桦褐孔菌菌丝体与子实体蛋白质营养价值评价

采用国际上通用的营养价值评价方法,对桦褐孔菌菌丝体与子实体蛋白质的营养价值进行全面评价。结果表明,桦褐孔菌子实体中蛋白质的氨基酸含量分别比菌丝体和鸡蛋模式高45．9％和49．1％,其菌丝体的氨基酸评分、化学评分、必需氨基酸指数、生物价、营养指数和氨基酸比值系数分分别为75．0,44．5,92．2,88．8,24．7和52．0,营养价值比子实体高。     

羽叶决明栽培鸡腿菇子实体营养价值评价

采用国际通用营养价值评价方法,研究了羽叶决明替代常规培养料木屑对鸡腿菇子实体蛋白质营养价值的影响。结果表明,羽叶决明替代木屑栽培鸡腿菇,其子实体蛋白质营养价值较高;在6种不同替代组合培养料栽培的鸡腿菇中,以羽叶决明100％替代木屑的蛋白质综合评价最优、营养价值最高,6项评价指标中,其中必需氨基酸指数（Essential AminoAcid Index）、生物价（Biological Value）、营养指数（Nutritional Index）3项分别为117．37、116．23和68．08,均居首位,且与对照间差异达到极显著水平（P〈0．01）,氨基酸评分（Amino Acid Score）和氨基酸比值系数分（Amino Acid Ratio Coefficient Score）分别为80．39和77．56,均居第2位,化学评分（Chemical Score）为69．84,居第3位。     

蛋白质的营养评价及其在食用菌营养评价上的应用

本文阐述了食用菌蛋白质营养评价的基本方法。在生物学评价法中主要介绍了真消化值、蛋白质功效比、净蛋白比、净蛋白利用率及生物价;在非生物学评价法中主要介绍了化学分、氨基酸分、必需氨基酸指数、营养指数及氨基酸比值系数分。文中分析了各种评价方法的优缺点,井探讨了菌类蛋白质营养评价的应用价值及其前景。     

牛舌菌蛋白质的营养评价

分析了牛舌菌蛋白质的营养价值。结果表明，牛舌菌蛋白质所含的9种人体必需氨基酸（包括酪氨酸和胱氨酸）占氨基酸总量的51.4% ,其氨基酸评分（AAS）、化学评分（CS）、必需氨基酸指数（EAAI）、生物价（BV）和氨基酸比值系数分（SRCAA）分别为71.4、45.4、85.1、81.1和70.18，营养指数（NI）为16.0。这些结果表明，牛舌菌具有较高的营养价值。     

牧草料栽培金顶侧耳蛋白质营养价值评价

应用国际上通用的营养价值评价方法，对牧草料栽培金顶侧耳（Pleurotus citrinopileatus Sing.）子实体蛋白质的营养价值进行全面评价，并与稻草料和棉籽壳料栽培金顶侧耳的子实体蛋白质进行比较。分析表明，牧草料栽培金顶侧耳的必需氨酸总量最高，占其氨基酸总量的40.8%，化学评分（CS）、氨基酸评分（AAS）、必需氨基酸指数（EAAI）、生物价（BV）、营养指数（NI）分别为38.2、60.0、79.8、75.2和35.5，均居三种参比培养料栽培金顶侧耳的第一位。氨基酸比值系数分（SRCAA）为77.9，居第二位，优于棉籽壳料栽培金顶侧耳的子实体蛋白质。这些结果证实，牧草料栽培金顶侧耳的子实体蛋白质具有较高营养价值，牧草是栽培金顶侧耳的优良培养料。     

以大米为基质栽培杏鲍菇的营养成分分析及评价

采用大米为基质栽培杏鲍菇,并将得到的子实体与木屑栽培的杏鲍菇的营养成分进行比较,结果米基杏鲍菇的碳水化合物含量提高164.71%,蛋白质含量提高68%,膳食纤维含量提高500%,氨基酸总量提高63.23%。米基杏鲍菇必需氨基酸指数(EAAI)、生物价(BV)分别为66.06、60.31,均高于木屑杏鲍菇。     

工厂化栽培金耳的氨基酸组成及蛋白质营养评价

为研究工厂化栽培金耳的氨基酸特征,评价其蛋白质营养价值,进行工厂化栽培金耳的氨基酸组成的测定,并基于现行国际模式,采用系统的模型评价其蛋白质品质。试验结果:金耳的蛋白含量为9.2 g/100g(干重),共检测到16种氨基酸(色氨酸未检测,半胱氨酸未检出),其中甜味氨基酸的含量高达4.4785 g/100g(干重),占总氨基酸的比率达41.5%,鲜味氨基酸的含量达4.9746 g/100g(干重),占总氨基酸的比率达46.09%;必需氨基酸占总氨基酸的比率为48.94%,必需氨基酸与非必需氨基酸的比值为95.85%,均高于FAO/WHO的标准;但氨基酸组成不平衡,蛋氨酸和苯丙氨酸异常丰富,赖氨酸和异亮氨酸含量相对缺乏。     

