光动力疗法防治损伤性血管内膜增生的实验研究

目的 :观察光动力疗法 (PDT)防治损伤性血管内膜增生的近期疗效 ,并筛选PDT参数。方法 :首先筛选PDT参数。兔 2 4只 ,按L4 ( 2 3)正交实验方案分为四组 ,处理因素为血啉甲醚 5或 10mg/kg、功率密度 30或 90mw/cm2 、照射时间 15或 30min。球囊法损伤髂总动脉内膜 ,随后进行PDT处理 ,2 1天后取材 ,以中膜与内膜面积的比值为指标分析各处理因素的作用。其次观察PDT疗效。兔 2 4只 ,随机分成 4组 :( 1)单纯损伤组 ;( 2 )PDT对照组 ,内膜损伤后先激光照射 ,后注射血啉甲醚 ;( 3)低PDT剂量组 ,内膜损伤后先注射血啉甲醚 5.0mg/kg ,然后以 30mw/cm2 照射 15min ;( 4 )高PDT剂量组 ,以 90mw/cm2 照射 ,其余同低剂量组。以内膜与中膜面积的比值为指标分析PDT疗效。结果 :预选参数的最优搭配为 :功率密度为 30mw/cm2 ,照射时间为 15min ,血啉甲醚剂量为 5mg/kg。低剂量组与高剂量组的S1/S2 比值分别比单纯损伤组低 71.9% (P <0 .0 1)和 59.4 % (P <0 .0 5) ,比PDT对照组低 63.1% (P <0 .0 1)和 4 6.5% (P <0 .0 5)。结论 :经皮血管内光动力疗法可能成为防治损伤性血管内膜增生的有效方法。     

低频超声对金黄色葡萄球菌生物膜的促渗作用研究

目的:探讨低频超声对血卟啉单甲醚(Hematoporphyrin monomerthyl ether,HMME)在金黄色葡萄球菌生物膜中渗透效果的影响。方法:将直径25 mm,孔径0.22μm的微孔滤膜平铺于LB琼脂培养基表面,再将纯种的金黄色葡萄球菌(ATCC6538)菌液,通过均匀涂布接种到滤膜表面,37℃恒温培养48 h获得实验所需的生物膜模型。将培养所得生物膜样本随机分为空白对照组,单独超声组,单独药物组和实验组,其中实验组按超声强度(0.5 w/cm~2或1 w/cm~2)和超声作用时间(1 min、2 min或3 min)不同又分为实验(A、B、C、D、E、F)组。所用渗透药物为20μg/m L的HMME溶液。对照组使用等量的生理盐水孵育,孵育时间为3 min,单独超声组也用生理盐水孵育并加0.5 W/cm~2的超声作用3 min。HMME在405 mm激发光下有特征性光谱,通过测量光谱曲线下积分面积,可间接反映生物膜中HMME渗入量。结果:实验A组(0.5 w/cm~2,1 min)的荧光光谱曲线下积分面积显著高于单独药物组(P0.05)。声强相同时,随作用时间增加,所得光谱曲线下积分面积显著增加(P0.01或P0.05);作用时间相同时,声强越大,光谱曲线下积分面积亦越大(P0.01)。结论:低频超声可以显著增加HMME渗透进入金黄色葡萄球菌生物膜的量,且渗透效果与超声作用时间和超声声强正相关。     

库克点百合的组织培养

培养条件:材料用0.1％HgCl_2消毒8分钟,无菌水冲洗3次,切成0.5～1.0cm长。诱导愈伤组织培养基:(1)MS十BA0.5mg/L(单位下同)十NAA0.2+2真4-D0.2;诱导芽分化培养基:(2)MS十BA1.5+NAA0.5;生根培养基:(3)MS+IBA0.2～1.0。蔗糖3％,琼脂0.8％,温度25士2℃,光     

低剂量三聚氰胺对小鼠精子质量影响的研究

采用灌胃法,研究了三聚氰胺在1.0 mg/kg、2.0 mg/kg、4.0 mg/kg浓度下染毒35 d时,小鼠体重、精子运动参数和精子形态的变化.结果显示,4.0 mg/kg 处理组小鼠部分精子运动参数显著性降低(P＜0.05),1.0 mg/kg 和2.0 mg/kg 浓度组各项精子运动参数与对照比较变化不显著(P＞0.05);在染毒一周后,2.0 mg/kg 浓度组小鼠体重增长显著性下降(P＜0.05),4.0 mg/kg 浓度组小鼠体重极显著降低(P＜0.01);3个处理浓度组对小鼠精子形态影响均不显著.总之,食物中低浓度三聚氰胺对小鼠精子活性和形态无明显的影响.     

