incident wave angle of 90° investigated in this study, attributable to the ‘4-kHz notch’ in patients with noise-induced hearing loss. According to the results, the pressure distribution patterns at the ear canal are very similar for both sinusoidal pressure waveform with the frequency of 2 kHz and blast wave. The ratio of the peak pressure value at the eardrum to that at the canal entrance increases from about 8% to 30% as the peak pressure value of the blast wave increases from 5 to 100 kPa for the incident wave angle of 90° investigated in this study. Furthermore, incorporation of the auricle to the ear canal model is associated with centerline pressure magnitudes of about 50% and 7% more than those of the ear canal model without the auricle throughout the ear canal for sinusoidal and blast waves, respectively, without any significant effect on pressure distribution pattern along the ear canal for the incident wave angle of 90° investigated in this study.     

Detection of Alloiococcus otitidis in the nasopharynx and in the outer ear canal

Alloiococcus otitidis has been recovered from the middle ear of children with otitis media with effusion, but its natural habitat is not known. To determine whether the nasopharynx and the outer ear canals are the natural habitats of A. otitidis, 145 swabs (50, nasopharynx; 95 outer ear canal) collected from 50 children were screened by polymerase chain reaction. A. otitidis DNA was detected in seven (4.8%) of the 145 specimens, of which four were nasopharynx, and three outer ear canal. These results indicate that the nasopharynx and outer ear canal may be the body sites for localization of A. otitidis.     

The Environment and the Microbial Ecology of Human Skin

Microbial flora of the skin of three human population groups representing different natural environments was examined quantitatively and qualitatively to determine whether environmental differences in temperature and humidity can influence the microbial flora of normal skin. Five anatomical skin sites - hands, back, axillae, groin, and feet - were sampled from 10 subjects working in a high-humidity, high-temperature environment, 10 subjects from a low-temperature, high-humidity environment, and 10 subjects working in a moderate-temperature and low-humidity environment. Bacterial populations were significantly larger from the back, axillae, and feet in individuals from the high-temperature and high-humidity environment as compared to the moderate-temperature, low-humidity environment. High humidity and low temperature had no significant effect on total populations, but this group showed a higher frequency of isolation of fungi, and gram-negative bacteria from the back and feet. Although there was an indication that increase in the environmental humidity could result in an increased frequency of isolation of gram-negative bacteria, there was no evidence that an increase in either temperature or humidity altered the relative proportions of gram-negative bacteria in the predominantly gram-positive microbial flora found on normal skin. It was concluded that, although climatic changes may cause fluctation in microbial populations from certain sites, they are not a major influence on the ecology of the microbial flora of normal skin in the natural environment. The variables introduced by studying individuals in their natural environment and the influence of these on the results are discussed.     

施用微生物制剂对育蚌水体微生物区系的影响

研究了育蚌水体分别施用微生物制剂利水素与鱼虾菌乐后水样中微生物区系的变化。试验结果表明:施用利水素的水样革兰氏阴性杆菌和嗜水气单胞菌含量分别下降26.67%、20%,芽胞杆菌和表皮葡萄球菌含量分别上升13.33%、26.67%;施用鱼虾菌乐的水样革兰阴性杆菌和嗜水气单胞菌含量分别下降20%、13.33%,芽胞杆菌和表皮葡萄球菌含量分别上升6.67%、13.33%;未施用微生物制剂的水样革兰阴性杆菌和嗜水气单胞菌含量均上升13.33%,芽胞杆菌消失,新增肠杆菌和链球菌。施用微生物制剂利水素和鱼虾菌乐,可有效优化养殖水体微生物种群结构,改良养殖水质环境。     

Evaluation of nongermicidal handwashing protocols for removal of transient microbial flora

A method has been described which can compare the efficacy of different nongermicidal handwashing protocols for removal of transient microbial flora without the necessity of establishing or relying on a previously determined baseline for an individual subject. The wash effluent is collected, and colony counts for the effluent reflect the number removed by the wash protocol. A second standard wash in a handwashing machine is performed, and the test criterion is the percent removed in the test wash based on the sum of total CFU recovered from the two washes. The method was used to compare an 8-s cycle for a newly developed handwashing machine with a conventional 15-s Ivory soap wash. When machine pressure was adequate (42 lb/in2), there was no statistically significant difference in the percent removal of transient flora by the two methods (48.8% from the machine versus 45.1% from the Ivory soap wash). At 32 lb/in2, the Ivory soap wash recovered 60.3%, whereas the machine recovered 45.1%.     

