对血平皿及双相培养基两种培养方法的评价

目的:通过实践了解血平皿及双相培养基两种培养方法的差异性,提高阳性率.方法:将体液标本同时接种血平皿及双相培养基,按<全国临床检验操作规程>,根据分离菌株形态、氧化酶、触酶,选择相应的生化/药敏鉴定板,用美国DADE Behring公司生产的MicroScan AS-4微生物分析仪进行菌种鉴定及药敏试验.结果:364例标本血平皿131例为阳性,阳性率为36.0%(131/364),双相培养基164例为阳性,阳性率为45.1%(164/364).两种培养方法经统计学处理χ2=6.26,P<0.05.两种培养方法总阳性率为53.3%(194/364).如将标本仅接种血平皿,漏检率达17.3%(63/364),标本只接种双相培养基,漏检率达8.2%(30/364).结论:将体液标本同时接种上述两种培养基进行培养,可使阳性率有较大的提高.     

229例血培养阳性结果分析

目的研究血液感染的微生物种类分布及病因。方法5 m l血标本在优快双相血培养瓶内于恒温(35±1)℃培养箱孵育,分离阳性标本菌落,采用“第2代15 e系统”进行细菌鉴定,并做药敏试验。结果2 310例血培养标本共分离出229株细菌,其中革兰阴性菌198株占86.46%,革兰阳性菌27株占11.79%,真菌4株占1.74%,阳性率从高到低为甲副伤寒菌、大肠埃希菌、洋葱假单胞菌、产碱杆菌属、金黄色葡萄球菌,分别是55.89%、6.55%、5.24%、3.93%、2.62%。沙门菌占65.06%,分离自感染内科及门、急诊科,大肠埃希菌主要见于感染内科及内分泌科,常见菌阳性标本在孵育<24、24~48、48~76、≥76 h的阳性率依次为5.29%、46.47%、85.88%、99.40%。结论血液感染病原菌在沿海地区以沙门菌属尤其以甲副伤寒菌为主,其次为非发酵菌和葡萄球菌。快速血培养技术仍是血液感染病原学诊断的必要手段。     

双向瓶1589份血培养结果分析

目的:了解双向瓶血培养细菌的分类情况.方法:用法国生物梅里埃公司生产的双向血培养瓶(Hemoline),对临床1 589份血液标本进行检测,阳性者用法国生物梅里埃公司生产的API系统进行鉴定.结果:分离出265株细菌共42种,阳性率为16.68%,较传统培养方法高.菌种涉及范围广,其中革兰阳性菌占56.98%,革兰阴性菌占31.70%,真菌占11.32%.结论:用Hemoline双向血培养瓶进行血培养提高了细菌检出的阳性率,增加了检出细菌的种类.     

栽培与野生化血丹不同部位中两种化学成分含量分析

为探讨并分析栽培与野生化血丹植株中不同部位中两种化学成分的含量差异,该研究采用超声法提取、高效液相色谱法(HPLC)测定栽培与野生化血丹根、茎、叶、花、混合样等部位中桃叶珊瑚苷和梓醇的含量,并进行比较。结果表明:(1)桃叶珊瑚苷在栽培与野生化血丹植株内均有分布,含量均以根中最高,其在栽培与野生化血丹植株内的含量表现分别为根>叶>混合样>茎>花、根>混合样>茎>花>叶,栽培化血丹不同部位中桃叶珊瑚苷的含量均高于野生化血丹。(2)梓醇在栽培化血丹的茎中未检出,在栽培与野生化血丹其他部位均有分布,含量均以叶中最高,其在栽培与野生化血丹植株内的含量分别表现为叶>花>混合样>根、叶>混合样>茎>花>根,野生化血丹不同部位中梓醇的含量均高于栽培化血丹。(3)桃叶珊瑚苷和梓醇在栽培和野生化血丹植株不同部位中的含量均存在显著差异(P<0.05),栽培与野生同一部位间总体上无显著差异,为该濒危药用植物资源药用部位选择和合理开发利用提供实验参考。     