5个枇杷晚熟品种果实氨基酸组成和含量分析

为了探讨白肉枇杷和红肉枇杷果实的风味和营养品质的差异,利用氨基酸自动分析仪测定了5个晚熟枇杷品种果实的氨基酸含量。结果表明,新白1号枇杷果实中天门冬氨酸和谷氨酸等鲜味类氨基酸含量占总氨基酸的53.9%,明显高于红肉枇杷中的含量;而红肉枇杷的甜味类氨基酸和芳香类氨基酸含量略高于白肉枇杷。白肉枇杷中药用氨基酸含量高于红肉枇杷,而红肉枇杷符合氨基酸模式谱的指标多于白肉枇杷。     

成熟度对‘红地球’葡萄氨基酸营养价值的影响

【目的】探明不同成熟度对‘红地球’葡萄氨基酸营养价值的影响。【方法】以不同成熟度(盛花期后88~123 d)的‘红地球’葡萄为试材,测定果实氨基酸含量。同时,以鸡蛋蛋白质为标准蛋白,WHO/FAO氨基酸参考模式为评价标准,进行氨基酸营养价值评估。【结果】‘红地球’葡萄至少包含18种氨基酸,包括8种必需氨基酸、2种儿童必需氨基酸。氨基酸含量随着采摘期变化而变化,其中必需氨基酸、非必需氨基酸、儿童必需氨基酸、药用氨基酸、支链氨基酸、增香与着色氨基酸、伯胺基氨基酸、特殊功效氨基酸含量(w,后同)分别为557.3~659.7、1 176.3~1 690.8、326.8~703.8、1 130.0~1 644.1、231.7~285.3、926.1~1 404.8、1 126.0~1 609.0、1 032.9~1 510.7 mg·kg^-1,组间差异显著(p <0.05),在完全成熟期Ⅰ达到峰值;精氨酸、天门冬氨酸和谷氨酸等风味氨基酸含量与其味觉阈值的比值分别介于2.62~6.31、4.50~5.47和4.64~6.09,随着采摘期的延长呈先升后降变化趋势。氨基酸评分、化学评分和氨基酸比值系数分依次为11.33~14.07、18.32~4.43、49.70~57.04。苯丙氨酸等8种氨基酸含量相对不足,赖氨酸和苏氨酸含量相对过剩。【结论】不同成熟度对‘红地球’葡萄氨基酸含量及其营养价值影响显著。     

甜瓜果实蔗糖代谢酶提取液中2种去糖方法的比较

以甜瓜鲁厚甜1号开花当天的果实为试材,分别测定蔗糖代谢相关酶—酸性转化酶(AI)、中性转化酶(NI)、蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)的活性。对酶提取液中干扰测定结果的可溶性糖采取2种方法进行去除(1)直接用SephadexG-25柱离心去糖;(2)酶提取液先经过硫酸铵沉淀,去糖,重新溶解后再用SephadexG-25柱脱盐。结果显示,方法2处理后AI和NI、SPS和SS的活性均高于方法1。这表明在研究甜瓜果实蔗糖代谢相关酶活性时,采用方法2处理酶提取液效果更好。     

Expression of endoplasmic reticulum-associated apoptosis protein CHOP in lung tissues of COPD model rats

