Characteristics of morphology and left ventricular function in the mouse with myocarditis

AIM:To determine the relationship between microhistology and cardiac contractility in myocarditis animal model. METHODS: Setting up myocarditis animal model by injecting Coxsackivevirus B₃ (CVB₃) into mice, then observed myocardial morphological changes and measured left ventricular function of mice at the time of first three days and two weeks after injecting CVB₃. RESULTS:Subcellular structure (mitochondria) changed at the first three days after injecting CVB₃. The left ventricular pressure (LVP) and the rate of intraventricular pressure development (dp/dt) which is the index of reflecting cardiac contractility depressed in this stage (14.2±0.8) kPa and (273.1±10.0)kPa/s, respectively. There were (17.1±0.7)kPa and (359.8±9.3)kPa/s in normal mice, respectively (P＜0.01). Myocardial lesions were more severe during immune response stage-two weeks after injecting CVB₃, including myocardial inflammation and necrosis. LVP was (11.8±0.2)kPa and dp/dt was (209.5±6.1)kPa/s in immune response stage. There was significant difference between mice with myocarditis at early stage and at immune response stage (P＜0.01).CONCLUSIONS:The factor of causing the depression of cardiac contractility in early stage (virus-induced damage) is mainly change of subcellular structure. Mitochondria cannot provide energy as normal. There were more severe myocardial lesions in later stage (cell-mediated autoimmune response)than in early stage. The depression of cardiac contractility is a consequence of multifactor.     

茄子株高性状鉴定与遗传分析

以长茄高代自交系125 和126 构建的茄子6 个不同世代的遗传群体〔P₁、P₂、F₁、F₂、 B₁（125×F₁）、B₂（126×F₁）〕 为试材，利用主基因+ 多基因混合数量性状遗传模型对茄子的株高性状进行多世代遗传联合分析。结果表明：供试亲本株高 性状差异显著，分离世代株高性状数值均呈单峰的偏正态分布，属于数量性状遗传。多世代遗传联合分析结果显示茄子株高 性状的最适遗传模型为C-0 模型，不存在主基因遗传效应，表现为多基因控制的加性- 显性- 上位性遗传模式。采用二阶 遗传参数进一步分析株高的多基因遗传效应，结果显示，茄子分离世代F₂、B₂ 的多基因遗传率分别为49.24%、22.77%，茄 子株高以多基因遗传为主。     

苦瓜单瓜种子数与种皮颜色的遗传分析

以苦瓜自交系29 和惠州大顶为亲本构建6 个世代（P₁、P₂、F₁、B₁、B₂、F₂）群体,分别对
苦瓜单瓜种子数和苦瓜种皮颜色进行遗传分析。结果表明,苦瓜单瓜种子数的遗传符合2 对等显性主基因+
加性-显性多基因混合遗传模型（E-6 模型）,由主基因和多基因共同控制,多基因显性效应较大,F₁ 表现
出超亲优势。苦瓜种皮颜色的黑色对棕黄色表现为1 对基因控制的显性遗传。     

番茄绿果与橙果间果实颜色及主要色素含量的遗传研究

对番茄组合绿樱(绿果)×金珠1号(橙果)的6个世代遗传群体(P1、P2、F1、BC1、BC2和F2)进行果色性状、番茄红素含量、叶绿素含量和胡萝卜素含量等的遗传规律分析。结果表明:正反交F1的果色性状无明显差异,而色素含量存在显著差异;说明番茄果色性状受核基因控制,而色素含量遗传除受核基因控制外还可能存在胞质效应。采用多世代联合分析法的分析结果表明,番茄绿果与橙果间的果色性状符合2对加性主基因+加性-显性多基因(MX2-A-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为73.42%、78.25%和61.41%,多基因遗传率分别为22.87%、15.35%和34.94%,即果色性状遗传的主基因遗传力较强;叶绿素含量符合1对负向显性主基因+加性-显性多基因(MX1-AEND-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为0、1.73%和0.65%,多基因遗传率分别为45.47%、0和37.82%,即主基因遗传力在BC2群体中最高,多基因遗传力在BC1群体中最高;番茄红素含量与胡萝卜素含量均符合2对加性-显性-上位性主基因+加性-显性多基因(MX2-ADI-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为75.74%、1.79%、84.26%和61.53%、87.21%、81.05%,多基因遗传率分别为20.32%、74.12%、12.68%和0.68%、0、0,表明番茄红素含量和胡萝卜素含量的主基因遗传力较强。     

