多药耐药肺炎克雷伯菌9种可移动遗传元件标记研究

目的了解多药耐药肺炎克雷伯菌可移动遗传元件的存在状况。方法收集连云港市第二人民医院2008年11月-2009年10月住院患者标本中分离的多药耐药肺炎克雷伯菌20株,采用PCR法检测接合性质粒遗传标记(traAt、rbC)、转座子遗传标记(tnpUt、np513)、插入序列遗传标记(IS26、ISEcp1)、整合子遗传标记(intⅠ1、intⅠ2i、ntⅠ3)9种可移动遗传元件的遗传标记。结果 20株多药耐药肺炎克雷伯菌9种可移动遗传元件标记检出阳性率traA 10.0%t、rbC 60.0%t、npU 45.0%t、np513 60.0%I、S26 100.0%I、SEcp1 60.0%i、ntⅠ95.0%、intⅡ0i、ntⅢ0。结论该组20株多药耐药肺炎克雷伯菌可移动遗传元件遗传标记的携带率较高,尤其是插入序列26和Ⅰ类整合子;肺炎克雷伯菌多药耐药的表型可能与携带多种可移动遗传元件导致耐药基因高表达相关。     

质粒型AmpC酶DHA基因在耐药肺炎克雷伯菌中的流行研究

目的调查对常用抗菌药物耐药肺炎克雷伯菌(DRK)获得性耐药基因及相关可移动遗传元件的携带状况及菌株间的亲缘关系。方法 20株耐药肺炎克雷伯菌均分离自医院2015年1-12月住院患者痰标本,用K-B法测定抗菌药物的敏感性,采用聚合酶链反应(PCR)及序列分析的方法分析24种β-内酰胺类获得性耐药基因、11种氨基糖苷类获得性耐药基因、10种可移动遗传元件遗传标记,阳性耐药基因测序后直接作BLAST比对,耐药基因检测结果作样本聚类分析(UPGMA法)。结果 20株耐药肺炎克雷伯菌对β-内酰胺类、氨基糖苷类、喹诺酮类均耐药,但对碳青霉烯类均敏感;20株菌均检出β-内酰胺类获得性耐药基因,阳性率为100.0%,共检出6种β-内酰胺酶基因,blaDHA群基因检出率为最高90.0%;每株也均检出氨基糖苷类修饰酶基因,阳性率为100.0%,共检出4种氨基糖苷类获得性耐药基因,ant(3″)-Ⅰ群基因检出率为最高90.0%,但16SrRNA甲基化基因未检出;可移动遗传元件标记基因每株也均有检出,共检出7种可移动遗传元件标记基因,其中tnp513、IS26、IS903、ISEcp1、intⅠ1阳性率均为100.0%,trbC阳性率95.0%;样本聚类分析显示,该组菌株有明显的聚集性,分A与B二个群,B群又可分为B-1、B-2两个亚群,B-2亚群存在二个克隆传播,其中5-6号株均携带10种基因,4-7-8-9-13-14-15-16-17-18-19-20号株均携带9种基因。结论 20株耐药肺炎克雷伯菌同时携带了β-内酰胺类获得性耐药基因、氨基糖苷类修饰酶基因和可移动遗传元件标记基因,是对β-内酰胺类和氨基糖苷类产生耐药的重要原因,该组菌检出的2个克隆高度疑似医院感染,同一克隆菌株携带相同基因。     

耐药肺炎克雷伯菌湖北襄阳分离株获得性耐药元件研究北大核心CSCD

目的调查耐药肺炎克雷伯菌获得性耐药元件基因的携带情况及菌株间的亲缘关系,为临床治疗提供参考依据。方法收集2016年1-10月医院住院患者痰液标本分离15株耐药肺炎克雷伯菌,用K-B法测定9种抗菌药物的敏感性,采用聚合酶链反应(PCR)及序列分析的方法分析35种β-内酰胺类获得性耐药基因,21种氨基糖苷类获得性耐药基因和10种可移动遗传元件遗传标记;阳性耐药基因测序后直接作BLAST比对,耐药基因检测结果作样本聚类分析。结果 15株耐药肺炎克雷伯菌对β-内酰胺类、氨基糖苷类、喹诺酮类耐药率高;15株菌每株均检出β-内酰胺酶基因,总阳性率达100.0%,共检出7种β-内酰胺酶基因,其中仅检出1种β-内酰胺酶基因1株,占6.7%,同时检出2种9株,占60.0%,同时检出3种5株,占33.3%;样本聚类分析可见有一定的聚集性,并可分为A与B二个群;除A群有2个菌株属克隆传播,其余菌株亲缘关系稍远。结论本组15株耐药肺炎克雷伯菌同时携带β-内酰胺类药物获得性耐药基因、氨基糖苷类药物获得性耐药基因和可移动遗传元件遗传标记,是对β-内酰胺类和氨基糖苷类产生耐药的重要原因。     