不同剂量氯胺酮复合顺式阿曲库铵预注对肌松效应的影响

目的比较顺式阿曲库铵预注、不同剂量氯胺酮复合顺式阿曲库铵预注对顺式阿曲库铵起效时间、血流动力学的影响。方法选择全麻下行择期手术的患者120例随机分成4组,即Ⅰ(生理盐水0.5ml)组、Ⅱ(顺式阿曲库铵0.01mg/kg)组、Ⅲ(顺式阿曲库铵0.01mg/kg预注复合氯胺酮0.5mg/kg)组、Ⅳ(顺式阿曲库铵0.01mg/kg预注复合氯胺酮1mg/kg)组,预注、预处理3min后,Ⅰ组静注插管剂量顺式阿曲库铵0.15mg/kg,Ⅱ、Ⅲ、Ⅳ组静注剩余插管剂量顺式阿曲库铵0.14mg/kg。使用四个成串刺激(TOF),待T1达最大抑制程度时行气管插管,记录肌松起效时间,同时观察HR、BP的变化。结果Ⅱ、Ⅲ、Ⅳ组肌松起效时间明显短于Ⅰ组(P0.05),且Ⅲ、Ⅳ组起效时间较Ⅱ组短(P0.05),但Ⅲ、Ⅳ组起效时间比较无统计学意义(P0.05);各组麻醉诱导期间均无明显心血管不良反应。结论氯胺酮0.5mg/kg复合顺式阿曲库铵0.01mg/kg、氯胺酮1mg/kg复合顺式阿曲库铵0.01mg/kg均能进一步缩短肌松起效时间,且无明显心血管不良反应。     

脂肪干细胞移植对野百合碱诱发的肺动脉高压大鼠肺动脉压的量效和时效作用

目的探索脂肪干细胞（ADSC）移植治疗野百合碱（MCT）诱导的肺动脉高压（PAH）大鼠的适宜细胞数和干预时间。 方法（1）MCT的建模时效和量效：雄性SD大鼠48只分为正常对照组,20 mg/kg、30 mg/kg、40 mg/kg MCT组分别予腹腔注射生理盐水、MCT 20 mg/kg、30 mg/kg、40 mg/kg,4和8周后,右心室插管法检测平均肺动脉压（mPAP）,称重法计算右心室肥厚指数（RVHI）。（2）ADSC的治疗量效作用：雄性SD大鼠分别予腹腔注射MCT（30只）和生理盐水（30只）,1周后通过颈静脉注射分别移植0.5×10⁶、1.0×10⁶、3.0×10⁶、5.0×10⁶ADSC,其他组予等量生理盐水。移植3周后检测mPAP和RVHI。（3）ADSC的治疗时效作用：雄性SD大鼠30只,分别注射40 mg/kg MCT（24只）和生理盐水（6只）。MCT腹腔注射1 d,1、2周后分别移植1.0×10⁶个ADSC。MCT注射4周后检测mPAP和RVHI。多组间比较采用单因素或双因素方差分析,两两比较采用LSD检验。 结果（1）腹腔注射4周后,30 mg/ kg或40 mg/kg MCT组mPAP和RVHI均升高[mPAP值（24.89±3.31）mmHg,（27.19±2.11）mmHg比（15.80±0.42）mmHg,差异有统计学意义（P均< 0.05）;RVHI值0.42±0.06,0.47±0.04比0.25±0.02,差异有统计学意义（P均< 0.05）]。8周后,20 mg/kg或30 mg/ kg MCT组mPAP和RVHI均恢复正常,而40 mg/kg MCT组大鼠全部死亡。（2）40 mg/ kg MCT诱导的PAH大鼠mPAP和RVHI均升高。移植1.0×10⁶个ADSC可降低PAH大鼠的mPAP[（17.24±0.66）mmHg比（27.19±1.73）mmHg,P < 0.05]。移植0.5×10⁶、3.0×10⁶、5.0× 10⁶个ADSC不能降低PAH大鼠的mPAP和RVHI。（3）MCT腹腔注射1周和2周后,移植1.0×10⁶个ADSC可降低PAH大鼠的mPAP。 结论40 mg/kg MCT造模4周可建立稳定的PAH大鼠模型;造模1或2周后移植1.0×10⁶个ADSC能有效降低PAH大鼠的mPAP。     

