Correction of abnormal cerebroside sulfate metabolism in cultured metachromatic leukodystrophy fibroblasts

Cultured fibroblasts derived from patients with late infantile metachromatic leukodystrophy incorporated arylsulfatase A from the growth medium. Upon exposure to cerebroside sulfate, they exhibited patterns of uptake and hydrolysis indistinguishable from cells derived from control subjects. Furthermore, inclusion granules formed in the metachromatic leukodystrophy fibroblasts upon exposure to sulfatides were cleared by subsequent supplementation of the growth medium with arylsulfatase A.     

Genotoxicity of formaldehyde in cultured human bronchial fibroblasts

Formaldehyde, a common environmental pollutant, inhibits repair of O6-methylguanine and potentiates the mutagenicity of an alkylating agent, N-methyl-N-nitrosourea, in normal human fibroblasts. Because formaldehyde alone also causes mutations in human cells, the compound may cause genotoxicity by a dual mechanism of directly damaging DNA and inhibiting repair of mutagenic and carcinogenic DNA lesions caused by other chemical and physical carcinogens.     

Diploid azaguanine-resistant mutants of cultured human fibroblasts

Two azaguanine-resistant clones of cultured, human fibroblasts were isolated from unrelated strains of karyotypically normal, male cells. The most resistant mutant has little hypoxanthine-guanine phosphoribosyltransferase activity, is virtually unable to incorporate hypoxanthine (a normal substrate of the enzyme), and resembles fibroblasts cultured from boys with the Lesch-Nyhan syndrome. The less resistant mutant has about one-third as much enzyme activity as its parent strain and is less able to utilize hypoxanthine. Both mutants are morphologically and karyotypically normal. These mutations may have occurred at the X-chromosomal, hypoxanthine-guanine phosphoribosyltransferase locus and may provide a realistic experimental model for studying mutation in human genetic material.     

Molecular mechanisms of the biological clock in cultured fibroblasts

In mammals, the central circadian pacemaker resides in the hypothalamic suprachiasmatic nucleus (SCN), but circadian oscillators also exist in peripheral tissues. Here, using wild-type and cryptochrome (mCry)-deficient cell lines derived from mCry mutant mice, we show that the peripheral oscillator in cultured fibroblasts is identical to the oscillator in the SCN in (i) temporal expression profiles of all known clock genes, (ii) the phase of the various mRNA rhythms (i.e., antiphase oscillation of Bmal1 and mPer genes), (iii) the delay between maximum mRNA levels and appearance of nuclear mPER1 and mPER2 protein, (iv) the inability to produce oscillations in the absence of functional mCry genes, and (v) the control of period length by mCRY proteins.     

Mutant enzymatic and cytological phenotypes in cultured human fibroblasts

Fibroblasts were cultured from the cells of two children who shared some characteristics of Hurler syndrome, but they did not show corneal clouding and excessive excretion of mucopolysaccharides. The fibroblasts differ from those of controls and of patients with typical Hurler syndrome or other mucopolysaccharidoses in that they have abundant cytoplasmic inclusions, striking diminutions in beta-glucuronidase, and elevations in acid phosphatase.     

鲟鱼硫酸软骨素制备工艺优化

为制备高纯度鲟鱼硫酸软骨素,利用鲟鱼头骨和脊骨为原料,经过硫酸软骨素提取专用酶与碱性蛋白酶联合酶解新工艺,并进一步经季铵盐纯化、乙醇沉淀,制得硫酸软骨素纯品。硫酸软骨素提取专用酶和碱性蛋白酶提取的最佳工艺为:硫酸软骨素提取专用酶用量15mg/g,酶解温度55℃,酶解时间4h;碱性蛋白酶用量50mg/g,酶解温度60℃,酶解时间2h。季铵盐-乙醇沉淀法纯化最佳工艺为:季铵盐质量浓度1g/100mL、作用温度20℃,作用时间1.5h;乙醇沉淀体积分数为75%、溶液pH为7。该工艺制得的鲟鱼头骨、脊骨硫酸软骨素纯品的产率分别为19.5%、22.0%,纯度分别为94.80%、95.32%,蛋白含量分别为0.287%、0.267%。     

5-hydroxytryptamine: a cytospecific growth stimulator of cultured fibroblasts

5-Hydroxytryptamine (serotonin) in micromolar amounts increased the growth of fibroblasts in culture while not affecting five other cell lines. Serotonin appeared to shorten the lag phase of cell growth. The effect was less when serotonin was added to the fibroblast culture after the initial 24-hour period. The two functional groups of the serotonin molecule were required for growth enhancement. Serotonin in millimolar concentrations was toxic to fibroblasts.     

