Effects of Different Nitrogen Application Methods on Enzyme Activities in Leaves at Late Growth Stage of Spring Maize

[Objective] This paper aimed at exploring the effects of different nitrogen application methods on the enzyme activities of leaves at ear position in late growth stage of maize. [Method] By pot experiment, the superoxide dismutase （SOD） activity, peroxidase （POD） activity, malonyl dialdehyde （MDA） content and soluble protein content were determined. [Result] At the filling stage and ripening stage, with the increasing of nitrogen application rate, MDA content gradually decreased, while SOD activity, POD activity and soluble protein content increased. MDA content with two topdressing nitrogen was lower than that with one top dressing nitrogen at the same nitrogen application rate, while SOD activity, POD activity and soluble protein content with two topdressing nitrogen were higher than that with one topdressing nitrogen. [Conclusion] Different nitrogen application methods have relatively significant effects on the MDA content, SOD activity, POD activity and soluble protein content, which is of certain directive significance for preventing spring maize prematuration.

pH Stability of Defibrase_ in Aqueous Solution Determined by En-zyme-linked Immunosorbent Assay

Stability of Defibrase in various pH buffer solutions was investigated. Enzyme-linked immuno-sorbent assay (ELISA) and coagulating time method were used to assess antigenic stability and coagulating stability, respectively. The change of antigenic activities and coagulating activities of Defibrase in the same buffer solutions (pH 6, 7 and 8, with the exception of pH 3.6) showed similar tendency to decline with the time. Concentrated Defi-brase was relatively stable at neutral pH 6~7, more than 95% of its initial activities (100BUmL-1) was kept after a 10-day storage at 40 oC, whereas in pH 3.6 and pH 9 buffer solutions, diluted Defibrase was very labile. Addition of Triton X-100 or bovine serum albumin could effectively prevent loss of Defibrase by minimizing adsorption of De-fibrase to plastic surface (P<0.005). Concentration of Defibrase could also affect its stability in aqueous solutions.

Telomerase Activity in Human Renal Cell Carcinoma with Reference to Clinicopathologic Features

Objective: To study the telomerase activity in human renal cell carcinoma and to evaluate the correlation with the clinicobiologic features of the neogrowth.Methods: The telomerase activity was studied by means of a modified telomeric repeat amplification protocol (TRAP) in 32 renal cell carcinoma tissues, 32 normal renal tissues and 32 paracancer tissues and its correlation with the clinicopathologic features of the tumor was evaluated.Results: Telomerase activity was strongly positive in 17, positive in 12 and negative in 3 cases of renal cell carcinoma tissues, the total positive rate being 91%. Telomerase activity was weakly positive (6%) in only 2 out of 32 samples of normal renal cortex tissues and positive in 6 paracancer tissues (19%), the difference was conspicuous (P<0.01).Conclusion: The positive rate of telomerase activity was significantly higher in renal cell carcinoma tissues and might serve as a prognostic marker for estimating the biologic characteristics of renal cell carcinoma.

油田排放污水水质分析化验技术研究

石油生产过程中需要进行大量的污水排放,为了降低污水对环境的污染,需要对污水水质进行化验。基于此,本文将从发光细菌法、呼吸速率法、色谱分析法、拉曼光谱法等方面对油田排放污水水质分析化验技术进行分析,从而提高油田排放污水水质的化验水平。

Effect of Cistanoside Compounds on Oxidative Stress and Immunity

Cistanoside compounds were studied as the scavengers of hydroxyl and superoxide anion free radicals with spin trapping ESR method in vitro. Low-temperature ESR technique, experimental technique of immunotoxicology and biochemical method were used to detect the level of reactive oxygen radicals in kidney tissue of rats and SOD level and GSH-Px activity in rat serum. The results indicated that cistanoside compounds could inhibit reactive oxygen free radicals in vitro and prevent and repair the free radical damages for diabetic nephropathy. The experimental data of 揷arbon-particle detection in mouse serum?showed that cistanoside compounds could improve the phagocytotis index of macrophages (Mj) in mice blood and increase the weights of immune organs of mice.

