新型战伤急救止血剂体内抗菌活性的实验研究

目的：探讨新型战伤急救止血剂对家兔急性感染伤口的抗菌作用。方法：选用兔感染创面模型,分新型战伤止血剂、5A沸石、Quiclot和空白对照组对创面进行治疗,通过组织学观察、组织内细菌计数等方法对各组抗菌性能进行研究。结果：肉眼和组织学观察新型战伤急救止血剂治疗组动物模型伤口,炎症反应均小于其他各组;新型战伤急救止血剂治疗组织内细菌计数为104,比其他3组显著减少（P〈0.01）。结论：新型战伤急救止血剂具有良好的体内抗菌性能。     

新型战伤急救止血剂体外细胞毒性研究

目的:研究新型战伤急救止血剂的体外细胞毒性,初步探讨其用于战伤急救止血时的生物安全性.方法:参照我国医疗器械生物学评价标准,选用小鼠L929细胞,应用MTT法、直接接触培养法、流式细胞检测细胞凋亡法、扫描电镜直接观察细胞生长状况等检测新型战伤急救止血剂的细胞毒性.结果:新型战伤急救止血剂细胞毒性为0-1级,符合我国医疗器械评价标准毒性分级标准,各浓度组与阴性对照组无差异,P＞0.05;L929细胞与沸石直接接触生长良好,流式细胞仪检测细胞凋亡率与阴性对照组无差异;扫描电镜观察在沸石表面生长良好.结论:复合新型战伤急救止血剂细胞相容性良好,符合我国医疗器械安全性评价标准,是一种安全、高效、多功能的战伤止血剂.     

Cu 型医用抗菌不锈钢在大鼠体内抗细菌感染研究

目的:比较普通304医用不锈钢和304-Cu型医用抗菌不锈钢的体内抗细菌感染性能。方法:将两种不锈钢片涂布浓度为1×107cfu/mL的金黄色葡萄球菌培养24 h,植入SD大鼠胫腓骨肌袋内,继续饲养并在术前及术后第1、4、7、14天进行大体观察、植入部位细菌培养、白细胞计数、组织学观察等检测。结果:普通304不锈钢组术后中度化脓,检测到较多细菌,白细胞增多且有大量的炎症细胞出现;而304-Cu型抗菌不锈钢术后只有轻微化脓,检测到的细菌较少,白细胞数稍有增多但无统计学差异,炎症细胞少,感染程度轻。结论:与医用304不锈钢相比,304-Cu型抗菌不锈钢有较好的抗细菌性能,有一定的抗感染作用。     

创面生物活性玻璃修复材料促进家猪创面愈合的实验研究

目的:观察创面生物活性玻璃修复材料对家猪皮肤创面的促愈合作用。方法:选择14头家猪,随机分成7组,每组2头,在每头猪的脊柱两旁制造3个4×4cm的全层皮肤缺损的创面模型,每头猪6个创面又分成实验组和空白对照组,于试验后每天观察创面愈合情况,第1、3、7、14、21、28、35天图像分析计算创面愈合率,并同时取创面组织行组织学染色,观察各组材料对家猪皮肤全层缺损创面愈合的影响。结果:在涂材料的实验组和空白对照组创面愈合时间分别是23.19±1.27d、29.52±1.54d两组组间比较,具有统计学意义(P<0.05);实验组的创面愈合率在各时间段均高于空白对照组,差异有统计学意义(P<0.05);组织学观察实验组的上皮化程度、表皮生长、成纤维细胞、毛细血管数量均好于空白对照组。结论:创面生物活性玻璃修复材料对家猪皮肤创面愈合具有促进作用,可作为一种新型的促愈合覆盖材料进一步研究。     