AIM: To investigate whether cigarette smoke (CS) promotes the expression of endoplasmic reticulum-associated apoptosis protein CCAAT/enhancer-binding protein homologous protein (CHOP) in rat lung tissues.METHODS: Adult male Wistar rats (n=40) were randomly divided into 4 groups with 10 rats in each group:control group, CS-2 group (exposed to CS for 2 months), CS-4 group (exposed to CS for 4 months) and ex-smoking (Ex-S) group (exposed to CS for 4 months and then quit smoking for 1 month). The percentage of forced expiratory volume in 0.3 second to forced vital capacity (FEV_0.3/FVC) and peak expiratory flow (PEF) were measured. TUNEL assay was used to detect the apoptotic cells. In situ hybridization and RT-PCR were used to determine the mRNA expression of CHOP. The methods of immunohistochemistry and Western blot were used to determine the protein expression of CHOP. Western blot was also used to determine the protein levels of protein kinase R-like endoplasmic reticulum kinase (PERK), p-PERK, eukaryotic initiation factor (eIF) 2α and p-eIF2α.RESULTS: The pulmonary function greatly decreased in the rats exposed to CS for 2 months in comparison with control group (P<0.05), markedly decreased in the rats exposed to CS for 4 months as compared with the rats after exposure to CS for 2 months (P<0.05), and was improved little in ex-smoking rats (P>0.05). The structural destruction of the lung was observed in the rats exposed to CS for 2 months, and more obvious changes were found in the rats exposed to CS for 4 months. However, the structural destruction of the lung remained obvious in ex-smoking rats. The apoptotic cells were markedly increased in the rats exposed to CS for 2 months and were even more in the rats exposed to CS for 4 months. The apoptotic cells were alveolar epithelial cell I (ACE I), ACE Ⅱ, vascular endothelial cells and bronchial epithelial cells. The protein levels of p-PERK, p-eIF2α and CHOP were remarkably increased in the rats after exposure to CS for 2 months compared with the control rats (P<0.05), significantly elevated in the rats exposed to CS for 4 months compared with the rats exposed to CS for 2 months (P<0.05), and slightly decreased in ex-smoking rats in comparison with the rats after exposure to CS for 4 months (P>0.05). The total protein levels of PERK and eIF2α did not change between the control rats and those exposed to CS.CONCLUSION: CS promotes the development of chronic obstructive pulmonary disease (COPD) by inducing the expression of endoplasmic reticulum-associated apoptosis protein CHOP via PERK/eIF2α/CHOP signaling pathway.     

Anti-inflammatory effects of PYNOD on lipopolysaccharide-stimulated BV2 microglial cells

AIM: To validate the anti-inflammatory effect of PYNOD on lipopolysaccharide (LPS)-stimulated BV2 microglial cells.METHODS: A specific GFP and PYNOD fluorescence expression vector pEGFP-C2-PYNOD driven by the promoter of CMV gene was constructed. The anti-inflammatory properties of PYNOD were studied using LPS-stimulated BV2 microglia model. The productions of nitric oxide (NO), inducible NO synthase (iNOS), interleukin-1β (IL-1β) which caspase-1 were evaluated as inflammatory parameters. RESULTS: Pretreatment with pEGFP-C2-PYNOD to BV2 microglia cells stimulated by LPS significantly inhibited the excessive productions of NO and IL-1β, which was associated with down-regulation of iNOS and caspase-1 at mRNA and protein levels. CONCLUSION: PYNOD might be useful for treating the inflammatory and deleterious effects of BV2 microglial cell activation in response to LPS stimulation.     

甜瓜抗旱性相关基因MeP5CS的克隆、序列分析及表达

用GenBank登录的抗旱相关基因序列进行比对,在保守区域设计一对引物,利用RT-PCR获得了一个甜瓜抗旱相关基因,命名为MeP5CS。生物信息学分析表明,该基因全长1000bp,开放阅读框(ORF)753bp,编码250个氨基酸;MeP5CS蛋白大小约82.18kD,理论pI值为4.90;MeP5CS编码蛋白与桐花树、猕猴桃和葡萄同源性较高,分别为94%、81%和73%。该蛋白含约40.2%的α-螺旋,25.2%的β-转角,34.6%的不规则卷曲,在第19～20氨基酸之间存在信号肽的剪切位点;MeP5CS编码蛋白是疏水性蛋白,有2个跨膜螺旋结构和13个磷酸化位点。半定量RT-PCR表明,MeP5CS在甜瓜根系中表达最高,茎中次之,叶片中表达较低。MeP5CS基因在甜瓜组织中的表达受丛枝菌根真菌(AMF)和水分胁迫双重诱导,与甜瓜的组织特异性和水分胁迫处理时间有关。水分胁迫条件下,接种AMF可以显著增加甜瓜幼苗脯氨酸的积累量,菌根甜瓜幼苗的脯氨酸积累与MeP5CS基因的表达呈正相关。     

Endogenous hydrogen sulfide regulates oxidant/antioxidant balance in rat with chronic obstructive pulmonary disease

