Effect of Nano red elemental selenium on GPx activity of broiler chick kidney cells<Emphasis Type="Italic">in vitro</Emphasis>

A new selenium source, Nano red elemental selenium (Nano-Se) was used to study the effect on the GPx activity of broiler chick kidney cells (BCKC)in vitro, Sodium selenite (Na₂SeO₃) and seleno-1-methionine (Se-Met) were used as the controls. The results showed that the effects of three kinds of Se forms on the GPx activity of BCKC were accordant (p>0.05) compared with each other at 0.01. 0.05 and 0.10 μmol/L Se concentrations treatments. In the range of 0.00–0.10 μmol/L Se concentrations, the GPx activity increased with elevation of Se concentrations in medium. For the three kinds of Se forms, the GPx activity reached the climax at 0.10 μmol/L Se concentration. At 0.20 and 0.30 μmol/L Se concentrations, the influnces of three kinds of Se forms were not accordant with one another. For Nano-Se, the GPx activity at 0.20 and 0.30 μmoi/L Se concentrations remained the same as that at 0.10 μmol/L Se concentration treatment. For Se-Met, the GPx activity at 0.20 μmol/L Se concentration treatment remained the same with 0.10 μmol/L treatment; the GPx activity at 0.30 μmol/L Se concentration treatment was declined significantly (p<0.05) compared with 0.10 or 0.20 μmol/L treatment. For Na₂SeO₃, the GPx activity falled gradually with Se concentration increasing from 0.10 μmol/L to 0.30 μmol/L, and at 0.30 μmol/L Se concentration treatment, the GPx activity was less than the original of BCKC. The results implicated, on the GPx activity of BCKCin vitro, the ranking of width range of the most suitable Se concentration for nutrition curve of the three Se formes is Nano-Se>Se-Met>Na₂SeO₃. Foundation item: Supported by the Key Item of the Science and Technology Bureau of Zhejiang Province (021122680) Biography: Xu Bao-hua (1966-), male, Associate professor, Ph. D. research direction: nanobiology and animal nutrition.     

Preparation of exine-detached pollen inNicotiana tabacum

A method of preparing exine-detached pollen inNicotiana tabacum was established. Anthers containing early-middle binucleate pollen were cold-pretreated at 4–6°C for 7–14 days, and were suspended in 0.3 mol/L sucrose solution for 2 days. During this process, the exine of most pollen grains dehisced. Then they were transferred into an enzyme solution containing 1% cellulase, 1% pectinase, 0.1% pectolyase, 1 mol/L mannitol, 0.3 mol/L sorbitol, 0.5% potassium dextran sulphate and K3 medium macro elements. After 15–20 min enzymatic maceration, the exine was detached resulting in the release of exine-detached pollen. Factors affecting preparation of exine-detached pollen were investigated, including cold-pretreatment, osmoticum concentration and enzymes used. Xia Huijun: born in Oct. 1963, Ph.D. Current research interest is in plant reproductive biology Supported by the National Natural Science Foundation of China     

Adrenomedullin inhibits the endothelin production induced by urotensin II in rat vascular smooth muscle cells

The aim of this study was to observe the effects of adrenomedullin (ADM) on endothelin (ET) production induced by urotensin Ⅱ (UⅡ) in rat vascular smooth muscle cells (VSMCs). Cultured VSMCs which were incubated with UⅡ (10^-8 mol/L) and with various concentrations of ADM were used to measure the VSMCs ³H-TdR incorpora- tion, the activity of extracellular signal-regulated kinase (ERK), the amount of ET mRNA and ET production in VSMCs. In this work we found that incubation with UⅡ(10^-8 mol/L) increased obviously the amount of ET mRNA in VSMCs and ET production in medium, however, coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ(10^-8 mol/L) reduced the amount of ET mRNA by 15%, 24% and 45% (P< 0.01) respectively, compared with UⅡ alone. The content of ET in medium was 14.13, 11.38 and 11.00 pg/mL. ADM alone (10^-8 mol/L) had no effect on ET production in VSMCs. UⅡ (10^-8 mol/L) promoted the 3H-TdR incorpo- ration and activity of ERK in VSMCs. ADM inhibited VSMCs 3H-TdR incorporation and activation of ERK in a concentration-dependent manner. Compared with UⅡ group, after coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ (10^-8 mol/L) the VSMCs 3H-TdR incorporation was decreased by 7% (P > 0.05), 32% (P < 0.05) and 41% (P < 0.01), respectively, and the activity of ERK was decreased by 24% (P > 0.05), 32% (P < 0.05) and 36% (P < 0.05), re- spectively, in a concentration-dependent manner. The results show that in cultured VSMCs ADM inhibits ET mRNA expression, ET production and proliferation stimulated by UⅡ, and that inhibitory effect of ADM on UⅡ bioaction could be mediated through inhibiting MAPK pathway.     