VSD联合金因肽治疗手足外科难愈性伤口的临床研究

目的:观察封闭式负压引流技术(VSD)联合金因肽治疗手足外科难愈性伤口的临床疗效,为临床联合运用提供依据。方法:86例手足外科难愈性伤口患者随机分为治疗组和对照组各43例,治疗组采用VSD联合金因肽治疗,对照组采用常规换药治疗,观察两组临床疗效和上皮化时间、愈合时间、创面愈合率、换药次数、伤口疼痛评分、住院费用差异。结果:治疗组总有效率90.70%;对照组总有效率72.09%;两组比较,有显著性差异(P0.05);治疗组上皮化时间、愈合时间、创面愈合率、换药次数、伤口疼痛评分、住院费用较对照组相比明显缩短,差异有统计学意义(P0.05)。结论:VSD联合金因肽治疗手足外科难愈性伤口疗效确切,能更好促进损伤组织的修复,加速创面愈合,值得临床推广应用。     

壳聚糖-三聚磷酸钠纳米粒对于肝组织严重出血模型的止血效果

目的:建立SD大鼠肝脏严重出血模型,考察壳聚糖-三聚磷酸钠(TPP)纳米粒的止血效果对于实质性器官严重出血的止血效果。方法:24只SD大鼠各分3组,每组8只,分为阴性对照组、壳聚糖-TPP组(NP组)和正常组,其中阴性对照组和NP组的大鼠建立肝脏严重出血模型。NP组,将壳聚糖-TPP纳米粒喷射到伤口,直至将整个伤口覆盖。阴性对照组,不使用任何止血材料处理创面。术后15天,用SEM和TEM观察NP组和阴性对照组的肝组织的微观结构。结果:通过组织学检查发现壳聚糖-TPP纳米粒在治疗严重伤口时能加速肉芽和大量胶原蛋白的生成,这正是伤口愈合初期的标志。结论:壳聚糖-TPP纳米粒对于严重出血的实质器官可发挥优良的止血性能,并能促使伤口愈合。     

解毒活血汤促进肛周脓肿术后创面愈合的效果分析

目的:研究解毒活血汤治疗肛周脓肿术后患者创面愈合的临床效果及可能机制。方法:选择2017年1月～2019年1月我院肛肠科三病区收治的300例肛周脓肿患者,根据其就诊的顺序,随机数字表法均分为两组,各150例,两组均采取一期肛周脓肿根治术治疗,对照组于术后采用高锰酸钾溶液坐浴疗法,观察组于术后联合服用解毒活血汤治疗。均治疗4 w后,比较两组创面愈合的有效率,肛周脓肿患者创面的愈合时间和视觉模拟评分(visual analogue scale,VAS)评分,治疗前后肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数。结果:治疗后,观察组创面愈合的总有效率明显高于对照组(90.67 vs. 71.33%,P0.05);观察组肛周脓肿患者创面愈合的时间明显短于对照组(P0.05)。对照组治疗前后的肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数无明显的差异(P0.05),观察组治疗后的肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数明显高于治疗前和对照组(P0.05)。观察组术后3 d、1 w和2 w的VAS评分均明显低于对照组(P0.05)。结论:解毒活血汤可能通过提升肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数有效促进肛周脓肿术后的创面愈合,减轻术后疼痛。     

烧烫伤创面感染的小鼠模型构建

目的探讨一种简单、稳定的烧烫伤创面感染的小鼠模型构建方法,以便进行相关烧烫伤创面修复研究。方法取30只BALB/c小鼠,采用自制木质烫伤板,沸水浴法烫取直径8 mm的圆形创面,烫伤时间分别为5 s、10 s、15 s。伤后48 h,取创面组织进行HE染色观察,筛选最佳创面烫伤时间。另取72只小鼠制成深Ⅱ度烫伤创面,采用擦刮法分别接种20μL菌浓度为1×106、l×107、1×108CFU/mL金黄色葡萄球菌标准菌株ATCC 25923的菌液。接种细菌后72 h,取创面组织HE染色观察创面炎症反应情况,并测定3、7、14 d的皮肤菌负荷,筛选最佳的细菌接种浓度。最后,按最佳条件建模后,观察创面的完全愈合时间以及创面愈中、愈后的组织学变化,以确定此创面感染模型是否建立成功。结果组织学结果表明,10 s为深Ⅱ度创面的理想致伤时间,最佳接种菌浓度为l×108CFU/mL,此时期,14 d内菌负荷均高于l×105CFU/g。该模型的创面愈合时间(21±0.95 d)较正常创面愈合时间(15.92±0.34 d)明显延长(P<0.01),炎性反应明显,愈后不佳。结论烧烫伤创面感染的小鼠模型构建成功,可作为感染创面防治研究的实验动物模型。     