Decrease in free cystine content of cultured cystinotic fibroblasts by ascorbic acid

The 100-fold increase in free cystine content characteristic of cultured skin fibroblasts from patients with nephropathic cystinosis was decreased more than 50 percent by addition of L-ascorbic acid to the culture medium at concentrations of 0.29 to 2.9 millimolar. Fresh ascorbic acid must be added to the culture medium daily to produce a progressive decrease of the free cystine content of the cells over a 3-day period. Upon removal of ascorbic acid from the medium, the free cystine content returns to its initial value.     

Hurler and Hunter syndromes: mutual correction of the defect in cultured fibroblasts

The biochemical defect of cultuired skin fibroblasts from Hurler or Hunter patients (faulty degradation of sulfated mucopolysaccharide, resulting in excessive intracellular accumulation) may be corrected if cells of these two genotypes are mixed with each other or with normal cells. The effect is mediated by substances released into the medium.     

Reactivity in vivo of the carcinogen N-hydroxy-2-acetylaminofluorene: increase by sulfate ion

Injections of sulfate ion in rats given the carcinogen N-hydroxy-2-acetylaminofluorene increased (i) the formation of 1-and 3-(methion-S-yl)-2-acetylaminofluorene bound to protein in the liver, (ii) the formation of fluorenyl derivatives bound to total protein, ribosomal RNA, and DNA in the liver, and (iii) the toxicity of the carcinogen. These data provide evidence that the highly reactive ester 2-acetylaminofluorene-N-sulfate, previously suggested as an ultimate reactive and carcinogenic metabolite of N-hydroxy-2-acetylaminofluorene, is formed in the rat liver in vivo.     

Sanfilippo disease type B: enzyme replacement and metabolic correction in cultured fibroblasts

alpha-Acetylglucosaminidase, purified from human placent, corrected the defect in mucopolysaccharide degradation when added to culture fibroblasts from patients with Sanfilippo disease type B. A small cellular concentration of enzyme gave a large corrective effect. The half-life of disappearance of enzyme activity was 4 to 7 days.     

波长585 nm脉冲染料激光对成纤维细胞增殖以及其前胶原基因mRNA表达的影响

以不同能量密度的脉冲染料激光照射体外培养的正常皮肤的成纤维细胞,照射后继续培养24h,分别以MTT法检测其增殖功能,以定量PCR(LightCycler)法检测其细胞内不同类型的前胶原基因及SMADs mRNA的表达.结果显示能量密度为5 J/cm2和7 J/cm2的脉冲染料激光能抑制体外培养的正常皮肤的成纤维细胞的增殖功能(P＜0.05),在能量密度为5 J/cm2时能显著抑制Ⅰ型前胶原基因α1(P＜0.01)以及SMAD 2,3,4,7 mRNA的表达(P＜0.05或P＜0.01),下调Ⅰ型前胶原基因α1与Ⅲ型前胶原基因mRNA的比率.在能量密度为6,7 J/cm2时能抑制Ⅲ型前胶原基因α1mRNA的表达(P＜0.05),而对Ⅰ型前胶原基因α1,α2的表达与对照无显著性差异.提示波长585 nm脉冲染料激光对成纤维细胞的增殖及其胶原产生的某些相关基因的表达具有直接的作用,且与能量密度有关.     

Hurler's syndrome: demonstration of an inherited disorder of connective tissue in cell culture

Skin fibroblasts from three patients with Hurler's syndrome were grown in tissue culture and shown to contain metachromatic granules when stained for mucopolysaccharides with toluidine blue O. Similar inclusions were observed in cultures of fibroblasts from other members of the families, who appeared to be clinically normal but who were, judged from studies of pedigree, heterozygous or hemizygous for the abnormal gene.     

