家蚕、蓖麻蚕、樗蚕和樟蚕酸性磷酸酶同工酶遗传分析

根据垂直不连续聚丙烯酰胺凝胶电泳法研究家蚕、蓖麻蚕、樗蚕和樟蚕体液酸性磷酸酶(ACP)同工酶的结果指出:1)家蚕A染色区的4条ACP酶带由2个基因座上的4个等位基因支配;2)多化性品种和中国系统二化性品种的ACP同工酶具有明显的起源同源性;3)蓖麻蚕的ACP同工酶是2个基因座上3个等位基因编码的单链分子,樗蚕和蓖麻蚕具有相似的酶谱类型;4)种蚕的ACP同工酶活性存在显著差异.

应用酸性磷酸酶同工酶测定杂交菜心种子纯度的研究

用等电聚焦电泳（ＩＥＦ）方法对杂交菜心种子子一代杂种及其亲本早期幼苗的酸性磷酸酶同工酶（ＡＣＰ）进行分析，发现供试的两个菜心子一代杂种与其亲本的酸性磷酸酶同工酶有明显稳定的区别。在此基础上，建立用酸性磷酸酶同工酶电泳分析检测菜心一代杂种纯度技术，检查结果与田间鉴定结果基本一致。

不同施药周期DDVP对德国小蠊酶活性的影响

目的 探讨德国小蠊在敌敌畏 (DDVP)不同施药周期作用下的酶活性变化规律。方法 采用分光光度法测定德国小蠊乙酰胆碱酯酶、酸性磷酸酯酶、羧酸酯酶及α-乙酸萘脂 (1-NA)和 β -乙酸萘酯 (2 -NA)酶的活性 ,应用SAS软件进行统计分析。结果 随着施药周期的延长 ,乙酰胆碱酯酶酶活性受抑制 ,酶活性降低 ,随后受抑制程度缓慢降低 ,酶活性逐渐上升 ;酸性磷酸酯酶的活性呈上升趋势 ;羧酸酯酶、1-NA和 2 -NA酶都存在随施药周期的延长 ,酶活性上升后又降低的趋势。结论 乙酰胆碱酯酶是DDVP的主要作用靶部位之一 ,受DDVP作用影响最大。酸性磷酸酯酶和 1-NA是DDVP的重要代谢酶。

Analysis on Activity of Immunity-related Enzymes in First Familiar Generation and Self Cross F1 of Platichthys stellatus Pallas

[Objective] The aim was to study the activity of immunity-related enzymes in each tissue of self cross F1 of Platichthys stellatus Pallas and first familiar generation of P.stellatus ♀×Verasper variegates ♂.[Method] The activity difference of three important non-specific immunity-related enzymes：acid phosphatase （ACP）,alkaline phosphatase （AKP） and superoxide dismutase （SOD） existed in muscle,liver,spleen and intestine of first familiar generation and self cross F1 of P.stellatus was compared.[Result] The activity of ACP and AKP in muscle,liver and intestine of first familiar generation was higher than that of self cross F1,while the activity of ACP and AKP in liver of first familiar generation showed no significant difference with that of self cross F1; the SOD activity decreased significantly in tissues expected for muscle.[Conclusion] The result had provided theoretic basis for the breeding of P.stellatus.

Phosphorus Starvation-induced Expression of Leaf Acid Phosphatase Isoforms in Soybean

Leaf acid phosphatase (APase) activities of 274 soybean genotypes were surveyed under field conditions with two levels of P supplies, and a nutrient solution culture experiment with eight selected genotypes was subsequently conducted under greenhouse conditions to further characterize APase activity and its isoform expression induced by P starvation. Results from the field experiment showed that there was a great genotypic variation for leaf APase activity among the tested soybean genotypes from different origins, and APase activity in many of the tested genotypes (about 60%) was generally increased in the treatment without P fertilizer addition. Results from the nutrient solution culture experiment showed that APase activity in all the eight tested genotypes was generally enhanced by P starvation. Six isoforms of APases were detected in isoelectric focusing gels with samples from both young and old leaves. The activity of all the six isoforms was increased by P starvation, but no new APase isoform was induced. Our results suggest that leaf APase activity could serve as an enzymatic indicator of P starvation for soybean; the increase in leaf APase activity under low P stress was mainly caused by the increase in the activity of existing isoforms but not by the induction of new isoforms.