Evaluation of nongermicidal handwashing protocols for removal of transient microbial flora.

A method has been described which can compare the efficacy of different nongermicidal handwashing protocols for removal of transient microbial flora without the necessity of establishing or relying on a previously determined baseline for an individual subject. The wash effluent is collected, and colony counts for the effluent reflect the number removed by the wash protocol. A second standard wash in a handwashing machine is performed, and the test criterion is the percent removed in the test wash based on the sum of total CFU recovered from the two washes. The method was used to compare an 8-s cycle for a newly developed handwashing machine with a conventional 15-s Ivory soap wash. When machine pressure was adequate (42 lb/in2), there was no statistically significant difference in the percent removal of transient flora by the two methods (48.8% from the machine versus 45.1% from the Ivory soap wash). At 32 lb/in2, the Ivory soap wash recovered 60.3%, whereas the machine recovered 45.1%.     

应用PCR对尖锐湿疣和外耳道乳头状瘤中HPV DNA的分型检测

从手术切除的24例女性和12例男性尖锐湿疣新鲜标本中,以及42例女性尖锐湿疣、16例男性外耳道乳头状瘤和4例女性假性湿疣的石蜡包埋标本中,提取组织的基因组DNA,用人工合成的人乳头瘤病毒6.11和16型E6区特异性寡聚核苷酸引物,通过PCR进行HPV DNA的分型检测。结果66例女性尖锐湿疣中,感染HPV6型者4例,感染11型者12例,6+11型混合感染者49例;阴性1例,总检出率达98.4％。4例女性假性湿疣中1例为HPV6型感染,阳性率25％。16例男性外耳道乳头状瘤中HPV6+11型感染者5例,6+16型感染者3例,6+11+16型多重感染者8例,阳性率100％。12例男性尖锐湿疣中,HPV11型感染者7例,6+11型4例,阴性1例,总阳性率91.6％。还对细胞学上空泡化和非典型空泡化尖锐湿疣标本的HPV感染做了比较,未发现差异。     

肠道菌群变化对实验小鼠肠黏膜免疫的影响

目的探讨肠道菌群变化对肠黏膜相关淋巴组织的影响。方法通过变性梯度凝胶电泳（Denatu-ring gradient gel electrophoresis,DGGE）法研究了三种不同级别实验小鼠即清洁级小鼠、SPF小鼠和普通小鼠肠道菌群的组成,并用免疫组织化学（immunohistochemistry,IHC）方法研究了此三种不同级别的实验小鼠肠黏膜相关淋巴组织sIgA阳性细胞分布情况。结果普通小鼠肠道细菌种类最多,其sIgA阳性细胞分布最多,肠道不同部位之间sIgA分布情况差异有显著性（P〈0.05）,小肠和大肠之间的阳性细胞分布差异极显著（P〈0.01）;其次是清洁级小鼠,其肠道不同部位之间菌种组成差异无显著性,小肠和大肠之间的阳性细胞分布差异有显著性（P〈0.05）;SPF小鼠肠道细菌种类最少,故其sIgA阳性细胞分布最少,且其肠道不同部位之间菌种组成差异无显著性,小肠和大肠之间的阳性细胞分布差异无显著性（P〉0.05）。结论随着动物微生物控制级别的增高,肠道微生物多样性递减;sIgA阳性细胞与肠道细菌种类正相关。     

In Vitro Communities Derived from Oral and Gut Microbial Floras Inhibit the Growth of Bacteria of Foreign Origins

