胎羊体外循环中胎盘一氧化氮合酶的变化

目的 探讨胎羊体外循环中胎盘一氧化氮合酶（NOS）的变化。方法孕120-140d母羊8只，随机分为对照组和转流组，每组4只。对照组假手术，转流组运用离心泵和胎盘建立胎羊体外循环，转流30min。记录转流前、后胎羊平均动脉压、脐动脉流量和动脉血气值，计算胎盘血管阻力，检测胎羊血浆内皮素-1（ET-1）、一氧化氮（NO）和胎盘组织NOS活性，RT-PCR半定量分析eNOSmRNA转录状况。结果转流组胎羊体外循环结束后胎盘血管阻力上升，转流后2h胎盘NOS活性增强，胎盘eNOS转录水平增高（P〈0．05），与对照组比较差异有统计学意义（P〈0．05）。转流组胎羊血浆ET-1进行性增加，与对照组之间的差异也有统计学意义（P〈0．05）。两组之间血浆NO的变化没有显著差异。结论胎羊体外循环中胎盘组织NOS活性增强，不能降低胎盘血管阻力，NOS的变化可能是胎盘血管阻力增高的一种代偿。     

全层组织工程皮肤修复猪糖尿病体表溃疡的实验研究

目的：观察有活性细胞成分的组织工程皮肤在修复猪糖尿病体表溃疡中的效果。方法：体外构建有活性细胞成分的组织工程皮肤；采用6只2．5～3月龄约克猪建立糖尿病溃疡模型；彻底清创形成48个创面(直径50mm)，随机分为三组：A(组织工程皮肤)组，B(自体皮肤)组，c(空白)组；术后1、2、3、4、6、8、12周观察皮片存活率、创面收缩率及组织学变化。结果：A、B组移植后皮片与创面紧密贴合、生长良好；A、B组皮片存活率无明显差别(P=O．667)；A、B组收缩率与C组有明显差别(P＜O．01)；A组皮片可见表皮和真皮两种成分，且两种成分不断改建，12周时其组织学表现接近正常皮肤。结论：全层组织工程皮肤修复糖尿病体表溃疡是可行的。     

根据添加剂选择化妆品

在选购美容护肤品时,常会看到产品说明中注有添加各种特殊成分的内容,这些添加成分就是美容护肤品中的添加剂。添加剂的使用是使美容品具有某种营养作用或治疗效果,可以根据不同的添加成分制成不同功效、品种繁多的产品。比如维生素、人参皂甙、胎盘素等是人们耳熟能洋的添加剂,但还有许多添加剂以及它们的功效却鲜为人知。只有了解美容品中不同添加剂的不同功效,消费者才能根据自身的特殊需要选择适合自己的产品,下面介绍几种常见的化妆品添加剂。丝肽(Sile Peptide)是丝蛋白的分解物,由真丝中单独存在的氨基酸组成,因而主要由天然真丝经水解获取。它具有较强的保温作用并含有人体所需的氨基酸,能使皮肤和毛发保持光     

超脉冲CO2激光与美容外科手术治疗睑黄瘤的疗效比较

目的：比较超脉冲CO2激光与美容外科手术治疗睑黄瘤的美容效果.方法：2003年1月至2004年3月采用美容外科手术和超脉冲CO2激光治疗106例（161个）睑黄瘤,其中手术治疗65例（98个）,激光治疗41例（43个）.分别于术后3个月及6～9个月,根据伤口愈合后的残存瘤体、瘢痕增生、色素异常三个指标进行评分比较.结果：美容手术组的复发率和瘢痕增生低于激光治疗组（均为P＜0.05）,而术后色素异常的差异无显著性（P＞0.05）.术后3个月及6～9个月的总满意率分别为：激光治疗组71.43%,65.22%;美容手术组90.82%,93.10%.激光治疗组与美容手术组的总满意率比较：3个月x2=8.9848,P＜0.05;6～9个月x2=4.7519,P＜0.05.同组病例在不同时期的总满意率比较差异无显著性（均为P＞0.05）.结论：美容外科手术比超脉冲CO2激光治疗睑黄瘤更能使近期美容疗效达到最佳的效果.     