6 Kinds of Isozymes after Long-term Subculture of Emmenopterys henryi Oliv.

[Objective] The aim was to study the changes of zymography in 6 kinds of isozymes after long-term subculture of Emmenopterys henryi Oliv.. [Method] Non-denaturing polyacrylamide gel electrophoresis was used to analyze isozyme patterns such as esterase (EST),acid phosphatase (ACP),ATP enzyme (ATPase),amylase (AMY),superoxide dismutase (SOD) and peroxidase (POD) in long-term subculture callus of Emmenopterys henryi Oliv. [Result] The research showed that there were differences among the 6 kinds of isozymes in embryogenic callus and non-embryogenic callus of Emmenopterys henryi Oliv.,and both levels could be taken as the basis for identification,the EST,ACP,and POD of non-embryogenic callus were significantly higher than embryogenic callus. The browning of non-embryogenic callus was non-level in the AMY,SOD and POD isozymes when was compared with normal non-embryogenic callus,while the EST,ACP and ATPase isozymes decreased; When the browning of embryogenic callus was contrasted with normal embryogenic callus,EST isozyme increased and the other 5 kinds of enzymes decreased. [Conclusion] The study provided theoretical basis for research morphological difference and browning of long-term tissue culture of Emmenopterys henryi Oliv..

Research on Sex and Tissue Differences in Isozymic Phenotype of Varicorhinus macrolepis through Isozyme Electrophoresis

Polyacrylamide gel electrophoresis (PAGE) and biochemical staining method were used in this study for the analysis on malate dehydrogenase (MDH,E.C. 1.1.1.37) isozymes zymogram in 11 different types of tissues of male and female Varicorhinus macrolepis. It had been found for the first time that the phenotype of malate dehydrogenase (MDH),acid phosphatase (ACP) and superoxide dismutase (SOD) showed difference between male and female V. macrolepis,and there was no difference among different individuals in the same sex. Therefore,the electrophoresis band of malate dehydrogenase,acid phosphatase and superoxide dismutase could be used as an indicator for the identification of gender and tissues of V. macrolepis,which would provide basic data for the developmental genetics,variety improvement and directed breeding of V. macrolepis groups,thus facilitating the development and protection of this valuable fish species.

Biochemical response of the mussel Mytilus coruscus(Mytiloida:Mytilidae) exposed to in vivo sub-lethal copper concentrations

Many aquatic organisms are negatively affected by exposure to high copper concentrations. We investigated the biochemical response of the mussel Mytilus coruscus （Mytiloida： Mytilidae） to copper exposure. In vivo bioassays using M. coruscus and different copper concentrations were conducted. The activity of six biomarkers, namely superoxide dismutase （SOD）, catalase （CAT）, acid phosphatase （ACP）, alkaline phosphatase （AKP）, glutamic-oxaloacetic transaminase （GOT） and glutamic-pyruvic transaminase （GPT） were measured. Survival rates decreased with increased copper concentrations and exposure times. The LCs0 values at 48, 72, and 96 h exposure were 0.48, 0.37, and 0.32 rag/L, respectively. Within digestive glands, CAT activity increased with increasing Cu concentrations. The activity of AKP showed no significant change, while the remaining four enzymes showed decreasing activity with increasing Cu concentrations. Within the gills, AKP activity increased when the Cu concentration was 0.05 mg/L, but showed no significant changes at higher concentrations. Activity of CAT and ACP within gills tended to decrease with increasing Cu concentration. The activity of SOD and GPT decreased at an exposure concentration of 0.2 mg/L. GOT activity within gills decreased at 0.1 mg/L and increased at an exposure concentration of 0.2 mg/L. Within the adductor muscle, AKP activity increased at 0.05 mg/L but did not change at higher exposure concentrations. ACP activity within adductor muscle tissue showed no change, while activities of CAT, GOT and GPT decreased with increasing Cu concentrations. SOD activity within the adductor muscle tissue significantly decreased at the 0.02, 0.05 and 0.2 mg/L exposure concentrations. Our results show tissue specific differences for the six biomarkers in for M. coruscus. Our findings provide the basis for the establishment of reference activity levels against which biomarker changes can be estimated, and are essential preliminary steps in development of in vivo bioassays.

