Co-immobilized aerobic/anaerobic mixed cultures in stirred tank and gaslift loop reactors

Co-immobilized Aspergillus awamori and Zymomonas mobilis cultures were investigated in a stirred tank reactor on synthetic medium with starch as substrate at various dissolved oxygen concentrations. In a gaslift loop reactor, freely suspended and immobilized A. awamori were cultivated on synthetic medium and soluble potato starch. In the same reactor, the growth and ethanol production of freely suspended and immobilized Z. mobilis cultures were studied on synthetic medium and glucose. Co-immobilized A. awamori and Z. mobilis were cultivated in batch and continuous operations in the gaslift loop reactor on synthetic medium with starch substrate at different dissolved oxygen concentrations. The interrelations between the different process variables are discussed.     

Acetoin and Phenylacetylcarbinol Formation by the Pyruvate Decarboxylases of Zymomonas Mobilis and Saccharomyces Carlsbergensis Stephanie Bringer-Meyer

Cell suspensions of Zymomonas mobilis and Saccharomyces carlsbergensis and the pyruvate decarboxylases from the two organisms were compared with respect to their efficiencies of acyloin formation. Although Z. mobilis contained five times more pyruvate decarboxylase activity than yeast, sugar-fermenting suspensions of Z. mobilis produced, in the presence of benzaldehyde, 4-5 times less phenylacetylcarbinol than the yeast. The pyruvate decarboxylases of both organisms catalyzed acetoin and phenylacetylcarbinol synthesis from pyruvate and acetaldehyde or benzaldehyde, but the affinity of the Z. mobilis pyruvate decarboxylase towards the aldehyde reactants was lower than that of the yeast enzyme. Because of the limited solubility of benzaldehyde, neither enzyme could be saturated with this substrate for phenyl-acetylcarbinol synthesis. Studies with 2-toluidinonaphthalene-6-sulfonate and substrate analogues showed that the catalytic sites of pyruvate decarboxylase from Z. mobilis were less lipophilic than those of the enzyme from yeast. This difference could explain the lower affinity for benzaldehyde of the Z. mobilis enzyme.     

Hydrophobicity of bacteria Zymomonas mobilis under varied environmental conditions

Changes in the cell surface hydrophobicity (CSH) of bacteria Zymomonas mobilis 113S were examined in response to varied environmental conditions (temperature and phase of growth, concentration or type of carbon source, the presence of amphiphilic compounds). The values of CSH were elevated with a decreased growth rate over the time of cultivation up to 20–22% at the stationary phase. CSH values increased proportionally with the growth of cultivation temperature and concentration of carbon source (glucose or sucrose) or amphiphilic compound (aliphatic alcohols, Tween80) in the medium. Replacement of sucrose by glucose and the presence of Tween20 in the growth medium resulted in reduced values of CSH. An inverse relationship was detected between the number of attached cells to the hydrophilic glass surfaces and the CSH values of Z. mobilis whereas direct linear relationship was observed for hydrophobic surfaces. Permeation rates of the fluorescent probe (NPN) into the cells were directly proportional to the concentration of extracellular protein in the medium and to the values of CSH indicating the impaired barrier function for more hydrophobic cells. The multiple correlation between the CSH values and absorption indices of FT-IR spectra at the fingerprint region (866–1088 cm⁻¹) suggests the possible contribution of carbohydrates and/or lipopolysaccharides in observed changes of Z. mobilis hydrophobicity.     