负压创面治疗肝源性糖尿病并发糖尿病足的研究

目的:讨论创面负压治疗促进肝硬化合并糖尿病并发糖尿病足的临床疗效。方法:将30例慢性肝硬化合并糖尿病并发糖尿病足患者随机分为创面负压治疗组和敷料包扎组,观察两组创面细菌负荷及创面微循环血流量变化。结果:清创即刻,负压创面治疗组和敷料包扎组细菌负荷与创面微循环血流量无显著差异(P>0.01);创后3天、6天和9天时,负压创面组创面细菌负荷低于敷料包扎组(P<0.01),创面微循环血流量高于敷料包扎组(P<0.01)。负压创面组创面愈合时间少于敷料包扎组(P<0.01)。结论:负压创面治疗有助于慢性肝硬化并发糖尿病合并糖尿病足的创面愈合。     

家蝇幼虫抗菌肽对铜绿假单胞菌感染小鼠创面的抗菌活性

通过超声处理诱导家蝇幼虫产生抗菌肤,观察抗菌肤对铜绿假单胞菌感染小鼠创面的抗菌作用.超声处理诱导家蝇幼虫产生抗菌肽,经分离纯化后,体外试验分析抗菌肽的抑菌特性.结果表明,术后对照组小鼠相比其他两组活动减少,且3组白细胞计数均减少.14 d后对照组存活率为30%,明显少于磺胺米隆组90%(P＜0.05)和抗菌肽组80%(P＜0.05).对照组小鼠的重量在第6 d开始下降,明显少于其他两组(P＜0.05).而无菌对照组无死亡,重量持续增加.因此,超声处理能诱导家蝇幼虫产生抗菌肽,其对铜绿假单胞菌感染的ICR小鼠创面有较强抗感染作用.能明显增加其存活率.     

Induction of antibacterial activity in larvae of the blowfly Lucilia sericata by an infected environment

Maggot debridement therapy (MDT) is a method for the treatment of intractable, infected and necrotic wounds. In MDT, sterile larvae of Lucilia sericata Meigen (Diptera: Calliphoridae) are applied to infected wounds, where they exert antibacterial effects. Once the larvae are placed in the wound, they are no longer germ-free. This study analysed the influence of infected environments on larval antibacterial activities. Sterile larvae were mixed in a test tube containing a bacterial suspension of Staphylococcus aureus or Pseudomonas aeruginosa, transferred to liver puree agar, and incubated at 25 °C for set periods. To collect the larval extracts, the incubated larvae were transferred to a test tube containing phosphate buffered saline (PBS), cut into multiple pieces with scissors, and centrifuged. The supernatant was used to test antibacterial activities. The results showed that infected larvae had better antibacterial capacities than sterile larvae. Antibacterial activities were induced by pretreatment with a single bacterial species, S. aureus or P. aeruginosa, within 24 h and 12 h, respectively, and disappeared after 36 h. The activities were effective against S. aureus, but not against P. aeruginosa. This natural infection model is very similar to the clinical wound context in MDT and will be a powerful tool with which to study the antibacterial activities of L. sericata larvae in MDT.     