Regenerating plants from cryopreserved adventitious buds of haploids in rice

A large number of adventitious buds were induced fromin vitro cultured young inflorescences of haploids of rice. Having been subcultured on solidified subculture media at 26°C for 7 days, the adventitious buds were loaded into 1.8 mL plastic cryotubes with cryoprotectant and kept on ice for 45–60 min. After cooled at a rate of 1.0°C/min down to −40°C, the samples, were kept in liquid nitrogen. The adventitious buds which have been cryopreserved for about 30 days were thawed rapidly in 38–40°C water and then plated on solidified MS medium containing 3% sucrose, 0.5 mg/L NAA and 2.0 mg/L kinetin. After plated, 23%–32% of adventitious buds resumed growth and 15%–22% regenerated plantlets. The results of this work indicated that the adventitious buds derived fromin vitro cultured young inflorescences is a critical factor for the success and subculturing adventitious buds on MS medium containing 3% sucrose and 4% sorbitol or 20% potato extract is essential to the procedure. The effective cryoprotectant is 10% DMSO (dimethyl sulfoximide)+0.5 mol/L sorbitol. Supported by the Natural Science Foundation of Hubei Province Zhang Zhihong: born in 1963, Lecturer     

Effect of Cl- on photosynthetic bicarbonate uptake in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803

Photosynthetic inorganic carbon utilization was investigated in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803 grown in standing culture. Photosynthetic rates for the two algae reached about 10 times the theoretical CO₂ supply rate at low dissolved inorganic carbon (DIC) of 100 μmol/L, and the rates were unaffected by the addition of 20 mmol/L Na⁺, indicating that the two algae possessed Na+-independent HCO^-₃ utilization for photosynthesis under low DIC. Their photo- synthetic rates at low DIC were inhibited by higher Cl^- and the degrees of inhibition were increased with the rise of CI^- concentration, and in the presence of Diphenylamine-2-carboxylate (DPC), a reported Cl^- channel inhibitor, the rates decreased by 74%－82%, implying that putative DPC-sensitive Cl^- channels participate in Na+-independent HCO₃^- uptake for photosynthesis. The experiment of intracellular 14C-DIC accumulation for photosynthesis showed that internal DIC pools decreased by about 80% with 200 μmol/L DPC and by 64%－70% with 100 mmol/L Cl^-. The experiment of chlorophyll a fluorescence quenching showed that initial rates and extents of fluorescence quenching obviously decreased with 200 μmol/L DPC or 100 mmol/L Cl^-. The two experiments gave further evidence that putative DPC-sen- sitive Cl^- channels participate in Na+-independent HCO^-₃ uptake for photosynthesis in the two algae grown in standing culture.     

HPLC determination of ascorbic acid using luminol-Cu (II) chemiluminescence-application to the analysis of Juice beverage

The chemiluminescence (CL) of luminol-Cu (II) was applied to HPLC determination of ascorbic acid, which was separated by a C₁₈ reverse-phase column with a mobile phase of 0.25 mol/L HAc. The eluted ascorbic acid was mixed with 0.3 mmol/L luminol and 0.05 mol/L CuSO₄. The light emission from the reaction of Cu(II) oxidized ascorbic acid and luminol was detected by a modified luminometer. The detection limit was 3.6×10⁻⁶ mol/L for ascorbic acid at aS/N ratio of 3, and the linear calibration range was 2×10⁻⁴–2×10⁻³ mol/L. The relative standard deviation for 5 replicate injections of 1×10⁻³ mol/L ascorbic acid was calculated as 4.3%. The method was successfully applied to determination of ascorbic acid in juice beverage. Foundation item: National Natural Science Foundation of China (29605001) Biography: Wu Feng-wu(1963-), male, research direction: analytical chemistry.     