Cloning and expression of the Zymomonas mobilis pyruvate kinase gene in Escherichia coli

The homotetrameric pyruvate kinases (PK) constitute a fine example of allosteric enzymes subjected to sophisticated regulatory mechanisms. We have cloned and sequenced the Zymomonas mobilis structural gene for the first prokaryotic dimeric PK, as an initial step toward understanding the peculiar properties of this enzyme. The deduced amino acid sequence of the pyk gene consists of 475 residues with a calculated molecular mass of 51.4 kDa and exhibits up to 50% sequence identity with other PKs. Heterologous expression in Escherichia coli was not obtained from the native promoter, but only when the pyk gene was under the control of a strong inducible promoter when a ribosome-binding site was present upstream of the putative TTG start codon of the pyk gene. Kinetic characterization of PK in concentrated crude cell extracts showed that the enzyme is not activated by sugar phosphates or AMP but is slightly inhibited by ATP. Thus, PK of Z. mobilis is unique among the characterized prokaryotic PKs due to its high activity in the absence of any allosteric activator. Amino acid sequence alignments revealed that glutamate 381 may play a role in ineffective binding of the usual PK activator, fructose-1,6-bisphosphate.     

Lysozyme for capture of microorganisms on protein biochips

Lysozyme placed on the SiO₂ surfaces that have previously been derivatized with C₁₈ coating will capture both Escherichia coli and Listeria monocytogenes cells from PBS buffer at pH 7.2. This phenomenon is of significance for the design and fabrication of protein biochips that are designed to capture bacteria from buffer or water so that these can be further interrogated with respect to possible pathogenicity. Fluorescent microscopy shows that two types of bacteria (gram-negative E. coli and gram-positive Listeria spp.) will be adsorbed by lysozyme placed on the surface of the biochip but that strong adsorption of the bacteria is reduced but not eliminated when Tween 20 is present (at 0.5%) in the PBS buffer in which the cells are suspended. In comparison, Tween 20 and Bovine Serum Albumin (BSA) almost completely block adsorption of these bacteria on C₁₈ coated surfaces. The combination of a lysozyme surface with Tween 20 gives a greater degree of adsorption of L. monocytogenes than E. coli, and hence suggests selectivity for the more hydrophobic E. coli may be reduced by the Tween 20. This paper presents protocols for preparing protein-coated, SiO₂ surfaces and the effect of buffer containing Tween 20 on adsorption of bacteria by SiO₂ surfaces coated with C₁₈ to which BSA, lysozyme or C11E9 antibody is immobilized at pH 7.2 and ambient temperature.     

大叶藻对氮磷营养盐的吸收动力学特征

营养盐是影响海草生长的关键因子, 目前有关海草不同组织对不同形式氮和磷的吸收特征尚不明确。该研究通过利用海草地上和地下组织分隔培养装置, 设置不同的氨态氮、硝态氮和磷酸盐浓度, 探究了大叶藻(Zostera marina)植株及其地上和地下组织对氮磷营养盐的吸收动力学特征。结果显示: (1)大叶藻对氮磷的吸收符合饱和吸收动力学特征, 吸收速率和水体氮磷浓度可用米式方程描述; (2)大叶藻植株对NH₄⁺-N的最大吸收速率(V_max, 52 μmol·g^-1·h^-1)显著高于其对NO₃^--N的V_max (39 μmol·g^-1·h^-1); (3)大叶藻地上组织和地下组织均可吸收氮磷, 但地上组织对氨态氮、硝态氮、磷酸盐的V_max分别为43.1、30.5和15.6 μmol·g^-1·h^-1, 为地下组织的2.6、1.2和6倍。结果表明, 大叶藻对氨态氮的吸收能力高于硝态氮, 且对氮磷的吸收可能主要依赖地上组织(叶片)。结果为查明大叶藻对氮磷的吸收利用机制及评估大叶藻的海洋生态效应提供了理论依据。     

Studies on the continuous production of (R)-(−)-phenylacetylcarbinol in an enzyme-membrane reactor

