瘦素对肝细胞三酰甘油沉积的影响

目的观察瘦素对人L-02脂肪变肝细胞三酰甘油(TG)沉积的影响,探讨瘦素在非乙醇性脂肪性肝病发生、发展中的作用。方法人肝L-02细胞培养接种于50 m l培养瓶中,用50%胎牛血清1640培养基造模,时间24 h。设立正常组、模型组、瘦素处理1,2,3组,处理组瘦素浓度分别为10-7mol/L,10-6mol/L,10-5mol/L,分别处理6 h;再以10-6mol/L浓度瘦素建立时效关系分别处理(6,12及24 h)。每组均行油红“O”染色观察脂肪化情况,高效液相色谱测细胞内TG含量变化。结果油红“O”染色显示50%的胎牛血清的1640培养基24 h能成功地制造肝细胞脂肪变性模型,加入瘦素处理后脂肪化程度减轻。高效液相色谱显示细胞内TG明显减少,且与瘦素呈剂量、时间依赖性关系。结论肝细胞脂肪变性时,瘦素能使肝细胞脂肪化程度减轻,能减少肝细胞TG的沉积,对脂肪肝防治有一定的作用。     

微嵌合体表达与移植肾免疫耐受的实验研究

目的探讨肾移植术后受者微嵌合体表达与免疫耐受的相关性。方法采集接受男性供体肾脏移植术后女性受者不同生存期外周血及尿沉渣标本,采用Y染色体三对引物SRY1、DYZ11st和DYZ12nd,应用PCR和RT-PCR方法检测标本中微嵌合体特异性代表Y染色体DNA和mRNA的表达,同时测定移植肾功能。结果在130例供者为男性的女性肾移植受者检测到嵌合阳性者98例,占75.39%(n=130),嵌合阴性者32例,占24.61%。嵌合阳性与阴性两组比较,移植肾平均存活时间分别为(8.7±3.5)年和(5.4±3.3)年,P<0.05;嵌合阳性者发生过排斥反应的有11例,占11.22%,而嵌合阴性者为9例,占28.13%,P<0.05;血清肌酐在嵌合阳性者(74.3±32.5)mol/L,而嵌合阴性者为(113.6±37.8)mol/L,P<0.05。结论受者体内嵌合阳性与阴性者相比具有更好的移植肾功能和低排斥率,肾移植术后受者生存期愈长嵌合阳性率愈高,提示肾移植术后受者体内的嵌合状态与免疫耐受具有相关性。     

An alkylpolyglucoside surfactant as a prospective pharmaceutical excipient for topical formulations: The influence of oil polarity on the colloidal structure and hydrocortisone in vitro/in vivo permeation

There is a growing need for research into new skin- and environment-friendly surfactants. This paper focuses on a natural surfactant of an alkylpolyglucoside type, which can form both thermotropic and lyotropic liquid-crystalline phases. The aim of this study was to relate some physicochemical properties (characterised by polarisation and transmission electron microscopy, thermal analysis and rheology) of the three formulations based on cetearyl glucoside and cetearyl alcohol, to the results of in vitro and in vivo bioavailability of hydrocortisone (HC). The three formulations contained oils of different polarity (medium chain triglycerides: MG, isopropyl myristate: IPM and light liquid paraffin: LP), respectively. In vitro permeation was followed through the artificial skin constructs (ASC), while the parameters measured in vivo were erythema index: EI (using instrumental human skin blanching assay), transepidermal water loss (TEWL) and stratum corneum hydration (SCH). The vehicles based on cetearyl glucoside and cetearyl alcohol showed a complex colloidal structure of lamellar liquid-crystalline and lamellar gel-crystalline type, depending on oil polarity. Rheological profile of the vehicle was directly related to the in vitro profile of the HC permeation. In vivo results suggested that the vehicle with MG retarded the HC permeation, whereas less polar IPM and non-polar LP enhanced it. It is suggested that the enhancement is achieved either by a direct interaction with lipid lamellae of the SC or indirectly by improving skin hydration.

There were no adverse effects during in vivo study, which indicates a good safety profile of this alkylpolyglucoside surfactant.     