2008-2012年昆明市儿童医院50199例患儿血培养结果的变迁及耐药状况分析

目的分析近5年(2008-2012年)本院儿童血培养结果,了解儿童血培养病原菌分布及耐药情况,为临床合理用药提供参考。方法采用BD BACTEC9240全自动血培养仪及其配套的树脂儿童血培养瓶,对5年来50 199份患儿静脉血样本进行培养,采用Sensititre Aris2X全自动微生物分析系统,对细菌进行鉴定及药敏试验。结果近5年共检测血培养50 199例,分离出1 713株病原菌,阳性检出率为3.41%。其中革兰阳性球菌1 209株,占70.58%;革兰阴性菌492株,占28.72%;真菌12例,占0.70%。其中检出频次最高的5种病原菌分别为凝固酶阴性葡萄球菌属、大肠埃希菌、屎肠球菌、肺炎克雷伯杆菌及洋葱伯克霍尔德菌。革兰阴性菌检出率有逐年增高趋势,而革兰阳性球菌检出率则呈现逐年下降趋势。凝固酶阴性葡萄球菌属对青霉素、苯唑西林、红霉素和头孢唑林敏感性低,对万古霉素敏感,未发现耐万古霉素的葡萄球菌及肠球菌。大肠埃希菌和肺炎克雷伯杆菌对第3、4代头孢菌素耐药率高,对亚胺培南、美洛培南敏感。结论凝固酶阴性葡萄球菌属是引起儿童菌血症或败血症的首要病原菌,并且耐药率高;但据该研究发现,近年来革兰阴性菌感染率逐年升高,并且对3、4代头孢菌素耐药率高。因此,临床治疗用药需参考药敏试验结果及儿科用药毒副作用合理选择抗菌药物。     

两种方法诊断女性需氧菌阴道炎的对比分析

摘要：目的 对比分析两种方法诊断女性需氧菌阴道炎的效果。方法 使用Donders湿片高倍镜检法、Tempera临床和微生物诊断法对2014年5月至2015年4月妇科门诊8 326例就诊患者需氧菌感染情况进行对比分析。结果 在8 326例就诊患者中,采用Donders湿片高倍镜检法检出需氧菌阴道炎患者1 819例,检出率为21.8%;采用Tempera临床和微生物诊断法检出需氧菌阴道炎患者1 825例,检出率为21.9%。通过SPSS13.0统计学分析,两种方法在需氧菌阴道炎检出率方面不存在显著差异（χ2=20.4,P>0.05）。结论 这两种方法诊断需氧菌阴道炎具有很好的一致性,医院可根据自身实际情况选择其中一种。南宁地区女性需氧菌阴道炎检出率仅次于细菌性阴道病,针对需氧菌阴道炎应采取科学有效的治疗方案,提升需氧菌阴道炎治疗效率。     

中山地区小儿血培养病原菌分布及耐药性分析

目的了解中山地区小儿血培养病原菌分布及其常见病原菌对抗生素的耐药情况。方法对2008年至2009年南方医科大学附属中山市博爱医院新生儿科、儿科住院患儿进行血培养,应用BacT/ALERT 3D全自动血培养仪进行检测,阳性菌株用VITEK-32全自动微生物分析仪进行菌株鉴定及药敏试验。结果 4 647例小儿血培养共检出病原菌316株,总阳性率为6.8%;革兰阳性菌258株(81.6%),其中凝固酶阴性葡萄球菌(CNS)209株(66.1%);革兰阴性菌55株(17.4%),以大肠埃希菌14株(4.4%)和肺炎克雷伯菌12株(3.8%)为主。革兰阳性菌对青霉素G耐药率高达93.4%,其次为氨苄西林/舒巴坦(82.9%)、苯唑西林(82.3%),耐药率均在80%以上。耐甲氧西林金黄色葡萄球菌(MRSA)占25%,耐甲氧西林凝固酶阴性葡萄球菌(MRSCN)占82.3%。未检出对万古霉素、利奈唑胺和呋喃妥因耐药的菌株。革兰阴性菌中大肠埃希菌、肺炎克雷伯菌ESBLs的检出率分别为50%和33%,这些菌株对头孢哌酮/舒巴坦、丁胺卡那霉素、左旋氧氟沙星和亚胺培南的敏感率分别为91.7%、100%、92.9%、100%,表现了较低耐药率。结论本地区小儿血培养病原菌以革兰阳性菌为主,且表现为多重耐药。     

两种方法诊断女性需氧菌阴道炎的对比分析

目的对比分析两种方法诊断女性需氧菌阴道炎的效果。方法使用Donders湿片高倍镜检法、Tempera临床和微生物诊断法对2014年5月至2015年4月妇科门诊8 326例就诊患者需氧菌感染情况进行对比分析。结果在8 326例就诊患者中,采用Donders湿片高倍镜检法检出需氧菌阴道炎患者1 819例,检出率为21.8%;采用Tempera临床和微生物诊断法检出需氧菌阴道炎患者1 825例,检出率为21.9%。通过SPSS13.0统计学分析,两种方法在需氧菌阴道炎检出率方面不存在显著差异(χ2=20.4,P0.05)。结论这两种方法诊断需氧菌阴道炎具有很好的一致性,医院可根据自身实际情况选择其中一种。南宁地区女性需氧菌阴道炎检出率仅次于细菌性阴道病,针对需氧菌阴道炎应采取科学有效的治疗方案,提升需氧菌阴道炎治疗效率。     

