中草药添加剂对异育银鲫肌肉生化成分的影响

设计中草药配方Ⅰ和Ⅱ，各以0．5％、1％、2％剂量添加到基础饲料中，饲喂异育银鲫（体重1．6－2．2g），2个月后，测定鱼体肌肉的水分，蛋白质、脂肪、灰分，脂肪酸，氨基酸的含量，结果表明，异育银鲫肌肉中水分、蛋白质，各种脂肪酸，水解和游离氨基酸的含量均无显著变化，复方中草药添加剂Ⅰ和Ⅱ对异育银鲫肌肉蛋白质的营养价值和呈味氨基酸的含量无不良影响（P＞0．05），其中，配方Ⅱ的2％添加量可显著地提高异育银鲫鱼肉的脂肪含量，从而改善鱼的肉质。     

Farrerol inhibits nicotine-induced proliferation of pulmonary vascular smooth muscle cells by enhancing Kv1.5 and Kv2.1 expression

AIM:To study the effect of farrerol (Far) on nicotine-induced proliferation of rat pulmonary smooth muscle cells (PASMCs), and further to explore its relationship with voltage-dependent potassium channels (Kv) 1.5 and Kv2.1. METHODS:Firstly, the effect of nicotine on the proliferation of PASMCs was detected by cell counting method, and the optimal concentration of nicotine was selected. Primary cultured PASMCs were randomly divided into 5 groups:normal control group, nicotine (1 μmol/L)group, nicotine (1 μmol/L) + Far (10^-6 mol/L, 10^-5 mol/L and 10^-4 mol/L) Far group. The activity of caspase-3 was measured by apoptosis kit, the cell viability was measured by CCK-8 assay, the apoptotic rate was analyzed by flow cytometry. The expression of Kv1.5 and Kv2.1, and apoptosis-related factors Bcl-2 and Bax at mRNA and protein levels was determined by RT-qPCR and Western blot respectively. RESULTS:Nicotine at 1 μmol/L increased the number of PASMCs to the maximum extent (P<0.01). Nicotine at 1 μmol/L significantly reduced the caspase-3 activity and enhanced the cell viability of the PASMCs (P<0.01). Farrerol at 10^-6~10^-4 mol/L eliminated the effect of PASMCs induced by nicotine in a concentration dependent manner. Compared with control group, nicotine at 1 μmol/L significantly increased the proliferation and inhibited the apoptotic rate of rat PASMCs (P<0.01). The apoptotic rate of PASMCs in farrerol intervention group was significantly higher than that in nicotine group (P<0.01). Nicotine at 1 μmol/L significantly inhibited the expression of Kv1.5, Kv2.1 and Bax but increased the expression of Bcl-2 in PASMCs (P<0.01). Farrerol at 10^-5 mol/L obviously inhibited the effect of PASMCs induced by nicotine. CONCLUSION:Farrerol eliminates nicotine-induced inhibition of caspase-3 and Bax, and enhancement of Bcl-2 in PASMCs by enhancing Kv1.5 and Kv2.1 expression.     

Effect of xeroderma pigmentosum group D gene on proliferation of human umbilical arterial smooth muscle cells induced by Ox-LDL

