利用LC／MS检测中药总成分中各大类成分含量的方法学探讨

目的:利用液相-质谱(LC-Ms)技术检测总成分,并对其含量计算进行研究.方法:以感冒中药复方YL2000为例,利用LC-MS测定其中的总黄酮、总生物碱和总香豆素等主要大类成分,以HPLC测定的其中单体成分含量作参照,对LC-MS中各大类成分百分比进行含量计算.结果:以此方法计算的总黄酮、总生物碱、总香豆含量以及总含量与所用紫外分光光度法所测的含量基本一致.结论:利用LC-MS对成分结构的归属来确定各大类成分的构成比例,以所测代表成分HPLC含量结果作参照来计算各大类成分的含量具有一定的可行性.     

Biophysical, histological and biochemical changes after non-ablative treatments with the 595 and 1320 nm lasers: a comparative study

BACKGROUND/PURPOSE: The objective was to compare the efficiencies of the 595 nm pulsed dye and the 1320 nm Nd : YAG laser non-ablative rejuvenation. METHODS: KM mice were irradiated with the 595 nm pulsed dye and the 1320 nm Nd : YAG lasers. Histological changes were evaluated immediately, 1, 7, 21, 30 and 60 days after the two laser treatments. Skin hydration and hydroxyproline content were measured to quantify the degree of improvement of the skin's water-holding capacity and the rate of hydroxyproline synthesis. RESULTS: Although not statistically significant, the 1320 nm Nd : YAG laser treatment induced 9.7% greater improvement of skin hydration than the 595 nm laser while the 595 nm pulsed dye laser treatment led to a thicker dermis and 8.7% greater increase of hydroxyproline than the 1320 nm laser. More than 50% increase of collagen type I was observed in 75% of 595 nm laser-treated sites and 42% of 1320 nm laser-treated sites, and more than 25% increase of collagen type III was observed in 75% of 595 nm laser-treated sites and 50% of 1320 nm laser-treated sites. The 595 nm laser treatment was better in increasing the amount of collagen fibers, especially collagen type I (P < 0.05). CONCLUSION: Our results demonstrated that the 595 nm laser appeared to be more effective in increasing new collagen formation, while the 1320 nm laser was superior to the 595 nm laser in improving the skin's water-holding capacity.     

Photodynamic therapy for treatment choroidal neovascularization in angioid streaks--case report

The aim of this paper is to present the effects of photodynamic therapy for treatment of bilateral macular choroidal neovascularization in angioid streaks during 16 months period. 50 years old man has one PDT in the right eye and three in the left eye. In the left eye conversion from choroidal neovascular membrane (CNV) to a fibrous disciform lesion following photodynamic therapy, was observed. In the right eye vision decreased from 1.0 to 0.2, but the leakage was minimal and stabilization of CNV size after the progression was noted.     

Stage specificity of novel growth factor expression during development of proliferative vitreoretinopathy

OBJECTIVE: To compare the relative levels of connective tissue growth factor (CTGF), platelet-derived growth factor alpha (PDGF-AA), and hepatocyte growth factor (HGF) in glial and retinal pigment epithelial (RPE) cells of epiretinal membranes from proliferative vitreoretinopathy (PVR). METHODS: A total of 37 PVR membranes, of various stages, underwent fluorescent immunohistochemisty and confocal laser scanning microscopy to localize CTGF, HGF, and PDGF-AA in RPE and glial cells. RESULTS: Numerous RPE, and relatively fewer glial cells, were found in all stages of PVR. CTGF immunoreactivity increased from early to late stage PVR and was principally expressed by RPE cells in early stage, and by glial cells in late stage PVR. HGF, expressed by both RPE and glial cells, was principally expressed in mid-stage PVR. PDGF-AA, expressed by both cell types, demonstrated a uniform level of staining throughout all stages of PVR. CONCLUSIONS: RPE and glial cells contribute to the expression of CTGF, HGF, and PDGF-AA during PVR, but with specific developmental patterns. PDGF-AA is expressed uniformly throughout all stages of PVR, while HGF expression peaks during mid stage, and CTGF expression is highest during late stage PVR. These results allow for the development of stage-specific therapeutics for PVR that may allow targeting of the early proliferative and/or the late tractional stages of PVR.     