两种不同内皮祖细胞的分离培养与鉴定

目的:探讨从小鼠骨髓中分离、培养、诱导分化及鉴定两种内皮祖细胞的方法,为进一步研究和临床应用奠定基础。方法:密度梯度离心法分离小鼠骨髓单个核细胞,接种于内皮祖细胞条件培养基,通过贴壁培养法培养出早期内皮祖细胞和晚期内皮祖细胞,并在0 d、6 d、10 d流式鉴定早期内皮祖细胞,在第8周流式鉴定晚期内皮祖细胞。结果:通过体外贴壁扩增培养,从小鼠骨髓细胞中成功培养出EEPC(早期内皮祖细胞)和EOC(晚期内皮祖细胞),表达CD34+/CD133+/VEGFR2+的EEPC比例从最初的0.08%能够增长至70%;EOC大约出现于3-4周,5-8周时呈现指数增长,具有典型的内皮细胞鹅卵石样形态,表达CD31、VEGFR2等内皮细胞表面标志而不表达CD34、CD133等干细胞表面标志。结论:确立了内皮祖细胞体外分离培养和诱导分化的实验方法,为进一步研究奠定基础。     

14种蜂花粉的DNA和RNA分析

本文使用紫外光吸收法,分析了芝麻、葵花、泡桐等１４种蜂花粉中核酸（ＤＮＡ和ＲＮＡ）的含量,以进一步探讨花粉的抗衰老作用。结果说明不论那一种花粉,均含有丰富的核酸,但种类不同的花粉其中核酸的含量是不完全相同的。     

痰涂片和细菌培养结果符合性探讨

为了探讨工作中痰涂片和细菌培养结果符合性,对株州市第二医院2009年1月至2009年12月痰标本的1 780份样本进行了痰涂片和细菌培养,观察其两种方法的符合率。结果显示,1 780份痰标本中较理想标本404例,占22.7%;可接受标本B组524例,占29.5%;不可接受标本C组,852例,占47.8%。A组标本涂片与培养结果符合率为67.8%,B组为65.6%,C组为49.1%,A组和B组符合率之间没有统计学差异(P>0.05),A组、B组样本涂片与培养结果符合率显著高于C组。痰涂片镜检在标本培养前与培养结果具有很大的相关性,可排除不合格标本,提高致病菌的检出率。     

BacT／Aler3D全自动血培养仪的临床应用及评价

目的评价全自动血培养仪BacT／Aler3D的临床应用情况。方法对BacT／Aler3D全自动血培养仪检测的1853份标本,其检出的阳性率、病原菌种类及阳性检出时间进行了评估。结果1853份血培养分离出病原菌189株,阳性率为10．2％,分离出病原菌23种,最快检出时间为2h,24h以内阳性率为68．5％,48h内阳性率为87．3％,72h以内阳性率为92．3％。结论BacT／Aler3D全自动血培养仪提高血培养的阳性率,检出的细菌种类多,污染机会少,缩短阳性检出时间,而且操作简便,结果快速、准确。     

血培养分离出212株念珠菌的菌种分布及耐药性分析

目的分析血培养分离出的念珠菌的菌种分布及耐药分析,为临床念珠菌血症的诊治提供理论依据。方法回顾性分析2017年1月—2020年12月期间,本院血培养分离出的念珠菌的菌种分布、药敏试验结果及患者血培养分离出念珠菌前后96 h内的G试验结果。结果血培养中分离出念珠菌314例,阳性率2.1%,其中非重复分离株212例。检出率最高的是近平滑念珠菌(72株,34.0%),其次是白念珠菌(55株,25.9%)和光滑念珠菌(28株,13.2%)。念珠菌检出率最高的科室为ICU(62株,29.2%),其次是新生儿科(39株,18.4%)和血液科(20株,9.4%)。检出的212株念珠菌除一株近平滑念珠菌为两性霉素B的非野生株,其余均为两性霉素B的野生株。白念珠菌对唑类药物的敏感率超过90%。但光滑念珠菌和热带念珠菌对唑类药物的敏感性较低。血培养分离出念珠菌的前后96 h内,G试验的阳性率为73.7%。结论本院念珠菌检出率前3位分别是近平滑念珠菌、白念珠菌和光滑念珠菌。光滑念珠菌、热带念珠菌对唑类药物敏感性比较低,在经验用药时需要合理选择抗真菌药物。G试验对念珠菌血症有较高的价值,需要对结果进行动态监测。     