AIM: To investigate the role of hydrogen sulfide (H₂S) in the protection against oxidative stress in rats with chronic obstructive pulmonary disease (COPD).METHODS: The rat model of COPD was established by cigarette smoking (CS) combined with lipopolysaccharide (LPS) instillation. Thirty-two healthy male Sprague-Dawley rats were randomly divided into 4 groups: control group, CS+LPS group, CS+LPS+NaHS (H₂S donor) group and CS+LPS+PPG (DL-propargylglycine, an inhibitor of cystathionine-γ-lyase) group. After 30 days, the lung functions of the rats were measured, the histological changes of lungs were observed under light microscope and the pathological scores were calculated. The H₂S level in plasma and the protein expression of cystathionine-γ-lyase (CSE) in the lung tissues were measured. The content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and catalase (CAT) were detected to reflect oxidative stress.RESULTS: Compared with control group, the peak expiratory flow (PEF) decreased by 24% and intra-pressure (IP) increased by 66% in CS+LPS group. The pathological scores of the lung tissues also increased. Compared with CS+LPS group, no change in the lung function was observed after given NaHS or PPG, but the pathological scores decreased in CS+ LPS+ NaHS group. Compared with control group, the content of H₂S in plasma was increased by 26% on day 16. Compared with CS+LPS group, the content of H₂S in plasma of CS+LPS+PPG group was decreased by 22% after 30 days. Compared with control group, the protein expression of CSE increased, and no statistical difference among CS+LPS group, CS+ LPS+ NaHS group and CS+LPS+PPG group was observed. Compared with control group, MDA content in the lung tissues was increased by 24% in CS+LPS group, the activity of SOD was increased by 47% and the activity of CAT was increased by 52%. Compared with CS+LPS group, the MDA content in CS+LPS+NaHS group was decreased by 21%, and no statistical difference in the activity of SOD and CAT was observed. The activity of SOD decreased by 33% after given PPG.CONCLUSION: H₂S plays a role as anti-oxidant in the rats with COPD. The CSE/ H₂S pathway may be involved in the development of COPD.     

Changes in sugar content and relative enzyme activity in grape berry in response to root restriction

Root restriction often depresses photosynthetic capacity and reduces shoot growth, but the photosynthate accumulation in fruit under restriction is higher than in control fruit. However, changes in metabolism responsible for these differences are unclear. To identify the metabolic mechanism by which root restriction affects sugar accumulation in fruit of ‘Kyoho’ grape (Vitis vinifera × V. labruscana), sugar metabolism and related enzyme activities in grape berries produced with and without root restriction were compared. Total sugar content of fruit produced under root restriction was higher than that of control fruit. Acid invertase (AI, EC 3.2.1.26) activity, which increases with berry development, was significantly higher in root-restricted berries than in control berries. Neutral invertase (NI, EC 3.2.1.26) activity showed a similar trend to AI, but the amount of NI activity was lower than AI in both treatments. Sucrose phosphate synthase (SPS, EC 2.4.1.14) and sucrose synthase (SS, EC 2.4.1.13) activity changed slightly with berry development, and there was no significant difference in SS and SPS activity between root restriction and control treatments. Therefore, AI appears to be the key enzyme induced by root restriction that explains the higher sugar content found in grape berry produced under root restriction.     

Effect of acupuncture on mitophagy of skeletal muscle in rats with exercise-induced skeletal muscle damage

AIM: The effect of acupuncture on mitophagy-related protein expression in skeletal muscle of rats after heavy-load exercise was investigated to explore the role of acupuncture in the repairment of exercise-induced skeletal muscle damage. METHODS: Male adult Sprague-Dawley rats (n=128) were randomly divided into 4 groups:control (C, n=8) group, exercise (E, n=40) group, acupuncture (A, n=40) group, and exercise and acupuncture (EA, n=40) group. The rats in E group and EA group performed an eccentric exercise, and the rats in A group and EA group immediately after exercise received acupuncture treatment. The rats in the latter 3 groups were further divided into 0 h, 12 h, 24 h, 48 h and 72 h sub-groups (n=8), and soleus muscle was collected at each time point. The transmission electron microscopy was used to observe the ultrastructural changes of the mitochondria in skeletal muscle. The content of citrate synthase (CS) was measured by ELISA. The protein expression of skeletal muscle PTEN-induced putative kinase 1 (PINK1), parkin and microtubule-associated protein 1 light chain 3 (LC3) was determined by Western blot. RESULTS: After the heavy-load exercise, the mitochondria swelled and accumulated under cell membrane. The number of mitophagosomes was increased, and the content of CS was significantly decreased (P<0.05). The expression of PINK1, parkin and LC3 was significantly elevated (P<0.05). However, the acupuncture intervention after exercise promoted the recovery of mitochondrial ultrastructure, attenuated mitophagolysosome formation, maintained CS content and down-regulated the expression of PINK1, parkin and LC3 (P<0.05). CONCLUSION: Heavy-load exercise causes the damages of mitochondrial structure and function in the skeletal muscle and activates PINK1/parkin pathway to induce excessive occurrence of mitophagy. Acupuncture intervention after exercise is able to alleviate the damage of mitochondria in the skeletal muscle through decreasing the expression of mitochondrial outer membrane protein PINK1, reducing the recruitment of downstream cytoplasmic protein parkin, thereby affecting the combination of LC3 and mitochondria to inhibit the overactivation of mitophagy.     

