甘肃省某三甲医院糖尿病足患者创面感染病原菌及相关因素分析

目的探讨甘肃省某三甲医院收治的糖尿病足患者创面感染病原菌情况,分析创面感染的相关因素,为后期临床救治提供理论依据。 方法选择2017年1月至2021年12月甘肃省人民医院收治的符合入选标准的糖尿病足患者,收集并分析患者的人口信息(年龄、性别)、糖尿病足创面特征(糖尿病病程、创面持续时间、创面部位)、抗生素应用数量、实验室检测指标(红细胞计数、白细胞计数、血红蛋白、白蛋白、肌酐、胆固醇、甘油三脂、高密度脂蛋白、低密度脂蛋白、空腹血糖、糖化血红蛋白、白细胞介素-6、降钙素原)、创面细菌培养结果及药物敏感试验结果。数据比较采用χ²检验、单因素分析、多因素Logistic回归分析。 结果(1)本研究共纳入糖尿病足患者173例,其中51~60岁人数最多,有53例,占比30.6%,其次为61~70岁、41~50岁、71~80岁,分别占24.3%、20.2%、16.8%;男性总体多于女性,男性138例,占比79.8%,女性35例,占比20.2%。(2)糖尿病足患者糖尿病病程10~14年人数最多,38例,占比为22.0%,创面持续时间中人数最多为15~30 d,53例,占比为30.1%。173例患者共185个创面,其中位于足趾部位最多,有68例,占比36.8%,其次为足底52例,占比28.1%。(3)患者的实验室检测指标结果显示,红细胞计数低于正常值范围的患者占比60.1%,白细胞计数高于正常值范围的患者占比34.7%,血红蛋白低于正常值范围的患者占比42.8%,白蛋白低于正常值范围的患者占比86.1%,肌酐高于正常值范围的患者占比20.8%,胆固醇高于正常值范围的患者占比11.6%,甘油三酯高于正常值范围的患者占比23.7%,高密度脂蛋白低于正常值范围的患者占比73.4%,低密度脂蛋白高于正常值范围的患者占比12.7%,空腹血糖高于正常值范围的患者占比82.1%,糖化血红蛋白高于正常值范围的患者占比77.5%,白细胞介素-6高于正常值范围的患者占比77.5%,降钙素原高于正常值范围的患者占比68.2%。(4)173例患者共送检标本257份,其中阳性210份,阴性47份,阳性率81.7%;其中革兰阳性菌共120株,占比57.1%,革兰阴性菌共89株,占比42.4%,真菌1株,占比0.5%;革兰阳性菌中金黄色葡萄球菌65株,占比最高,为31.0%,其次是粪肠球菌16株,占比7.6%;革兰阴性菌中阴沟肠杆菌24株,占比最高,为11.4%,其次是大肠埃希菌21株,占比10.0%;药物敏感试验结果显示,金黄色葡萄球菌对甲氧苄啶/磺胺甲恶唑、万古霉素、利奈唑胺等抗生素较为敏感,敏感率均为100.0%;其对青霉素耐药率高,耐药率为89.2%,其次为克林霉素和红霉素,耐药率分别为78.5%、76.9%;阴沟肠杆菌对美洛培南、厄他培南、头孢哌酮/舒巴坦、左旋氧氟沙星等较为敏感,敏感率均为100.0%,其次为阿米卡星、亚胺培南、头孢吡肟,敏感率均为95.8%;大肠埃希菌对美洛培南、厄他培南、亚胺培南、替加环素、头孢西丁、阿米卡星等较为敏感,敏感率均为100.0%。173例患者中,抗生素应用数量为0的有19例,占比11.0%;1种及2种的均有61例,占比均为35.3%;3种的有25例,占比14.4%;4种以上7例,占比4.0%。(5)对糖尿病足患者感染的17个相关因素进行单因素分析,结果显示,创面持续时间、抗生素应用数量、白细胞计数、血红蛋白、白蛋白、高密度脂蛋白、肌酐、糖化血红蛋白、白细胞介素-6、降钙素原等相关因素差异均有统计学意义(P<0.05);进一步行多因素Logistic回归分析结果显示,与创面感染相关的因素有创面持续时间、抗生素应用数量,高密度脂蛋白(OR＝1.530、1.923、2.587,P<0.05)。 结论糖尿病足患者创面病原菌培养中,革兰阳性菌中以金黄色葡萄球菌、粪肠球菌为主,而革兰阴性菌以阴沟肠杆菌、大肠埃希菌为主;糖尿病足患者感染的独立危险因素为创面形成时间长、抗生素滥用,高密度脂蛋白低。     

