乙醇脱氢酶应用研究现状

乙醇脱氢酶(ADH)是广泛分布于人和动物肝脏、植物及微生物细胞之中的含锌金属酶,具有广泛的底物特异性.ADH作为生物体内主要短链醇代谢的关键酶,近年来已经受到了国内外研究者的普遍关注,并对其同功酶的结构、理化性质、生物学功能、遗传学特性、分离提取技术等进行了较多的基础性研究工作,为其在科学研究和工农业生产中的应用奠定了良好基础.文章主要对ADH在工业分析、化工生产、食品和医药研究以及生物科学中的应用进行了概述,以便为其进一步研究开发提供参考.     

Evaluation of Thermal Environment of Cowshed and Correlation Analysis Between Thermal Parameter and Physiological Parameter of Dairy Cows in Central Plain of Hebei

The purpose of this study was to evaluate thermal environment of cowshed in four seasons in central plain of Hebei,and analyze the correlation between the temperature parameters and physiological indexes of dairy cows.Three dairy cow houses with different building structures were selected,the thermal parameters (ambient temperature (Ta) and relative humidity (RH)) and physiological parameters (respiratory rate,rectal temperature and body surface temperature) were detected.The results showed that the Ta,RH and index of temperature and humidity (THI) changed significantly in four seasons (P<0.05),the average daily temperature in summer was the highest (28.59 ℃),and the average daily temperature in winter was the lowest (1.55 ℃).In summer,dairy cows were suffering from mild heat stress for 15.5 h every day,and from moderate heat stress for 6.0 h.In winter,dairy cows were under mild cold stress for an average of 12.0 h per day.The Ta and THI of three cowsheds in each season had no significant differece (P>0.05),except for that in winter.Compared with the cowshed with low wall or roller blind,the average Ta in the cowshed with only roof was 0.80-1.27 ℃ higher in summer and 1.36-1.84 ℃ lower in winter.In addition,the physiological parameters of dairy cows were extremely significantly higher in summer than those in other seasons (P<0.01).The respiration frequency or rectal temperature of cows among cowsheds in summer were significantly different (P<0.05),and the seasonal difference in body surface temperature was significant (P<0.05).Correlation analysis of thermal parameters and cow physiological parameters showed that the respiratory frequency,rectal temperature and body surface temperature were positively correlated with THI and Ta (P<0.05).However,there was no significant correlation between physiological parameters and RH (P>0.05).The results provided scientific basis for environment evaluation of cows,and physiological status of dairy cows were inferred according to environmental thermal parameters,which provided data for occurrence and early warning of stress.     

微卫星标记与考力代羊体重体尺性状遗传关系研究

本试验用单标记法研究考力代绵羊1号染色体ILSTS004~CSSM004和BM6506~MCM130之间的12个微卫星位点ILSTS004、MCMA008、MNS0094、CSSM004、BM6505、BMS4008、CSRD0298、TGLA415、BM8246、RM0509、URB0038、MCM0130,与其绵羊初生重、断奶重、体重(8月龄)、体高、体长、胸围、管围的遗传关系,结果ILSTS004位点不同基因型对初生重影响极显著(P<0·01)、对体重影响显著(P<0·05),CSSM004位点对体长影响显著(P<0·05),其余位点对体重等性状无显著影响(P>0·05)。其中,ILSTS004位点:基因型101099个体的初生重显著高于基因型095093(P<0·05),基因型101099个体的体重显著高于基因型103103(P<0·05);CSSM004基因型196196个体的体长显著大于基因型202202(P<0·05)。提示在1号染色体微卫星位点ILSTS004附近可能存在影响绵羊初生重、体重的数量性状位点(QTL),ILSTS004可作为其遗传标记之一;在CSSM004附近可能存在影响体长的QTL,CSSM004可作为其遗传标记之一。     

Effect of nitric oxide on urotensin-Ⅱ expression in pulmonary arterioles of rats chronically exposed to hypoxia-hypercapnia