免疫系统与青光眼

青光眼是一组以视神经凹陷性萎缩和视野缺陷为共同特征的疾病,病理性眼压增高是其主要危险因素。多种因素决定了青光眼视神经节细胞的最后结局。而免疫因素在青光眼的视网膜视神经损害中所起的作用应该是双重的。所以探明免疫系统在青光眼损伤修复中的作用,对青光眼的治疗具有重要的意义。     

虹膜识别技术在准分子激光原位角膜磨镶术治疗近视性散光中的临床研究

目的评价虹膜识别技术应用于准分子激光原位角膜磨镶术（LASIK）治疗近视性散光的准确性、稳定性及可预测性。方法采用虹膜识别引导的LASIK治疗近视散光患者97例（183只眼）,按术前柱镜度数分为3组：1组（-0．50～-1．00D）79只眼,2组（-1．25～-2．00D）70只眼,3组（-2．25～4．00D）34只眼;按术前柱镜轴向分为组A（循规散光）106只眼、组B（逆规散光）43只眼、组C（斜轴散光）34只眼。术前采集散瞳前后的虹膜数据和波阵面像差数据,经过虹膜识别后形成ate文件,将该数据同Orbsesn角膜地形图系统产生的ore文件相结合设计手术方案形成tls文件,导入准分子激光系统,激光器对平卧位时术眼再次进行虹膜识别,确定瞳孔中心偏移量和眼球旋转角度,在治疗时加以补偿,术中三维眼球自动跟踪系统监测眼球运动。观察手术前后不同时期的视力、散光度及轴向变化。结果术中检测出瞳孔中心总体的偏移量为X轴方向（0．41±98．90）μm、Y轴方向（109．15±141．35）μm、眼球旋转偏移角度0．83°±3．40°,术中对其加以补偿。术后6个月裸眼视力≥0．5者183只眼（100．0％）,≥1．0者169只眼（92．3％）,散光度由术前（-1．54±0．65）D减少为术后6个月的（-0．26±0．25）D,对术后各时间点的样本总体散光度进行单因素方差分析,差异有统计学意义（F=5．74,P〈0．01）。各组间两两比较采用SNK检验,术后1周与1、3、6个月比较差异有统计学意义（P〈0．05）;术后1、3、6个月之间差异无统计学意义（P=0．88）。术后6个月顺规散光下降为45只眼（24．6％）,逆规散光下降为31只眼（16．6％）,斜轴散光上升为38只眼（21．0％）,术后6个月有69只眼（37．8％）成为无散光眼。结论虹膜识别引导的LASIK治疗近视散光效果良好,准确性及可预测性较高,是目前精确、先进、有效的散光治疗方法之一。     

紧密连接蛋白及神经胶质原纤维酸性蛋白在糖尿病大鼠视网膜的表达变化及其与血-视网膜屏障功能的关系

目的研究糖尿病鼠视网膜内皮细胞间紧密连接蛋白Occludin及神经胶质原纤维酸性蛋白（GFAP）的表达改变及其与血-视网膜屏障（BRB）的关系。方法链脲佐菌素腹腔注射建立大鼠糖尿病模型1、3、6个月后行伊凡思蓝（EB）注射评价血．视网膜屏障破坏的形态学改变。并行免疫荧光组织化学观察Occludin及GFAP的表达变化。结果1个月时大鼠视网膜中神经纤维层及节细胞层中GFAP表达明显增高,Occludin网线状荧光排列紊乱,但未见荧光强度减弱及中断。3—6个月GFAP阳性的Mtiller细胞逐渐增多,Occludin表达减弱且连续性中断的范围不断扩大。EB注射显示血-视网膜屏障损害呈现同步的发展趋势。结论在糖尿病视网膜病变早期星形胶质细胞的活化可能在维持BRB功能中起重要作用,病情发展Mtiller细胞活化使BRB的完整性进一步破坏。     