革兰阴性菌引起的血培养阳性标本直接细菌鉴定和药敏的应用

目的 评估血培养阳性标本直接细菌鉴定和药敏可行性.方法 将血培养瓶放入Bact/Alert 3D 60血培养系统进行培养筛选.选取78份含革兰阴性杆菌的阳性血培养瓶进行试验.抽取培养液,用BD真空分离管离心血细胞.在收集到足量菌液后,用VITEK-32革兰阴性菌鉴定药敏卡做直接鉴定药敏试验.用标准方法及亚培养后的鉴定药敏试验对直接鉴定药敏试验进行评估.结果 VITEK-32直接鉴定试验,78株中的74株(94.9％)准确鉴定,直接药敏试验标准符合率95.6％.KB法血标本直接药敏试验标准符合率96.2％,但微小错误率高于VITEK-32直接药敏法.结论 Bact/Alert血培养阳性标本直接VITEK-32细菌鉴定和药敏对革兰阴性菌是切实可行的,可大幅度缩短时间,为临床及时修正用药提供依据,具有较高的应用价值.     

低能量激光与集落刺激因子对人脐带血造血细胞的增殖作用

本研究用铜蒸气激光照射人脐带带血造血细胞,观察低能量激光与集落因子对造血细胞的增殖作用。结果显示激光加ＣＳＦ对造血细胞ＧＭ－ＣＦＵｃ有协同增殖作用,与单用激光照射组,ＣＳＦ刺激组及对照组相比,有非常显著性差异。     

Cage culture effects on mullets (Mugilidae) liver histology and blood chemistry profile

A comparative study of blood chemistry and histology was conducted on two groups of mullets (Mugilidae) living under different conditions with different feed sources. The aquaculture influenced mullet group (AIM), was collected near fish farms and the control group of mullet (CM) was caught in the waters without any aquaculture activities. Histological and biochemical procedures were employed to study liver histomorphology, plasma aspartate and alanine aminotransferase (AST, ALT), triglyceride (TRIG), cholesterol (CHOL), glucose (GLU) and total protein (TP) of both AIM and CM. Moderate histological changes (lipid infiltration) were observed in the liver of AIM. Significant changes in plasma variables were observed in AIM. Blood chemistry variables measured proved to be good indicators of artificial feed effects. Classical statistical approaches were applied to the blood chemistry and histopathology data. For the first time machine learning techniques were used to generate comprehensible classification models and to explore blood chemistry variable importance, strength, their mutual interactions or dependencies, and to investigate reliability of particular variables within the groups.     

The utilization of glycogen accumulating organisms for mixed culture production of polyhydroxyalkanoates

Production of polyhydroxyalkanoates (PHAs) by an open mixed culture enriched in glycogen accumulating organisms (GAOs) under alternating anaerobic–aerobic conditions with acetate as carbon source was investigated. The culture exhibited a stable enrichment performance over the 450‐day operating period with regards to phenotypic behavior and microbial community structure. Candidatus Competibacter phosphatis dominated the culture at between 54% and 70% of the bacterial biomass throughout the study, as determined by fluorescence in situ hybridization. In batch experiments under anaerobic conditions, PHA containing 3‐hydroxybutyrate (3HB) and 27 mol‐% 3‐hydroxyvalerate (3HV) was accumulated up to 49% of cell dry weight utilizing the glycogen pool stored in the SBR cycle. Under aerobic and ammonia limited conditions, PHA comprising only 3HB was accumulated to 60% of cell dry weight. Glycogen was consumed during aerobic PHA accumulation as well as under anaerobic conditions, but with different stoichiometry. Under aerobic conditions 0.31 C‐mol glycogen was consumed per consumed C‐mol acetate compared to 0.99 under anaerobic conditions. Both the PHA biomass content and the specific PHA production rate obtained were similar to what is typically obtained using the more commonly applied aerobic dynamic feeding strategy. Biotechnol. Bioeng. 2009; 104: 698–708 © 2009 Wiley Periodicals, Inc.     