Negative-Pressure Wound Therapy Enhances Local Inflammatory Responses in Acute Infected Soft-Tissue Wound

Clinical studies found that negative-pressure wound therapy (NPWT) displayed significant clinical benefits in the healing of infected wounds. However, the effect of NPWT on local inflammatory responses in acute infected soft-tissue wound has not been investigated thoroughly. The purpose of this study was to test the impact of NPWT on local expression of proinflammatory cytokines, amount of neutrophils, and bacterial bioburden in wound from acute infected soft-tissue wounds. Full-thickness wounds were created on the back of rabbits, and were inoculated with Staphylococcus aureus strain ATCC29213. The wounds were treated with sterile saline-moistened gauze dressings and NPWT with continuous negative pressure (?125 mmHg). Wound samples were harvested on days 0 (6 h after bacterial inoculation), 2, 4, 6, and 8 at the center of wound beds before irrigation for real-time PCR analysis of gene expression of IL-1β, IL-8, and TNF-α. Wound biopsies were examined histologically for neutrophil quantification in different layers of tissue. Quantitative bacterial cultures at the same time point were analyzed for bacterial clearance. Application of NPWT to acute infected wounds in rabbits was compared with treatment with sterile saline-moistened gauze, over an 8-day period. NPWT-treated wounds exhibited earlier and greater peaking of IL-1β and IL-8 expression and decrease in TNF-α expression over the early 4 days (P < 0.05). Furthermore, histologic examination revealed that significantly increased neutrophil count was observed in the shallow layer in wound biopsies of NPWT treatment at day 2 (P < 0.001). In addition, there was a statistically significant decrease of bacteria load from baseline (day 0) at days 2 and 8 in NPWT group (P < 0.05). In conclusion, this study demonstrates that NPWT of acute infected soft-tissue wounds leads to increased local IL-1β and IL-8 expression in early phase of inflammation, which may trigger accumulation of neutrophils and thus accelerate bacterial clearance. Meanwhile, the success of NPWT in the treatment of acute wounds can attenuate the expression of TNF-α, and the result may partly explain how NPWT can avoid significantly impairing wound healing.     

Anti-bacterial effects of poly-N-acetyl-glucosamine nanofibers in cutaneous wound healing: requirement for Akt1

BACKGROUND: Treatment of cutaneous wounds with poly-N-acetyl-glucosamine nanofibers (sNAG) results in increased kinetics of wound closure in diabetic animal models, which is due in part to increased expression of several cytokines, growth factors, and innate immune activation. Defensins are also important for wound healing and anti-microbial activities. Therefore, we tested whether sNAG nanofibers induce defensin expression resulting in bacterial clearance. METHODOLOGY: The role of sNAG in defensin expression was examined using immunofluoresence microscopy, pharmacological inhibition, and shRNA knockdown in vitro. The ability of sNAG treatment to induce defensin expression and bacterial clearance in WT and AKT1-/- mice was carried out using immunofluoresent microscopy and tissue gram staining. Neutralization, using an antibody directed against β-defensin 3, was utilized to determine if the antimicrobial properties of sNAG are dependent on the induction of defensin expression. CONCLUSIONS/FINDINGS: sNAG treatment causes increased expression of both α- and β-type defensins in endothelial cells and β-type defensins in keratinocytes. Pharmacological inhibition and shRNA knockdown implicates Akt1 in sNAG-dependent defensin expression in vitro, an activity also shown in an in vivo wound healing model. Importantly, sNAG treatment results in increased kinetics of wound closure in wild type animals. sNAG treatment decreases bacterial infection of cutaneous wounds infected with Staphylococcus aureus in wild type control animals but not in similarly treated Akt1 null animals. Furthermore, sNAG treatment of S. aureus infected wounds show an increased expression of β-defensin 3 which is required for sNAG-dependent bacterial clearance. Our findings suggest that Akt1 is involved in the regulation of defensin expression and the innate immune response important for bacterial clearance. Moreover, these findings support the use of sNAG nanofibers as a novel method for enhancing wound closure while simultaneously decreasing wound infection.     