The optimization of a continuous enzymatic reaction yielding (R)-(−)-phenylacetylcarbinol ((R)-PAC), a key intermediate of the (1R,2S)-(−)-ephedrine synthesis, is presented. We compare the suitability of different mutants of the pyruvate decarboxylase (PDC) from Zymomonas mobilis with respect to their application in biotransformation using pyruvate or acetaldehyde and benzaldehyde as substrates, respectively. Starting from 90 mM pyruvate and 30 mM benzaldehyde, (R)-PAC was obtained with a space time yield of 27.4 g/(L·day) using purified PDCW392I in an enzyme-membrane reactor. Due to the high stability of the mutant enzymes PDCW392I and PDCW392M towards acetaldehyde, a continuous procedure using acetaldehyde instead of pyruvate was developed. The kinetic results of the enzymatic synthesis starting from acetaldehyde and benzaldehyde demonstrate that the carboligation to (R)-PAC is most efficiently performed using a continuous reaction system and feeding both aldehydes in equimolar concentration. Starting from an inlet concentration of 50 mM of both aldehydes, (R)-PAC was obtained with a space-time yield of 81 g/(L·day) using the mutant enzyme PDCW392M. The new reaction strategy allows the enzymatic synthesis of (R)-PAC from cheap substrates free of unwanted by-products with potent mutants of PDC from Z. mobilis in an aqueous reaction system.     

Uptake kinetics of nitrogen and phosphorus by Zostera marina

Aims Nutrient availability is an external factor that affect the growth of seagrasses. However, the demand for and absorption of different forms of nitrogen and phosphorus by different tissues of seagrasses are unclear. In this study, the uptake of nitrogen and phosphorus by Zostera marina was studied to determine the nutrient uptake kinetics. The objectives of this research are to: 1) investigate the absorption characteristics of ammonia nitrogen, nitrate nitrogen and phosphorus in Z. marina; 2) evaluate the differences in absorption between the different forms of nitrogen; and 3) analyse the differences in absorption between the different tissues of Z. marina.Methods Equipment was used to separate the aboveground and belowground tissues of Z. marina. Six concentration levels of ammonia nitrogen, nitrate nitrogen and phosphorus were established to experimentally test the uptake kinetics of nutrients by Z. marina. The nutrient concentrations in different parts of seawater column were measured to determine the nutrient changes and calculate the kinetic characteristics of nutrient uptake.Important findings 1) The absorption of ammonia nitrogen, nitrate nitrogen and phosphorus by Z. marina was consistent with the characteristics of saturated absorption kinetics. The relationship between the absorption rate and the nutrient concentrations in water could be described by the Michaelis-Menten equation. 2) The maximum absorption rate (V_max) of ammonia nitrogen by Z. marina (52 μmol·g^-1·h^-1) was significantly higher than that of nitrate nitrogen (39 μmol·g^-1·h^-1). 3) Both aboveground and belowground tissues of Z. marina could absorb nutrient, but the V_max of leaves (aboveground tissues) for ammonia nitrogen, nitrate nitrogen and phosphorus absorption were 43.1, 30.5 and 15.6 μmol·g^-1·h^-1, respectively, which were 2.6-fold, 1.2-fold and 6-fold higher than the corresponding V_max of belowground tissues. The results show that the absorption capacity of Z. marina for ammonia nitrogen is higher than that for nitrate nitrogen, and the absorption of nitrogen and phosphorus may depend primarily on the aboveground tissues (leaves). The results provide a theoretical basis for the study of the mechanisms of nitrogen and phosphorus uptake and utilization by Z. marina and the evaluation of marine ecological impacts.     

Synthesis and biological activity of cis-dichloro mono- and bis(platinum) complexes with N-alkyl-ethylenediamine ligands

Mono- and bis(platinum) complexes containing N-alkyl-ethylenediamine units of the type {cis-PtCl₂[H₂NCH₂CH₂NH(CH₂)_nCH₃]} (n=8, 9, 11, 15) and [{cis-PtCl₂(H₂NCH₂CH₂NH)}₂(CH₂)_n] (n=6, 8, 10, 12) and their corresponding dihydroxo-platinum(IV) complexes were synthesized. The structures of the metal chelates were derived from elemental analyses and their ¹H, ¹³C, IR spectra. The length of the aliphatic chains has been varied systematically, in order to increase the lipophilicity. Enlargement of the linker could also lead to more flexibility of one platinum sphere in reference to the attached DNA species. Using in vitro cytotoxicity tests it is shown that the biological activity of the bis(platinum) complexes increased, up to n=12, with the length of the linker. The longest linker in the ligands resulted in the most effective bis(platinum) complexes against L1210 murine leukemia cells.     

