原位再生医疗技术与微粒皮种植术在双溪毛鲁医院的临床应用经验

总结了接受微粒皮种植术的12 例患者的治疗经验.随机选择2010 年1 月到2012 年2 月间接受此疗法治疗的12 例患者,年龄13 ~68 岁,创面按致伤原因分为：3 例全层皮肤烧伤创面、3 例创伤创面、3 例术后创面、3 例糖尿病溃疡创面.对于这些难愈性创面,在用湿润烧伤膏（MEBO） 培养创面组织后,进行微粒皮种植术治疗,治疗后10 例患者创面愈合,2 例患者由于微粒皮种植术后只有50% 创面上皮化而再次实施微粒皮种植术,创面愈合后随访3 周~2 年.优点：是不适合手术患者的理想治疗方法,供皮区相对较小,可促进创面较快愈合,手术操作简便,可在门诊实施,患者耐受性好,节省手术和随访时间,是一种临床治疗难愈性创面的实用方法.     

Prevalence of Toxocara Spp. eggs in Public Parks in Tehran City,Iran

Background

The objective of the present research was to determine the frequency of Toxocara spp. eggs in soil samples of public parks, in the city of Tehran, Iran.

Methods

A total of 600 soil samples were taken from 120 parks between Aprils to November, 2008. Soil samples were collected from 5 distinct sites in the parks. The samples were washed with saline solution and the collected sediment from each park were equally divided and examined by floatation and Petri dish methods for Toxocara eggs.

Results

Ten percent were contaminated with Toxocara spp. eggs. The number of observed Toxocara eggs in each microscopic field was varied from 1-3. No significant differences were observed between floatation and Petri dish methods.

Conclusion

Our public parks showed a high risk of toxocariasis and the need for preventive studies.     

IL-23 and IL-27 Levels in Macrophages Collected from Peripheral Blood of Patients with Healing Vs Non-Healing Form of Cutaneous Leishmaniasis

Background

In this study the level of IL-23 and IL-27 produced by macrophages derived from peripheral blood mononuclear cell culture collected from patients with healing or non-healing form of cutaneous leishmaniasis lesion were compared before and after treatment with live Leishmania to explore whether IL-23 or IL-27 plays any role in healing process of cutaneous lesions induced by L. major.

Methods

Twenty patients resident in Isfahan Province, with healing or non-healing form of cutaneous leishmaniasis lesion caused by Leishmania major participated in this study. In vitro productions of IL-23 and IL-27 by peripheral blood derived macrophages, before and after stimulation with live L. major (MRHO/IR/75/ER) promastigotes were evaluated using ELISA method. Patient with healing form of lesion received no treatment and patient with non-healing form of lesion received at least 2 courses of glucantime.

Results

The mean production of IL-23 and IL-27 from macrophages of patients with healing form of lesion was significantly higher than patients with non-healing form of lesion. The levels of IL-23 and IL-27 in culture supernatants before and after stimulation in healing form of CL was significantly higher than non- healing form of CL (P < 0.001).

Conclusion

IL-23 and IL-27 might play a role in human leishmaniasis and further studies are needed to understand the role of IL-23 and IL-27 in leishmaniasis.     

手术及经皮自体骨髓移植治疗骨延迟愈合和骨不连

观察手术及经皮自体骨髓移植治疗四肢骨折骨不连、骨延迟愈合的临床效果。选择1997-11/2004-02山东省聊城市第二人民医院应用经皮自体骨髓移植法治疗骨不连、骨延迟愈合的患者23例,患者均了解相关治疗目的、方法并同意治疗方案。①17例骨不连患者采用手术治疗。15例存在骨质缺损者在骨折端间隙及周围自体髂骨植骨10例,收集术中骨屑及骨髓原位植骨4例,异体植骨1例;2例单纯外固定架重新固定未植骨并行自体骨髓移植。术后2周抽取自体骨髓注入骨折部位,注射10～20mL/次,每2周1次,需注射2～4次。②6例骨延迟愈合患者仅行自体骨髓移植,未改变固定方式。术后定期随访,观察患者骨折愈合情况。骨折愈合标准:X射线片显示连续骨痂通过折线,骨折处无疼痛及压痛,能够全部负重。23例骨不连、骨延迟愈合患者全部进入结果分析,无脱落。23例患者术后平均随访12.5个月,其中随访7～9个月11例,10～12个月6例,13～15个月5例,16个月1例。23例患者均达到骨性愈合,愈合率100%,平均愈合时间为5.6个月。坚强固定加必要的植骨修复骨缺损是治疗骨折术后骨不连的基础和关键,自体骨髓具有成骨作用,可进一步促进骨折愈合。     