"东湖早"枇杷的组织培养(简报)

以“东湖早”枇杷的幼胚为外植体,在B5 2,4-D 0.5 mg/L 6-BA 1.0 mg/L培养基上诱导愈伤组织,经继代培养基B5 2,4-D 0.1 mg/L 6-BA 0.5 mg/L培养后,置于B5 6-BA 1.5 mg/L培养基上诱导分化,产生不定芽。将幼苗切段转接到MS 6-BA 2.0 mg/L NAA 0.5 mg/L培养基上进行增殖培养,繁殖系数为3~5。待芽长2~3cm时再分切成单株于1/2MS NAA 0.5 mg/L培养基上,可正常生根。     

羟基喜树碱脂质体多次静脉滴注对Beagle犬的毒性

为了研究羟基喜树碱脂质体连续4周静脉滴注给予Beagle犬后产生的重复给药毒性作用,采用薄膜分散法制备羟基喜树碱脂质体,将30只Beagle犬按照体质量和性别随机分为生理盐水对照组、羟基喜树碱脂质体(0.3、0.6和1.2 mg/kg剂量)组、市售羟基喜树碱注射液1.2 mg/kg对照组,每组6只,雌雄对半,静脉滴注,每日1次,连续给药2 d停药2 d,共连续滴注4周,恢复期为2周。临床观察,检测体质量、摄食量、心电图、临床病理及组织病理学各项指标,全面评价Beagle犬重复给药后产生的毒性反应。结果发现：制备的羟基喜树碱脂质体重现性较好,其剂量相关性地引起Beagle犬食欲不振、呕吐、流涎、体质量减轻、稀便腹泻等消化道毒性反应;在0.6和1.2 mg/kg剂量下引起Beagle犬血液学、血生化部分指标的异常改变,对红细胞和粒细胞有明显的抑制作用;病理结果显示,1.2 mg/kg羟基喜树碱脂质体对给药部位(皮肤)、腺垂体、大肠、胸骨、睾丸有一定损伤作用。同等剂量的注射用羟基喜树碱脂质体和市售羟基喜树碱注射液毒性表现及程度相当。注射用羟基喜树碱脂质体毒性靶器官主要为消化道和睾丸,但毒性可...     

应用基因芯片技术检测SDS对人表皮细胞TNF信号通路基因差异表达的影响

目的:应用基因芯片技术,检测十二烷基硫酸钠(SDS)处理人表皮细胞不同时间对肿瘤坏死因子(TNF)信号通路基因差异表达的影响。方法:通过MTT试验确定表皮细胞的铺板密度和SDS的处理浓度,保证细胞的存活率;将人皮肤表皮细胞用SDS分别处理0.5、1.0、1.5、2和4 h后,提取其总RNA,反转录后通过基因芯片技术检测,比较不同处理与空白组TNF信号通路中基因表达量的差异。结果:接种细胞密度19 000/cm2、SDS在处理体系中的浓度为0.550μg/m L;SDS处理人皮肤表皮细胞0.5、1.0、1.5、2和4 h后,TNF信号通路基因差异表达的数量依次为4、10、12、8和3个。结论:SDS处理人皮肤表皮细胞1.5 h可以作为考察其对TNF信号通路影响的时间;通过基因芯片技术可以筛选SDS皮肤刺激性差异表达基因。     