Biochemical and Economical Aspects of Levan Synthesis by Zymomonas Mobilis

With flocculent cells of Zymomonas mobilis levan was produced in a continuous upflow-tower fermentation with a productivity of up to 16 gl^-1h^-1. A large-scale process basing on whole Zymomonas cells is thought to be economical, if levan and ethanol production can be carried out simultaneously. Levan sucrase as the enzyme responsible for levan biosynthesis was purified and partially characterized.     

干旱对不同花椒种植模式下土壤微生物和线虫群落的影响

随着全球气候变暖, 我国岷江上游干旱区面积呈现增加的趋势。花椒(Zanthoxylum bungeanum)是岷江上游重要的经济树种之一, 对当地经济和社会发展起着重要作用, 提高花椒生态系统应对干旱干扰已成为迫切的问题。本研究设置了花椒单作、花椒-苜蓿(Medicago sativa)间作和花椒-大豆(Glycine max)间作3种种植模式, 在2015年8月对每种种植模式模拟干旱30 d, 每种种植模式包括干旱和对照处理, 在模拟干旱结束后、恢复15 d、30 d和45 d后分别采集土壤样品, 分析土壤化学性质、土壤微生物和线虫群落, 以探究花椒林下豆科植物能否缓和干旱的遗留效应对土壤化学性质和土壤生物的影响。重复测量方差分析表明: 在花椒单作模式下, 干旱恢复45 d后土壤硝态氮含量显著高于对照, 微生物量和真菌/细菌比与对照无显著差异, 线虫密度与对照无显著差异, 但线虫功能团没有恢复到对照水平; 在花椒-苜蓿间作模式下, 干旱恢复45 d后土壤含水量、铵态氮、硝态氮、溶解性有机碳、溶解性有机氮、微生物量、真菌/细菌比、线虫密度和线虫功能团组成与对照无显著差异, 但植食性线虫属Boleodorus相对多度显著高于对照; 在花椒-大豆间作模式下, 干旱恢复45 d后土壤含水量、铵态氮、硝态氮、溶解性有机碳、溶解性有机氮、微生物量和真菌/细菌比与对照无显著差异, 但线虫密度和功能团组成与对照有显著差异。在3种花椒种植模式中, 花椒-苜蓿间作模式下干旱的遗留效应对土壤养分和生物的影响最小。因此, 在干旱背景下, 花椒林下间作豆科植物可以加快土壤养分、土壤微生物和线虫群落的恢复, 进而有利于目标作物生长。     

Kinetic modeling of immobilized-lipase catalyzed transesterification of n-octanol with vinyl acetate in non-aqueous media

Organic esters are employed as solvents, fragrance, flavors, and precursors in a variety of industries. Particularly, aliphatic esters are greatly used in flavor industry, mainly as fixatives and modifiers, and aromatic esters in fragrance compositions. Esters are produced by a variety of methods among which esterification and transesterification with acid catalysts under reflux conditions are prominent. The use of biocatalysts provides an opportunity for carrying out reactions under milder conditions leading to better quality products suitable in fragrance and flavor industry. Transesterification of n-octanol with vinyl acetate was studied at 30 °C as a model reaction by employing different lipases as catalysts such as Psedomonas species lipase immobilized on diatomite, free Candida rugosa lipase. Novozym 435 (lipase B from Candida antarctica; immobilized on macro-porous polyacrylic resin beads) and Lipozyme IM 20 (Mucor miehei lipase immobilized on anionic resin). Novozym 435 was found to be the most active catalyst in heptane as a solvent. A conversion of 82% with 100% selectivity of n-octyl acetate was obtained at 30 °C in 90 min using equimolar quantities of the reactants with 0.833 g l⁻¹ of Novozym 435. Transesterification of other alcohols such as n-decanol, benzyl alcohol, cinnamyl alcohol, 2-ethyl-1-hexanol, 1-phenyl ethyl alcohol, and 2-phenyl ethyl alcohol was also studied with vinyl acetate. The analysis of the initial rate data and progress curve data showed that the reaction obeys the ternary complex bi–bi mechanism with inhibition by n-octanol. The experimental and theoretical values matched very well.