AIM: To investigate the effects of xeroderma pigmentosum group D (XPD) gene on the proliferation of human umbilical arterial smooth muscle cells (HUASMCs) induced by oxidized low-density lipoprotein (Ox-LDL). METHODS: The recombinant plasmid pEGFP-N2/XPD was transfected into HUASMCs by liposome. The cells were divided into blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, Ox-LDL group, Ox-LDL+pEGFP-N2 group and Ox-LDL+pEGFP-N2/XPD group. The proliferation rate of the cells was detected by MTT and EdU assays. The apoptotic rate and cell cycle distribution were analyzed by flow cytometry. The protein levels of XPD, caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS: Compared with blank control group, the expression of XPD was increased in pEGFP-N2/XPD group (P<0.05). According to the results of MTT and EdU assays, the cell proliferation in pEGFP-N2/XPD group was reduced compared with blank control group (P<0.05). Compared with Ox-LDL group, the cell proliferation in Ox-LDL+pEGFP-N2/XPD group was significantly inhibited (P<0.05). According to the results of flow cytometry, the cell proportion of S phase decreased and the G₀/G₁-phase cell proportion increased significantly in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group compared with blank control group and Ox-LDL group, repectively (P<0.05). Compared with blank control group and Ox-LDL group, the protein level of Bcl-2 decreased and the protein levels of Bax and cleaved caspase-3 increased in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group, respectively (P<0.05). CONCLUSION: XPD inhibits the proliferation of HUASMCs and promotes their apoptosis, and reduces the promoting effect of Ox-LDL on the proliferation of HUVSMCs. XPD may be the target for treatment of atherosclerosis.     

Model construction of rat coronary artery smooth muscle cell endoplasmic reticulum stress induced by thapsigargin

AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress (ERS) model in CASMCs of SD rats. METHODS: CASMCs were cultured by tissue explant method. The morphological characteristics were observed under optical microscope. The marker proteins of CASMCs, including α-SMA and SM-MHC, were identified by immunofluorescence technique. The protein expression levels of BiP and CHOP, the marker molecules of ERS, were determined by Western blot. RESULTS: The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical "hill and valley" growth pattern of CASMCs at 9~10 d. The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed. The results of Western blot showed that the protein expression of BiP and CHOP in TG (1 and 2 μmol/L) treatment groups was increased compared with control group. Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION: CASMCs can be successfully cultured by tissue explant method. ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.     

夹竹桃花挥发油的GC-MS分析及β-葡萄糖苷酶对夹竹桃花的增香和豚鼠离体子宫平滑肌的作用

目的:建立夹竹桃花挥发油GC-MS的色谱分离鉴定方法,分析夹竹桃花挥发油的化学成分和对豚鼠离体子宫平滑肌的作用。方法:采用挥发油提取器提取夹竹桃花挥发油,以GC-MS法进行分析鉴定,建立了豚鼠离体子宫模型。结果:分别从无酶和有酶的夹竹桃花挥发油提取物中确证了58和57个化合物,挥发油对豚鼠离体子宫平滑肌有收缩作用。结论:对夹竹桃的挥发油化学成分进行了比较分析结果表明,β-葡萄糖苷酶对夹竹桃花有较弱增香作用,也能增加豚鼠离体子宫平滑肌收缩。     

患肌肉溃烂病中华倒刺鲃血细胞数量及形态变化

采集嘉陵江中网箱养殖的中华倒刺鲃(Spinibarbus sinensis Bleeker)患肌肉溃烂病病鱼和健康鱼,对比测定二者红细胞数量、白细胞数量、血红蛋白含量、红细胞大小等指标,以研究病鱼血细胞的形态和数量变化。结果表明病鱼和健康鱼鱼体的多项指标存在显著差异。相对于健康鱼,病鱼外周血中红细胞数目显著减少,白细胞数目极显著增加,红细胞有变小趋势,表明肌肉溃烂对中华倒刺鲃红细胞有一定破坏作用;相对于健康鱼,病鱼淋巴细胞比例下降,单核细胞比例升高,表明在病理情况下,鱼体的特异性免疫功能降低,吞噬功能和非特异性免疫功能有所增强。     