Mammalian genotoxicity assessment of methylene blue in plasma: implications for virus inactivation

BACKGROUND: The risk of adverse consequences of virus-inactivation procedures for plasma and cellular blood components must be less than the risk of transfusion-associated viral disease. Previous studies demonstrated that methylene blue, which is currently used in Europe for virus inactivation in fresh-frozen plasma, can elicit mutations in bacterial test systems. This study investigates the potential for methylene blue genotoxicity in two mammalian test systems. STUDY DESIGN AND METHODS: Different concentrations of methylene blue were prepared in plasma (heat-treated at 56 degrees C for 1 hour to reduce cytotoxicity) and used, without illumination, in an in vitro mouse lymphoma cell assay designed to detect forward mutations in the gene encoding thymidine kinase. The assay was performed in the presence or absence of rat liver S9 microsomal fraction. Similarly prepared samples of methylene blue in heat-treated plasma were used in an in vivo mouse micronucleus assay. Each system included a negative vehicle control (heat-treated plasma without methylene blue) and a positive control consisting of a known genotoxic agent. RESULTS: Intravenous administration to mice of 62 mg per kg of methylene blue did not increase the frequency of micronuclei in polychromatic red cells harvested from bone marrow. However, methylene blue concentrations of 10 micrograms per mL (with S9 activation) and 30 micrograms per mL (without S9 activation) significantly increased the thymidine kinase mutation frequency of mouse lymphoma cells to approximately 110 × 10(- 6), from a spontaneous frequency of 28 × 10(-6). CONCLUSION: Methylene blue is mutagenic in cultured mammalian cells. In contrast, results from the mouse micronucleus assay suggest that the genotoxicity is not expressed in vivo. Considerably more investigation will be required to assess the genotoxic potential of intravenously administered methylene blue used in virus-inactivation procedures, because of the likelihood of the formation of methylene blue photoproducts or the impact of metabolic conversion of methylene blue to leukomethylene blue in vivo.     

Enhanced nitric oxide release during cortical spreading depression following infusion of glyceryl trinitrate in the anaesthetized cat

Intravenous infusion of glyceryl trinitrate (GTN) into migraineurs induces an immediate headache followed by migraine. We studied the effect of GTN 10.25 g kg¹ min ¹) on local cerebrovascular laser Doppler flux (rCBF_LDF), artery diameter and NO concentration (selective NO microelectrode) in the pial middle cerebral artery perfusion territory of the anaesthetized cat, at rest and during cortical spreading depression (SD). GTN infusion induced a significant increase in pial artery diameter, rCBF_LDF, and NO concentration. Following termination of infusion, NO concentrations remained significantly elevated above controls for 60 min, other parameters returned to baseliae within 10 min ( p 0.05, ANOVA, post hoc Dunnett's multiple comparison procedure). Two hours after termination of infusion KCl-evoked SD was initiated. GTN-treated animals exhibited significantly ( p 0.05, Kruskal-Wallis) elevated SD-induced NO release compared to controls. All other parameters remained unaffected. Our results demonstrate that GTN induces a prolonged increase in local NO concentrations and enhances SD-induced NO release.     

Approaches to preventing or reversing platelet alloimmunization using animal models

Animal transfusion models were established to assess treatment programs for preventing or reversing platelet alloimmunization. Five control baboons given weekly transfusions of radiolabeled platelets from a single unrelated donor became immunized after an average of 2.4 +/- 2.1 transfusions. Similarly, 18 of 21 (86%) dogs given up to eight platelet transfusions from a single unrelated donor became immunized after an average of 2.3 +/- 1.7 transfusions. In six of seven baboons, prednisone or antithymocyte globulin alone or in combination effectively delayed platelet alloimmunization. In contrast, only two of 12 (17%) dogs given prednisone or antithymocyte serum (ATS) resisted alloimmunization. Neither splenectomy nor cyclophosphamide prevented alloimmunization in the baboon. In addition, attempts to reduce the immunogenicity of transfused platelets by inactivating the contaminating leukocytes with gamma radiation or by giving leukocyte-poor platelets were of no benefit in dogs. Reversal of platelet alloimmunization was achieved in two of three dogs treated with ATS and procarbazine hydrochloride. However, neither splenectomy, cyclophosphamide, ATS plus prednisone, nor vincristine sulfate produced any improvement. These studies show that the highly immunogenic nature of platelet transfusions in animals makes feasible the study of the prevention and reversal of platelet alloimmunization.     