NaCl胁迫对不同品种向日葵幼苗生理特性的影响

对46天苗龄的盆栽向日葵幼苗分别用0．05mol／L、0．10mol／L、0．20mol／L的中性盐NaCl进行胁迫处理，检测3个不同品种向日葵的脯氨酸、丙二醛含量和保护酶活性的变化．从而比较研究不同品种向日葵对NaCl的耐受程度，结果表明：中性盐对津引6号伤害较轻，而对津引1号和津引2号伤害比较严重，津引1号和津引2号伤害程度相差不大．就生物膜透性伤害程度而言，津引6号明显低于津引1号和2号。其脯氨酸含量显著高于津引1号和2号，丙二醛含量则明显低于津引1号和2号．在抗氧化保护酶活性变化方面，津引6号POD，CAT活性较津引1号和2号增加要快得多，而SOD活性上升更明显．简言之。津引6号有较强的耐盐性．     

Study of the voltammetric behavior of heme and its direct analysis application

The electroanalytical method of ferriheme was studied by linear sweep voltammetry in medium of 0.05 mol/L Tris+0.05 mol/L NH₃−NH₄Cl buffer at hang mercury drop electrodes (HMDE). Heme exhibits two pair reversible redox peaks and one irreversible peak. The cathodic peak potentials are−;0.236 V, −0.422 V and −1.408 V respectively. The first and the third peaks can be used for directly quantitative determination of heme concentrations. The peak currents are good linear relationship with heme concentration in ranges of 3×10⁻⁶–6×10⁻⁵ mol/L and 3×10⁻⁷–1.5×10⁻⁵ mol/L respectively. Supported by the National Natural Science Foundation of China Lju Huihong: born in 1965, Lecturer. Visiting scholar from Xiangyang Teacher’s College, Hubei, 441053     

Effect of iron deficiency on heterocyst differentiation and physiology of the filamentous cyanobacterium<Emphasis Type="Italic">Anabaena</Emphasis> sp. PCC 7120

The effect of iron deficiency on heterocyst differentiation and some physiological properties of the filamentous cyanobacteriumAnabaena sp. PCC 7120 was investigated. Under moderate iron limitation conditions, achieved by addition of iron chelator 2, 2′-Dipyridyl (<80 μmol/L) led to delayed heterocyst differentiation, no heterocyst differentiation was observed under severe iron limitation conditions, when the concentration of 2,2′-Dipyridyl in the medium was more than 100 μmol/L. it seemed that there are certain iron-regulated genes or operons whose function is to control heterocyst development. In addition, iron deficiency impaired the growth. Low iron cells had a decrease in the quantities of pigment content (chlorophyll and phycocyanin content), the whole cellin vivo absorbance spectra confirmed the decrease, the protein electrophoretic profiles revealed that iron-deficient cells had less protein bands, with the increase of 2,2′-Dipyridyl, the protein bands was more and more less. And differently, iron deficiency also caused an increase of ROS (Reactive Oxygen Species) and SOD activity, it suggests that iron deficiency led to oxidative stress, which generally occured under high-iron conditions. Foundation item: Supported by the National Natural Sciences Foundation of China (30070154), the Frontier Science Projects Programme of the Institute of Hydrobiology, the Chinese Academy of Sciences (220316), State Key Project on Cyanobacterial Bloom Control in Lake Danchi (K9905-35-01) Biography: Xu Wenllang (1974-), male, Ph. D. research direction: molecular genetics of eyanobacteria.     

Submerged culture of Magnetospirillum gryphiswaldense under N2-fixing condition and regulation of activity of nitrogen fixation

A submerged culture technique for Magnetospirillum gryphiswaldense under the nitrogen-fixing condition (microaerobic and N-limited) was set up. In N-limited medium with Na-lactate as a sole carbon source, the optical density (A_600nm) and activity of nitrogen fixation of cells were 1.3 and 217 nmol of ethylene produced per hour per A_600nm respectively within 21 h by three times of feeds. The pH and temperature were controlled at 7.2 and 30℃ respectively, and the oxygen concentration was controlled by sparging with N₂ containing 0.4%—0.8% of O₂. The activity of nitrogen fixation of cells was obviously inhibited by oxygen and ammonium. It indicated that the posttranslational regulation of nitrogenase existed in M. gryphiswaldense.     