玉米幼芽提取物对小鼠皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达的影响

分离培养小鼠皮肤成纤维细胞,用2种富含腺嘌呤类衍生物的玉米幼芽提取物(20 mg/L)提前保护细胞后,体外构建H2O2急性氧化损伤细胞模型,应用RT-PCR对小鼠皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA的表达进行半定量检测。结果表明,损伤对照组(Ⅱ组)小鼠皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA的表达量分别为空白对照组(I组)的24%(P<0.01)和20%(P<0.01),而药物保护组(Ⅲ组、Ⅳ组)小鼠皮肤成纤维细胞Ⅰ型前胶原mRNA的表达量均高于损伤对照组,分别是损伤对照组的3.5倍(P<0.01)和1.8倍,Ⅲ型前胶原mRNA的表达量也均高于损伤对照组,分别是损伤对照组的4.6倍(P<0.01)和3倍(P<0.01)。说明在H2O2致小鼠皮肤成纤维细胞急性氧化损伤过程中,这2种玉米幼芽提取物能够保护Ⅰ、Ⅲ型前胶原mRNA的表达,且0501提取物对小鼠皮肤成纤维细胞Ⅰ、Ⅲ型前胶原表达的保护作用优于0502提取物。     

微生物法测定维生素饮料中的维生素B_6

[目的]建立维生素饮料中维生素B6的微生物检测法。[方法]利用卡尔斯伯酵母菌(ATCC 9080)对维生素B6极高的灵敏性和特异性,定量测定出试样中维生素B6含量。即以不同浓度标准溶液的吸光度相对于各浓度水平标准物质的浓度绘制标准曲线,再根据标准曲线计算出试样中维生素B6的含量。[结果]试验方法精密度分别为1.5%、1.1%及1.5%,相对标准偏差为0.788 7%,回收率在96.5%以上。[结论]微生物法可以作为维生素饮料中维生素B6的检测方法,且灵敏度高、测定结果准确。     

硫酸钾在农作物上的施用技术

推广应用硫酸钾施用技术,是实施平衡施肥技术的重要内容之一。一是提高农作物产量改善农产品品质,保持和提高土壤肥力的重要措施;二是有利于优化化肥资源配置,提高肥料利用率;三是降低生产成本,提高经济效益;四是可以避免和减轻因单一和不合理施肥带来的环境污染。九五期间我市     

维生素E在反刍动物上的应用

维生素E具有很强的抗氧化作用，也是细胞呼吸的必需因子，参与体内DNA、维生素C和辅酶Q的合成。此外，维生素E还与动物的生殖机能、免疫机能密切相关。日粮中添加维生素E可以缓解鲜肉和冻肉失色速度，维持展示期内肉的持水力，从而延长牛肉与羊肉的货架期；同时维生素E可以控制奶牛乳房炎的发生，延长牛奶的贮藏时间，改善牛奶的质量维生素E还可以提高羔羊的成活率。     

HPLC法检测复合维生素片中叶酸和维生素B_(12)的含量

[目的]建立应用高效液相法测定复合维生素片中叶酸和维生素B12含量的方法。[方法]色谱柱用菲罗门lunaC18(4.6×250 mm,5μm),流动相为甲醇-0.028 mol/L磷酸氢二钠水溶液(用磷酸调pH到3.5)(21∶79),流速1.0 ml/min,检测波长为361 nm,进样量为20μl,柱温为25℃。[结果]维生素B12在6.65～66.50 ng范围内呈现良好的线性关系,回归方程是y=0.495 1x+1.342 8(R2=0.999 4,n=6),平均回收率为96.83%,RSD为0.82%(n=6)。叶酸在80～800 ng范围内呈现良好的线性关系,回归方程是y=0.021 1 x+0.661 1(R2=0.998 8,n=6),平均回收率为95.56%,RSD为1.54%(n=6)。[结论]高效液相色谱法灵敏度高,操作简便,重现性好,结果准确,可以用于复合维生素片中叶酸和维生素B12的含量测定。     

硫酸头孢喹肟混悬液在猪体内的药物动力学研究

以健康猪为试验动物,按随机交叉设计试验,肌肉注射头孢喹肟混悬液和静脉注射头孢喹肟溶液,研究其在猪体内的药物动力学。结果表明:肌肉注射的主要药物动力学参数,分布半衰期(T1/2α)为1.33 h,消除半衰期(T1/2β)为4.92 h,表观分布容积(Vd)为0.34 L.kg-1,曲线下面积(AUC)为17.22μg.h.mL-1,达峰时间(tmax)为0.83 h,峰浓度(cmax)为3.36μg.mL-1,生物利用度(F)为93.87%。静脉注射的主要药物动力学参数,T1/2α为0.30 h,T1/2β为2.32 h,Vd为0.13 L.kg-1,AUC为18.35μg.h.mL-1。硫酸头孢喹肟混悬液在健康猪体内的药物动力学特征可归纳为:肌注吸收迅速,达峰时间短,生物利用度高,有效血药浓度可维持24 h。