当归水溶性多糖的分离、纯化及结构初步分析

采用80℃热水提取，得到当归水溶性多糖W-ASP。经阴离子交换柱层析、凝胶过滤柱层析对其进行纯化分级。实验结果表明：W-ASP11主要由葡萄糖组成，相对分子质量约为380000；W-ASP12主要由半乳糖和阿拉伯糖组成，相对分子质量约为19000；W-ASP2和W-ASP3主要舍半乳糖、阿拉伯糖和鼠李糖以及少量葡萄糖和甘露糖，并含有较高的糖醛酸，由单糖组成及红外图谱初步分析，可判断是一种果胶类多糖。当归多糖主要组分W-ASP3经凝胶色谱（GPC）鉴定为均一组分，其相对分子质量约为62000。     

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目的：比较肠外营养与肠内营养对胃癌病人术后细胞免疫功能的影响。方法：45例胃癌病人随机分为2组，其中肠外营养（PN）组23例，肠内营养9EN）组22例。术后第1天开始行肠外或肠内营养支持，时间1周，检测术前和术后第8天外周血T淋巴细胞亚群（CD3^ 、CD4^ 、CD8^ ）和NK细胞活性等细胞免疫指标。结果：PN组术后CD3^ 、CD4^ 水平和NK细胞活性明显下降，CD8^ 水平明显升高；而EN组术后CD3^ 、CD4^ 、CD8^ 水平和NK细胞活性与术前相比无显著性变化。结论：肠外营养对胃癌病人术后细胞免疫功能无明显改善作用，而肠内营养有明显的细胞免疫增强作用，可有效地纠正胃癌病人术后细胞免疫抑制状态。     

Bcl-2过度表达对大鼠成骨细胞Bax、Caspase-3表达的影响

目的：探讨凋亡相关基因Bcl-2过表达对大鼠成骨细胞Bax蛋白以及活性Caspase-3表达的影响。方法：将含有Bcl-2cDNA的逆转录病毒表达载体pDOR-SB质粒转染大鼠成骨细胞使之过表达，用免疫组化ABC法检测Bax、Caspase-3活性蛋白表达变化。结果：转染筛选后的阳性克隆细胞表现为Bcl-2表达显著增强。同时Bax蛋白表达下降接近显著性差异，活性Caspase-3表达无明显变化。结论：Bcl-2过度表达对活性Caspase-3表达无影响，但对Bax蛋白表达起下调作用，提示Bcl-2家族成员之间可以相互影响。     

基于网络药理学和分子对接技术探讨益气养阴汤治疗甲状腺癌术后的作用机制

目的：利用网络药理学和分子对接技术探讨益气养阴汤治疗甲状腺癌术后的作用机制。方法：通过本草组鉴（HERB）数据库检索益气养阴汤10味药物化学成分,通过PubChem、SwissADNE筛选药物活性成分,Swiss target prediction预测活性成分作用靶点。在Gene Cards、OMIM、DisGeNET、Drug Bank数据库检索获得甲状腺癌术后的疾病靶点。Venny2.1.0平台获得药物与疾病的交集靶点,使用String数据库构建交集靶点蛋白互作网络,并导入Cytoscape3.9.1筛选得到核心靶点蛋白;借助Cytoscape3.9.1构建药物-活性成分-靶点网络图,并获取关键活性成分;使用Metascape数据库进行GO和KEGG富集分析;对关键靶点和关键成分进行分子对接验证。结果：得到益气养阴汤活性成分157种,预测活性成分作用靶点507个,甲状腺癌术后相关疾病靶点1817个,药物活性成分—疾病交集靶点154个。PPI分析得到SRC、HSP90AA1、PIK3R1、PIK3CA、AKT1为关键靶点基因,槲皮素、山柰酚、木犀草素为关键活性成分,分子对接显示关键靶...     