SSR标记辅助芥蓝×甘蓝型油菜种间杂交后代的遗传背景筛选

为筛选芥蓝×甘蓝型油菜种间杂交后代的遗传背景,加速回交转育进程,采用蕾期授粉结合胚挽救手段进行远缘杂交,获得芥蓝和甘蓝型油菜的种间杂种F1和BC1群体。利用已有的220对SSR引物对双亲进行多态性筛选,获得多态性引物51对。挑选均匀分布在甘蓝9条染色体上、扩增稳定、条带清晰的33对多态性SSR引物,对3株F1单株和35株BC1单株进行遗传背景筛选。NTSYSpc2.11a分析结果表明:F1植株的遗传背景与亲本甘蓝型油菜更为接近,遗传相似系数为0.74;而BC1植株的遗传背景差异较大,与亲本芥蓝的遗传相似系数在0.26~0.65之间。在35株BC1植株中,单株14Y1与芥蓝的遗传背景最为相近,形态观察结果进一步验证了该单株的遗传背景与回交亲本芥蓝更相似,可用于下一步回交转育。     

甜瓜果实酸性性状的遗传分析

以果实口感无酸味的甜瓜材料60 和酸甜味的材料61 为亲本，构建P₁、P₂、F₁、BC₁、BC₂ 及F₂ 六世代群体，利用主基因+多基因混合遗传模型的六世代联合分析法，分析甜瓜果实柠檬酸含量、可滴定酸值（TA）和pH 值的遗传效应。结果表明：柠檬酸含量的遗传模型为1 对加性-显性主基因+加性-显
性-上位性多基因模型，F₂ 群体的主基因遗传率为31.06%，多基因遗传率为30.48%；TA 值的遗传模型为2 对加性-显性-上位性主基因+加性-显性-上位性多基因模型，F₂ 群体的主基因遗传率为78.06%，多基因遗传率为0；pH 值的遗传模型为1 对加性-显性主基因+加性-显性-上位性多基因模型，F₂ 群体的主基因
遗传率为84.07%，多基因遗传率为12.90%。     

茄子果顶形状数据采集方法比较和遗传分析

以圆顶茄子和尖顶茄子为亲本构建6世代群体(P_1、P_2、F_1、BC_1P_1、BC_1P_2、F_2),在春露地和秋大棚两个茬口分别种植6世代群体,利用目测法和Tomato Analyzer软件分析法采集茄子果顶形状数据,应用植物数量性状主基因+多基因混合遗传模型多世代联合分析方法,对茄子果顶形状进行遗传分析。结果表明:目测法和软件法都可以用来描述茄子果顶形状,Tomato Analyzer软件分析法获取的果顶数据计算出的AIC值完整性好、适应性检测中的统计量差异显著个数少,对茄子果顶形状的数据测量更为准确,更加适用于茄子果顶形状的采集。茄子果顶形状表现为数量性状,受2对加性-显性-上位性主基因控制,同时也受环境条件的影响。在分离群体中F_2群体主基因遗传率高、稳定性好,主基因遗传率达到72%以上,育种过程中适合在F_2群体进行果顶形状的选择。     

苦瓜植株卷须发生的初始节位及其分叉的遗传分析

以栽培苦瓜自交系佛沥112 和野生苦瓜自交系THMC170 为亲本，构建4 个世代（P₁、P₂、F₁、F₂）遗传群体，采 用主基因+ 多基因混合遗传模型对苦瓜植株卷须发生的初始节位进行遗传分析；以栽培苦瓜自交系佛沥734 和半野生苦瓜自 交系AVBG1602 为亲本，构建4 个世代（P₁、P₂、F₁、F₂）遗传群体，对苦瓜植株卷须的分叉性进行遗传分析。结果表明， 苦瓜植株卷须发生的初始节位遗传符合2 对等显性主基因+ 加性- 显性多基因遗传模型（E-6 模型），其中主基因遗传率为 15.81%，多基因遗传率为7.83%。苦瓜植株卷须的分叉对不分叉受1 对显性基因控制，其中卷须分叉表现为显性，卷须不分 叉表现为隐性。研究结果为深入解析苦瓜植株卷须发生及发育的遗传机制奠定了基础。     