Dynamic alteration of telomerase expression and its diagnostic significance in liver or peripheral blood for hepatocellular carcinoma

AIM： To investigate the dynamic alteration of telomerase expression during development of hepatocellular carcinoma （HCC） and its diagnostic implications in liver tissues or peripheral blood mononuclear cells for HCC. METHODS： Dynamic expressions of liver telomerase during malignant transformation of hepatocytes were observed in Sprague-Dawly （SD） rats fed with 0.05% of 2-fluoenyacetamide （2-FAA）. Total RNA and telomerase were extracted from rat or human liver tissues. The telomerase activities in livers and in circulating blood were detected by a telomeric repeat amplification protocol-enzyme-linked immunosorbent assay （TRAP- ELISA）, and its diagnostic value was investigated in patients with benign or malignant liver diseases. RESULTS： The hepatoma model displayed the dynamic expression of hepatic telomerase during HCC development. The telomerase activities were consistent with liver total RNA levels （r = 0.83, P 〈 0.01） at the stages of degeneration, precancerosis, and cancerization of hepatocytes. In HCC patients, the telomerase levels in HCC tissues were significantly higher than in their adjacent non-cancerous tissues, but liver total RNA levels were lower in the former than in the latter. Although the circulating telomerase of HCC patients was abnormally expressed among patients with chronic liver diseases, the telomerase activity was a non-specific marker for HCC diagnosis, because the incidence was 15.7% in normal control, 25% in chronic hepatitis, 45.9% in liver cirrhosis, and 85.2% in HCC, respectively when absorbance value of telomerase activity was more than 0.2. If the value was over 0.6, the incidence was 60% in HCC group and 0% in any of the others （P 〈 0.01） except in two cases with liver cirrhosis. However, the combination of circulating telomerase with serum alpha-fetoprotein level could increase the positive rate and the accuracy （92.6%, 125 of 135） of HCC diagnosis. CONCLUSION： The overexpression of telomerase is associated with HCC development, and its abnormality in liver tissues or in peripheral blood could be a useful marker for diagnosis and prognosis of HCC.

Deletion of a Non-Catalytic Region Increases the Enzymatic Activity of a β-Agarase from Flammeovirga sp. MY04

A Glycoside hydrolase （GH） typically contains one catalytic module and varied non-catalytic regions （NCRs）. However, effects of the NCRs to the catalytic modules remain mostly unclear except the carbohydrate-binding modules （CBMs）. AgaG4 is a GH16 endo-β-agarase of the agarolytic marine bacterium Flammeovirga sp. MY04. The enzyme consists of an extra sugar-binding peptide within the catalytic module, with no predictable CBMs but function-unknown sequences in the NCR, which is a new characteristic of agarase sequences. In this study, we deleted the NCR sequence, a 140-amino acid peptide at the C-terminus and expressed the truncated gene, agaG4-T140, in Escherichia coli. After purification and refolding, the trtmcated agarase rAgaG4-T140 retained the same catalytic temperature and pH value as rAgaG4. Using combined fluorescent labeling, HPLC and MS/MS techniques, we identified the end-products of agarose degradation by rAgaG4-T140 as neoagarotetraose and neoagarohexaose, with a final molar ratio of 1.53：1 and a conversion ratio of approximately 70%, which were similar to those of rAgaG4. However, the truncated agarase rAgaG4-T140 markedly decreased in protein solubility by 15 times and increased in enzymatic activities by 35 times. The oligosaccharide production of rAgaG4-T140 was approximately 25 times the weight of that produced by equimolar rAgaG4. This study provides some insights into the influences of NCR on the biochemical characteristics of agarase AgaG4 and implies some new strategies to improve the properties of a GH enzyme.