过氧化氢对人晶状体上皮细胞内细胞质膜微囊蛋白的影响

目的探讨过氧化氢（H2O2）对人晶状体上皮细胞的细胞质膜微囊蛋白（CP）及磷酸化CP-1分布与表达的影响。方法给予人晶状体上皮细胞（SRA01／04）不同浓度及不同时间点的H2O2刺激。用激光共聚焦显微镜和免疫荧光显微镜观察CP及磷酸化CP-1的分布。通过免疫印迹试验观察CP表达水平的变化以及磷酸化CP—1的表达。结果通过激光共聚焦显微镜和荧光显微镜,观察到人晶状体上皮细胞的质膜和细胞质内含有丰富的CP;当给予细胞H2O2刺激后,细胞质内CP的分布增多;当刺激时间达到1h,细胞膜被破坏,但仍可观察到CP的分布情况。此外,H2O2的刺激可以使CP-1发生磷酸化。免疫印迹试验发现,随着H2O2作用时间的延长和刺激浓度的升高,细胞质膜和细胞总蛋白质的CP表达水平呈下调趋势。结论H2O2刺激人晶状体上皮细胞后,CP重新分布并可能破坏了细胞质膜微囊（caveolae）的结构,使得细胞的CP表达下调。细胞质膜微囊和CP可能在人晶状体上皮细胞中具有重要作用。     

色素上皮源性因子对缺血-再灌注视网膜神经节细胞的保护作用

目的:研究色素上皮源性因子(pigment epithelium derived factor,PEDF)对高眼压诱导的大鼠视网膜缺血-再灌注后视网膜神经节细胞的保护作用.方法:经眼角膜进行前房平衡盐水(BSS)灌注,维持眼内压110 mmHg,以阻止视网膜正常血液灌注.60 min后取出灌注针头,恢复视网膜正常血流,从而建立大鼠视网膜缺血-再灌注模型.实验分为正常非缺血组和视网膜缺血-再灌注组,后者又分为生理盐水注射对照组和PEDF注射实验组,再灌注模型建立后立即向实验组大鼠玻璃体腔内注射0.2 g/L PEDF 2 μL.实验对照组用同样方法注射等量生理盐水.分别于注射后2 d和7 d进行眼球摘除,对视网膜进行光学显微镜形态学观察和Fas原位杂交免疫学分析,探讨PEDF对缺血-再灌注视网膜神经节细胞的保护作用.结果:缺血-再灌注2 d时生理盐水注射组和PEDF注射组视网膜神经节细胞明显少于正常对照组(P＜0.01)和(P＜0.05),PEDF注射组视网膜神经节细胞数较生理盐水注射组明显较多,相比有显著性差异(P＜0.05);视网膜神经节细胞计数再灌注7 d后结果与2 d时类似.再灌注2 d生理盐水注射组Fas阳性染色细胞比PEDF注射组明显较多(P＜0.05),生理盐水组比PEDF注射组阳性细胞百分率明显较高(P＜O.01);再灌注7 d时两组Fas阳性细胞计数无明显差异.结论:视网膜缺血-再灌注后即刻行玻璃体腔内PEDF注射可以改善视网膜神经节细胞的损伤并有一定保护作用.     

1CU可调节人工晶状体眼拟调节力临床分析

目的:探讨1CU可调节人工晶状体植入眼内后术眼拟调节能力.方法:通过对2003-06至今我院行白内障超声乳化联合可调节人工晶状体1CU术后对术眼裸眼远、近视力,矫正视力,矫正远视力下的近视力进行测量,并运用主觉近点法、离交法、动态视网膜检影法以及A超测量10 g/L匹罗卡品点眼前后前房深度,检测术眼拟调节能力,同期以单焦人工晶状体Acrysof作为临床对比,评价可调节人工晶状体1CU临床应用效果.结果:1CU组的裸眼近视力和矫正远视力下的近视力好于Acrysof组;带状光检影测量1CU组的调节力为0.99±0.45D,Acrysof组的调节力为0.25±0.22D,两组差异有显著性意义(t=5.623,P＜0.05);主觉近点法测量1CU组的调节力为1.60±0.55D,Acrysof组的调节力为0.42±0.22D,两组差异有显著性意义(t=2.147,P＜0.05);离焦法测量1CU组的调节力为1.46±0.52D,Acrysof组的调节力为0.52±0.35D,两组差异有显著性意义(t=2.647,P＜0.05);滴用匹罗卡品眼药水后分别测量1CU组和Acrysof组前房深度(ACD)变化,两组比较有显著性差异(t=4.374,P＜0.05).结论:1CU可调节人工晶状体可以明显提高近视力,使白内障患者术后获得一定的调节力.