辣椒果皮颜色的遗传分析

以商品成熟期果皮紫色(245)、绿色(246)、乳白色(247)的3份辣椒品系为亲本,配制杂交组合245×246和245×247,构建6世代遗传群体,通过目测分级、色差仪分析测定P1、P2、F1、B1、B2、F2各单株的辣椒商品成熟期果皮颜色L值、C值和颜色级值,并应用6个世代联合分析法研究辣椒商品成熟期果皮颜色的遗传规律。结果表明,辣椒商品成熟期果皮颜色遗传为细胞核遗传,紫-绿组合的辣椒商品成熟期果皮颜色性状符合1对加性主基因+加性-显性混合多基因模型,即D-2模型,其L值、C值和颜色级值F2主基因遗传率分别为22.39%、91.39%和82.26%;紫-白组合的辣椒商品成熟期果皮颜色性状符合2对加性-显性-上位性主基因+加性-显性-上位多基因模型,即E-0模型,其L值、C值和颜色级值F2主基因遗传率分别为87.02%、94.34%和97.54%。表明主基因遗传力较强,多基因遗传力和环境效应影响弱,在育种时对辣椒商品成熟期果皮颜色的选择应在早期分离世代进行。     

Characteristics of energy metabolism in brain mitochondria of rats exposed to hypoxia

AIM:To explore the characteristics of energy metabolism in brain mitochondria of rats exposed to acute and chronic hypoxia. METHODS: Animal grouping: Wistar rats were randomized into acute hypoxic group (AH), chronic hypoxic group (CH) and the control. Respiratory function, F₀F₁-ATPase activity, mitochondrial ATP, ADP and AMP contents and ATP production rate were measured respectively. RESULTS: In AH, brain mitochondrial respiratory state IV (ST₄) was increased, while respiratory control rate (RCR), mitochondrial ATP content, ATP production rate and F₀F₁-ATPase activity were decreased respectively. In CH, ST₄, RCR, mitochondrial ATP content and F₀F₁-ATPase activity were reversed partially.CONCLUSION: Acute hypoxia may impair brain mitochondria energy metabolism by way of depressing mitochondrial oxidative phosphorylation and ATP production and these parameters gain partial reablement during chronic hypoxia.     

番茄绿果与红果颜色性状遗传的研究

以绿果番茄‘绿樱’和红果番茄‘TTD1003A’为亲本材料,构建4个世代P_1、P_2、F_1和F_2遗传群体,采用标准比色卡,对成熟果实的果色、果皮色、果肉色和胎座胶状物质颜色进行观察分析。结果表明:在F_2代分离群体中,果色分离比例为,红︰棕︰黄︰绿=9︰3︰3︰1;果皮色为,黄色︰透明=3︰1;果肉色为,红︰浅黄︰浅绿=12︰3︰1,即果色、果皮色和果肉色的遗传符合孟德尔遗传规律,且分别由两对、一对和两对核基因控制;果实绿色相对果实红色为隐性,果皮透明相对果皮黄色为隐性,果肉浅绿色相对果肉红色为隐性,果皮与果肉颜色独立遗传。同时,运用色差仪测定果实表面颜色的L值、a值和b值,计算色光值后,运用植物数量性状主基因+多基因遗传分析法分析得出:番茄果实绿色对红色的遗传可能符合两对加性—显性—上位性主基因+加性—显性多基因遗传(MX2-ADI-AD),其中两对主基因均以加性效应为主,第一对主基因的加性作用更为明显。在F_2代中,色差仪测定指标的主基因遗传率为76%~89%,而多基因遗传率接近0,即该组合控制果色性状的主基因遗传力很高,多基因遗传力很低,对番茄果色的选择应在分离早期世代进行。     