AIM: To investigate the roles of nitric oxide/L-arginine (NO/L-Arg) pathway and urotensin-Ⅱ (UⅡ) in the development of pulmonary hypertension induced by chronic hypoxia-hypercapnia in rats.METHODS: Forty male Sprague-Dawley rats were randomly divided into four groups (n=10): normal control group (A), hypoxia-hypercapnia+saline group (B), hypoxia-hypercapnia+L-Arg liposome group (C) and hypoxia-hypercapnia+N-nitro-L-arginine methyl ester (L-NAME) group (D).Contents of UⅡ, UⅡ mRNA and receptor of UⅡ (UT) mRNA in pulmonary arterioles were measured with immunohistochemistry analysis and in situ hybridization, respectively.Change of small pulmonary vascular microstructure was also investigated.RESULTS: (1) The mean pulmonary artery pressure (mPAP) and the weight ratio of right ventricle to left ventricle plus septum ［RV/(LV+S)］ in B and D groups were all higher than those in A group (respectively, P<0.05), with C group significantly lower than those in B group (respectively, P<0.01).(2) Light microscopy showed that the ratio of vessel wall area to total area (WA/TA) and the media thickness of pulmonary arterioles (PAMT) in B group were higher than those in A group (P<0.05), with C group significantly lower than those in B group.(3) The contents of UⅡ, UⅡ mRNA and UT mRNA in pulmonary arterioles in B and D groups were all higher than those in A group (respectively, P<0.01), while the expression of UⅡ and UⅡ mRNA in C group were lower than those in B group (P<0.01).CONCLUSION: The pathological process of pulmonary hypertension induced by chronic hypoxia-hypercapnia might be related to upregulation of UⅡ located in pulmonary arterioles, which might be partially inhibited by exogenous NO in rats.     

Correlation between the signal pathway IKK/NF-κB and the anti-oxidant activity in asthma rats and the modulation of Ginkgo biloba extract

AIM:To investigate the correlation between the signal pathway of IKK/NF-κB and the anti-oxidant activity in asthmatic rats and the modulation of Ginkgo biloba extract (Egb). METHODS:Thirty-six male SD rats were randomly divided into three groups:control group (group C), asthmatic group (group A) and Egb group(group E). Asthma in rats was established by ovalbumin (OVA) challenge methods. The mRNA of IKKβ and the protein of NF-κB P65 in lung tissue were assessed by using in situ hybridization with oligonucleotide probe and immunohistochemisty, respectively. RESULTS:The expression of IKKβ mRNA and NF-κB P65 protein in group A were significantly increased when compared with group C (P＜0.01, respectively), but those data in group E were significantly decreased when compared with group A (P＜0.01, respectively). The concentrations of MDA in serum and BLAF in group A were significantly higher than those in control group (P＜0.01，respectively), but the concentrations of MDA in group E were significantly lower than those in group A (P＜0.01, respectively). The concentrations of GSH in group A were significantly lower than those in group C (P＜0.01, respectively), but the concentrations of GSH in group E were significantly higher than those in group A (P＜0.01, respectively). CONCLUSION:The expression of IKKβ mRNA and NF-κB P65 protein were increased significantly in asthmatic rats. This was possibly attributed to the decreased anti-oxidant activity. Egb increased the anti-oxidant activity and inhibited the activity of IKK/NF-κB.     

条斑型玉米圆斑病病原鉴定及其生物学特性研究

 本研究通过致病性测定、形态学和分子生物学鉴定方法,首次明确陕西省发生的玉米条斑型叶斑病是玉米圆斑病的一种症状类型,由玉米生离蠕孢(Bipolaris zeicola)3号生理小种引起。供试品种豫玉22、农大108、农大3138、东单7号、沈单10号、沈单16号、户单4号、新陕资1号、高农1号和陕单9号都对3号生理小种表现感病,品种间抗病性差异不显著。此菌在PDA、PSA、OMA、CMA、WA和Czapek上均可生长,最适生长培养基为PDA。菌落生长温度为5~35℃,最适菌落生长温度为25℃。分生孢子在5~40℃均可萌发,低于5℃高于40℃萌发率极低,萌发的最适温度为28℃。孢子在pH 3~11均可萌发,萌发最适pH为5。在自来水中分生孢子可以萌发,蔗糖和葡萄糖溶液可以明显促进孢子萌发。     

陈皮对肉鸡生产性能及回盲肠微生物菌群的影响

选用健康、体重一致的1日龄肉仔鸡450羽,随机分为5组,对照1和2为对照组,分别饲喂不添加抗生素和添加抗生素的基础日粮;A、B、C为试验组,饲喂无抗基础日粮,并分别在日粮中添加0.1%,0.2%和0.4%的陈皮,研究了陈皮对肉鸡生产性能和回盲肠乳酸菌及大肠杆菌菌落数的影响。结果表明,日粮中添加不同比例陈皮对肉鸡生产性能和回盲肠的乳酸菌及大肠杆菌菌落数产生的影响不同,这种差异在饲养的前期尤为明显,其中以添加0.2%陈皮组效果最好。     