银杏雌配子体发育及原生质体分离与培养的研究

经观察,银杏雌配子体在4月下旬-5月下旬为游离核时期,5月上旬采集的雌配子体在0.5%纤维素酶(Onzuka R—10)与0.5%果胶酶(Serva)混合酶液中酶解4—5h,原生质体密度为6×10~5—8×10~5/ml,活性87.3%。原生质体在去掉NH_4NO_3的MT培养基中,附加BA1.0mg/L,NAA3.0mg/L,谷氨酰胺1000mg/L,Vc5mg/L,采用液体浅层培养获得了肉眼可见的多细胞团。     

原子力显微镜观测血卟啉单甲醚对细菌光动力杀伤作用

[目的]探讨血卟啉单甲醚(Hematoporyrin monomethyl Ether,HMME)对革兰氏阳性(G )、阴性(G-)菌的光动力杀伤作用.[方法]通过平板菌落计数法和原子力显微镜(AFM),观察细菌与HMME作用前后形貌的变化.[结论]当HMME浓度为50 μg/mL,可见光(光功密度为200 mW/cm2)光照30min时90%以上的金黄色葡萄球菌(Staphyrlococcus aureus)能被杀死,无光照时对S.aureus杀灭效果显著.同等条件下,无论光照还是无光照,HMME对大肠杆菌(E.coli)无明显的杀伤作用.AFM图像显示,S.aureus细菌表面破坏严重,完全碎裂成鱼鳞状的片状堆积.对HMME作用后的E.coli扫描可见,菌体原来光滑的表面变成网格状的裂纹排列.[讨论]HMME对G 有明显的光失活效应,而对G-效果不明显.AFM的超微图像显示HMME对细菌细胞的攻击位点主要在细胞膜上.AFM为我们研究光敏剂对细菌的光动力损伤作用机制的可视化提供了依据.     

Effects of hyperthermia and photodynamic therapy on BALB/c mice bearing subcutaneous tumors

The therapeutic effects of photodynamic therapy and hyperthermia on mice bearing subcutaneous tumors were investigated. Ehrlich ascites tumor cells (1 x 10(7)) were implanted subcutaneously into the femoral area of BALB/c mice. A total of 134 tumor-bearing mice were treated with photodynamic therapy, i.e., administration of laser irradiation (514.5 nm, 112.5 mW/cm2 for 11.12 min with a total energy 75 J/cm2) after injection (i.p.) of hematoporphyrin derivative (HPD, 7.5 and 10.0 mg/kg body weight) and/or hyperthermia (by electric heating needles to 44 and 45 degrees C for 30 min) once a day for three successive days. The results revealed that the therapeutic effects of the combination of photodynamic therapy and hyperthermia were improved when compared with photodynamic therapy or hyperthermia alone. A combination of photodynamic therapy (10.0 mg HPD/kg body weight and 75 J/cm2 of total laser irradiation energy) and hyperthermia (44 degrees C for 30 min) had the best therapeutic effect, indicating that the mortality rate within 120 days (MR120) was 12.5% and the mean survival time (MST120) was 113.8 days.     

光敏剂HMME对人骨肉瘤细胞U-2OS的光动力作用及其作用机制的初步研究

目的：探讨光敏剂（HMME）介导的光动力疗法对人骨肉瘤细胞U-2OS的杀伤效应及机制研究。方法：使用不同浓度（0、10、20、30、40μg.ml-1）的光敏剂,采用不同光照能量（0、3、6、9 J.cm-2）照射人骨肉瘤细胞,与空白对照组、药物对照组（无光照但加光敏剂）和光照对照组（不加光敏剂但加光照）进行比较,MTT法检测细胞的存活率,选择半数有效量药物浓度和光照能量,作为实验组。以空白对照组为对照,采用Hoechst33342染色法,观察细胞凋亡情况。用western blot方法检测细胞凋亡蛋白caspase-7、caspase-9和PARP-1。结果：MTT结果显示,空白对照组、药物对照组和光照对照组对细胞存活率在96.7%和100%之间,药物的半数有效量为40.1μg.ml-（16 J.cm-2）和25.0μg.ml-（19 J.cm-2）。Hoechst33342染色法观察到实验组细胞明显凋亡。westen blotting检测结果,实验组与对照组相比,caspase-7、caspase-9和PARP-1表达明显增高。结论：HMME-PDT对人骨肉瘤细胞U-2OS有显著的杀伤效应,且呈光敏剂浓度和光照强度依赖性,其杀伤效应与caspase途径相关。     

Response to ALA-based PDT in an immortalised normal breast cell line and its counterpart transformed with the Ras oncogene.

