粗糙脉孢菌漆酶基因的克隆及在毕赤酵母中的初步表达

采用连续延伸PCR方法克隆到粗糙脉孢菌（Neurospora crassa）漆酶基因，并将其克隆到表达载体pPIC9k，重组质粒经线性化、电激转化Pichia pastoris KM71，部分阳性克隆的PCR结果表明：漆酶基因已整合到巴斯德毕赤酵母染色体上，重组菌经甲醇诱导后3～5d产漆酶量最高，为2-3U／mL。     

络合碘在预防阑尾炎术后切口感染中的作用

目的络合碘预防阑尾切除术后切口感染。方法0．05％络合碘冲洗切口65例。结果络合碘冲洗的化脓、坏疽或穿孔性阑尾炎切口无感染，与传统组感染率9．1％（6／66）比较p〈0.05（x^2=4．266）。结论0．05％络合碘冲洗切口是预防阑尾切除术后切口感染的有效方法。     

Opioid peptides and receptors in joint tissues: study in the rat.

Using immunohistochemical and biochemical techniques, the occurrence of endogenous opioid peptides and their receptors in normal rat bone and joint tissues was investigated. Opioid receptors were detected, quantified, and characterized in homogenates from capsule/synovium and periosteum using radioligand binding assays. Receptor binding of the nonselective opioid [3H]naloxone to tissue homogenates was stereospecific and saturable, showing similar characteristics to that of brain tissue, although with lower binding capacities. By immunohistochemistry, the neuronal occurrence of four different enkephalins was demonstrated in synovium, bone marrow, periosteum, and juxta-articular bone, whereas no neuronal dynorphin immunoreactivity was detected. Double-staining studies disclosed that enkephalins coexisted with substance P in primary afferent fibers. The applied techniques can be used to assess changes in the distribution of endogenous opioids and their receptors in joint tissues in conditions associated with pain and inflammation. The endogenous opioid system now demonstrated might be targeted and exploited therapeutically to obtain peripheral control of symptoms in joint disorders.     

No evidence for association between the −112G/A polymorphism of UGRP1 and childhood atopic asthma

BACKGROUND: Susceptibility to asthma is known to involve genetic factors. Genome-wide screens have indicated that the chromosome 5q31-q34 region is linked to and/or associated with asthma. A new gene, named UGRP1 and reported by Niimi et al., encodes uteroglobin-related protein and is expressed in the lung and trachea. Niimi et al. showed the -112G/A polymorphism of the UGRP1 gene to be associated with asthma in a case-control study. OBJECTIVE: The objective of the present study was to replicate this association and confirm the possible role of the UGRP1-112G/A polymorphism in the aetiology of childhood asthma in a Japanese population. METHODS AND RESULTS: We conducted a transmission disequilibrium test (TDT) in 131 families identified through paediatric patients being treated for asthma. A case-control study was also carried out by comparing the probands and 137 unrelated non-atopic non-asthmatic Japanese children and 211 unrelated healthy Japanese adults. The -112G/A polymorphism was genotyped by the PCR-RFLP method. The TDT revealed that the -112A allele was not preferentially transmitted to asthma-affected children (P=0.85). Neither the presence of at least one A allele in an individual's genotype (sum of the G/A and A/A genotypes) nor the -112A allele was more prevalent among the asthma subjects than among the control subjects. CONCLUSION: Our findings indicate that the UGRP1-112G/A polymorphism does not play a substantial role in genetic predisposition to childhood asthma in this Japanese population.     

参芪胶囊的鉴别与含量测定方法研究

对参区胶囊中黄芪、人参、当归、知母、黄连等味药进行了薄层鉴别。并对黄连中小檗碱进行了薄层光密度法测定，方法灵敏，重现性好，回收率１００％，相对标准偏差ＲＳＤ％＝２．０（ｎ＝６）。     

Additive effects of earthworm extract with HpD-laser on the enhancement of reactive oxygen production and the inhibition of[~3H]—thymidine.uptake in mouse ascitic tumor cells

The effects of combined use of earthworm extract(912)and HpD-laser on the produc-tion of reactive oxygen and the biosynthesis of DNA in S_(180) tumor cells were studied throughchemiluminescence measurement and[~3H]-TdR incorporation assay.The results showed that as com-pared with the control,the intensity of chemiluminescence emitted by tumor cells treatedsimultaneously with 912 and HpD-laser was enhanced more than ten-folds,while that treated with912 or HpD-laser alone was increased only 2～4 folds.The[~3H]-TdR incorporation into tumorcells of the former group was inhibited upto 74.1%,and that of the latter groups decreased onlyby 42.2% and 40.0%,respectively.In accordance with these biochemical changes,the ultrastructuraldamage of tumor cells of the former,combinedly treated group appeared to be the most serious.This suggests an additive effect of 912 with HpD-laser on tumor cells.In addition,if free radicalscavengers,such as catalase and superoxide dismutase,were added to the reaction systembefore chemiluminescence assay,the luminescent enhancement effect mentioned above was dramaticallyalleviated,implying the presence of O_2~ and H_2O_2 in the system.Therefore,as to the toxic effecton tumor cells,912 and HpD-laser are not only additive in efficiency,but also similar in theunderlying mechanism of action.     