力竭运动大鼠血红蛋白及血液生化指标变化与槲皮素的影响

将40只雄性SD大鼠随机分为安静组8只、运动组16只、槲皮素运动组16只。以跑台训练建立动物模型，在最后一次训练时．将运动组和槲皮素运动组分别分成2个亚组：对照组和力竭运动组．每组均8只。实验期间按每天槲皮素50mg／kg体质量的剂量，于运动前3h灌胃给槲皮素运动组动物，其余组按体质量灌胃给相应量的生理盐水。通过测定力竭运动大鼠血液中血红蛋白含量、血清乳酸脱氢酶、血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶活性。结果显示．服用槲皮素的大鼠在运动力竭即刻血红蛋白含量下降幅度明显小于运动力竭组．同时血清乳酸脱氢酶、血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶活性的上升幅度明显降低。     

肺纤维化鼠Ⅰ型前胶原和Ⅲ型前胶原mRNA表达及芪丹颗粒剂的干预效应

目的:观察芪丹颗粒对博莱霉素致肺纤维化鼠的Ⅰ型前胶原和Ⅲ型前胶原mRNA表达的影响,探讨其作用机制。方法:实验于2004-10/2006-10在山东省医学科学院基础所病理实验室完成。实验材料:清洁级雄性SD大鼠160只,体质量180～210g。氢化可的松琥珀酸钠50mg/支;芪丹(颗粒剂)是由黄芪、丹参、川芎等中草药加工提纯后的粗提取物制成的颗粒剂,10g/包(1g干粉相当于原生药8g)。实验分组:160只SD大鼠按随机区组设计分为正常组20只、模型组40只、芪丹颗粒剂50只、氢化可的松50只。实验干预:正常组20只气管内灌注和灌胃均用生理盐水。其余大鼠经气管内一次性灌注博莱霉素A5按0.25mL左右(5mg/kg体质量)诱导大鼠肺间质纤维化。随机取40只为模型组。取芪丹颗粒剂组和氢化可地松组大鼠各30只,分别从造模后第2天灌注芪丹颗粒剂(3125mg/kg)和腹腔注射氢化可的松(25mg/kg),药物干预后第7,14,28天麻醉下处死动物。两组各余20只分别从造模14d后灌注芪丹颗粒剂和腹腔注射氢化可的松(用量同前),于第28、42天分别处死动物。实验评估:用苏木精-伊红评价肺组织病理学变化和原位杂交方法检测各组大鼠肺Ⅰ型和Ⅲ型前胶原mRNA表达。结果:160只大鼠全部进入结果分析。①大鼠肺脏大体标本观察:对照组肺组织各观察时间点无明显改变,模型组肺组织表面凸凹不平,部分肺叶体积缩小,表面见灰白色结节。氢化可的松组与模型组相似。芪丹颗粒剂组见部分肺叶表面不光滑及大小不等结节。②肺组织病理学观察:模型组7d肺泡腔内大量巨噬细胞淋巴细胞中性粒细胞浸润,肺间质成纤维细胞增殖,28d肺泡结构破坏肺泡内见大量胶原纤维和成纤维细胞。芪丹颗粒剂组肺泡炎及肺纤维化程度均明显轻于模型组和氢化可的松组(P<0.05)。③肺间质纤维化形成中Ⅰ型、Ⅲ型前胶原mRNA表达:原位杂交显示两种前胶原mRNA表达呈动态变化,早期肺泡炎以Ⅲ型前胶原mRNA大量增生为主,晚期纤维化期以Ⅰ型前胶原mRNA增生为主。芪丹颗粒剂组Ⅲ型前胶原mRNA的表达在第14天处于最高,至28d仍维持较高的水平,芪丹颗粒剂组Ⅲ型前胶原mRNA的表达高于模型组和氢化可的松组(P<0.05)。28d,Ⅰ型前胶原mRNA的表达在第二天给药芪丹颗粒剂组和氢化可地松组及第14天给药芪丹颗粒剂组和氢化可的松组组间差异均有显著性(P<0.05)。结论:大鼠肺纤维化的早期以Ⅲ型前胶原mRNA的表达为主,晚期纤维化期以Ⅰ型前胶原mRNA的大量表达为主。芪丹颗粒可减轻博莱霉素诱导的大鼠肺泡炎及肺纤维化的程度,其机制可能通过影响了Ⅰ型和Ⅲ型前胶原mRNA的代谢和表达,从而减慢肺间质纤维化的进程。芪丹颗粒对肺间质纤维化有治疗作用,且优于氢化可的松。