耐药肺炎克雷伯菌湖北襄阳分离株获得性耐药元件研究

目的调查耐药肺炎克雷伯菌获得性耐药元件基因的携带情况及菌株间的亲缘关系,为临床治疗提供参考依据。方法收集2016年1-10月医院住院患者痰液标本分离15株耐药肺炎克雷伯菌,用K-B法测定9种抗菌药物的敏感性,采用聚合酶链反应(PCR)及序列分析的方法分析35种β-内酰胺类获得性耐药基因,21种氨基糖苷类获得性耐药基因和10种可移动遗传元件遗传标记;阳性耐药基因测序后直接作BLAST比对,耐药基因检测结果作样本聚类分析。结果 15株耐药肺炎克雷伯菌对β-内酰胺类、氨基糖苷类、喹诺酮类耐药率高;15株菌每株均检出β-内酰胺酶基因,总阳性率达100.0%,共检出7种β-内酰胺酶基因,其中仅检出1种β-内酰胺酶基因1株,占6.7%,同时检出2种9株,占60.0%,同时检出3种5株,占33.3%;样本聚类分析可见有一定的聚集性,并可分为A与B二个群;除A群有2个菌株属克隆传播,其余菌株亲缘关系稍远。结论本组15株耐药肺炎克雷伯菌同时携带β-内酰胺类药物获得性耐药基因、氨基糖苷类药物获得性耐药基因和可移动遗传元件遗传标记,是对β-内酰胺类和氨基糖苷类产生耐药的重要原因。     

耐碳青霉烯类肺炎克雷伯菌耐药元件分析研究

目的探讨一组耐碳青霉烯类肺炎克雷伯菌获得性耐药基因及可移动遗传元件遗传标记的携带情况及关联性。方法收集2013年1-12月宁波市第二医院住院患者中分离到的20株耐碳青霉烯类肺炎克雷伯菌,用gyrA与parC测序确认菌种,再采用聚合酶链反应(PCR)及序列分析的方法分析β-内酰胺类、氨基糖苷类、喹诺酮类共56种获得性耐药相关基因和12种可移动遗传元件遗传标记,最后对检测结果作指标聚类分析。结果 20株耐碳青霉烯类肺炎克雷伯菌每株均检出获得性耐药基因与可移动遗传元件标记,20株菌共检出6种β-内酰胺类获得性耐药基因、4种氨基糖苷类获得性耐药基因、1种16SrRNA甲基化酶基因、8种可移动遗传元件遗传标记,指标聚类分析提示bla KPC与ISKpn6相强关联,bla OXA-1与tnp513相强关联,17株bla KPC-2阳性菌bla KPC-ISKpn6连锁检测均为阳性。结论肺炎克雷伯菌中携带的耐药基因和耐药表表型相对应,携带bla KPC-2和bla IMP-4是该组菌耐碳青霉烯类的重要原因。     

携带blaCTX-M-55基因的大肠埃希菌氨基糖苷类耐药相关元件分析

目的调查携带blaCTX-M-55基因的大肠埃希菌中氨基糖苷类耐药相关元件的携带状况,以及了解本组菌株间的亲缘关系。方法收集2015年1-12月宁波大学医学院附属医院住院患者痰液标本中分离30株携带blaCTX-M-55基因的大肠埃希菌,用K-B法测定9种抗菌药物的敏感性,采用聚合酶链反应(PCR)及序列分析的方法分析7种氨基糖苷类修饰酶基因,2种16SrRNA甲基化酶基因,7种可移动遗传元件遗传标记,阳性耐药基因测序后直接作BLAST比对,耐药基因检测结果作样本聚类分析(UPGMA法)。结果 30株携带blaCTXM-55基因的大肠埃希菌对头孢类、氨基糖苷类、喹诺酮类抗菌药物均耐药,但对碳青霉烯类抗菌药物均为敏感;30株菌均检出氨基糖苷类修饰酶基因和可移动遗传元件遗传标记,另有5株检出16SrRNA甲基化酶基因,30株菌共检出5种氨基糖苷类修饰酶基因、1种16SrRNA甲基化酶基因、7种可移动遗传元件遗传标记,且阳性率较高;样本聚类分析显示,该组菌株有明显的聚集性,有6个克隆传播。结论本组30株携带blaCTX-M-55基因的大肠埃希菌同时携带了氨基糖苷类修饰酶基因、16SrRNA甲基化酶基因和可移动遗传元件遗传标记,是对头孢类和氨基糖苷类产生耐药的重要原因;本组菌检出的6个克隆高度疑似医院感染,同一克隆菌株携带相同基因。     