Growth, Metabolism and Physiological Response of the Sea Cucumber, Apostichopus japonicus Selenka During Periods of Inactivity

The growth, metabolism and physiological response of the sea cucumber, Apostichopus japonicus, were investigated during periods of inactivity. The body weight, oxygen consumption rate (OCR), activities of acidic phosphatase (ACP), alkaline phosphatase (AKP), catalase (CAT) and superoxide dismutase (SOD), and content of heat shock protein 70 (Hsp70) in the body wall and coelomic fluid of A. japonicus were measured during starvation, experimental aestivation and aestivation. The results showed that the body weight of sea cucumber in the three treatments decreased significantly during the experimental period (P < 0.05). The OCR of sea cucumber reduced in starvation and experimental aestivation treatments, but increased gradually in natural aestivation treatment. The activities of ACP and AKP of sea cucumber decreased gradually in all treatments, whereas those of SOD and CAT as well as Hsp70 content decreased in the starvation and experimental aestivation treatments and increased in natural aestivation treatment. The sea cucumber entered a state of aestivation at 24℃. To some extent, the animals in experimental aestivation were different from those in natural aestivation in metabolism and physiological response. These findings suggested that the aestivation mechanism of A. japonicus is complex and may not be attributed to the elevated temperature only.

Effect of E. coli coli on Anti-disease Activities of Scailops: Argopecten irradians

The effect of acute E. coli challenge on the anti-disease activity of scallops Argopecten irra-dians is examined. The treatments of scallop from which hemolymph samples were taken for study included (1) control scallops, (2) sham-injected scallops, (3) PSW-injected scallops and (4) E. coli-injected scallops. From the beginning, the anti-disease activities of scallops are deter -mined at 12 hr and 24 hr.The concentrations of circulating hemocytes, the total serum protein concentrations and the activities of alkaline phosphatase, acid phosphatase and superoxide dismutase in the scallops Argopecten irradians are determined.Injection with E. coli results in a significant elevation in the concentration of circulating hemocytes and in the alkaline phosphatase activity and a significant decline in the total serum protein concentration and in the superoxide dismutase activity at 24 hr postchallenge. It shows that metabolism of bay scallop is expedited to adopt the challenge.

Hepatoprotective evaluation of Anogeissus latifolia:In vitro and in vivo studies

AIM： To evaluate the hepatoprotective activity of a hydroalcoholic extract of the bark of Anogeissus latifolia; in vitro in primary rat hepatocyte monolayer culture and in vivo in the liver of Wistar rats intoxicated by carbon tetrachloride （CCl4）. METHODS： In the in vitro study, a primary hepatocyte monolayer culture was treated with CCh and extract of Anogeissus latifolia. Hepatoprotective activity was demonstrated in the CCh damaged primary monolayer culture. In the in vivo study, the hepatoprotective activity of a hydroalcoholic extract ofAnogeissus latifolia was analyzed in liver injured CCh-treated rats. Biochemical parameters including serum transaminases [aspartate aminotransferase （AST） and alanine aminotransferase （ALT）] and alkaline phosphatase （ALP） in serum wereanalyzed. The biochemical findings were supplemented with histopathological examination of rat liver sections. RESULTS： In vitro： primary hepatocyte monolayer cultures were treated with CCh and extract of Anogeissus latifolia. A protective activity could be demonstrated in the CCh damaged primary monolayer cultUre. In vivo： Hydroalcoholic extract of Anogeissus latifolia （300 mg/kg） was found to have protective activity in rats with CCh-induced liver damage as judged from serum marker enzyme activity. CONCLUSION： The above findings lead to the conclusion that the hydroalcoholic extract of Anogeissus latifol/a is hepatoprotective. Hence, we suggest that the inclusion of this plant in the management of liver disorders is justified.