甜瓜白粉病抗病基因的定位及候选基因分析

以甜瓜高抗白粉病自交系MR-1为母本,易感白粉病自交系Top Mark为父本,构建BC_1P_2和F_2群体,经13个国际通用的甜瓜白粉病生理小种鉴别寄主鉴定,2015年东北农业大学设施园艺工程中心的白粉病发病病菌为P.xanthii生理小种1,甜瓜MR-1对P.xanthii生理小种1的抗性由单显性基因控制。通过对266个F2分离群体的抗病性鉴定和CAPS标记分析,采用复合区间作图法对甜瓜抗白粉病性状进行QTL分析,最终构建了1张包含203个CAPS标记的甜瓜遗传连锁图谱,并将抗病基因PXR定位在第12号染色体上M12-GH和M12-TE 2个标记之间,该基因与两侧翼标记间的距离分别为0.63 c M和0.42 c M,两侧翼标记间在甜瓜参考基因组上对应的物理距离为303 kb,该区域内含有60个预测基因,其中10个基因含有非同义单碱基突变。10个基因荧光定量分析结果显示,在抗病与感病甜瓜表达量存在较大差异的4个基因MELO3C002434、MELO3C002437、MELO3C002441、MELO3C002457为甜瓜白粉病抗病候选基因。     

二倍体月季F_1群体的SSR鉴定与遗传分析

以中国古老月季品种‘月月粉’(Rosa chinensis‘Old Blush’,用OB表示)为母本,‘无刺光叶蔷薇’(Rosa wichuriana‘Basye’s Thornless’,用W表示)为父本,构建F_1代共296个单株。从146对SSR标记中筛选出亲本间多态性好且条带清晰的23对SSR标记,对随机选择的94株F_1进行杂种鉴定和遗传分析。结果表明:筛选到3个纯合显性标记分别为Fv512、Fv609和305,可单独一次性鉴定全部杂种真实性;随机筛选的94株子代均为真杂种;20个SSR标记用于基因型分析,有9个标记出现了偏分离,并在统计上达到显著或极显著水平,偏分离率45%,说明该F_1群体基于SSR位点的基因型偏分离率较高,在进行高密度遗传图谱构建时应重视偏分离标记对作图的影响;UPGMA聚类分析显示,94株F_1的遗传变异大,且遗传多样性丰富,可划分为2个大类7个亚类。     

Effect of sodium selenite on proliferation of endometrial cancer cells

AIM: To investigate the effect and mechanism of sodium selenite (Na₂SeO₃) on the proliferation of endometrial cancer cells. METHODS: Endometrial cancer Ishikawa cells and HEC-1A cells were treated with Na₂SeO₃. The effect of Na₂SeO₃ on cell proliferation was determined by MTT assay. The effects of Na₂SeO₃ on cell cycle distribution and apoptosis were tested by flow cytometric analysis. The expression of cyclin A was detected by Western blotting. RESULTS: Na₂SeO₃ inhibited the proliferation of Ishikawa cells and HEC-1A cells. For Ishikawa cells, IC₅₀ was 3.26 μmol/L, and for HEC-1A cells, IC₅₀ was 4.77 μmol/L. After treated with Na₂SeO₃, the cells in G₀/G₁ phase were reduced and the cells in S phase and G₂/M phase were increased. Na₂SeO₃ also increased the percentage of apoptosis cells. The result of Western blotting showed that the expression of cyclin A was increased. CONCLUSION: Na₂SeO₃ inhibits the proliferation of endometrial cancer Ishikawa cells and HEC-1A cells via up-regulating the expression of cyclin A, arresting cell cycle and inducing apoptosis.     

Role of calcium ion in the cyotoxicity and DNA fragment induction in HL-60 cells by 6F isolated from Pteris semipinnata L.

AIM: To investigate the changes of cytosolic free calcium concentration([Ca²⁺]_i) and expression of Bcl-2 in HL-60 cells treated by 6F isolated from Pteris semipinnata L.(PSL), and to discuss the relations between calcium ion and cytotoxicity and DNA fragment induction effects of 6F. METHODS: HL-60 cells were used as in vitro model. [Ca²⁺]_i was measured on fluorescent spectrophotometry using Fura-2/AM as Ca²⁺ indicator. Bcl-2 expressing level was measured by flow cytometry. Tetrazolium salt(MTT) and diphenylamine staining methods were applied for cytotoxicity assay and DNA fragmentation detection, respectively. RESULTS: [Ca²⁺]_i increased obviously in a dose and time dependent manner after treated HL-60 cells with 6F. 6F decreased the expressing level of Bcl-2. Adding 2 mmol/L Ca²⁺ to the medium, or 1 mmol/L EDTA to chelate Ca²⁺, or 4 μmol/L calcium ionophore A 23187 to increase the concentration of cytosolic Ca²⁺, the DNA fragment induction by 6F was not affected, whereas the cytotoxicity of 6F was enhanced. 250 μmol/L Zn²⁺ attenuated the DNA fragment induction, and the cytotoxicity of 6F against HL-60 cells was enhanced significantly. CONCLUSION: It was speculated that the decreased expressing of Bcl-2 by compound 6F was related to increased [Ca²⁺]_i in HL-60 cells, and DNA fragment induction was possibly catalyzed by Ca²⁺ - independent DNase.     