Aminolevulinic acid (ALA)-based photodynamic therapy (PDT) has been successfully employed in the treatment of certain tumours. Porphyrins endogenously generated from ALA induce tumour regression after illumination with light of an appropriate wavelength. The aim of this work was to compare porphyrin production from ALA and sensitivity to photodynamic treatment in a tumour/normal cell line pair. We employed the HB4a cell line from normal mammary luminal epithelium and its counterpart transfected with the oncogen H-Ras (VAL/12 Ras). After 3 h of exposure to ALA, HB4a-Ras cells produce a maximum of 150 ng porphyrins per 10(5) cells whereas HB4a produce 95 ng porphyrins per 10(5) cells. In addition, HB4a-Ras cells show a plateau of porphyrin synthesis at 1 mM whereas HB4a porphyrins peak at the same concentration, and then decrease quickly. This higher porphyrin synthesis in the tumorigenic cell line does not lead to a higher response to the photodynamic treatment upon illumination. Lethal doses 50, LD(50), determined by MTT assay were 0.015 J cm(-2) and 0.039 J cm(-2) for HB4a and HB4a-Ras respectively after 3 h exposure to 1 mM ALA. The conclusion of this work is that a tumour cell line obtained by transfection of the Ras oncogene, although producing higher porphyrin synthesis from ALA, is more resistant to ALA-PDT than the parental non-tumour line, however the mechanism is not related to photosensitiser accumulation, but very likely to cell survival responses.     

光动力疗法诱导增生滑膜细胞凋亡的初步研究

目的：观察光动力疗法（PDT）诱导类风湿性关节炎（RA）动物模型的增生滑膜细胞的凋亡情况。方法：兔AIA模型在关节类出现第七天进行PDT治疗,随机治疗一侧膝关节,另一侧作自身对照,免耳静脉注入HMME10mg/Kg,20分钟后,膝关节用金蒸气激光照射,激光波长627.8nm,功率密度100mW/cm^2,能量密度100J/cm^2,24小时后取膝关节滑膜作病理检查,并用脱氧核苷酸末端转移酶介导的缺口末端标记法（TUNEL）原位检测凋亡结论。结果：PDT治疗组凋亡阳性细胞较对照组明显增多,图像分析结果单位视野内阳性细胞数和面密度PDT治疗组高于对照组,统计学检验差异有显著性,凋亡细胞核直径PDT治疗组较小,与对照组相比,统计学检验差异有显著性。结论：PDT有可能通过诱发滑膜细胞的凋亡,使增生的滑膜细胞减少。     

血啉甲醚在鼻咽癌细胞中的浓度分布

利用激光诱导荧光方法实验测定了新型光敏剂血啉甲醚（Hematoporphyrin Monomethyl Ether, HMME）在鼻咽癌细胞株CNE中的时间分布和浓度变化规律.实验结果表明,鼻咽癌细胞株CNE的荧光光谱中的两个特征峰615 nm和675 nm证实了鼻咽癌细胞对HMME的选择性特异吸收.HMME在鼻咽癌细胞中的潴留浓度在混合培养后的140分钟达到最大值.当用于培养鼻咽癌细胞的HMME药物浓度低于32 μg/ml时,测量得到的相对荧光光谱强度与HMME浓度呈现出很好的线性关系,通过拟合方程可以间接确定在给定注射剂量条件下鼻咽癌细胞中所潴留HMME的浓度.结果可以为HMME在光动力学诊断与治疗中的临床应用提供理论依据和剂量参考.     