Dietary supplementation of some antioxidants against hypoxia

The present study aims to clarify the protective effect of supplementation with some antioxidants,such as idebenone(200 mg/kg,ip),melatonin(10 mg/kg,ip) and arginine(200 mg/kg,ip) and their combination,on liver function(T.protein,albumin,alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase),energetic parameters(adenosine triphosphate,adenosine diphosphate,adenosine monophosphate,inorganic phosphate,total adenylate,adenylate energy charge and potential phosphate).The effect on glycolytic and glycogenolytic enzymes(glucose,glycogen,glycogen phosphorylase,pyruvate kinase and phosphofructokinase against hypoxia) was also studied.The drugs were administered 24 and 1 h prior sodium nitrite intoxication.All biochemical parameters were estimated 1 h after sodium nitrite injection.Injection of sodium nitrite(75 mg/kg,sc) produced a significant disturbance in all biochemical parameters of liver function,energetic parameters and glycolytic and glycogenolytic enzymes.Hepatic damage was confirmed by histopathological examination of the liver as compared to controls.The marked changes in hepatic cells induced by sodium nitrite were completely abolished by pretreatment with the drug combination,suggesting potential protection against sodium nitrite-induced hypoxia.It could be concluded that a combination of both idebenone and melatonin or idebenone and arginine provides potential protection against sodium nitrite-induced hypoxia by improving biochemical parameters and preserving liver histology.

Temperature Effects on Seed Dormancy Relieving of Fritillaria thunbergii Miq

The seed dormancy relieving of Fritillaria thunbergii Miq required 50 days at 8～10℃and 80 days at 3～5℃ In order to study the physiological basis for seed dormancy relieving,peroxidase,acid phosphatase and proteins were assayed by spectrophotometric analysis and electrophoresis The results showed that peroxidase activity and acid phosphatase activity increased and new bands were detected in protein pattems as the seeds were treated with low temperature,these changes resulted in the seed dormancy relieving

Severe chronic diarrhea and weight loss in cholesteryl ester storage disease: A case report

AIM: An inherited deficiency of human lysosomal acid lipase (LAL)results in the rare conditions of Wolman disease and cholesteryl ester storage disease (CESD). We want to present the rare case of CESD in an adult.METHODS: We report about an adult female patient with severe chronic diarrhea and weight loss as a consequence of CESD. Clinical examination revealed signs of malabsorption and slightly elevated liver enzymes.RESULTS: Histopathologic changes in the liver tissue and DNA sequence analysis confirmed the diagnosis of CESD due to homozygosity for the most common CESD mutation,a G934A splice site defect encoded by exon 8 of the lysosomal acid lipase (LIPA) gene.CONCLUSION: It is the first case in the literature with diarrhea as a putative symptom of CESD in adult patients.