产ESBL肺炎克雷伯菌的β-内酰胺类与氨基糖苷类和磺胺类耐药元件分析

目的调查一组20株产ESBL肺炎克雷伯菌中β-内酰胺类、氨基糖苷类与磺胺类获得性耐药元件基因的携带状况,以及菌株间的亲缘关系。方法收集2015年1月-6月医院住院患者标本中分离的20株产ESBL肺炎克雷伯菌,采用聚合酶链反应(PCR)及序列分析的方法分析23种β-内酰胺类耐药基因、11种氨基糖苷类耐药基因、5种磺胺类耐药基因、8种可移动遗传元件遗传标记,阳性耐药基因测序后直接作BLAST比对,耐药基因检测结果作样本聚类分析(UPGMA法)。结果 20株产ESBL肺炎克雷伯菌每株均检出β-内酰胺类耐药基因、磺胺类耐药基因和可移动遗传元件遗传标记基因,阳性率达100.0%,有19株检出氨基糖苷类药物获得性耐药基因,阳性率95.0%;20株菌共检出6种β-内酰胺酶类耐药基因、4种氨基糖苷类耐药基因、3种磺胺类耐药基因、6种可移动遗传元件遗传标记,且阳性率较高;样本聚类分析显示菌株有聚集性,共有12-14号株、7-20号株、15-18-19号株、8-9号株4个克隆传播。结论本组20株产ESBL肺炎克雷伯菌携带的β-内酰胺类耐药基因、氨基糖苷类耐药基因、磺胺类耐药基因和可移动遗传元件遗传标记是对β-内酰胺类、氨基糖苷类、磺胺类产生耐药的重要原因,本组菌检出的四个克隆高度疑似医院感染。     

耐碳青霉烯类肺炎克雷伯菌耐药相关基因分析

目的探讨耐碳青霉烯类肺炎克雷伯菌耐药基因及可移动遗传元件遗传标记的携带,与菌株间的亲缘性。方法收集2014年1-12月住院患者中分离20株耐碳青霉烯类肺炎克雷伯菌,经gyrA测序比对确认菌种,再采用聚合酶链反应(PCR)及序列分析的方法分析β-内酰胺类、氨基糖苷类、喹诺酮类共57种耐药相关基因与12种可移动遗传元件遗传标记,最后对检测结果作样本聚类分析。结果 20株耐碳青霉烯类肺炎克雷伯菌每株均检出β-内酰胺酶基因、氨基糖苷类修饰酶基因、喹诺酮类作用靶位基因gyrA突变以及可移动遗传元件标记;20株菌共检出4种β-内酰胺类耐药基因、4种氨基糖苷类耐药基因、1种16SrRNA甲基化酶基因、1种喹诺酮类耐药基因、6种可移动遗传元件遗传标记,阳性率非常高,且阳性基因可分为6种阳性模式;样本聚类分析发现,20株菌可区分为A、B两个家族,A家族中1号株为孤独株,A1(2和9号株)与A2(10、11、12、13、15、16、17和18号株)子家族均为克隆传播,A3子家族有2株菌;B家族(3、4、5、6、7、8和20号株)亦为克隆传播,并已构成暴发流行。结论本组肺炎克雷伯菌中携带的耐药基因与耐药表表型相对应,样本聚类分析提示本组菌有克隆传播,有的已构成暴发流行。     

多药耐药肺炎克雷伯菌抗菌制剂外排泵基因研究

目的了解20株多药耐药肺炎克雷伯菌(MDR-KPN)的灭菌剂、消毒剂、抗菌药物等抗菌制剂外排泵基因存在情况。方法运用PCR法对20株多药耐药肺炎克雷伯菌mdfA、tehA、smr、qacE△1等抗菌制剂外排泵基因进行检测。结果检出mdfAt、ehA、smr、qacE△1等4种外排泵基因,阳性率分别为:85.0%、5.0%、20.0%、75.0%;mdfA、qacE△1阳性率高,多数菌株能同时检出多种外排泵基因;在肺炎克雷伯菌中检出mdfA、tehA、smr基因为国内首次报道。结论多耐药肺炎克雷伯菌易检出多种外排泵基因,与医院多药耐药肺炎克雷伯菌耐药机制相关的可移动遗传元件主要有mdfA、qacE△1,耐药菌外排泵携带率高,开发抑制剂显得十分重要。     