Synergistic effect of trace elements and flavonoids from Epimedium koreanum Nakai on primary osteoblasts

Several trace elements, particularly, manganese （Mn） and zinc （Zn）, are essential in bone metabolism as cofactors for specific enzymes. It has been reported that there exists the relationship between osteoporosis and trace element-deficiency and the efficacy of Ca, Mn and Zn supplementation on spinal bone mineral density in postmenopausal women. Traditional Chinese medicines （TCM）, such as Herba epimedii, were proved to be effective for prevention of osteoprosis in vivo; however, the efficacy of the main constituents and/or crude extract was not ideal in vitro, which suggested that they may work in another way. The purpose of the present study was to examine whether the combination of icariin and total flavonoids （TF） from Herba epimedii with mineral elements, which were abundant in Herba epimedii, would have a more beneficial effect on the viability and differentiation of primary osteoblasts than either agent alone, and to analyze the dada for a possible synergistic, additive or antagonistic effect. The combinations of 10 μmol/L Zn, Ca and Mn with icariin and total flavonoids greatly improved the cell viability and meanwhile dramatically enhanced the alkaline phosphatase activity as compared to each agent alone. On the other hand, an increased cell growth inhibition was also observed by combining 0.1 μmol/L, 1 pmol/L Zn with 10μmol/L icariin, and 10 μmol/L Mn with 0.06 μg/mL total flavonoids. Meanwhile a decreased alkaline phosphatase activity was also found in several icariin-Zn/Mn and total flavonoids-Zn/Ca/Mn combinations. These results suggested that mineral elements （Zn, Ca, Mn） greatly enhanced the efficacy of icariin and total flavonoids from Herba epimedii on the viability and differentiation of primary osteoblasts by certain combinations.     

Assaying poly(ADP-ribose) polymerase activity in plants by polarographic method

A new method has been developed to assay poly(ADP-ribose) polymerase (PARP) activity in plant tissues through determining the content of nicotinamide (NIC) produced by enzymatic reaction by linear sweeping polarographic method. The detection limit of NIC was 0.03μmol/L, the calibration graph was linear up to 5 Mmol/L ( r = 0.999). The recoveries were approximately in the range of 92% to 98% and the relative standard deviations were less than 6.6% . Moreover, NAD+ and other interference existing in the mixture after enzymatic reaction had been removed by simple pretreatment, thus PARP assays were not interfered. A rapid, simple, sensitive and reliable nonisotopic method is reported to assay PARP activity in plant tissues . The results show that the KmNAD+ value of PARP in maize ( Zea mays L.) seedlings is 59 and the optimum pH for PARP activity is 8.5. Moreover, physiological conditions affect PARP activity in plant tissues, which has not been reported previously. When tobacco ( Nico-tiana tobacum) suspension cells were stressed by NaCI at low concentrations (100, 200 mmol/ L), the PARP activity increased significantly; when the cells were stressed at high concentrations (400, 1 000 mmol/L), it decreased to or even below the control level. PARP activity in etiolated maize seedlings was higher than that in light-grown seedlings.     

Rice protoplasts isolated directly from mature-embryo-derived calli

Mature-embryo-derived calli of japonica rice (Oryza sativa L) Taipei 309 were used for replicated protoplast isolation experiments. Six out of nine callus lines produced protoplasts with satisfactory yield of 5.20×10⁶–8.96×10⁶ protoplasts/g FW (fresh weight). The remaining three callus lines initiated from seeds of cryopreserved-callus-derived plants had rooty calli, resulting in low yield of protoplasts and a large number of isolated banana-shape intact cells. Viability of protoplasts ranged 87.46%–94.15%. The average size of protoplasts was 207.49–379.04 μm² in different callus lines. Comparitive experiments were also carried out using both calli and suspension culture cells for protoplast isolation. The results demonstrated that protoplast isolation of calli was a substantially simplified and reliable method for preparing rice protoplasts. Jin Deming: born in Jan. 1959, Associated Professor     

Research of Inclusion Complexation of 4,4＇-Thiodiphenol Effected by β-Cyclodextrin in Aqueous Solutions and the Effect on Photodegradation