Correlation between thylakoid protein phosphorylation and molecular organization of the photosynthetic apparatus in a dynamic system

In-vitro thylakoid protein phosphorylation has been studied in synchronized cells of Scenedesmus obliquus at the 8- and 16-h of the life cycle, stages which are characterized by the maximum and minimum photosynthetic activities, respectively. The stage of maximum photosynthetic activity (8-h) is characterized by the highest protein phosphorylation in vitro and in vivo, by the largest proportion of the heavy subfraction of thylakoids, and by maximum oligomerization of the light-harvesting chlorophyll a/b-protein complex, altogether creating the highest energy charge of the thylakoid membranes. Protein phosphorylation in vitro decreases the amount of the heavy subfraction and increases the amount of oligomerization of the antenna of photosystem I (PSI) (increase of chlorophyll b in the light fraction). Concomittantly, PSII units become smaller (longer time for the rise in fluorescence induction) and photosynthetic efficiency increases (decrease of fluorescence yield). In-vivo protein phosphorylation is controlled mainly endogenously during the 8-h of the life cycle but is exogenously modulated by light to optimize the photosynthetic activity by redistribution of pigment-protein complexes. In-vitro protein phosphorylation seems to restore partially the conditions prevalent in vivo and lost during the preparation of membranes. The effect is greater in 16-h cells which have less-stable membranes. The regulatory mechanism between membrane stabilization and oligomerization on the one hand and redistribution of the light-harvesting chlorophyll a/b-protein complex from PSII to PSI on the other hand remains unexplained. We have confirmed that the mechanism of protein phosphorylation is regulated via plastohydroquinone, but experiments with the plastohydroquinone analogue 2,3,5,6-tetramethyl-p-benzoquinone demonstrated that plastohydroquinone is not solely responsible for the differences in protein phosphorylation of 8- and 16-h thylakoids. The inhibitory effect of ADP and the distinct rates of kinase reaction indicate that the adenylate energy charge and changes in the organization of the photosynthetic apparatus also contribute to the observed differences in protein phosphorylation. Phosphorylation in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea indicated that the 32-kDa phosphoprotein and the herbicide-binding Q_B protein may be the same. These experiments also indicated that 3-(3,4-dichlorophenyl)-1,1-dimethylurea-binding reduces kinase activity directly and not only by inhibiting electron transport.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - LHCP light-harvesting chlorophyll a/b-protein complex - PSI, II photosystem I, II - TMQ 2,3,5,6-tetramethyl-p-benzoquinone Dedicated to Professor Dr. W. Nultsch on the occasion of his 60th brithday     

Studies on the relationship between glycolysis and (Na++K+)-ATPase in cultured cells

In several tissues a coupling between glycolysis and (Na⁺+K⁺)-ATPase has been observed. We report here studies on the coupling of glycolysis and (Na⁺+K⁺)-ATPase in Rous-transformed hamster cells and Ehrlich ascites tumor cells. The rate of (Na⁺+K⁺)-ATPase was estimated by the initial rate of ouabain-sensitive K⁺ influx after K⁺ reintroduction to K⁺-depleted cells. Experiments were performed with cells producing ATP via oxidative phosphorylation alone (i.e., lactate sole substrate), glycolysis alone (i.e., glucose as substrate in the absence of oxygen or with antimycin A), or glycolysis and oxidative phosphorylation (i.e., glucose as substrate in the presence of oxygen). The cells produced ATP at approximately the same rate under all of these conditions, but the initial rate of K⁺-influx was approx. 2-fold higher when AtP was produced from glycolysis. Changes in cell Na⁺ due to other transport processes related to glycolysis, such as Na⁺-H⁺ exchange, Na⁺-glucose cotransport, and K⁺-H⁺ exchange were ruled out as mediators of this effect on (Na⁺+K⁺)-ATPase. These data suggest that glycolysis is more effective than oxidative phosphorylation in providing ATP to (Na⁺+K⁺)-ATPase to these cultured cells.     

Detection of filarial infection usingWuchereria bancrofti microfilariae culture antigen and filter paper blood samples in enzyme linked immunosorbent assay

Blood collected on filter paper by finger-prick gave results comparable to intravenous serum samples when analysed by enzyme-linked immunosorbent assay (ELISA). All the 100 microfilaraemia, 5 out of 100 endemic normals and none of the 10 nonendemic normal filter paper blood samples showed the presence of filarial antibody when tested by this method,using culture antigen and anti-immunoglubulins, class G, M and A — penicillinase conjugate. When the same samples were screened for the presence of IgM antibody, 91 out of 100 microfilaraemia, 13 out of 100 endemic normal and none of the 10 nonendemic normal samples showed a positive reaction. Enzyme linked immunosorbent assay, using culture antigen and filter paper blood samples, appears to work in large field studies for detection of filarial infection.