携带rmtB基因的大肠埃希菌β-内酰胺酶基因研究

目的调查一组携带rmtB基因的大肠埃希菌中β-内酰胺酶基因与可移动遗传元件的存在状况,了解该组菌株之间的亲缘关系。方法收集2016年1-10月湖北医药学院附属襄阳市第一人民医院住院患者痰液标本分离到的20株携带rmtB基因的大肠埃希菌,采用K-B法测定9种抗菌药物的敏感性,采用聚合酶链反应(PCR)及序列分析法分析16种β-内酰胺酶基因和7种可移动遗传元件遗传标记;阳性耐药基因测序后直接作BLAST比对,耐药基因检测结果作样本聚类分析(UPGMA法)。结果 20株携带rmtB基因的大肠埃希菌对头孢类、氨基糖苷类、喹诺酮类均耐药,但对碳青霉烯类均敏感;20株菌均检出β-内酰胺酶基因及可移动遗传元件遗传标记;20株菌共检出5种β-内酰胺酶基因和7种可移动遗传元件遗传标记,且阳性率较高;样本聚类分析显示20株菌有明显的聚集性,分A与B二个群,并有4个克隆传播。结论本组20株携带rmtB基因的大肠埃希菌同时携带了β-内酰胺酶基因和可移动遗传元件遗传标记,是对头孢类和氨基糖苷类产生耐药的重要原因;本组菌检出的4个克隆高度疑似医院感染,同一克隆菌株携带相同基因。     

Influence of vitamin K on anticoagulant therapy depends on vitamin K status and the source and chemical forms of vitamin K

Warfarin therapy requires close monitoring to avoid excessive bleeding and to maintain the effective therapeutic concentration assessed with the internationalized ratio (INR). High vitamin K intake can decrease the therapeutic effectiveness of warfarin, while poor vitamin K status appears to increase the sensitivity to small changes in vitamin K intake, especially from supplements. Very large amounts of vitamin K from a single meal with vegetables (400 g of vegetables with 700 to 1500 microg of vitamin K1) can measurably change INR, but occasional typical servings (<100 g) would probably have little lasting impact on INR. Warfarin requirements may change in those altering their intake of dark-green vegetables. The 2005 Dietary Guidelines for Americans recommends 3 cups/week of dark-green vegetables, which contain about 100 to 570 microg/serving of vitamin K1. Less well-known sources and chemical forms of vitamin K, such as MK-7 in natto (a fermented Japanese product), also measurably influence INR. Additional research is needed in warfarin-treated patients to fully quantify the interactions among various sources and chemical forms of vitamin K, age, genotype, and other factors.     

Vitamin K and Osteoporosis

Vitamin K acts as a coenzyme of carboxylase, catalyzing the carboxylation of several vitamin K dependent proteins. Beyond its well-known effects on blood coagulation, it also exerts relevant effects on bone and the vascular system. In this review, we point out the relevance of an adequate vitamin K intake to obtain sufficient levels of carboxylated (active form) vitamin K dependent proteins (such as Osteocalcin and matrix Gla protein) to prevent bone health. Another bone-related action of Vitamin K is being a ligand of the nuclear steroid and xenobiotic receptor (SXR). We also discuss the recommended intake, deficiency, and assessment of vitamin K. Furthermore, we review the few available studies that have as pre-specified outcome bone fractures, indicating that we need more clinical studies to confirm that vitamin K is a potential therapeutic agent for bone fractures.     

K3机用镍钛锉与K锉的临床应用比较

目的：比较K3机用镍钛锉与手用K型锉的临床应用。方法：选择285颗患牙（失查数5颗）,共526个根管,随机数字表法分为K3组和K锉组,分别用K3机用镍钛锉和K锉进行根管预备,比较两组患者根管预备时间、根管预备术后疼痛反应和根管充填质量。结果：K3组所用时间明显少于K锉组;K3组根管预备后疼痛发生率为（5.51％）,较K锉组的根管预备后疼痛发生率（10.24％）明显降低（P〈0.01）;K3组根管恰填率为（94.18％）,较K锉组（86.22％）明显提高（P〈0.01）。结论：K3机用镍钛锉预备根管具有效率高、成形能力强、根充效果好、明显减少术后疼痛等优点,是一种理想的根管预备器械,值得临床上广泛应用。     