血管重塑对大鼠血管成形术后再狭窄的影响

目的：建立大鼠颈动脉再狭窄模型，原位灌注固定取材。评价PTA后血管重塑（VR）的动态变化规律，定量分析血管重塑在血管再狭窄过程中的变化及作用。方法：制作70只SD雄性大鼠颈总动脉再狭窄模型，分原位灌注实验组、对照组，于术后1h、3、7、14、28和42天原位灌注固定取材，行HE染色、Masson染色，观察标本血管狭窄情况。结果：①血管重塑指数（VRI）在PTA后即刻最大，3天组明显降低，7天组稍有增大，其后不断减小，剩余血管腔面积百分比同VRI的变化曲线基本一致。②FFA后，血管腔面积总体呈逐渐缩小趋势，内弹力板围绕面积（IELA）1h组较对照组明显增大，3天组较1h组明显缩小，14、28、42天组较对照组明显缩小。外弹力板围绕面积（EELA）逐渐缩小。EELA、IELA的变化与血管腔面积变化呈正相关。③VRI与血管腔面积的变化呈正相关，新生内膜面积与剩余狭窄率、血管腔面积无直线相关。结论：再狭窄过程中存在扩张性重塑和收缩性重塑现象，管腔的狭窄与否取决于血管重塑指数的变化，而不是新生内膜的变化，新生内膜的形成是血管重塑过程中的一部分。IELA和EELA可作为判断管腔狭窄及评价血管重塑的指标。     

胰岛素样生长因子-Ⅰ对软骨细胞凋亡的抑制

目的:探讨胰岛素样生长因子-Ⅰ(IGF-Ⅰ)对软骨细胞增殖及白细胞介素-1(IL-1)诱导软骨细胞凋亡的影响,揭示其抗损伤作用机制,为关节软骨损伤治疗提供理论依据。方法:分离培养人胚胎关节软骨细胞,采用四氮甲基唑蓝(MTT)法测定不同含量软骨细胞增殖活性的变化,利用光镜、电镜、DNA电泳及流式细胞仪测定作为凋亡检测指标。结果:IGF-Ⅰ呈剂量依赖式促软骨细胞增殖,当IGF-Ⅰ含量达50μg/L时,促软骨细胞增殖作用达最大值。IL-1组光镜、电镜下可见典型的细胞凋亡形态学改变,琼脂糖凝胶电泳示特征性的DNA梯状条带,IGF-Ⅰ处理组未见明显凋亡征象。流式细胞仪检测发现,IGF-Ⅰ处理后软骨细胞凋亡率显著降低。结论:IGF-Ⅰ能促进软骨细胞增殖,对IL-1诱导的软骨细胞凋亡具有保护作用。     

THE EFFECT OF ACUPUNCTURE ON PREVENTION AND TREATMENT OF ISCHEMIC ARRHYTHMIA AND THE RELATION BETWEEN EFFECT OF ACUPUNCTURE AND CHOLINERGIC MUSCARINE-LIKE RECEPTORS

For investigating the effect of acupuncture on ischemic arrhythmia and its mech-anism, adult albino rats with ligated anterior descending branch of coronary artery as experimentalmodel were treated with or without acupuncture, and others with imitative operation but without bothcoronary artery ligation and acupuncture treatment were used as control. It was found in acupuncturegroup that the fibrillation-liability of ischemic myocardium was efficiently decreased, the affinity ofAch-M receptors on membranes of ischemic myocardium was markedly increased, and the tolerance ofischemic myocardium to atropine was elevated in the experiment of atropine inducing fibrillation.These results indicate that acupuncture may play a therapeutic role on ischemic arrhythmia throughactivating the activity of muscarine-like receptors of cholinergic nervous system.     

严重肺挫伤早期肺泡灌洗液表面活性蛋白变化与肺功能及形态学改变的关系

目的探讨严重肺挫伤及缺血再灌注后早期肺泡灌洗液（BALF）中表面活性蛋白（SP）-A、B、C、D动态变化与肺功能及肺形态学改变的关系。方法75只兔随机分为对照组（A）、肺挫伤组（B）及缺血再灌注组（C）。每组均为25只。检测各组0，1，2，3，4h肺功能变化及BAIF中SP—A、B、C、D含量百分比，进行组织病理学及电镜检查。结果与A组比较，B、C组sSP—A、B、C、D含量百分比明显减少（P〈0．01）；肺功能出现明显的损害且呈持续下降（P〈0．05）；肺泡Ⅱ型细胞电镜及病理学检查呈不同程度的炎症及变性改变。结论严重肺挫伤BALF中SP—A、B、C、D动态变化与肺功能及肺组织病理学改变一致。