The gastrointestinal (GI) tract is home to trillions of microbes. Within the same GI tract, substantial differences in the bacterial species that inhabit the oral cavity and intestinal tract have been noted. While the influence of host environments and nutritional availability in shaping different microbial communities is widely accepted, we hypothesize that the existing microbial flora also plays a role in selecting the bacterial species that are being integrated into the community. In this study, we used cultivable microbial communities isolated from different parts of the GI tract of mice (oral cavity and intestines) as a model system to examine this hypothesis. Microbes from these two areas were harvested and cultured using the same nutritional conditions, which led to two distinct microbial communities, each with about 20 different species as revealed by PCR-based denaturing gradient gel electrophoresis analysis. In vitro community competition assays showed that the two microbial floras exhibited antagonistic interactions toward each other. More interestingly, all the original isolates tested and their closely related species displayed striking community preferences: They persisted when introduced into the bacterial community of the same origin, while their viable count declined more than three orders of magnitude after 4 days of coincubation with the microbial flora of foreign origin. These results suggest that an existing microbial community might impose a selective pressure on incoming foreign bacterial species independent of host selection. The observed inter-flora interactions could contribute to the protective effect of established microbial communities against the integration of foreign bacteria to maintain the stability of the existing communities.     

Comprehensive Analysis of Secondary Dental Root Canal Infections: A Combination of Culture and Culture-Independent Approaches Reveals New Insights

Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12 samples with culture-dependent and -independent methods combined. The number of bacterial species ranged from 1 to 12 in one sample. The majority of the 26 taxa belonged to the phylum Firmicutes (14 taxa), followed by Actinobacteria, Proteobacteria and Bacteroidetes. One sample was positive for fungi, and archaea could not be detected. The results obtained with both methods differed. The cloning technique detected several as-yet-uncultivated taxa. Using a combination of both methods 13 taxa were detected that had not been found in root-filled teeth so far. Enterococcus faecalis was only detected in two samples using culture methods. Combining the culture-dependent and –independent approaches revealed new candidate endodontic pathogens and a high diversity of the microbial flora in root-filled teeth with periradicular lesions. Both methods yielded differing results, emphasizing the benefit of combined methods for the detection of the actual microbial diversity in apical periodontitis.     

胎膜早破与孕妇阴道微生态 及血清IL-10、IL-22、NF-κB的相关性

摘要：目的 探讨胎膜早破与孕妇阴道微生态及血清IL-10、IL-22、NF-kB的相关性。方法 选取我院妇产科收治的84例胎膜早破孕妇为胎膜早破组,100例正常孕妇为正常孕妇组及100例正常未怀孕妇女为正常妇女组。比较3组对象阴道分泌物各细菌阳性率、阴道微生态状态及各组间血清IL-10、IL-22、NF-kB水平。对胎膜早破孕妇阴道微生态失衡与血清IL-10、IL-22、NF-kB水平进行Pearson相关性分析。结果 胎膜早破组孕妇阴道乳杆菌（79.8%）、大肠埃希菌（48.8%）、溶血葡萄球菌（22.6%）检出率均显著高于正常孕妇组和正常妇女组（均P<0.05）。胎膜早破组患者阴道pH（6.87±0.75）、阴道微生态失衡率（40.5%）均显著高于正常孕妇组和正常妇女组,H2O2阳性率（11.9%）低于正常孕妇组（21.0%）和正常妇女组（23.0%）,差异均有统计学意义（均P<0.05）。胎膜早破组孕妇血清IL-10、IL-22、NF-kB水平[（51.28±6.58）pg/mL、（45.91±7.13）pg/mL、（30.47±3.27）ng/mL]均显著高于正常孕妇组和正常妇女组。经过Pearson相关性分析,IL-10、IL-22、NF-kB水平与胎膜早破孕妇阴道微生态失衡呈正相关（r=0.657、0.692、0.732,P<0.05）。结论 孕妇阴道微生态环境发生改变引起阴道菌群失调及血清免疫因子的变化,可能是引发胎膜早破的作用机制之一。     

不同地区人群咽部正常菌群抑制脑膜炎双球菌生长的调查

应用琼脂铺层技术检查了健康者咽部正常菌群对脑膜炎余瑟氏菌(Nm)生长的抑制作用。有些健康者咽部正常菌群对 A 群、B 群和 C 群 Nm 的生长均有抑制作用,而且这种抑菌作用较稳定.1989年冬～1990年春选择流脑发病高的江西宁都县青塘乡和该县未发病的东山坝乡与北京昌平县百善乡作为未发病地区的对照.在流脑发病率高的青塘乡只有8.5%的健康者咽部正常菌群具有抑菌作用;在东山坝乡和百善乡中分别有30%与50%的健康者咽部正常菌群可以抑制 A 群 Nm 生长。这种抑菌作用随年龄增长而增强。初步调查结果表明,健康者咽部正常菌群对 Nm 原发感染似乎有一定的影响。我们对调查中所收集的对 Nm 具有抑菌作用的细菌按 Bergey 氏手册进行了鉴定,现已确定它们是血液链球菌、缓症链球菌、米勒氏链球菌和唾液链球菌四种。     

The normal microbial flora of the baboon vagina.