血管内皮生长因子在糖尿病大鼠视网膜中的表达

目的 :利用糖尿病大鼠动物模型 ,通过大鼠视网膜中血管内皮生长因子 ( vascular en-dothelial growth factor,VEGF)的表达情况 ,探讨 VEGF在糖尿病视网膜病变 ( diabetic retinopa-thy,DR)发展过程中作用机理。方法 :复制链脲佐菌素 ( streptozocin,STZ)糖尿病大鼠动物模型 ,取不同组、不同时期大鼠视网膜 ,利用免疫组织化学及原位杂交方法 ,检测 VEGF蛋白质及 m RNA的表达情况。结果 :1正常对照组 ( M)、血糖恢复组及糖尿病 1个月组 ( M1)大鼠视网膜均无 VEGF蛋白质及 m RNA的阳性表达 ;2糖尿病 3个月组 ( M3 )未见 VEGF m RNA表达 ,而在内核层及节细胞层有 VEGF蛋白表达阳性 ( 34%) ,此表达与胶质纤维酸性蛋白 ( GFAP)染色阳性细胞相同 ;3糖尿病 5个月组 ( M5) VEGF m RNA表达阳性率为 67%,表达位于视网膜各层 ,内界膜中的表达与 GFAP免疫组化阳性分布相同 ,VEGF蛋白质表达强阳性 ( 89%) ,主要位于内核层、节细胞层的细胞及血管上。结论 :VEGF在 STZ大鼠视网膜中的表达与病程相关 ,而且 VEGF的产生存在着自分泌和旁分泌多种途径     

阶跃切应力对内皮细胞释放前列环素的影响

采用流室定量研究阶跃切应力对内皮细胞释放前列环素（ｐＧＩ＿２）的影响。对流室中的内皮细胞连续１００分钟施加１、１０、２０ｄｙｎｅ／ｃｍ￣２的阶跃剪切应力，用放免法测流室的８个时间段流出液的６－ｋｅｔｏ－ＰＧＦｉ＿（１ａ）。绘出切应力刺激内皮细胞释放ＰＧＩ＿２的曲线。结果表明：未施加切应力时，内皮细胞释放ＰＧＩ＿２的量在检测限以下，施加切应力后，ＰＧＩ＿２释放速率很快达极大值，且又在数分钟内减小．施加阶跃切应力越大，ＰＧＩ＿２释放速率曲线峰值越高，但衰减也越快，在８０～１００分钟时间内的前列环素释放速率平均值也越大。由此提示，在活体中血液流变学因素一血流切应力对血管内皮细胞释放前列环素有促进作用。     

慢性硬膜下血肿的微创治疗

目的：为减少慢性硬膜下血肿术后并发症，减轻患者经济负担．方法：对21例慢性硬膜下血肿行锥颅冲洗术．术中迅速置管，缓慢引流陈旧性血液，并用生理盐水冲洗至消亮．术后充分引流，早期复查头颅CT，配合高压氧治疗．结果：21例均痊愈出院，术后随访无1例复发．结论：此法不仅简便易行，经济有效，而且能有效防止钻孔引流术后并发症的发生．     

肝源性溃疡的临床观察

目的：研究乙肝后肝硬化患者消化性溃疡的临床特点及治疗转归，探讨内镜下止血及小剂量心得安对肝源性溃疡愈合的影响。方法：肝源性溃疡75例，观察其发病年龄、临床表现、溃疡部位、HP感染率、出血率及7d内止血率及溃疡愈合率等特点，随机分为治疗组37例，内镜下止血 心得安 泰胃美治疗，对照组38例，用维生素B6 泰胃美治疗；另外选单纯性消化性溃疡73例，随机对照2组36例，用维生素B6 泰胃美治疗，对照3组37例，内镜下止血 维生素B6 泰胃美治疗。结果：肝源性溃疡发病年龄较单纯性消化性溃疡晚10年，溃疡部位以复合性溃疡居多和并出血率高，内镜下止血 小剂量心得安可明显提高肝源性溃疡7d内止血率，促进肝源性溃疡愈合率，与对照组具有统计学意义(P＜0．01)。结论：肝源性溃疡难以治愈互易并出血，互以复合性溃疡多见，门静脉高压因素可能从中发挥作用，内镜下注射及喷洒止血剂及口服小剂量心得安及用制酸护肝治疗能有效止血，促进溃疡的愈合。     

