Diagnostic laparoscopy by physicians: we should do it

Only a few centres in the UK practise diagnostic laparoscopy and liver biopsy; in comparison, laparoscopy is widely practised by physicians in Europe, the Far East and in the US. We consider the role of diagnostic laparoscopy in the assessment of liver disease in the 1990s, and describe the technique of laparoscopy, including how it may be performed safely in the endoscopy suite, under local anesthesia with mild sedation and analgesia, enabling direct visualization of the liver. We examine potential complications and contraindications. Complication rate and mortality are similar to that for percutaneous liver biopsy. Finally, the invaluable role of laparoscopy in diagnosing and staging chronic hepatitis, cirrhosis, liver tumours, hepatic infiltration, infection and structural abnormalities is considered.      

药物在室内自然光照射下的贮存期预测方法研究

以己酸孕酮注射液为例,研究了药物在自然光和不同灯光照射下的含量变化规律,预测了药物在室内自然光照射下的贮存期。找出了不同光源对药物稳定性影响的等效数量关系,使在今后的研究中可以用灯光为光源预测药物在室内自然光照射下的贮存期。己酸孕酮注射液在光照试验中含量变化服从零级反应规律:C=C₀-K(Et),在室内自然光照射下的贮存期约为1.9年。     

The neural basis of visual skill learning: an fMRI study of mirror reading

The learning of perceptual skills is thought to rely upon multiple regions in the cerebral cortex, but imaging studies have not yet provided evidence about the changes in neural activity that accompany visual skill learning. Functional magnetic resonance imaging (fMRI) was used to examine changes in activation of posterior brain regions associated with the acquisition of mirror-reading skill for novel and practiced stimuli. Multiple regions in the occipital lobe, inferior temporal cortex, superior parietal cortex and cerebellum were involved in the reading of mirror-reversed compared to normally oriented text. For novel stimuli, skilled mirror-reading was associated with decreased activation in the right superior parietal cortex and posterior occipital regions and increased activation in the left inferior temporal lobe. These results suggest that learning to read mirror- reversed text involves a progression from visuospatial transformation to direct recognition of transformed letters. Reading practiced, relative to unpracticed, stimuli was associated with decreased activation in occipital visual cortices, inferior temporal cortex and superior parietal cortex and increased activation in occipito-parietal and lateral temporal regions. By examining skill learning and item- specific repetition priming in the same task, this study demonstrates that both of these forms of learning exhibit shifts in the set of neural structures that contribute to performance.      

Diffusion of sugars and carboxylic acids through human dental plaque in vitro

Diffusion through plaque was measured with an apparatus consisting of 3 identical diffusion cells contained in an aluminium alloy block thermostatically controlled at 35 °C. In two of the cells, measurements were made of diffusion through a composite membrane consisting of 18 h dental plaque contained between bacteriological grade filters and support screens. In the third cell, diffusion occurred through a reference membrane from which the bulk of the plaque was absent. Subtraction of the diffusion resistance of this reference membrane allowed calculation of apparent tracer diffusion coefficients in the bulk of the plaque alone. Diffusion of ¹⁴C-labelled sugars and carboxylic acids was compared with that of tritiated water using dual-channel liquid scintillation counting. The diffusion coefficient of tritiated water in the plaque samples varied from 0.81 to 1.02 × 10^?9 m²s^?1 depending on the wet wt used, with coefficients of variation of ~ 15 per cent. This is between $\text{1}\text{4}$ and $\text{1}\text{3}$ of the value in aqueous solution. Diffusion coefficients relative to that of tritiated water in plaque ranged from 0.154 ± 0.014 for sucrose to 0.363 ± 0.029 for acetate. Permselectivity was low, and diffusion coefficients of I he test molecules were all between $\text{1}\text{4}$ and $\text{1}\text{5}$ of their values in free aqueous solution.     