Histological and enzyme histochemical parameters for the age estimation of human skin wounds

Routine histological staining techniques form the basis of a forensic age estimation of human skin wounds and the determination of vitality is aided by the detection of neutrophilic granulocytes which appear earliest about 20–30 min after wounding. A clear granulocyte infiltration and a significant increase in the number of macrophages indicates a post infliction interval of at least several hours. Macrophages containing incorporated particles such as lipophages, erythrophages or siderophages appear earliest at a wound age of 2–3 days similarly to extracellular deposits of hemosiderin, whereas the rarely detectable iron-free pigment hematoidin and spot-like lymphocytic infiltrates in the granulation tissue appear approximately one week or more after wounding. A complete reepithelialization of surgically treated and primarily healing human skin lesions can be expected earliest 5 days after wound infliction and the absence of a complete new epidermal layer indicates a survival time of less than 21 days. Enzyme histochemical methods allow a wound age differentiation especially in the range of a few hours. An increase in nonspecific esterases can be observed earliest approximately 1 hour after wounding followed by other enzymes such as acid phosphatase ( 2 h), ATPase ( 4 h), aminopeptidase ( 4 h) or alkaline phosphatase ( 4 h). Positive results, however, cannot be regularly found. Therefore, the detection of reactive changes is useful for a wound age estimation whereas negative findings, which in general must be interpreted with caution, can provide information only in a limited number of histological parameters.Dedicated to Prof. Dr. W. Eisenmenger on the occasion of his 50th birthday     

Biological activity of tissue flaps in the treatment of complicated irradiation wounds

Within the last ten years, 79 patients were treated for 114 chronically contaminated, intractable irradiation wounds using various methods of the modern plastic surgery. Radical excision of the devitalised contaminated tissue has been impracticable in 25 cases due to the risk of life-threatening complications or significant functional loss. Different types of flaps such as cutaneous, fasciocutaneous, musculocutaneous, split muscle, isolated vascularised fascia and greater omentum have been used. Despite the incomplete excision, 84% of wounds healed primarily. The essential factor for good wound healing seems to be the biologic activity (BA) of the flap's deep tissue layer that directly contacts the wound bed. BA includes density of the vascular net, ability of neovascularisation, plasticity and specific immunological capacities. It seems to be possible to classify the flaps according to the BA level. Tissue defects in which the chances for radical debridement are poor need the highest BA level in the flap reconstruction.Presented to the European Congress on Wound Healing and Skin Physiology, Bochum, Germany, 1992     

Immunohistochemical analysis of markers for different macrophage phenotypes and their use for a forensic wound age estimation

A total of 117 vital skin wounds (post infliction intervals between a few seconds and 7 months), 20 postmortem wounds and skin specimens with beginning or advanced signs of putrefaction were investigated. Different markers for macrophage maturation (27 E 10, RM 3/1, 25 F 9, G 16/1) were analyzed by immunohistochemistry. The early stage inflammation marker 27 E 10 stained macrophages, but also monocytes and neutrophilic granulocytes localized in blood vessels or bleeding induced postmortem and therefore provided no further information for a forensic wound age estimation in comparison to the routine histological detection of macrophages. The antigens recognized by the RM 3/1- (intermediate stage inflammation marker) and 25 F 9-antibodies (late stage inflammation marker) were expressed exclusively by histiocytes and inflammatory cells that had migrated from the blood vessels as part of the acute inflammatory response associated with an intravital reaction. The morphometrical analysis revealed positive results (defined as at least a two-fold increase in number in 2 or more microscope fields when compared to the maximum value of histiocytes found in uninjured skin) for the RM 3/1- or 25 F 9-antibody earliest in wounds aged 7 or 11 days, respectively. Similarly to the 25 F 9-antibody, the chronic stage inflammation marker (G 16/1) reacted with a macrophage subpopulation first detectable 12 days after wounding but showed positive results in a comparably reduced percentage of cases. On the other hand, this marker did not stain a relevant number of resident macrophages thus facilitating the evaluation of the specimens. The markers 27 E 10, RM 3/1 and 25 F 9 are also useful for the evaluation of slightly - even though the staining intensity was considerably reduced - but not advanced putrefied skin. Therefore, the immunohistochemical analysis of the corresponding antigens can possibly contribute to an age estimation of wounds with advanced post infliction intervals obtained from corpses with longer - but limited - postmortem intervals.

