骨髓间质干细胞移植对急性心肌梗死模型香猪心室重构的影响研究

目的:研究自体骨髓间质干细胞(MSCs)移植至急性心肌梗死模型香猪心肌梗死区内对心室重构的影响。方法:抽取香猪骨髓,体外分离MSCs,并经培养、转化、标记后注入结扎的香猪冠状动脉左前降支及心肌内;对照组则注射等量的达尔伯克(氏)必需基本培养基。3、6wk分别行多普勒心脏超声检查。结果:实验组左心室射血分数、左心室短轴缩短率、左心室前侧壁增厚率以及室间隔增厚率均明显高于对照组;实验组心室腔则明显小于对照组。结论:MSCs移植可在急性心肌梗死区再生心肌组织,增加心肌厚度,减轻心肌变薄和心肌腔扩大程度,故可减缓心室重构的进程。     

激活Notch信号的骨髓间充质干细胞心肌移植促进血管新生

目的探讨干预Notch信号骨髓间充质干细胞(BMSCs)移植对心肌梗死(MI)大鼠心肌的治疗性血管新生作用及其机制。方法 60只Wistar大鼠经结扎冠状动脉前降支建立MI模型后2周进行相应处理后,随机分为激活Noth信号的BMSCs移植实验组(D组)、BMSCs移植对照组(E组)、培养液移植对照组(C组)、MI模型对照组(B组),每组15只;另选取10只为假手术对照组(A组)。4周后观察细胞生长及增殖情况,测定缺血心肌中血管内皮细胞生长因子(VEGF)蛋白的表达及缺血区心肌毛细血管密度的改变。结果 BMSCs在梗死区中可增殖分化为内皮细胞。与B组、C组及A组相比,D组、E组缺血心肌中VEGF蛋白的表达增多及毛细血管密度均明显增高(P<0.01),D组较E组更明显(P<0.05)。结论 Notch信号促进心肌梗死区BMSCs向内皮细胞分化,促进缺血心肌中毛细血管新生。     

骨髓间充质干细胞移植对小型猪急性心肌梗死模型治疗作用的初步研究

目的 观察自体骨髓间充质干细胞(MSC)心肌内移植对小型猪急性心肌梗死(AMI)的治疗效果。方法 30只小型猪制备AMI模型，实验组22只取骨髓分离MSC，体外培养扩增后注入至缺血坏死心肌周围，对照组8只输注等量生理盐水，1个月后比较两组的生存率、存活心肌来源和功能性蛋白质的表达。结果实验组存活率100％，存活心肌增加并检测到5溴脱氧尿嘧啶(BrdU)标记物，心肌钙蛋白I(cTnI)等心肌功能性蛋白质的表达。结论 AMI小型猪自体骨髓MSC心肌内移植后，梗死区出现MSC来源的新生心肌，并具有正常心肌工作特性。     

经冠脉移植自体骨髓间充质干细胞对急性心肌梗死后心功能的影响

目的研究骨髓间充质干细胞(MSC)植入幼猪急性心肌梗死(AMI)区对心肌灌注及心功能的影响.方法通过冠脉转运方法,将5-溴脱氧尿嘧啶(BrdU)标记的幼猪自体骨髓MSC植入心肌梗死区,8周后行99mTc-MIBI门控心肌灌注显像评价心肌灌注及心功能的变化.结果移植的MSC可通过毛细血管内膜并转化为心肌样细胞;8周后移植的MSC在心肌内微环境诱导下可心肌化及血管化;治疗组射血分数明显提高(32.75%vs49.86%,P＜0.001).结论MSC移植通过促进血管及心肌再生,改善心肌灌注及功能.     

超声联合一氧化氮微泡体外干预骨髓间充质干细胞安全性探讨

目的研究超声联合一氧化氮(NO)微泡体外干预大鼠骨髓间充质干细胞(BMSCs)的安全性。方法体外分离培养大鼠BMSCs,分为超声联合NO微泡组(包括1∶70,1∶50,1∶20浓度亚组)及空白对照组,体外干预后MTT法检测BMSCs增殖,PI染色流式细胞术分析细胞凋亡及细胞周期。结果与空白对照组比较,1∶20浓度组显著抑制BMSCs增殖(P<0.05);1∶50浓度组与1∶20浓度组明显诱导细胞凋亡(P<0.05)。结论超声联合NO微泡体外干预BMSCs,1∶70浓度对其是安全的。     

双龙方与自体骨髓单个核细胞经心导管移植对中国小型猪心肌梗死的影响

目的：研究中药双龙方与自体骨髓单个核细胞经心导管移植对中国小型猪心肌梗死的影响。方法：利用心导管技术建立中国小型猪冠状动脉栓塞心肌梗死模型，建立经心导管冠状动脉内骨髓单个核细胞移植的新方法，通过心电图、定量组织学、超声心动图、心肌酶及生化检查、心肌小血管密度、梗死区内骨髓单个核细胞观察等综合评价双龙方、自体骨髓单个核细胞经心导管移植及两者合用的疗效。结果：双龙方与自体骨髓单个核细胞经心导管移植可减小心肌梗死范围，超声心动图检查显示心功能明显改善，并可增加心肌梗死区小血管密度，对心肌酶及生化指标亦有不同程度改善作用。自体骨髓单个核细胞移植及合用双龙方后均有助于心肌侧枝循环的建立，心肌供血明显改善。自体骨髓单个核细胞可在移植区内存活，并部分分化为心肌细胞及心肌小血管，补充了心肌细胞的数量。结论：双龙方与自体骨髓单个核细胞移植对中国小型猪心肌梗死有明确的治疗作用。     