Optimization of conjugated linoleic acid triglycerides via enzymatic esterification in no-solvent system

We compared four esterifiable enzymes. The lipase Novozym 435 possessed the highest activity for the conjugated linoleic acid esterification during the synthesis of triglycerides. The triglycerides were synthesized by esterification of glycerol and conjugated linoleic acid （CLA） in a no-solvent system using lipase catalysis. We investigated the effects of temperature, enzyme concentration, water content, and time on esterification. Enzyme and water concentrations of up to 1% of the total reaction volume and a system temperature of 60℃ proved optimal for esterification. Similarly, when the esterification was carried out for 24 h, the reaction ratio improved to 94.11%. The esterification rate of the rotating screen basket remained high （87.28%） when the enzyme was re-used for the 5th time. We evaluated the substrate selectivity of lipase （NOVO 435） and determined that this lipase prefers the 10,12-octadacadienoic acid to the 9,11-octadecadienoic acid.

Optimization of Culture Components for Laccase Activity from Pleurotus Ostreatus P40 Using Response Surface Methodology

High enzymatic activity is required for laccase applications.Central composite design （CCD）-based response surface methodology （RSM） can effectively increase the enzymatic activity of Pleurotus ostreatus P40 in liquid substrate fermentation.Initial screening of the nutritional components was performed using a Plackett-Burman design.The variables,namely,bran,bagasse,Tween 80,and yeast extract,were found to have statistically significant effects on laccase activity.These variables were further optimized using CCD-based RSM.Optimal concentrations for the maximum laccase activity were 8.144 2 g/L bran,50 g/L bagasse,0.424 1 mL/L Tween 80,and 2.832 5 g/L yeast extract.Under optimized conditions,the maximum measured laccase activity reached 96 480 U/L,which was close to the predicted value （104 830 U/L） by RSM.Therefore,RSM can be used to optimize culture components for laccase activity from Pieurotus ostreatus P40.

Experimental Research of Tankyrase1 Antisense Oligodeoxynucleotides on the Proliferation of Lung Cancer Cell Nodules

OBJECTIVE To observe the effects of sense and antisense oligodeoxynucleotides of tankyrase 1 （TANK1-SODN and TANK1-ASODN） on murine tumor growth following intratumoral injection, investigate the actual suppressing result and mechanism of TANK1-ASODN on cancer cell proliferation, and discuss the possibility of using it in gene therapy on human lung cancer cells. METHODS After BALB/c nude mice had been subcutaneously inoculated with human lung cancer cell line CALU and it had grown into tumor nodules, we distributed these mice randomly into 3 groups： 4 in saline treatment group, and 5 each in TANK1-SODN group, and TANK1-ASODN groups. Then multiple direct intratumoral injections of synthesized TANK1-ASODN given continuously into tumor nodules for 16 days, this was compared with TANK1-SODN and saline control groups. During the experiment we measured the tumor volume every 5 days with vernier calipers; observed the histopathological characteristics of tumor tissues under microscope; went further to detect the minute changing of ultrastructure of cancer cells by electron microscope; tested the expression levels of ki67 and hTERT protein by means of SABC immunohistochemical method; and detected the lung cancer cells＇ hTERT mRNA expression level by hybridization in situ （ISH） in each group. RESULTS After 16 days of continuous injection, the tumor volume in TANK1-ASODN group was significantly smaller than the other 2 groups （both P 〈 0.01）; quite a lot of tumor cell degeneration and necrosis were observed in mice given TANK1-ASODN. The results of electron microscope also showed that TANK1-ASODN has the power to kill cancer cells in various ways. Moreover, statistically signi.cant decreases in the positive expression ratio of Ki67 Labeling Index （P 〈 0.01）, hTERT protein （P 〈 0.01）, and hTERT mRNA （P 〈 0.01） were consistently observed in the TANK1-ASODN group. CONCLUSION Human lung cancer cell line CALU expressed high telomerase activity. TANK1-ASODN had the ability to decline the high expression level of hTERT; inhibit the activity of telomerase, accelerate tumor cell degeneration and necrosis; and then suppress the proliferation of cancer cells.