小鼠宫颈癌腹水瘤U_(14)及其亚株侵袭特性的初步研究

本文将小鼠宫颈癌腹水瘤u_(14)和经本教研室活体筛选出的倾向于肺转移的u_(14)A-P_(11)和倾向于淋巴转移的u_(14)-AL_(10)两亚株,用鸡胚绒毛尿囊膜接种的体内方法和与经胰酶不同处理的BALB/C小鼠腹膜组织共同培养的体外方法,对三种瘤细胞的侵袭特性进行研究。结果证明,三种瘤细胞体内转移倾向的差异与其对鸡胚绒毛尿囊膜基质和小鼠腹膜基质的侵袭能力成正相关,表明瘤细胞转移的器官倾向性与瘤细胞的侵袭力有一定的关系。     

脉冲Nd：YAG激光照射对大鼠炎症牙龈组织的作用

患实验性牙龈炎的大鼠牙龈（６例）经脉冲掺钕钇铝柘榴石激光（Ｎｄ：ＹＡＧ激光）照射后进行光镜及超微结构观察。目的在于了解低能量Ｎｄ：ＹＡＧ激光照射对牙龈炎症的治疗作用。炎症牙龈经输出能量为１．５Ｗ，脉冲数为２０ＰＰＳ的激光照射后与对照组相比较，形态学上有如下变化：上皮细胞无变性；牙龈上皮层数较炎症时减少；上皮下炎细胞浸润明显减少；上皮细胞间隙变小；上皮下结缔组织内产生大量的排列致密的胶原纤维。结果提     

Cytochromes of the P450 2C subfamily are the major enzymes involved in the O-demethylation of verapamil in humans

The calcium channel blocker verapamil [2,8-bis-(3,4-dimethoxyphenyl)-6-methyl-2-isopropyl-6-azaoctanitrile] undergoes extensive biotransformation in man. We have previously demonstrated cytochrome P450 (CYP) 3A4 and 1A2 to be the enzymes responsible for verapamil N-dealkylation (formation of D-617 [2-(3,4-dimethoxyphenyl)-5-methylamino-2-isopropylvaleronitrile]), and verapamil N-demethylation (formation of norverapamil [2,8-bis(3,4-dimethoxyphenyl)-2-isopropyl-6-azaoctanitrile]), while there was no involvement of CYP3A4 and CYP1A2 in the third initial metabolic step of verapamil, which is verapamil O-demethylation. This pathway yields formation of D-703 [2-(4-hydroxy-3-methoxyphenyl)-8-(3,4-dimethoxyphenyl)-6-methyl-2-isopropyl-6-azaoctanitrile] and D-702 [2-(3,4-dimethoxyphenyl)-8-(4-hydroxy-3-methoxyphenyl)6-methyl-2-isopropyl-6-azaoctanitrile]. The enzymes catalyzing verapamil O-demethylation have not been characterized so far. We have therefore identified and characterized the enzymes involved in verapamil O-demethylation in humans by using the following in vitro approaches: (I) characterization of O-demethylation kinetics in the presence of the microsomal fraction of human liver, (II) inhibition of verapamil O-demethylation by specific antibodies and selective inhibitors and (111) investigation of metabolite formation in microsomes obtained from yeast strain Saccharomyces cerevisiae W(R), that was genetically engineered for stable expression of human CYP2C8, 2C9 and 2C18.In human liver microsomes (n=4), the intrinsic clearance (CL_int), as derived from the ratio of V _max/K_m, was significantly higher for O-demethylation to D-703 compared to formation of D-702 following incubation with racemic verapamil (13.9±1.0 vs 2.4±0.6 ml^*min^{-1 *}g^-1 mean±SD; p<0.05), S-Verapamil (16.8±3.3 vs 2.2±1.2 ml^* mini^*g^-1, p<0.05) and R-verapamil (12.1±2.9 vs 3.6 ±1.3 ml^*min^{-1 *} g^-1; p<0.05), thus indicating regioselectivity of verapamil O-demethylation process. The CL_int of D-703 formation in human liver microsomes showed a modest but significant degree of stereo selectivity (p<0.05) with a S/R-ratio of 1.41±0.17. Anti-LKM2 (anti-liver/kidney microsome) autoantibodies (which inhibit CYP2C9 and 2C19) and sulfaphenazole (a specific CYP2C9 inhibitor) reduced the maximum rate of formation of D-703 by 81.5±4.5% and 45%, that of D-702 by 52.7±7.5% and 72.5%, respectively. Both D-703 and D-702 were formed by stably expressed CYP2C9 and CYP2C18, whereas incubation with CYP2C8 selectively yielded D-703.In conclusion, our results show that enzymes of the CYP2C subfamily are mainly involved in verapamil O-demethylation. Verapamil therefore has the potential to interact with other drugs which inhibit or induce these enzymes.