骨髓间充质干细胞自体移植对心功能影响的超声观测

目的应用彩色多普勒超声心动图评价骨髓问充质干细胞（MSCs）自体移植对急性心肌梗死成年兔左心室形态及收缩功能的影响。方法22只日本大耳白兔。通过结扎左冠状动脉前降支，建立息性心肌梗死模型；随机分成实验组与对照组。将经过培养的MSCs经溴氮胞苷（BruD）标记后，直接注射到实验组动物左心室游离壁正常心肌组织与梗死组织交界处的心肌内。于术前1周，术后2、4周分别行彩色多普勒超声心动图检查，观测左心室内径及左心室收缩功能。术后4周处死动物，进行组织学与α-横纹肌动蛋白免疫组织化学染色。结果术后2周时，实验组与对照组比较心功能参数差异无显著性（P〉0．05），术后4周，与对照组比较，心功能改善（P〈0．05）。缺血心肌内可见部分BruD标记的移植细胞已与宿主心肌细胞相连接，并分化为具有典型的肌小节和表达α-横纹肌动蛋白阳性的心肌细胞样的横纹肌细胞，其排列方向与宿主心肌细胞排列方向平行。结论自体移植的MSCs可在急性梗死心肌内生长、发育、分化，并逐渐分化为心肌细胞取代瘢痕组织，影响左心室重构，改善心功能。     

不同途径移植人羊膜上皮细胞对心肌梗死大鼠心脏的保护作用

目的比较经舌下静脉和心外膜直视注射移植人羊膜上皮细胞(hAECs)对心肌梗死大鼠的心脏保护作用。方法雄性SD大鼠48只,随机分为假手术组(n=12)、模型组(n=12)、hAECs心外膜移植组(n=12)和hAECs静脉移植组(n=12),结扎大鼠左冠状动脉前降支制备心肌梗死模型。hAECs经体外分离、纯化、培养和标记后,在心肌梗死后1周,分别经心外膜和舌下静脉注射hAECs,4周后测定心功能,免疫组化法检测Ⅷ因子相关抗原(vWF)的表达,免疫荧光观察移植细胞的定植和存活。结果免疫荧光染色可见两种途径移植组心肌组织中均有DAPI标记细胞存在;免疫组化结果显示,200倍视野下hAECs静脉移植组和hAECs心外膜移植组的vWF染色阳性微血管计数较模型组明显增高[(22.00±2.61)和(19.80±2.32)vs.(9.80±1.17),P<0.01]。与模型组相比,两移植组左心功能显著改善(P<0.05)。结论 hAECs经静脉移植和心外膜移植均可归巢至心肌梗死区,并促进新生血管形成,对梗死大鼠的心脏具有保护作用。     

人胚胎生殖干细胞移植及心肌再生

目的 探讨直接注射移植人胚胎生殖(EG)细胞对大鼠心肌梗死的影响.方法 40只SO大鼠经缝扎左冠状动脉前降支,建立急性心肌梗死(AMI)模型,分干细胞移植组和对照组.取5～10周人胚胎牛殖腺嵴,组织块体外培养,生物学鉴定证实为人EG细胞,将其直接注入大鼠急性心肌梗死区边缘.移植后1 d,1、2、4周处死大鼠,采用免疫组化法观察心肌形成转录因子Nkx 2.5在移植细胞的表达.结果 移植组鼠抗人细胞核抗体MAB1281检测刚性,心肌形成转录因子Nkx2.5在移植细胞阳性表达.结论 人EG细胞胞接注入移植大鼠心肌梗死处,细胞能存活并呈现向心肌细胞分化的表型.     

脐血干细胞移植对梗死区心肌细胞及其分泌细胞因子的影响

目的观察脐血间质干细胞(UCB-MSCs)经冠脉移植入梗死心肌后对心肌细胞和内皮细胞增生情况。方法实验组将5-氮杂胞嘧啶核苷诱导分化后的UCB-MSCs经结扎的冠状动脉左前降支远端灌注移植入心肌梗死区域,对照组予以注射相同剂量的PBS液。1、2、4、8周后取心肌标本,应用放射免疫测定法检测心肌梗死区细胞因子IGF-1、IL-1β、IL-8的表达及含量。结果UCB-MSCs移植1、2、4、8周后,实验组梗死区心肌IGF-1、IL-1β含量明显高于对照组(P<0.01);IL-8含量也明显高于对照组(P<0.05)。结论UCB-MSCs移植入梗死心肌,不仅有心肌再生,尚可通过细胞因子分泌对梗死心肌起到血管再生起到促进作用。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.