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Zusammenfassung Insgesamt wurden 117 vitale Hautwunden (Überlebenszeit wenige Sekunden bis 7 Monate), 20 postmortal gesetzte Verletzungen sowie Haut mit leichten bzw. fortgeschrittenen Fäulnisveränderungen untersucht und verschiedene Marker der Makrophagen-Differenzierung (27 E 10, RM 3/l, 25 F 9 und G 16/1) analysiert. Der early stage inflammation marker 27 E 10 färbte neben Makrophagen auch Monozyten und neutrophile Granulozyten, die innerhalb von Blutgefäßen bzw. in postmortal gesetzten Blutungen lokalisiert waren und liefert somit keine Informationen zum Wundalter, die über die Möglichkeiten des Routine-histologischen Nachweises von Makrophagen hinausgingen. Die von den Antikörpern RM 3/1 (intermediate stage inflammation marker) und 25 F 9 (late stage inflammation marker) erkannten Antigene wurden ausschließlich von Histiozyten und reaktiv eingewanderten Makrophagen exprimiert. Die morphometrische Analyse ergab positive Ergebnisse (definiert als ein mindestens zweifacher Anstieg der Zellzahl in zwei oder mehr Gesichtsfeldern verglichen mit der maximal feststellbaren Zahl an Histiozyten in unverletzter Haut) bei Verwendung der Antikörper RM 3/1 bzw. 25 F 9 frühestens 7 bzw. 11 Tage nach Wundsetzung. Ab 12 Tagen Wundalter reagierte der chronic stage inflammation marker G 16/1 erstmals positiv. Das Antigen ließ sich insgesamt allerdings in einem geringeren Prozentsatz der untersuchten Wunden darstellen. Vorteilhaft ist jedoch das Fehlen einer relevanten Expression durch Histiozyten, wodurch die Auswertung der Präparate erleichtert wird. Die entsprechenden Antigene lassen sich zudem in leicht - wenn auch in einer deutlich geringeren Färbeintensität -, aber nicht forgeschritten fäulnisveränderter Haut nachweisen, so daß deren immunhistochemische Darstellung gegebensfalls auch zur Beurteilung von länger überlebten Verletzungen an Leichen mit etwas fortgeschrittener Liegezeit herangezogen werden kann.

     

Cellular dynamics of corneal wound re-epithelialization in the rat. I. Fate of ocular surface epithelial cells synthesizing DNA prior to wounding

The fate of ocular surface epithelial cells in response to injury of the cornea was examined. Corneal epithelial cells were labeled during DNA synthesis with [3H]thymidine 1 h prior to wounding. A 3-mm diameter epithelial defect was made in the center of the rat cornea, with the basement membrane remaining intact. Within 12 h of abrasion, labeled cells were detected in the regenerating surface. At 18 h, there was a 2.7- and 17-fold increase of labeled basal and suprabasal cells, respectively, in the epithelium adjacent to the wound, and at 24 and 30 h there was an excessive number of cell layers (up to 7) at the margin of the abrasion. Re-epithelialization progressed as a gradient of cell layers that became diminished towards the center of the wound. Completion of layers 1, 2, 3, and 4 were recorded at 24, 30, 36, and 72 h, respectively. No changes in the labeling index of the limbus or conjunctiva were noted. These results suggest that processes of centripetal and vertical migration, as well as events related to cell division, in the uninjured corneal surface are not impeded by wounding of the corneal epithelium. However, wound healing appears to require cells with a basal phenotype, presumably because of this cell type's migratory capability.     

The experience in the treatment of patients with extensive full-thickness burns

The aim of the present study is to analyze the changes of cure rate and the main causes of death in the last 40 years, and to summarize our experience in the treatment of extensive full-thickness burn patients. The clinical characteristics, cure rate and main causes of death of 73 cases with total burned area more than 90% TBSA and full-thickness burn area more than 70% TBSA were analyzed retrospectively. Among them, 21 cases (28.8%) were cured and 52 cases (71.2%) died. The cure rate increased significantly in the recent years, and the main causes of death changed from shock and sepsis in the time period 1959–1978 to sepsis and MODS in the past two decades. Due to the improvement of early comprehensive management of burn shock, aggressive surgical approach to full-thickness burn wound and potent systemic supporting measures, the survival rate increased significantly.     