5＇-一磷酸腺苷激活蛋白激酶对运动骨骼肌糖代谢的调节效应

目的：综述5＇-一磷酸腺苷激活蛋白激酶对运动中骨骼肌糖代谢的作用及其机制。 资料来源：应用计算机检索Medline和Highwire1986-01/2006-10有关运动、5＇-一磷酸腺苷激活蛋白激酶、骨骼肌糖代谢的文献,检索词“AMPK,glycometabolism,skeletal muscle,exercise”,限定文章语言种类为English。 资料选择：对资料进行初审,选取5＇-一磷酸腺苷激活蛋白激酶与运动骨骼肌糖代谢有关的文献,排除重复研究。 资料提炼：共收集相关文献120篇,排除重复研究,纳入28篇。 资料综合：运动激活骨骼肌5＇-一磷酸腺苷激活蛋白激酶（5＇-AMP activated protein kinase,AMPK）,AMPK激活后通过促葡萄糖转运和氧化、抑制糖原合成及促进糖原酵解而调节运动中的糖代谢。AMPK促葡萄糖转运的机制主要与促葡萄糖转运体4转位于胞膜有关,此过程涉及到AS160、一氧化氮、肌细胞增强因子2及过氧化物酶体增殖子γ辅激活因子1α等AMPK的下游靶：促糖原分解与抑制糖原合成的机制分别涉及到对糖原合成酶的抑制和磷酸果糖激酶的激活。 结论：AMPK在调控运动骨骼肌糖代谢中发挥着重要作用,阐明其机制有助于理解运动骨骼肌能量代谢的原理。     

银杏内酯和神经生长因子影响原代人胚肝细胞的增殖及分泌

目的:如何培养足够的肝细胞以及应用一些物质、因子从而使其增加分裂、减少凋亡?观察银杏内酯和神经生长因子对体外原代培养的人胚肝细胞增殖、凋亡及相关分泌功能的影响具有临床实用性意义。方法:实验于2005-07/2006-10在南通大学神经生物学研究所完成。①细胞来源:孕14～16周流产胎儿,由南通市妇幼保健院妇产科提供,产妇及其家属均知情同意,实验经医学伦理委员会批准。银杏内酯由中国药科大学提供,神经生长因子为Roche公司产品。②实验方法:用非灌流法从人胚肝中分离肝细胞,以DMEM作为培养基,接种于4块24孔培养板中,每两块培养板各接种36孔,12孔/组,细胞3×105/孔。银杏内酯组各孔加入含37.5g/L银杏内酯和体积分数为0.1小牛血清的DMEM培养液2mL,神经生长因子组各孔加入含100mg/L神经生长因子和体积分数为0.1小牛血清的DMEM培养液2mL,空白对照组各孔加入仅含体积分数为0.1小牛血清的DMEM培养液2mL。③实验评估:人胚肝细胞原代培养至9d,倒置显微镜下计数每视野的肝细胞数,同时行MTT试验,并用流式细胞仪检测肝细胞凋亡情况。分别于培养3,6,9d时收集肝细胞培养液进行白蛋白含量检测。结果:①原代肝细胞生长情况及肝细胞计数:接种12h后各组肝细胞均已贴壁,3d后可见由3～5个细胞组成的细胞集落,至9d时银杏内酯组和神经生长因子组肝细胞已基本铺满孔底,且呈立体生长倾向,空白对照组肝细胞较少,胞间有空隙存在。至培养9d,银杏内酯组、神经生长因子组、空白对照组每视野内肝细胞计数差异有显著性意义(F=12.97,P=0.00)。②MTT试验结果:原代肝细胞培养至9d时,银杏内酯组、神经生长因子组、空白对照组吸光度值差异有显著性意义(F=71.90,P=0.00)。③肝细胞培养液白蛋白含量:培养3d时各组细胞培养液的白蛋白含量方差分析无明显差异(F=0.23,P=0.79)。培养6,9d同一时间点组间比较差异有显著性意义(F=11.69,P=0.00;F=9.35,P=0.00)。④细胞凋亡情况:银杏内酯组、神经生长因子组、空白对照组凋亡肝细胞百分率分别为11.53%,9.29%,15.27%。结论:银杏内酯和神经生长因子对原代人胚肝细胞的增殖和分泌白蛋白功能均具有促进作用,并抑制细胞凋亡。     