梨贝壳杉烯氧化酶基因PpKO的克隆及生物信息学分析

以黄金梨（Pyrus pyrifolia Nakai）茎尖为试材,应用同源克隆和RACE方法从梨茎尖中克隆到赤霉素合成代谢的关键酶：内根-贝壳杉烯氧化酶（KO）基因cDNA的全长序列,命名为PpKO,GenBank登录号为：HM003112,其长度为1 752 bp。PpKO的开放阅读框（ORF）编码515个氨基酸,相对分子量为59.007 kD,等电点为7.19。氨基酸同源性分析表明,PpKO与已报道的其它植物的KO氨基酸序列具有59.4% ~ 95.9%相似性;氨基酸聚类分析表明,梨和苹果首先聚类,其次是草莓;生物信息学分析表明：PpKO含有细胞色素P450核心功能域FXXGXRXCXG和氨基端的跨膜结构域。     

凯氏定氮法测定灵芝破壁孢子粉中粗蛋白质含量的不确定度评定

对采用凯氏定氮法测定灵芝(Ganoderma lucidum)破壁孢子粉中粗蛋白含量的不确定度进行评定,结果表明合成相对标准不确定度为0.02%,扩展不确定度为0.04%。在本次孢子粉粗蛋白质测定中,引入不确定度的主要因素有盐酸标准溶液的浓度及样品测定中实际消耗的标准溶液体积。     

Propofol inhibits LPS-induced inflammatory response in microglia via TRPV1 ion channels

AIM:To investigate the effects of propofol (P) on the inflammatory response of microglia induced by lipopolysaccharide (LPS) and the mechanisms. METHODS:Mouse microglia BV2 cells were treated with LPS at 100 μg/L to establish a neuroinflammatory injury model. The BV2 cells were divided into 4 groups:control group (C group), model group (L group), L+P group and LPS+AMG517 group (L+A group). The level of tumor necrosis factor-α (TNF-α) in the cell culture supernatant was measured by ELISA. The mRNA expression of transient receptor potential cation channel subfamily V member 1 (TRPV1) was detected by real-time PCR. The protein levels of TRPV1, TNF-α, interleukin-1β (IL-1β), interleukin-6 (IL-6) and phosphorylated calcium/calmodulin-dependent protein kinase Ⅱ (p-CaMKⅡ) were determined by Western blot. The content of free Ca²⁺ in the microglia BV2 cells was detected by Fluo-3 AM assay. RESULTS:Compared with C group, the level of TNF-α was significantly increased in L group (P<0.01), but that in P group was not changed. Compared with L group, the level of TNF-α was significantly lower than that in L+P group within 4 h (P<0.01). Compared with C group, the mRNA expression of TRPV1 was significantly increased in L group (P<0.01). Compared with L group, the mRNA expression of TRPV1 was significantly down-regulated in L+P group (P<0.01).Compared with L group, the protein levels of TNF-α, IL-1β, IL-6 and p-CaMKⅡ and intracellular Ca²⁺ concentration were significantly lower than those in L+P group and L+A group (P<0.01). CONCLUSION:Propofol inhibits the inflammatory response of microglia by reducing the expression of TNF-α, IL-1 and IL-6, which may be related to the down-regulation of TRPV1 and p-CaMKⅡ and the reduction of intracellular Ca²⁺ concentration.     

多年生黑麦草P5CS基因的cDNA克隆、表达及亚细胞定位

以多年生黑麦草(Lolium perenne)‘Derby'为试材,采用RT-PCR结合RACE技术,克隆获得1个Δ1–吡咯啉–5–羧酸合成酶基因(P5CS)的cDNA序列,全长2528bp,推断其编码716个氨基酸,命名为LpP5CS。序列分析表明:LpP5CS基因与小麦TaP5CS和水稻OsP5CS基因核苷酸序列的相似性分别为92.05%和85.82%,氨基酸序列的相似性分别为93.99%和87.99%。半定量PCR结果表明,LpP5CS基因在多年生黑麦草根、茎,叶均有表达。盐处理下,表达量高于对照,其中叶片中表达量最高,根中最低。200mmol·L-1NaCl处理下,LpP5CS基因表达量随处理时间延长,有先升高后降低的趋势。洋葱鳞茎表皮细胞的瞬时表达显示,LpP5CS蛋白定位于细胞膜和细胞核。