Morphometrical analysis of hemosiderin deposits in relation to wound age

A morphometrical analysis of the extent of hemosiderin deposits in 71 human skin wounds with post-infliction intervals between 2 days and 7 months was performed. Earliest positive findings were detectable in a lesion aged 3 days, and with increasing wound age an increase in the amount of hemosiderin occurred. A value of more than 20% of the microscopic field with hemosiderin deposits was found earliest 8 days after wounding and therefore the detection of considerable amounts of hemosiderin (arbitrarily defined as 20% or more of the evaluated area) indicates a minimum wound age of approximately 1 week. Since the extent of hemosiderin formation depends upon the extent of the initial hemorrhage and a physiological reduction in the amount of this pigment with advanced wound age, slight or absent hemosiderin deposits cannot provide information on the post-infliction interval.     

Immunohistochemical localization of fibronectin as a tool for the age determination of human skin wounds

Summary We analyzed the distribution of fibronectin in routinely embedded tissue specimens from 53 skin wounds and 6 postmortem wounds. In postmortem wounds a faint but focal positive staining was exclusively found at the margin of the specimens which dit not extend into the adjacent stroma. Vital wounds were classified into 3 groups. The first comprising lesions with wound ages ranging from a few seconds to 30 min, the second comprising those with wound ages upt to 3 weeks, and the third group with lesions more than 3 weeks old. Ten out of 17 lesions with a wound age up to 30 min showed a clear positive reaction within the wound area. Three specimens in this group were completely negative, while in 4 additional cases the result was not significantly different from postmortem lesions. These 7 cases were characterized by acute death with extremely short survival times (only seconds). In wounds up to 3 weeks old fibronectin formed a distinct network containing an increasing number of inflammatory cells corresponding to the wound age. In 2 cases with a survival time of 17 days and in all wounds older than 3 weeks fibronectin was restricted to the surface of fibroblasts and to parallel arranged fibers in the granulation tissue without any network structures. We present evidence that fibronectin is a useful marker for vital wounds with a survival time of more than a few minutes. Fibronectin appears before neutrophilic granulocytes migrate into the wound area. Since a faint positive fibronectin staining is seen in postmortem lesions and bleedings, we propose that only those wounds which show strong positive fibronectin staining also extending into the adjacent stroma should be regarded as vital.This study was supported by a grant from the Deutsche Forschungsgemeinschaft (grant Ei 209/3-1) and by a grant from the Friedrich-Baur-Stiftung University of Munich     

Time-dependent RNA synthesis in different skin layers after wounding. Experimental investigations in vital and postmortem biopsies

Summary Incision wounds were made on the outer ear of rats and two biopsies were taken for examination after different survival times. In each case a biopsy was made of vital tissue and a second of postmortem tissue after refrigeration for 24 h. The biopsies were exposed to a solution containing the RNA precursor³H-cytidine for 1 h, washed and fixed in formalin. Sections 5 m thick were then autoradiographically prepared and automatically evaluated using Quantimet 920. The intravital specimens showed a significant increase in³H-cytidine incorporation in the basal cell layer after survival times of 10–24 h. No increase was seen in the stratum corneum, corium or cartilage tissue. The investigated distance from the wound margin did not have any significant bearing on the results. The³H-cytidine incorporation rate in postmortem tissue was practically identical with that of vital tissue, but no increase was observed in the rate of RNA synthesis in the basal cells as a function of the age of the wound. It may therefore be assumed that this method provides no additional information as to the age of wounds in postmortem examination.Dedicated to Professor O. Pribilla on the occasion of his 70th birthday     

Effect of electrolyzed water on wound healing

Electrolyzed water accelerated the healing of full-thickness cutaneous wounds in rats, but only anode chamber water (acid pH or neutralized) was effective. Hypochlorous acid (HOCl), also produced by electrolysis, was ineffective, suggesting that these types of electrolyzed water enhance wound healing by a mechanism unrelated to the well-known antibacterial action of HOCl. One possibility is that reactive oxygen species, shown to be electron spin resonance spectra present in anode chamber water, might trigger early wound healing through fibroblast migration and proliferation.