应用表皮干细胞构建组织工程皮肤及移植实验

目的:探讨应用人表皮干细胞和猪脱细胞真皮构建组织工程皮肤修复全层皮肤缺损的可行性。方法:实验于2004-06/2005-12在南昌大学医学院烧伤研究所完成。①取环状切除后幼儿包皮(家属知情同意)用中性蛋白酶与胰蛋白酶混合消化液消化,收集细胞,接种在已包被Ⅳ型胶原的培养皿中,快速黏附,保留快速黏附细胞继续培养,培养体系为低钙高糖DMEM培养基,待融和状态时,再次消化,离心收集细胞,再次经过Ⅳ型胶原快速黏附,保留黏附细胞,培养20d左右,用作构建表皮干细胞并进行细胞鉴定。②将第2代细胞接种于铺好Ⅳ型胶原的猪无细胞真皮上,每3天换液。10d后将其转移至不锈钢网架上,行气液界面培养5d左右,构建复合皮。③取SD大鼠20只,随机分2组。于动物背部尾侧正中作2.5cm×2.5cm的全层皮肤至肌筋膜的切除。实验组10只创面覆盖表皮干细胞构建的复合皮移植物,对照组10只覆盖无细胞真皮移植物。④移植后7,10,14,21和35d,两组动物均活检取材,进行创面大体观察、组织学观察及免疫组织化学、免疫荧光鉴定等。结果:①表皮干细胞的分离、扩增及鉴定:在扩增的细胞拟接种于无细胞真皮前,进行角蛋白19免疫组织化学鉴定、以流式细胞术进行α6、CD71鉴定。经胶原Ⅳ黏附的细胞培养后形成较大集落,拟接种的种子细胞中α6briCD71dim细胞占总细胞的(39±7)%;鼠抗人角蛋白19阳性细胞率为(53±6)%。②组织工程皮肤的建立结果:表皮干细胞种植于无细胞真皮上1d后,细胞便贴壁并开始生长增殖,8d左右透过间隙见无细胞真皮上表皮干细胞渐增殖融合成片;2周时无细胞真皮表面可见菲薄皮片状生长的细胞膜片物。复合皮组织学检查可见无细胞真皮表面覆盖多层融合的表皮层细胞,无细胞真皮内细胞成分去除完全。③创面愈合情况:术后2周,实验组移植物成活,创面上已上皮化,移植物柔软,而对照组无细胞真皮表面未上皮化,较干燥,色暗、触之较硬,两组创面均未见明显收缩。4周,实验组创面愈合,未见明显排斥反应。对照组无细胞真皮干燥脱落而创面暴露,创面收缩明显,创面边缘因自体表皮细胞爬行而缩小创面。④组织学检查及免疫组织化学染色结果:实验组2周时创面已全部上皮化,但细胞分层少;3,4周表皮层分化良好,多层结构,真皮纤维内见成纤维细胞长入,见少许炎症细胞,大鼠自体正常皮肤见毛囊及毛发结构。对照组2周创面尚未上皮化,无细胞真皮干燥变性,3,4周后创面仍未愈合,可见大量炎症细胞。实验组愈合创面抗人人类白细胞抗原-Ⅰ型抗原直接免疫荧光染色呈阳性,证明新生表皮由移植的人表皮细胞形成。而对照组阴性。实验组愈合创面广谱角蛋白免疫组织化学染色见整个表皮层细胞胞浆染呈棕黄,为阳性反应。结论:利用体外纯化扩增的表皮干细胞及制备的猪脱细胞真皮可体外联合构建具有复层结构的组织工程皮肤,该组织工程皮肤可用于修复动物全层皮肤缺损。     

当归补血汤对血细胞及相关免疫因子的作用

目的:当归补血汤对正常及免疫功能低下机体的免疫功能具有增强作用。文章就近年来当归补血汤免疫药理方面的研究作一综述。资料来源:应用计算机检索中国期刊全文数据库1979-01/2007-03期间的相关文章,检索词为“当归,黄芪”,并限定文章语言种类为中文。资料选择:对资料进行初审,并查看每篇文献后的引文。纳入标准:文章所述内容应与当归补血汤免疫药理研究相关。排除标准:重复研究。资料提炼:共收集到78篇相关文献,30篇文献符合纳入标准,排除的48篇文献为内容陈旧或重复。资料综合:当归补血汤是补益气血经典方剂,该方由黄芪和当归以5:1比例组成,重用黄芪大补脾肺之气,以滋生血之源,主治劳倦内伤,气弱血虚之证。该方在免疫药理作用方面,国内学者对正常及免疫功能低下机体进行了较多实验研究。既有体内实验,又有进一步的离体观察;既有对免疫器官形态的观察,又有对不同免疫细胞数量和功能的测定,并对其作用分子机制进行探讨。研究结果表明当归补血汤可以增强机体红细胞的免疫黏附及清除免疫复合物的能力,增强吞噬细胞的吞噬功能,增强NK细胞的杀伤功能,不同程度的促进T、B淋巴细胞的增殖和相应的免疫功能。当归补血汤还具有促进造血和血管生成作用。上述作用的发生机制可能与其调节基因活性和活性分子作用相关。结论:当归补血汤组方简单,配伍配比精当,能够增强正常及免疫功能低下机体的免疫功能,临床应用广泛。但药理研究大多集中在动物整体、器官、细胞等,分子水平研究还有欠缺。