A Mew Technique to Rapidly Test Agrochemicai Residues In Fruits and Vegetables

Extraction of AchE, relationship between substrate and enzyme concentration, and inhibition effects of the agrochemicals to AchE are discussed in this paper. Through the re-search, the proper AchE concentration for hydrolysis of 1 ml 1mmol/L substrate and I50 val-ues of the agrochemicals to AchE are decided. It is proved that Asch-DTNB method is a rapid test tool for agrochemical residues in fruits and vegetables.A rapid test card has been developed with sensitivity of 0.05mg/L.

Telomerase activity analysis of esophageal carcinoma using microdissection-TRAP assay

OBJECTIVES: To investigate telomerase activity in esophageal squamous cell carcinoma (SCC) and its preneoplasia lesions, and to study the relationships between telomerase activity and cancer differentiation, cancer invasiveness, and lymphatic metastasis. METHODS: Telomerase activity in esophageal SCC tissues, adjacent dysplasia tissues and normal epithelia from the surgical edge were assessed by microdissection-TRAP (telomeric repeat amplification protocol)-silver staining assay. RESULTS: Telomerase activity was detected in 37 (82.2%) of 45 esophageal tumors, 23 (79.3%) of 29 dysplasias, and 2 (5%) of 40 normal epithelia. There was a significant difference in activity between dysplasia and normal epithelium, as well as between tumor and normal epithelium. Twenty-six (92.9%) of 28 tumors with lymphatic metastasis had detectable telomerase activity compared to 11 (64.7%) of 17 non-lymphatic metastasis tumors. These relationships were statistically significant (P

Total synthesis of dansyl and biotin functionalized ganglioside GM3 by chemoenzymatic method

Ganglioside GM3, as well as other gangliosides, offers a variety of modifications in its sialic acid and ceramide moieties GM3 exhibits various types of important biological activities, due to the inability to effectively observe the trafficking of ganglioside GM3, developing sensitive research tools for specific monitoring of GM3 expression and activity is thus desirable. The total synthesis of a dansyl and biotin bifunctionalized fluorescent ganglioside GM3 were reported in this article. From lactose after 13 reaction steps, the compound of 2′ -biotinoylaminoethyl-6-N-dansylamido-6-deoxy-β-D-galatopyranosyl-(1→4)-β-D-glucopy-ranoside was obtained in total yield of 16.2%. Sialylation of dansyl and biotin functionalized lactose by enzymatic method gave dansyl and biotin labeled ganglioside GM3. The fluorescent property of this compound was also investigated.

A colorimetric method for α-glucosidase activity assay and its inhibitor screening based on aggregation of gold nanoparticles induced by specific recognition between phenylenediboronic acid and 4-aminophenyl-α-D- glucopyranoside

A colorimetric method has been established for a-glucosidase activity assay and its inhibitor screening. The method is based on the specific recognition between 1,4-phenylenediboronic acid （PDBA） and 4-aminophenyl-a-D-glucopyranoside （pAPG）, which may induce aggregation of pAPG-functionalized gold nano- particles （AuNPs） to achieve color change of the test solution. Because pAPG is the substrate of α-glucosidase, the aggregation of AuNPs will be influenced by α-glucosidase since there is no coordination reactivity between PDBA and 4-aminobenzene, the hydrolyzed product of pAPG catalyzed by the enzyme. Therefore, a simple and easily-operated colorimetric method for the assay of a-glucosidase activity can be developed. Under the optimized experimental conditions, the ratios of absorbance at a wavelength of 650 nm to that at 520 nm vary linearly with the α-glucosidase activity within a range from 0.05 to 1.1 U/mL with a lowest detection limit of 0.004 U/mL. Moreover, using the proposed method, the inhibition effect of gallic acid and quercetin on a-glucosidase activity can be tested with IC50 values of 1.16 mM and 1.82 μM, respectively. Thus, the method has a great potential not only for the detection of a-glucosidase activity, but also for the screening of its inhibitors.