The inclusion behavior of 4, 4＇ -Thiodiphenol （TDP）, a typical bisphenol and endocrine disruptor, reacts with β-cyclodextrin （β-CD） in aqueous solutions has been investigated by means of UV absorption spectrum and quantum-chemical calculation with Gaussian 98 software. The results show that the inclusion behavior of TDP is quite different in acidic solutions （pH 5.9） from that in alkaline solutions （pH 10.0）. This behavior difference is attributed to the different formula structures in aqueous solutions at acidic and alkaline pH values that are demonstrated by quantum- chemical modeling and calculation. TDP forms a 1 ： 1 fl-CD inclusion complex in aqueous solutions. The equiiibrium constant K was calculated to be 553.49 L/mol at pH 5.9 and 1 318.20 L/mol at pH 10.0 respectively for the inclusion complex reaction by using the modified Benesi-Heldbrand equation. After inclusion TDP＇s structure is changed especially at the inclusion part with the bond order becoming larger, which results in inhibitive photodegradation during direct photooxidation and H2O2 assisted photooxidation.     

Effect of CO<Subscript>2</Subscript> concentrations on the activity of photosynthetic CO<Subscript>2</Subscript> fixation and extracelluar carbonic anhydrase in the marine diatom <Emphasis Type="Italic">Skeletonema costatum</Emphasis>

The growth and activity of photosynthetic CO2 uptake and extracellular carbonic anhydrase (CAext) of the marine diatom Skeletonema costatum were investigated while cultured at different levels of CO2 in order to see its physio-logical response to different CO2 concentrations under either a low (30 靘ol·m-2·s-1) or high (210 靘ol·m-2·s-1) irradiance. The changes in CO2 concentrations (4—31 靘ol/L) affected the growth and net photosynthesis to a greater extent under the low than under the high light re-gime. CAext was detected in the cells grown at 4 mol/L CO2 but not at 31 and 12 靘ol/L CO2, with its activity being about 2.5-fold higher at the high than at the low irradiance. Photo- synthetic CO2 affinity (1/ K1/2(CO2)) of the cells de-creased with increased CO2 concentrations in culture. The cells cultured under the high-light show significantly higher photosynthetic CO2 affinity than those grown at the low-light level. It is concluded that the regulations of CAext activity and photosynthetic CO2 affinity are dependent not only on CO2 concentration but also on light availability, and that the de-velopment of higher CAext activity and CO2 affinity under higher light level could sufficiently support the photosyn-thetic demand for CO2 even at low level of CO2.     

Influence of PrP 106―126 on expression of laminin and fibronectin in astrocyte

Astrogliosis is a hallmark of prion disease, but the metabolic alterations of astrocytes remain poorly documented. A synthetic peptide corresponding to amino acid 106-126 of the human prion protein （PrP） has been shown to be toxic to neurons. In this study, the effects of PrP 106-126 on astrocytes were investigated in vitro. The proliferation of astrocytes was significantly （P 〈 0.05） increased when grown in media conditioned with PrP 106-126 （80 μmol/L） from microglia. The expression of laminin （LN） and fibronectin （FN） was examined at both mRNA and protein levels. The results showed that exposure of astrocytes to PrP 106-126 enhanced the expression of LN and FN. The increase of FN in astrocyte cultures required cytokines previously released by activated microglia. This study reveals the expression of LN and FN affected by PrP106-126.     

Bis(7)-Tacrine, a promising anti-Alzheimer's agent, attenuates glutamate-induced cell injury in primary cultured cerebrocortical neurons of rats

The effects of bis(7)-tacrine, a novel dimeric acetylcholinesterase (AChE) inhibitor, on glutamate-induced cell injury were investigated in primary cerebral cortical neurons of rats. Exposure of cultured neurons (12 days after plating) to 0.5 mmol/L glutamate for 30 min resulted in significant cell damage. Pretreatment with bis(7)-tacrine (0.03–1.0 μmol/L) reduced the glutamate-induced neurotoxicity in a concentration dependent manner and the maximal response was seen at 1 μmol/L with approximately, 30% protection. A receptor binding assay showed that bis(7)-tacrine can completely displace MK-801 binding to rat cortical membrane with an IC₅₀ of 0.57 μmol/L. These findings suggest that bis(7)-tacrine can directly interact with N-methyl-D-aspartate receptor channel complex, which may contribute to the inhibitor's protective effects against glutamate-induced excitotoxicity. Thus, it is possible that anti-glutamate/anti-AChE synergism is responsible for potentially better Alzheimer's therapy of bis(7)-tacrine relative to tacrine. Biography: Zhang Bai-fang (1974-), female, Ph. D. candidate, research direction: Alzheimer's disease and ischemia     