玉米须生物活性成分的初步研究──玉米须多糖及其免疫增强作用

经过系统的筛选首次从玉米须中分离提取了具有免疫增强作用的生物活性成分。运用离子交换、分级沉淀、Ｓｅｐｈｒｏｓｅ柱层析等手段对此成分进行了进一步的分离和纯化。初步的结构研究结果表明，此成分属多糖类化合物。     

整合素β1在微粒皮混合移植中的异位表达及定量分析

目的 了解自、异体微粒皮混合移植促进创面愈合的机制. 方法 建立自、异体微粒皮混合移植大鼠模型.第1部分实验分为3组,每组9只大鼠:(1)自体皮组,移植面积扩张比为10:1的自体微粒皮;(2)混合1组,移植自、异体微粒皮,面积扩张比均为10:1;(3)混合2组,移植自、异体微粒皮,面积扩张比分别为10:1和10:3.第2部分实验亦分为3组,每组6只大鼠:(1)自体皮组,移植面积扩张比为20:1的自体微粒皮;(2)混合1组,移植自、异体微粒皮,面积扩张比分别为20:1、20:3;(3)混合2组,移植自、异体微粒皮,面积扩张比分别为20:1和20:6.术后从各组大鼠愈合创面取样,第1部分实验于术后2、3、4周取样,第2部分实验于术后3、4周取样,每组大鼠各时相点检测3个样本.行常规组织学与免疫组织化学染色,测定整合素β1的表达.显微镜下测量表皮层厚度. 结果 (1)HE染色显示,术后各组大鼠创面的表皮层厚度明显增加,真皮内有不同程度的血管扩张和单个核细胞浸润.(2)术后2～4周,第1部分实验中2个混合组大鼠皮肤表皮层均明显厚于自体皮组(P＜0.05或P＜0.01);术后3、4周,第2部分实验中2个混合组大鼠皮肤表皮层厚度均明显厚于自体皮组(P＜0.05或P＜0.01).(3)免疫组织化学染色显示,术后各组新生表皮中均可见整合素β1阳性细胞,以棘层和颗粒层为主.术后2周,第1部分实验中混合组整合素β1的阳性表达明显强于自体皮组(P＜0.01),且混合1组的表达(10 982±2169)明显强于混合2组(4240±512,P＜0.01);术后3～4周,混合1组的表达仍明显强于自体皮组和混合2组(P＜0.01).第2部分实验仅在术后3周时混合2组整合素β1的阳性表达(1618±171)明显高于自体皮组(1060±146,P＜0.05). 结论 自、异体微粒皮混合移植中,整合素β1的异位表达和表达增强与表皮细胞的增殖分化、创面冉上皮化以及表皮层增厚有密切关系,在促进创面愈合过程中,整合素β1阳性表达细胞很有可能发挥了重要作用.     

大叶紫薇叶中降血糖活性成分的筛选

为筛选大叶紫薇叶中具有降血糖活性的成分,采用3T3-L1细胞葡萄糖消耗模型作为检测手段,对大叶紫薇叶提取物采用HP-20树脂吸附、溶剂萃取、制备薄层分离和制备高效液相分离,导向筛选具有降血糖作用的各分离组分.结果发现,大叶紫薇叶中corosolic acid、熊果酸和总三萜具有降血糖活性.     

ISOLATION AND PURIFICATION OF A HIGH MOLECULAR WEIGHT SUBSTANCE FROM HUMAN URINE WHICH INHIBITS CALCIUM OXALATE CRYSTAL GROWTH

A high molecular weight inhibitor of calcium oxalate crystal growth in human urine was investigated. Three inhibitors were isolated by DEAE-Sephacel ion-exchange chromatography and, of these, the substance we named Peak 3 protein seemed to be the main inhibitor in human urine. Peak 3 protein was purified by fast protein liquid chromatography and polyacrylamide gel electrophoresis. This substance, with a molecular weight of 30 kDa, did not contain uronic acid and its inhibitory activity decreased after digestion with proteinase. The difference between Peak 3 protein and several inhibitors previously reported was investigated but no clear difference could be found. The fact that it was the protein structure which was responsible for the inhibitory activity and the fact that Peak 3 protein probably possessed many side-chains which did not contribute to the inhibitory activity influenced the outcome of the investigation.     