球虫(菌)清缓释剂防治家兔球虫病和大肠杆菌病效果分析

目的寻求一种高效、优质药物防治家兔球虫病和大肠杆菌病。方法(1)60只健康兔分4组,第1、2组分别注射球虫清缓释剂、球菌清缓释剂,第3、4组分别用伯氨喹啉、地克珠利 氟哌酸拌料日常饲喂,于d 2、5、7、15、30口服球虫孢子化卵囊,隔10 d取粪查卵囊,观察保护率。(2)球虫病兔20只分为第1、3组,球虫病及大肠杆菌病兔20只分为第2、4组,分别用上述方法给药比较治愈率。(3)日常饲养兔150只分3组,分别为注射球虫清缓释剂、伯氨喹啉拌料日常饲喂、对照组,1、2、3个月后查卵囊,观察转阴率、OPG值、死亡率等。(4)幼兔100只分5组,前4组用方法(1)给药,第5组为对照组,比较增重率、饲料转化率、节省防治费率。结果(1)兔注射球虫清缓释剂、球菌清缓释剂后,在d 2~20时,攻击球虫,其保护率均为100%(3/3)。在d 30时,攻击球虫,其保护率均为66.7%(2/3);而口服伯氨喹啉,地克珠利 氟哌酸防治的兔,在d 2~5时攻击球虫,保护率为0,在d 10时,其保护率均为33.3%(1/3),在d 20~30时,其保护率均为66.7%(2/3)。(2)球虫病和大肠杆菌病兔(1、2组)注射球虫清缓释剂、球菌清缓释剂后d 5~60,球虫病、大肠杆菌病的治愈率均为100%;口服伯氨喹啉、地克珠利 氟哌酸(3、4组)后d 5,治愈率均为0,d 20~60时,治愈率为70.0%~90.0%。(3)球虫清缓释剂组在1~2个月时,球虫卵囊减少率及转阴率均为100%,在3个月时,球虫卵囊减少率为98.9%,转阴率为94.0%,存活率为100%。伯氨喹啉组1~3个月,球虫卵囊减少率及转阴率分别为64.4%~89.8%及33.0%~91.2%,死亡率为22.0%~28.0%,存活率为72.0%~78.0%。对照组1~3个月,球虫卵囊增加78.0%~126.0%,死亡率为46.0%~62.0%,存活率为38.0%~54.0%。(4)注射球虫(菌)清缓释剂比口服伯氨喹啉、地克珠利 氟哌酸及对照组,兔的增重率分别提高71.7%、76.1%、374.6%和69.8%、74.1%、369.3%,节省防治费75%。应用球虫(菌)清缓释剂防治球虫病、大肠杆菌病兔10.5万只,获得经济效益108.67万元(包括增重效益、挽回死亡兔损失、减少饲料费和节省防治费四项)。结论球虫(菌)清缓释剂能安全、高效、经济、方便地控制家兔球虫病和大肠杆菌病的发生,保护幼兔安全度过球虫病和大肠杆菌病高危期。     

南方地区中国荷斯坦牛乳中体细胞数变化规律的研究

采用最小二乘法分析了南方地区某奶业集团2006年7个牛场2 063头中国荷斯坦牛22 377次乳中体细胞数(SCC)测定日记录,以揭示SCC的变化规律.结果表明乳中SCC为459.49×103士924.66×103个/mL,不同牛场、月份、泌乳月和胎次对乳中体细胞数的影响均达到极显著水平(P＜0.01),而产犊季节对乳中SCC的影响未达到显著水平(P＞0.05).SCC随着胎次和泌乳月的增加有升高的趋势.     

Sequence analysis and prokaryotic expression system construction of PIA genes isolated from Neisseria gonorrhoeae

AIM: To analyze the nucleotide and putative amino acid sequences of PIA genes isolated from N.gonorrhoeae and to construct the prokaryotic expression system of PIA gene.METHODS: The entire PIA genes from 9 strains of N.gonorrhoeae were amplified by using high fidelity PCR.The target amplification fragments were sequenced after T-A cloning.Homology comparison of the nucleotide and putative amino acid sequences of PIA genes from the isolates with the reported sequences in GenBank was then performed.A prokaryotic expression system of PIA gene was constructed.Different dosages of IPTG were applied to induce the expression of the target recombinant protein (rPIA) and 10% SDS-PAGE plus Bio-Rad Agarose Image Analysor was used to determine the expression level of rPIA.rPIA was extracted using Ni-NTA affinity chromatography and the purified effect was detected by SDS-PAGE.RESULTS: In comparison with the reported PIA gene sequences (GenBank No: L19962),the homologies of nucleotide and putative amino acid sequences of PIA genes from the isolates were 99.6%-100% and 99.1%-100%,respectively,which indicated that all the isolates were belonging to serovars IA6.Output of rPIA was as high as 50.1% of the total bacterial proteins.The purified rPIA only showed a single target protein fragment in gel.CONCLUSION: Serovar IA6 is dominant in the local N.gonorrhoeae isolates and sequences of the encoding gene are relatively conserved.The constructed prokaryotic expression system is able to express rPIA with high efficiency,which may lay a foundation for further development of serological detection kit and vaccine of N.gonorrhoeae.