低氧胁迫对光棘球海胆Mesocentrotus nudus致死性及生理机能的影响

全球近海底层水体低氧现象日趋频繁，严重影响到海洋生态系统的稳定性。为掌握光棘球海胆Mesocentrotus nudus对低氧胁迫的耐受能力，通过室内生态模拟实验，分析3种规格的光棘球海胆，即大规格海胆平均体质量(91.41±3.31) g、中规格(60.79±3.42) g和小规格(4.15±0.18) g，在3个溶解氧(Dissolved Oxygen，DO)浓度水平(0.5 mg/L、2.0 mg/L、3.0 mg/L)胁迫下的行为、存活、半致死时间(Median Lethal Time，LT₅₀)以及超氧化物歧化酶(Superoxide Dismutase，SOD)活力。结果表明，在DO 0.5 mg/L和DO 2.0 mg/L低氧胁迫下，海胆行为表现异常，低氧初期海胆表现为管足活力下降、吸附力降低，移动能力变弱，同时伴随棘脱落，随着低氧暴露时间延长，海胆各项活动能力进一步减弱，直至死亡；在DO 3.0 mg/L胁迫下，中、大规格海胆无死亡现象。3种规格海胆死亡率随低氧暴露时间延长都有所增加，其中在DO 0.5 mg/L胁迫下，3种规格海胆的最终死亡率都达到100%，但在DO 2.0 mg/L胁迫下，随低氧胁迫时间的延长，3种规格海胆都对低氧胁迫表现出不同程度的适应，很少死亡。3种规格海胆的LT₅₀在DO 0.5 mg/L和DO 2.0 mg/L两种低氧胁迫水平下存在差异，DO 0.5 mg/L胁迫下，大规格海胆的LT₅₀为70.50 h，中规格为34.64 h，小规格为34.02 h；DO 2.0 mg/L胁迫下，中规格海胆的LT₅₀延长至120.88 h，小规格海胆延长至107.15 h。此外，中规格海胆在两种低氧胁迫水平下，体内的超氧化物歧化酶活力均发生了明显变化，说明低氧胁迫已影响到光棘球海胆的抗氧化系统。本研究初次掌握了光棘球海胆在不同环境低氧胁迫下的生存和低氧适应情况，可为低氧海域光棘球海胆种群维持以及海胆养殖业的健康发展提供参考。     

Primary Study on Phytodegradation of Bisphenol A by Elodea nuttallii

In a microcosm system where Elodea nuttallii and Bisphenol A （BPA） coexist, with the help of Liquid chromatography-mass spectrometer （LC-MS） and gas chromatography-mass spectrometer （GC-MS）, the dynamics of phytodegradation of BPA with concentration 1-20 mg/L and the products of phytodegradation was studied. Antioxidation activity and phospholipids fatty acids were measured to evaluate the effects of BPA on E.nuttallii. The results showed that the half life period of phytodegradation of BPA was less than 15 days. 2-（4-hydroxypheny）-2-（3,4-o-dihydroxypheny） propane and 2, 2-bis（4-hydroxypheny） propyl alcohol were identified to be as two possible products of BPA. The peroxidase （POD） activity of E.nuttallii decreased by 50% to 100% compared with that controlled at the end of the experiment. The fatty acid methanol esters （FAMEs） changed obviously too. It showed that oxidation stress and membrane damage would be the two effcting aspects of BPA on E.nuttallii.     

Organic solvents enhanced spectrofluorimetric method for determination of laccase activity

A novel fluorimetric method for determination of laccase activity in organic solvents is proposed, based on the oxidation ofo-phenylenediamine (1,2-diaminobenzene, OPDA) catalyzed by laccase yielding 2,3-diaminophenazine. The optimal conditions for laccase in organic media areT=55°C, pH=6.5, 1.0×10⁻²mol/L OPDA, 1.25 mL ethanol, 1.25 mL 1,4-dioxane and 1.25 mL acetone. The linear range of the method proposed in ethanol, 1,4-dioxane and acetone media were 0.44–19.33, 0.11–20.85, 0.38–21.05 U with the detection limit of 0.088, 0.022, 0.076 U, respectively. The proposed method has been applied to the analysis of laccase activity of real samples with more accurate and sensitive than that of the previous method reported. Supported by the National Natural Science Foundation of China and the Special Founds of State Education Committee for Doctorate Research Huang Zuyun: born in 1963, Ph.D