Identification of a prostate inhibitory substance in a pollen extract

Recently, much attention has focused on the treatment of BPH with the pollen extract, Cernilton. The present investigation was designed to identify the active component in this agent which might be responsible for the symptomatic relief of BPH as previously reported [1,2]. Sequential purification of the active component present in the pollen extract was carried out by a combination of dialysis, gel filtration, and reverse phase chromatography. To monitor the biological activity of each of the purified fractions, a biological assay employing the human prostate cancer cell line DU145 was undertaken. While we have identified a number of constituent components in the pollen extract, only one fraction designated V-7 (FV-7) maintained a strong inhibitory effect on the growth of DU145 cells. The inhibition was time- and dose-dependent, and the concentrations of FV-7 required to reduce the cell numbers by 50% (IC₅₀) after 2 days of exposure was 5 μg/ml. FV-7 was also inhibitory towards the primary culture of prostate stroma and epithelial cells, with the stroma/fibroblast showing greater sensitivity towards the HPLC-purified component. However, it should be noted that this inhibitory activity measured in the primary culture cells was only achieved at higher concentrations of FV-7. Preliminary characterization of the active ingredient identified FV-7 as DIBOA which is a cyclic hydroxamic acid. FV-7 and DIBOA induce similar inhibitory effects on the growth of DU145 cells.     

Purification and biochemical characterization of reno-ferredoxin from bovine kidney mitochondria

Bovine reno-ferredoxin was purified from kidney mitochondria by an improved method that included hydrophobic and ion-exchange chromatography on Toyopearl gels. The optical absorption spectrum of the oxidized reno-ferredoxin revealed two peaks, at 414 and 455 nm in the visible region. The minimum molecular weight of the ferredoxin was 12,900 Da by SDS-polyacrylamide gel electrophoresis. The amino acid residues of the NH2-terminal sequence of the ferredoxin were investigated using by a gas-phase sequencer. Bovine reno-ferredoxin and adreno-ferredoxin showed almost identical NH2-terminal amino acid sequences. Reconstitution of the 25-hydroxyvitamin D3-1 alpha -hydroxylase system was performed with the following three components: NADPH-ferredoxin reductase from bovine kidney mitochondria, reno-ferredoxin, and cytochrome P-450(D1 alpha) from bovine kidney mitochondria. The results demonstrated that the reno-ferredoxin was essential for the 1 alpha-hydroxylase activity of 25-hydroxyvitamin D3.     

人生长激素基因的克隆、高效表达及纯化

目的 研究人生长激素(hGH)基因在原核中的高效表达、纯化及鉴定。方法 设计带双酶切位点引物,采用逆转录-聚合酶链反应(RT-PCR)方法获得hGH cDNA片段,并亚克隆入pUC19质粒中进行 DNA序列测定。将 hGH cDNA片段克隆入原核表达载体 pBV220中进行表达。表达产物经 7mol/L 盐酸胍裂解变性、复性、层析等一系列纯化研究,纯化产物经SDS-PAGE电泳,高压液相色谱法(HPLC)纯度分析及活性测定,并通过N末端氨基酸序列测定鉴定表达产物。结果 RT-PCR扩增所得片段大小与预期值一致。pBV220-hGH表达产物经SDS-PAGE电泳显示,分子量为 22×10~3与预计结果相符。rhGH表达量占菌体总蛋白量的 40％。表达产物纯化后,纯度达 97％以上,比活性大于 3.0 IU/mg。纯化产物经 N末端 15个氨基酸测定验证,为重组人生长激素。结论 成功构建了 pBV220-hGH原核表达质粒,并在大肠杆菌中高效表达,表达产物经纯化得到纯度≥97％的 rhGH,比活性大于 3.0 IU/mg。该研究为 rhGH的中试生产奠定了基础。     

Resolution of proteins in the kidney stone matrix using high-performance liquid chromatography

The organic matrix accounts for 2-3% of the total stone weight and has been considered to play a role in stone formation and growth. Thus far, fractionation of the matrix proteins has been insufficient due to low resolution and reproducibility. In this report the matrix proteins of 22 stones were resolved by means of high-performance liquid chromatography. Following pulverization, the organic matrix was obtained by dialysis against EDTA. The average content of nondialyzable extractable proteins was 1.6% of the total stone weight. Analysis of the matrix proteins with high-performance gel permeation liquid chromatography and high-performance ion-exchange liquid chromatography has indicated that the protein composition of the stone matrix is identical regardless of the mineral composition. The major component of the matrix proteins was identified as glycoprotein and/or proteoglycan from their absorption to a concanavalin A Sepharose column. Higher molecular weight matrix proteins seem to be polymers or condensation products, since they have been degraded into lower molecular weight subfractions by sodium dodecyl sulfate treatment.     