The order of transesterification reactivity of vinyl acetate with various alcohols in presence of Novozym 435 under otherwise identical conditions at 30 °C was found to be as follows:

n-octanol>n-decanol>benzylalcohol>cinnamylalcohol>2-ethyl-1-hexanol>2-phenylethylalcohol>1-phenylethylalcohol.

     

Colocalization of NO and VIP in neurons of the submucous plexus in the rat intestine

Since very few previous studies have carried out the quantitative analysis for the colocalization of nitric oxide (NO) and vasoactive intestinal peptide (VIP) in the submucous neurons in the rat digestive tract, we applied in vivo treatment of colchicine to enhance the immunoreactivity and examined the colocalization of NO synthase (nNOS) and VIP in neurons of the submucous plexus throughout the rat digestive tract. The density of nNOS-containing neurons in the submucous plexus in the stomach corpus (103±25 cells/cm², n=3) and that in the antrum (157±9 cells/cm², n=3) were significantly lower than those in small and large intestine. However no difference was detected in the cell density among duodenum (1967±188 cells/cm², n=3), jejunum (2640±140 cells/cm², n=3), ileum (2070±42 cells/cm², n=3), proximal colon (2243±138 cells/cm², n=3) and distal colon (2633±376 cells/cm², n=3). The proportion of nNOS-immunoreactive (IR), nNOS/VIP-IR and VIP-IR neurons to the total number of submucous neurons was examined. nNOS/VIP-IR neurons comprised 45–55% of total number of submucous neurons from the duodenum to the proximal colon, however those comprised 66.4±5.1% in the distal colon. The results showed that the dense distribution of nNOS-containing neurons was found in the submucous plexus throughout the small and large intestine, and large population of submucous neurons co-stored nNOS and VIP.     

菰属系统与演化研究——外部形态

本文研究菰属Zizania L.及其有关属、种的外部形态后,揭示出菰属应在禾本科Gramineaee稻族Oryzeae中独立成菰亚族Zizaniinae。其与水稻属Oryza L.或拟菰属Zizaniopsis Doel&Aschers。在远古时代都可能起源于一个现已灭迹的共同祖先,而后平行向前演化。     

Phycomyces blakesleeanus car B mutants: Their use in assays of phytoene desaturase

Strains of car B (phytoene-accumulating) mutants of Phycomyces blakesleeanus have been characterized with respect to their carotene contents, in vitro formation of isoprenoids from [2-¹⁴C] mevalonic acid and their ability to produce [¹⁴C]phytoene in situ for use in coupled assays of phytoene desaturase activity. All strains produced predominantly (15-Z)-phytoene both in vivo and in vitro. Other isoprenoids were produced by cell extracts including squalene, sterols, prenyl diphosphates and prenyl alcohols. The addition of 1% Tween 60 to crude cell extracts of the mutants partially restored wild type carotenogenic activity and also altered the proportions of other isoprenoids formed. However, in a cytosolic fraction of the car B mutant, the addition of 1% Tween 60 did not result in the production of any carotenoid from phytoene. This fraction was the most effective source of [¹⁴C] phytoene for use in coupled assays of phytoene desaturase activity.     