The microbial flora of the upper vagina and cervix was examined in 38 adult baboons at various stages of the menstrual cycle. The mean number of different species isolated from each baboon was 9.5, with species of Bacteroides, Corynebacterium and group D streptococci predominating. Lactobacilli and mycoplasmas were found in 47.4 and 44.7% of the animals, respectively. No ureaplasms were isolated. Cyclical variations in the microbial flora were minimal.     

腋臭患者腋窝皮肤需氧微生物群的研究

对１６例女性腋患者腋窝皮肤菌群从微生态学的角度进行定性、定量分析。并将结果同２４例正常女性腋窝皮肤菌群做了比较。结果表明,美白喉杆菌及表皮葡萄球力为腋臭患者腋窝皮肤的主要菌生机制及用生态制剂等调整菌群,改善微环境,,达到防治目的有所帮助     

次氯酸钠和洗必泰作为牙根管消毒冲洗液对根管内的感染物质的影响

摘要 目的：探究次氯酸钠和洗必泰作为牙根管消毒冲洗液的对根管内感染物质的影响。方法：选取76例接受根管治疗的患者根据随机数字表法分为观察组和对照组2组,其中对照组患者采用次氯酸钠治疗,观察组患者则采用次氯酸钠和洗必泰联合治疗。对比分析两组患者根管内细菌菌落计数、根管内感染物质、治疗效果、远期有效率和不良反应发生率。结果：根管消毒冲洗后观察组根管内细菌菌落计数和根管内感染物质显著低于对照组,差异具有统计学意义（P<0.05）;观察组患者治疗总有效率和不良反应的发生率分别为100.00%和13.16%,对照组患者治疗总有效率和不良反应的发生率分别为88.00%和65.79%,差异具有统计学意义（P<0.05）。另外,观察组患者第3个月、第6个月和1年后的治疗有效率均大于对照组。结论：次氯酸钠和洗必泰作为牙根管消毒冲洗液能够有效的减少根管内的感染物质的数量,提高治疗有效率,降低不良反应的发生率,值得临床推广使用。     

生物质调理剂对川西北高寒草地沙化土壤养分和微生物群落特征的影响

将农牧废弃物进行资源化处置制成生物质调理剂,用于沙化土壤改良是目前川西北沙化草地生态治理的有效途径之一.为了阐明不同原料调理剂在川西北高寒沙化草地上的实际应用效果,本研究以不施用调理剂为对照(CK),设置施用量均为12 t·hm-2的菌渣(JZ)、秸秆(JG)和生物炭(SWT)3种调理剂,分析了调理剂施用对土壤养分和微...     

大棚蔬菜轮/连作系统土壤肥力与微生物因子综合评价

从土壤肥力与微生物因子探索连茬障碍机理,以期为其提供科学依据。研究草莓番茄轮作(RST)、番茄连作4年(CT4)和番茄连作10年(CT10)3种蔬菜种植模式根际与非根际土壤微生物区系及生理菌群,并对土壤肥力与微生物生物因子进行主成分分析。结果表明根际土壤微生物三大类群和生理菌群数量均高于非根际,根际效应显著。番茄连作根际与非根际土壤细菌和放线菌数量呈现先增加后减少趋势;真菌数量呈线型增加趋势,CT4和CT10在根际与非根际较RST分别增加9.09%和2.11%、75.48%和57.72%。番茄连作根际土壤硝化细菌和好气性自生固氮菌数量的减少,氨化细菌与好气性纤维素分解菌在短期连作表现为增加长期减少的变化趋势;解钾菌、无机磷和有机磷细菌数量在根际与非根际土壤均减少。在6种研究的种植模式中,RST根际土壤状况最好,其次为CT4的根际与RST非根际土壤,CT10的根际土壤、CT4与CT10非根际土壤状况最差。结论是蔬菜连作造成土壤质量下降,连作年限越长下降越显著。