孕妇抗磷脂抗体、胰岛素样生长因子-I与胎儿生长受限的相关性研究

目的：探讨胎儿生长受限（FGR）孕妇抗心磷脂抗体（ACA）与胎盘组织胰岛素样生长因子-I（IGF-I）表达水平间的关系及两者在FGR发生中的作用。方法采用酶联免疫吸附法（ELISA法）测定63例正常足月妊娠孕妇（对照组）及63例胎儿生长受限孕妇（研究组）血清中ACA-IgG、IgM;利用免疫组织化学方法及多媒体彩色病理图像分析技术测定胎盘组织IGF-I表达水平。结果研究组抗心磷脂抗体阳性率为19.0%,对照组为3.2%,两组比较,有显著性差异（P＜0.05）;研究组胎盘组织IGF-1表达水平明显低于对照组（P＜0.05）;ACA阳性组与ACA阴性组两组比较,胎盘组织IGF-I表达水平无显著差异（P＞0.05）,胎盘重量、胎儿体重有显著差异（P＜0.05）。结论孕妇血清ACA阳性或胎盘组织IGF-I表达水平下降是导致FGR的危险因素之一,ACA是否也会导致胎盘组织IGF-I表达水平下降有待进一步研究。     

Enhancement of new bone formation by hematoma at fracture site.

In order to study the osteogenetic potential of fracture hematoma, pellets which contain fluid components extracted from human fracture hematoma were transplanted to the subperiosteum of the rat parietal bone. Furthermore we have examined the effect of the fluid extract of the fracture hematoma and factors partially purified from this extract on DNA synthesis of cultured osteoblast-like cell line MC3T3E1. Implantation of the pellets containing the fluid extract did not promote new bone formation regardless of the presence of periosteum. This extract, in contrast, enhanced DNA synthesis of osteoblast-like cell line MC3T3E1 in a concentration dependent fashion. When this fluid extract was partially purified by ion exchange chromatography, the highest peak in DNA synthesis of MC3T3E1 was attained at 38mM NaCl, and the second highest peak at 290mM. These results suggest that the soluble extract from fracture hematoma contains growth factors which promote osteoblast proliferation, but does not possess osteogenetic potential.     

Specific growth factor activity identifies and predicts murine mammary tumor

In milk from substrains of mice with varying incidences of developing mammary tumor, we have isolated a specific mitogenic activity, which is unique in that it identifies mice with mammary tumor and predicts those mice that will eventually develop mammary tumor. None of the milk samples from control mice, who never developed mammary tumor, contained this specific predictive mitogenic activity. Chemical characterization has shown this specific mitogenic activity to be acid- and heat-stable and resistant to reducing agents. Partial purification, by ion-exchange, high-performance liquid chromatography size-exclusion, and isoelectrofocusing techniques, of this specific mitogenic activity from milk of mice that had or eventually developed mammary tumor identifies several peptide growth factor components in a 6-10 kDa molecular weight range. Of known growth factors, radioassay techniques identify an insulin-like growth factor-1-like peptide as a major component. Small amounts of platelet-derived growth factor and transforming growth factor-beta activities also were present. Our results suggest that a subset of growth factors that are diagnostic of the presence of murine mammary tumor and predictive of eventual tumor development may be early indicators of the transition of a competent cell to a progressively malignant state. Similar studies of a secreted body fluid from women at risk for breast cancer may lead to the identification of a specific biologic tumor marker for breast cancer.     

Systematic purification of free and matrix-bound phosphophoryns of bovine dentin: Presence of matrix-bound phosphophoryn as a distinct molecular entity

Summary Free and matrix-bound phosphophoryns, both highly phosphorylated proteins in dentin, were prepared from EDTA extract and CNBr-digests of bovine dentin. The two components were purified by DEAE-cellulose, SP-Sephadex, and gel filtration chromatography. The matrix-bound component was eluted as a distinct peak from the free component in the above chromatographic systems. Amino acid composition of the purified matrixbound component indicated that this component consisted of phosphophoryn and collagen in the ratio of 2:3 based on the number of the residues. The matrix-bound component could not be reconstituted by mixing phosphophoryn with collagen CNBr peptides. Artificial crosslink products of free phosphophoryn and collagen CNBr-peptides by the carbodiimide method showed similar properties to the physiological matrix-bound phosphophoryn. The bond between phosphophoryn and collagen of the matrix-bound component is assumed to be a covalent crosslink.