高原鼢鼠发情周期阴道细胞变化特征及血清促性腺激素含量动态

In order to determine the change characteristics of vaginal cells in the estrus cycle and the duration of the estrus cycle, vaginal smears and HE staining methods were used to observe and count the types, morphological changes, and proportions of vaginal cells in plateau zokor (Eospalax baileyi). The hormone secretion dynamics of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the serum of plateau zokor in the estrus cycle were determined by ELISA. The results showed that the main vaginal cells of plateau zokor were leukocytes, nuclear epithelial cells, incompletely cornified epithelial cells, and complete cornified epithelial cells. The proportion of nuclear epithelial cells in the proestrus was significantly higher than that during the other three periods (48. 4 ±3. 09)% (n=12, P < 0. 05). The proportion of complete cornified epithelial cells in the oestrus was the highest and significantly higher than that in the other three periods (59. 73 ±7. 59)% (n=15, P < 0. 05). The proportion of leukocytes in the metestrus and dioestrum was significantly higher than that in the proestrus and oestrus (P < 0. 05). The expression level of LH was the highest in the metestrus (4. 709 5 ±1. 094 0) ng/mL (n=6, P < 0. 05), which showed an increasing trend in proestrus and oestrus, and a decreasing trend in the metestrus and dioestrum. There was no significant difference in the expression level of FSH among the four periods (P > 0. 05); the estrus cycle duration of plateau zokor was 16 to 19 d, the characteristics of vaginal cell types in different stages of the estrus cycle were obvious, and the relative number was statistically significant. The study provided a basis for vaginal smear identification in the estrous cycle and reproductive physiology study of plateau zokor.     

丁苯酞对哮喘小鼠气道黏液高分泌及IL-13、TNF-α的影响*

目的: 研究丁苯酞对哮喘小鼠气道黏液高分泌及白介素-13(IL-13)、肿瘤坏死因子-α(TNF-α)的影响。方法: 小鼠随机分为空白对照组、模型对照组、阳性对照组和丁苯酞高、中、低剂量(100、50、25 mg/kg)组(n=12)。实验第1日、8日、15日通过注射卵清白蛋白(OVA)致敏,第22日吸入OVA连续激发5周复制哮喘模型,同时在激发前给予丁苯酞20 mg/kg进行干预,观测哮喘行为学、气道杯状细胞及黏蛋白5ac(Muc5ac)的分泌,测定支气管肺泡灌洗液(BALF)粘度,采用ELISA法测定BALF中Muc5ac、IL-13及TNF-α的水平。结果: 与空白对照组比较,模型对照组小鼠打喷嚏、抓鼻及哮喘的程度显著加重(P＜0.01),小鼠气道上皮杯状细胞增生及Muc5ac的分泌显著增加(P＜0.01),BALF的粘度及其中的Muc5ac、IL-13和TNF-α的含量显著升高(P＜0.01);与模型对照组比较,25、50、100 mg/kg丁苯酞干预后哮喘行为学评分明显下降(P＜0.01);小鼠气道上皮杯状细胞增生、Muc5ac的分泌、BALF的粘度及其中的Muc5ac、IL-13和TNF-α的含量均明显降低(P＜0.05, 0.01)。结论: 丁苯酞具有抑制哮喘气道黏液高分泌而平喘的作用,缓解IL-13、TNF-α异常高表达是其抑制气道黏液高分泌的作用机制之一。     