光敏剂特性影响光动力治疗鲜红斑痣的数学仿真研究

目的：通过建立光动力治疗鲜红斑痣中激光、光敏剂、氧的分布及其相互作用关系的数学模型,对表皮、真皮、血管中单线态氧的产生过程进行仿真,了解光敏剂的药代动力学和扩散特性对单线态氧产生的影响,进而了解光敏剂特性在光动力治疗鲜红斑痣中的作用和意义。方法：用’Monte Carlo方法描述光在组织中的分布;用药代动力学描述光敏剂在血管中的变化规律;用Fick定律描述光敏剂和氧在组织中的扩散和分布;用与氧含量有关的二级动力学描述光敏剂的漂白;用Lambert—Beer定律和单线态氧的量子产率来计算各层组织中单线态氧的产生。结果：光敏剂药代动力学的变化,使注射光敏剂后开始照光的时间对各层组织中单线态氧产量有明显的影响。光敏剂扩散特性的改变,对真皮和表皮中单线态氧的产生有较大影响,对血管中单线态氧的产生没有影响。结论：光敏剂的特性对光动力治疗鲜红斑痣有明显的影响,数学仿真能较全面地反应这种作用的特点和意义。     

Dose-dependent effects of FK506 on neuroregeneration in a rat model

This study explored the effects of different doses of FK506 on peripheral nerve regeneration, to determine whether neuroregeneration could be enhanced without the toxicity of systemic immunosuppression. In the first part of the study, subimmunosuppressive doses of FK506 were determined by examining skin allograft survival in a rat model. Full-thickness skin grafts (2 cm2) from Wistar rats were grafted to recipient Lewis rats. The procedure was performed for six groups (n = 6). The control group received no FK506, and the other five groups received daily doses of FK506 of 0.125, 0.25, 0.5, 1.0, or 2.0 mg/kg. Animals that received 2.0 mg/kg FK506 per day exhibited complete skin graft take, whereas all other groups demonstrated complete rejection. After determination of the immunosuppressive dose of FK506, the neuroregenerative effects of different doses of FK506 were explored by assessing nerve regeneration in 80 rats after tibial nerve transection and repair. The control group received no FK506, whereas the other four groups were given daily doses of FK506 of 0.25, 0.5, 1.0, or 2.0 mg/kg. Rats were euthanized at three time points (25, 30, and 35 days), to fully investigate the effects of different FK506 dosing regimens on neuroregeneration. Histomorphometric analyses performed on postoperative days 30 and 35 demonstrated statistically significant improvements in neuroregeneration with subimmunosuppressive FK506 doses of 0.5 and 1.0 mg/kg per day. Therefore, the study demonstrated that neuroregeneration was enhanced at low doses of FK506 that were not sufficient to prevent skin allograft rejection.     

Effect of low-level laser (Ga-Al-As 655 nm) on skeletal muscle fatigue induced by electrical stimulation in rats.

We investigated whether low-level laser therapy (LLLT) can reduce muscular fatigue during tetanic contractions in rats. Thirty-two male Wistar rats were divided into four groups receiving either one of three different LLLT doses (0.5, 1.0, and 2.5 J/cm2) or a no-treatment control group. Electrical stimulation was used to induce six tetanic muscle contractions in the tibial anterior muscle. Contractions were stopped when the muscle force fell to 50% of the initial value for each contraction (T50%). There was no significant difference between the 2.5 J/cm2 laser-irradiated group and the control group in mean T50% values. Laser-irradiated groups (0.5 and 1.0 J/cm2) had significantly longer T50% values than the control group. The relative peak force for the sixth contraction in the laser-irradiated groups were significantly higher at 92.2% (SD 12.6) for 0.5 J/cm2, 83.2% (SD 20.5) for 1.0 J/cm2, and 82.9% (SD 18.3) for 2.5 J/cm2 than for the control group [50% (SD 15)]. Laser groups receiving 0.5 and 1.0 J/cm2 showed significant increases in mean performed work compared with both the control group and their first contraction values. Muscle damage was indirectly measured by creatine kinase levels in plasma. A distinct dose-response pattern was found in which 1.0 and 2.5 J/cm2 LLLT groups had significantly lower creatine kinase levels than the 0.5 J/cm2 LLLT group and the control group. We conclude that LLLT doses of 0.5 and 1.0 J/cm2 can prevent development of muscular fatigue in rats during repeated tetanic contractions.