Effects of different enzymatic hydrolysis methods on the bioactivity of peptidoglycan in Litopenaeus vannamei

The effects of different hydrolysis methods on peptidoglycan （PG） were assessed in terms of their impact on the innate immunity and disease resistance of Pacific white shrimp, Litopenaeus vannamei. PG derived from Bifidobacteriurn thermophilum was prepared in the laboratory and processed with lysozyme and protease under varying conditions to produce several different PG preparations. A standard shrimp feed was mixed with 0.05% PG preparations to produce a number of experimental diets for shrimp. The composition, concentration, and molecular weight ranges of the soluble PG were analyzed. Serum phenoloxidase and acid phosphatase activity in the shrimp were determined on Days 6-31 of the experiment. The protective activity of the PG preparations was evaluated by exposing shrimp to white spot syndrome virus （WSSV）. Data on the composition of the PG preparations indicated that preparations hydrolyzed with lysozyme for 72 h had more low-molecular-weight PG than those treated for 24 h, and hydrolysis by protease enhanced efficiency of hydrolysis compared to lysozyme. SDS-PAGE showed changes in the molecular weight of the soluble PG produced by the different hydrolysis methods. Measurements of serum phenoloxidase and acid phosphatase activity levels in the shrimp indicated that the PG preparations processed with enzymes were superior to the preparation which had not undergone hydrolysis in enhancing the activity of the two serum enzymes. In addition, the preparation containing more low-molecular-weight PG enhanced the resistance of the shrimp to WSSV, whereas no increased resistance was observed for preparations containing less low-molecular-weight PG. These findings suggest that the immunity-enhancing activity of PG is related to its molecular weight and that increasing the quantity of low-molecular-weight PG can fortify the effect of immunity enhancement.

Impacts of Forest Gaps on Soil Properties After a Severe Ice Storm in a Cunninghamia lanceolata Stand

During January–February 2008, a severe ice storm caused significant damages to forests in southern China, creating canopy gaps and changing soil nutrient availability and enzyme activity. To understand the relationships between gap size, changes in the soil environment and the effects that these changes have on soil processes, we investigated the effects of gap size on soil chemical and biological properties in the forest gaps in a Cunninghamia lanceolata stand in northern Guangdong Province, southern China. Ten naturally created gaps, five large(80–100 m^2) and five small(30–40 m^2), were selected in the stand of C.lanceolata. The large gaps showed a significant increase in light transmission ratio and air and soil temperatures and a decline in soil moisture, organic matter,N and P compared with the small gaps and the adjacent canopy-covered plots in the 0–10 cm soil. The differences in organic matter and nutrient levels found between the large and small gaps and the canopy-covered plots may be related to changes in environmental conditions. This indicated rapid litter decomposition and increased nutrient leaching in the large gaps. Moreover, the lowest levels of catalase, acid phosphatase and urease activities occurred in large gaps because of the decline in their soil fertility. Large forest gaps may have a region of poor fertility, reducing soil nutrient availability and enzyme activity within the C.lanceolata stand.

Effect of Lead on Soil Enzyme Activities in Two Red Soils

Enzyme activities have the potential to indicate biological functioning of soils. In this study, soil urease, dehydrogenase, acid phosphatase and invertase activities and fluorescein diacetate(FDA) hydrolysis were measured in two red soils spiked with Pb2+ranging from 0 to 2 400 mg kg-1to relate the enzyme activity values to both plant growth and the levels of available and total Pb2+concentrations in soils, and to examine the potential use of soil enzymes to assess the degrees of Pb contamination. Soil samples were taken for enzyme activities assaying during 3 month's incubation and then after planting of celery(Apium graveolens L.) and Chinese cabbage(Brassica chinensis L.). Enzyme activities in the red soil derived from arenaceous rock(RAR) were generally lower than those in the red soil developed on Quaternary red earths(REQ). At high Pb2+loadings, in both incubation and greenhouse studies, urease activity and FDA hydrolysis were significantly inhibited. But there were no significant relationships between soil dehydrogenase, acid phosphatase or invertase activity and soil Pb2+loadings in both RAR and REQ soils. The growth of celery and Chinese cabbage increased soil urease activity and FDA hydrolysis, but had minimal effect on dehydrogenase and invertase activities. There were positive correlations between celery biomass and soil urease activity and FDA hydrolysis. These results demonstrate that urease activity and FDA hydrolysis are more sensitive to Pb2+than acid phosphatase, dehydrogenase and invertase activities in the RAR and REQ soils.