On the mechanism of silylformylation catalyzed by Rh-Co mixed metal complexes

Possible mechanisms for the silylformylation of 1-alkynes catalyzed by Rh₂Co₂(CO)₁₂ are investigated. Novel Rh-Co mixed metal complexes, (PhMe₂Si)₂Rh(CO)nCo(CO)₄ (n = 2 or 3) (3) and RhCo(HC≡CBuⁿ)(CO)₅ (5), are found to play important roles in this catalysis. The reaction of 3 with 1-hexyne and HSiMe₂Ph at ambient temperature and pressure of CO gives n-BuC(CHO)=CHSiMe₂Ph (1a, Z/E = 95/5), (PhMe₂Si)₂Rh(CO)₃Co(CO)₄ (3-B) and an Rh-Co mixed metal butterfly complex, h₂Co₂(HC≡CBuⁿ)(CO)₁₀ (4). The reaction of 5 with 1-hexyne and HSiMe₂Ph under the same ambient conditions affords 1a (100% Z) very cleanly as the sole reaction product. The crossover experiments u sing RhCo(DC≡CBuⁿ)(CO)₅(5-d), 1-hexyne-1d and DSiMe₂Ph strongly support the mixed metal bimetallic catalysis and involvement of bis(alkyne)-Rh-Co species. The most plausible catalytic cycle of silylformylation which can accommodate all the observed results is proposed.     

Role of NAD(P)H:quinone oxidoreductase polymorphism at codon 187 in susceptibility to lung, laryngeal and oral/pharyngeal cancers

NAD(P)H:quinone oxidoreductase (NQO1) has been proposed to play a protective role against the toxic effects of benzo[a]pyrene quinones. The C⁶⁰⁹T base change in the NQO1 gene, resulting in a Pro¹⁸⁷Ser amino acid change in the protein, has been associated with deficient enzyme activity. We examined whether this polymorphism modified the risks of smoking-related cancers in a case-control study involving patients with lung cancer (n = 150), laryngeal cancer (n = 129), oral/pharyngeal cancer (n = 121) and control individuals (n = 172), all Caucasian smokers. No statistically significant associations were observed between the NQO1 genotypes and smoking-related cancers, although the Ser/Ser genotype was associated with a tendency towards increased risk for lung cancer (odds ratio [OR] = 2.2, 95% confidence interval [CI] 0.7-6.7) and for oral/pharyngeal cancer (OR = 2.3, 95% CI 0.6-8.2). No significant interaction between the NQO1 genotype and either smoking exposure or GSTM1 genotype was found. Our results are consistent with the hypothesis that lack of NQO1 activity may be involved in some smoking-related cancers. However, they were based on small numbers of individuals with the putative atrisk genotype, and the associations did not reach statistical significance. Moreover, these results contrast with those observed in some other ethnic populations, where a protective effect of the NQO1 Ser allele was found. Further studies are therefore clearly needed for a better understanding of the potential role of NQO1 activity in tobacco-related cancers.     

厚朴酚联合吉非替尼对A549非小细胞肺癌细胞的干预作用*

目的: 探讨厚朴酚与吉非替尼协同影响非小细胞肺癌A549细胞的作用。方法: 以浓度为6.25～500 μmol/L厚朴酚、0.625～100 μmol/L吉非替尼分别处理A549细胞24 h,CCK-8实验检测细胞活力 (n=3),选24 h及100 μmol/L厚朴酚与5 μmol/L吉非替尼作后续处理(n=3);采用对照组、厚朴酚组、吉非替尼组和厚朴酚+吉非替尼组的析因分析设计;克隆形成检测细胞增殖;蛋白印迹测蛋白表达;流式细胞术检测细胞凋亡及分选CD44⁺和CD133⁺细胞。结果: 与对照组比,厚朴酚和吉非替尼组的克隆形成率显著降低(P＜0.05);凋亡率显著升高(P＜ 0.05);CD44⁺和CD133⁺细胞数量显著减少(P＜0.05);Ki67和PCNA及干细胞标记蛋白SOX2和OCT4表达显著下调(P＜0.05);Bax/Bcl-2表达比例显著上调(P＜0.05)。与厚朴酚组或吉非替尼组比较,厚朴酚+吉非替尼组进一步促进了上述改变(P＜0.05),且凋亡率、Bax/Bcl-2、SOX2和OCT4等指标都存在厚朴酚和吉非替尼的交互作用(P＜ 0.05)。结论: 厚朴酚与吉非替尼促进A549细胞凋亡和抑制其干细胞样特性,且联合用药效果优于单一给药。二者对A549细胞的抑癌作用有交互影响。