急性呼吸道感染患儿白细胞计数、内毒素、C 反应蛋白水平变化及 临床意义

目的:探讨急性呼吸道感染患儿白细胞(WBC)计数、血清内毒素(ET)、C反应蛋白(CRP)水平变化及临床应用价值。方法:对急性呼吸道感染组106例患儿(细菌感染组62例、病毒感染组44例)进行WBC计数、血清ET、CRP水平等指标进行检测,并与正常对照组的40例健康儿童进行比较分析。结果:急性呼吸道感染组患儿WBC计数、血清ET、CRP水平均显著高于正常对照组,差异具有统计学意义(P<0.05或P<0.01);血清ET、CRP水平对诊断脓毒症的敏感性差异无显著性(P>0.05),均显著高于WBC计数,差异具有统计学意义(P<0.05);WBC计数特异性显著高于ET、CRP水平,ET水平特异性显著低于CRP,差异均具有统计学意义(P<0.05);细菌感染组患儿血清ET、CRP水平显著高于病毒感染组,差异具有统计学意义(P<0.05),而两组间WBC计数差异不显著(P>0.05)。结论:测定WBC计数、ET、CRP对急性呼吸道感染患儿具有一定的诊断价值。     

炎症因子、生长因子以及凋亡因子在压疮慢性难愈合性创面中的表达及作用

目的：探讨炎症反应、生长因子及凋亡因子在压疮慢性创面中的表达及作用。方法：选取2013年10月至2015年7月河南大学第一附属医院收治的患者,其中临床Ⅲ、Ⅳ期压疮患者共20例,急性创面10例,正常皮肤组织6例。通过HE染色观察不同创面组织的形态学特征;免疫组织化学法检测组织中细胞凋亡因子Caspase-3的分布规律;荧光定量PCR法定量分析IL-1β、IL-6、TNF-α、VEGF、bFGF及其受体KDR、FGFR1基因水平的变化特征。结果：Ⅲ、Ⅳ期压疮创面中可见炎性细胞浸润;凋亡信号因子caspase-3在压疮组中的表达高于其他两组,差异有统计学意义;IL-1β、IL-6、TNF-α表达高于急性创面组和正常皮肤组;VEGF和bFGF生长因子及其受体KDR和bFGFR1表达分别低于对照组。结论：炎症因子和凋亡因子在压疮慢性创面中持续长时间的高表达、生长因子及其受体显著的低表达可能是压疮慢性难愈合性创面形成的机制和难以彻底治愈的重要因素。     

A morphological evaluation of the action of collagenase from the king crab Paralithodes camtschatica on the experimental wound process]

The paper presents the results of experimental morphological evaluation of the effect of a new proteolytic enzyme collagenase of crab Paralithodes camtschatica on wound healing in infected and aseptic rabbit wounds. The enzyme was applied on wounds using gauze and gelevin. The findings show that this protease is highly effective for debridement of infected wounds, the effect increasing at gelevin addition. To reach maximal therapeutical effect and diminish the inhibition effect of collagenase on granulation tissue, it is recommended to reduce the dose of protease during debridement process. Clinical application of crab collagenase must be individual as well as duration of wound enzymotherapy.     

Temporin A is effective in MRSA-infected wounds through bactericidal activity and acceleration of wound repair in a murine model

We investigated the effect of topical temporin A in the management of methicillin-resistant strain of Staphylococcus aureus (MRSA)-infected experimental surgical wounds in mice. The wound, cut through the panniculus carnosus of BALB/c mice, was inoculated with 5x10(7) colony-forming units of MRSA. Mice were treated with Allevyn, temporin A-soaked Allevyn, Allevyn and daily intraperitoneal teicoplanin (7mg/kg), temporin A-soaked Allevyn and daily intraperitoneal teicoplanin. Main outcome measurements were: quantitative bacterial culture, histological examination with assessment of micro-vessel density and of vascular endothelial growth factor (VEGF) expression in tissue sections, and VEGF plasma levels alike. Treatment with temporin-A associated with teicoplanin injection significantly reduced bacterial load to 0.85 x 10(1)+/-0.1 x 10(1)CFU/ml. Histological examination showed that infected mice receiving temporin A-soaked Allevyn (with or without teicoplanin) had a higher degree of granulation tissue formation and collagen deposition compared to the other treated groups. A significant increase in serum VEGF expression was observed in mice receiving temporin A topically and temporin A topically associated with intraperitoneal teicoplanin. In conclusion our results demonstrated that temporin A is effective in the management of infected wounds, by a significant bacterial growth inhibition and acceleration of wound repair process.