鱼游泳能力对体长的响应及其在鱼道设计中的应用

为了探讨鱼类体长对游泳能力的影响并为鱼道水流的设计提供参考,该研究在封闭水槽中使用"递增流速法"测试了海南省某水利枢纽鱼道目标对象的游泳能力,并用Origin软件进行了数据统计分析,得到了试验鱼感应流速、临界游泳速度和爆发游泳速度的直线回归方程和Kaplan-Meier曲线。结果表明:1)随着试验鱼体长增大,相对感应流速、临界游泳速度和爆发游泳速度(体长/s)均减小,体长和鱼类速度的相关关系可用直线方程表示,且数据经过对数变换后的直线方程拟合效果比未经过对数变换的拟合效果更好,其中R2由0.664～0.725提高至0.907～0.933。2)根据鱼道设计规范、导则及文献,结合本工程目标过鱼对象的感应流速、临界游泳速度和爆发游泳速度(m/s),建议本工程鱼道进口诱鱼流速控制在0.35～0.47 m/s,池室流速控制在0.21～0.59 m/s,竖缝流速控制在0.57～0.74 m/s,出口断面至下一个池室之间的流速控制在0.21～0.50 m/s。鱼类体长对相对游泳速度(体长/s)产生了负面影响,鱼类游泳速度及其变化规律可对鱼道水流设计值提供参考。     

饲料蛋白质水平对克氏原螯虾幼体消化酶活性和肌肉成分的影响

用4种等能不同质量分数的蛋白质饲料(30%、35%、40%和45%)饲喂克氏原螯虾(Procambarus clarkii Girard)幼虾(5.46 g±0.51 g )28 d后,分析不同蛋白水平的饲料对克氏原螯虾幼虾肠道和肝胰脏消化酶活性和体成分的影响.结果表明:(1)饲料蛋白质浓度对脂肪酶和蛋白酶活性均有显著性影响(P<0.05);并随着饲料蛋白水平的提高,两酶活性都呈现先升高后降低的趋势.其中肠道蛋白酶和肝胰脏蛋白酶活性分别在饲料蛋白质浓度为40%和35%时最高;肠道和肝胰脏脂肪酶活性均在饲料蛋白质浓度为40%时最高,且各饲料组肝胰脏脂肪酶活性差异显著(P<0.05),而35%组和40%组之间的肠道脂肪酶活性无显著性差异(P>0.05).克氏原螯虾幼体的肠道和肝胰脏淀粉酶活性随饲料蛋白质水平的升高均呈波浪式变化;35%组与45%组之间的肠道淀粉酶活性和30%与45%组之间的肝胰脏淀粉酶活性均无显著性差异(P>0.05).(2)随着饲料中蛋白水平的升高,克氏原螯虾肌肉中蛋白质含量只有45%组显著升高(P<0.05),其他饲料组肌肉蛋白含量虽有上升趋势,但差异不显著(P>0.05).随着饲料蛋白浓度的增加,肌肉中脂肪含量反而下降,除30%组外,各组间差异显著(P<0.05).对克氏原螯虾肌肉灰分的测定表明,饲料蛋白浓度对肌肉中灰分含量的影响不显著(P>0.05).     

野生和池塘养殖花鲭肌肉营养组成的比较分析

通过对野生花鱼骨和池养花鱼骨肌肉中常规营养组成、18种氨基酸含量及脂肪酸组成的比较分析。结果表明,野生鱼肌肉的水分及灰分含量显著高于池养鱼,而粗脂肪含量显著低于池养鱼(P<0.05),两者粗蛋白含量无显著差异(P>0.05);两者肌肉中氨基酸的含量,除胱氨酸外的其余17种氨基酸的含量无显著差异(P>0.05);野生鱼肌肉脂肪酸组成中,饱和脂肪酸总量和多烯酸总量的百分比显著高于池养鱼,单烯酸总量的百分比显著低于池养鱼(P<0.05)。在多烯酸中,野生鱼的C18∶2、C18∶3和C20∶2占脂肪酸总量的百分比显著低于池养鱼,而C20∶3、C20∶4、C20∶5、C22∶4、C22∶5、C22∶6、Σn-6及Σn-3占脂肪酸总量的百分比显著高于池养鱼(P<0.05)。但池塘养殖花鱼骨单位重量肌肉组织所含的人体所需必需脂肪酸的量,除EPA略低外,n-6系列PUFA总和、n-3系列PUFA总和及DHA均高于野生鱼。