Vitamine K, antivitamine K et alimentation

Subclasses of vitamin K, their origins, their differential characteristics of absorption and metabolism, their relative effects on gammacarboxylation of various proteins implicated in hemostasis and coagulation, in bone calcification are not well known even by experts in these fields. These misunderstandings explain errors in recommendations for public and for patients. This review tries to give an update view on the actual stage of knowledge and on its practical implications especially on the management of oral anticoagulant treatments since until recently vitamin K antagonist was the only available type of such a treatment. Several examples illustrate the need for a better understanding of this subject. The fear that diet vitamin K could deregulate the equilibrium of oral vitamin K antagonist treatment leads to recommend a quite total suppression of vitamin K containing components in the diet of anticoagulated patients. This leads to an opposite effect: a high sensitivity to vitamin K and to disequilibrium of the anticoagulant treatment while a comprehensive moderate and regular diet intake of vitamin K first facilitates the food choice of the patients but also helps to stabilise the treatment of chronically anticoagulated patients. Vitamin K plays a role in bone calcification and in osteoporosis prevention. Until recently, the food supplementation with vitamin K in view of preventing osteoporosis in the general population was strongly limited due to fear of affecting the treatment equilibrium in anticoagulated patients. While an understanding that the effects of moderate supplementation in vitamin K has no or limited effect on anticoagulation and on the long run could at the opposite help to stabilize the daily level of anticoagulation in patients chronically treated with vitamin K.     

Vitamin K content of foods and dietary vitamin K intake in Japanese young women

Several reports indicate an important role for vitamin K in bone health as well as blood coagulation. However, the current Adequate Intakes (AI) might not be sufficient for the maintenance of bone health. To obtain a closer estimate of dietary intake of phylloquinone (PK) and menaquinones (MKs), PK, MK-4 and MK-7 contents in food samples (58 food items) were determined by an improved high-performance liquid chromatography method. Next, we assessed dietary vitamin K intake in young women living in eastern Japan using vitamin K contents measured here and the Standard Tables of Food Composition in Japan. PK was widely distributed in green vegetables and algae, and high amounts were found in spinach and broccoli (raw, 498 and 307 microg/100 g wet weight, respectively). Although MK-4 was widely distributed in animal products, overall MK-4 content was lower than PK. MK-7 was observed characteristically in fermented soybean products such as natto (939 microg/100 g). The mean total vitamin K intake of all subjects (using data from this study and Japanese food composition tables) was about 230 microg/d and 94% of participants met the AI of vitamin K for women aged 18-29 y in Japan, 60 microg/d. The contributions of PK, MK-4 and MK-7 to total vitamin K intake were 67.7, 7.3 and 24.9%, respectively. PK from vegetables and algae and MK-7 from pulses (including fermented soybean foods) were the major contributors to the total vitamin K intake of young women living in eastern Japan.     

Vitamin K and bone health

Vitamin K, originally recognised as a factor required for normal blood coagulation, is now receiving more attention in relation to its role in bone metabolism. Vitamin K is a coenzyme for glutamate carboxylase, which mediates the conversion of glutamate to gamma-carboxyglutamate (Gla). Gla residues attract Ca2+ and incorporate these ions into the hydroxyapatite crystals. There are at least three Gla proteins associated with bone tissue, of which osteocalcin is the most abundant and best known. Osteocalcin is the major non-collagenous protein incorporated in bone matrix during bone formation. However, approximately 30% of the newly-produced osteocalcin stays in the circulation where it may be used as an indicator of bone formation. Vitamin K deficiency results in an increase in undercarboxylated osteocalcin, a protein with low biological activity. Several studies have demonstrated that low dietary vitamin K intake is associated with low bone mineral density or increased fractures. Additionally, vitamin K supplementation has been shown to reduce undercarboxylated osteocalcin and improve the bone turnover profile. Some studies have indicated that high levels of undercarboxylated osteocalcin (as a result of low vitamin K intake?) are associated with low bone mineral density and increased hip fracture. The current dietary recommendation for vitamin K is 1 microg/kg body weight per d, based on saturation of the coagulation system. The daily dietary vitamin K intake is estimated to be in the range 124-375 microg/d in a European population. Thus, a deficiency based on the hepatic coagulation system would be unusual, but recent data suggest that the requirement in relation to bone health might be higher.