大白菜紫心性状遗传规律分析及其基因初步定位

以大白菜[Brassica rapa(Lour.)ssp.pekinensis]紫心纯系‘14S839’和橙色纯系‘14S162’为亲本,构建F_2群体,对大白菜紫色基因(BrPur)进行遗传连锁分析。性状分离调查结果显示,在F_2群体中紫心与非紫心单株符合3︰1分离比例,说明大白菜紫心性状的有、无符合单基因显性遗传模式,紫心性状是一种显性性状。采用改良的BSA方法对紫色基因BrPur进行定位,经过对白菜全基因组297个SSR标记的筛选,得到了BrPur的两个侧翼标记A710和A714,同时结合白菜基因组信息,将BrPur定位于大白菜A07连锁群上。结合前人关于花青素代谢相关基因的研究报道,通过分析Br4CL3和Bra004214的序列、开发新的标记,得到了两个特异共显性标记CL-12和B214-87。利用F_2群体扩增验证筛选到的所有标记,结果表明,经Join Map 4.0作图得到的遗传连锁图与非紫色交换单株"基因型矩阵"显示的结果完全一致,标记CL-12和B214-87位于BrPur两侧,遗传距离分别是3.1 cM和3.5 cM。     

甜瓜抗蔓枯病基因Gsb-4 的分子标记

 以甜瓜（Cucumis melo L.）抗蔓枯病自交系PI482398（含抗蔓枯病基因Gsb-4）和感病自交系‘白皮脆’以及它们的F₁、BC₁P₁、BC₁P₂、F₂ 群体为材料,苗期进行蔓枯病菌（Didymella bryoniae）接种鉴定,结果表明甜瓜抗蔓枯病基因Gsb-4 为单显性遗传。利用集团分离分析法（bulked segregant analysis,BSA）对89 对SSR 引物进行筛选,引物CMTA170a 在抗性材料中可扩增出约为120 bp 的条带,并与抗性基因Gsb-4 表现出连锁关系。统计了CMTA170a 在118 个F₂ 单株上的多态性,并利用MAPMAKER/Exp version 3.0b 软件进行了计算,其与Gsb-4 的遗传连锁距离为5.14 cM。     

Effects of edible-medicinal fungus crude extracts on vascular endothelial cells with high glucose-induced injury

AIM: To investigate the effects of crude extracts of Cordyceps gunnii (CGE), Lepista lentinus (LLE), Cordyceps sinensis (CSE) and Lentinus striguellus (LSE) on the proliferation of high glucose-treated human umbilical vein endothelial cells (HUVECs). METHODS: The cultured HUVECs were divided into normal control group (treated with M199 culture medium alone), high glucose group (treated with M199 culture medium containing 33 mmol/L glucose) and 4 crude extracts of edible-medicinal fungi (CGE, LLE, CSE and LSE) intervention groups (treated with the crude extract of edible-medicinal fungus at concentrations of 12.5, 25, 50, 100 mg/L in high glucose M199 culture medium). The cell proliferation was evaluated by MTT assay. The cell cycle and ROS level were measured by flow cytometry. RESULTS: Compared with normal control group, the MTT absorbance value and the percentage of G₀/G₁ stage of the HUVECs in high glucose group were significantly decreased (P<0.05), while the percentage of S+G₂/M and ROS level were significantly increased (P<0.05). Compared with high glucose group, treatment with the crude extracts of Lepista lentinus and Lentinus striguellus decreased the cell absorbance value (P<0.05), and the inhibitory effect was enhanced in a dose-dependent manner. However, Cordyceps gunnii had no effect (P>0.05). The crude extracts of Cordyceps sinensis (12.5~50 mg/L) significantly enhanced the proliferative activity of HUVECs, decreased the percentage of G₀/G₁ stage of HUVECs, increased the percentage of S+G₂/M of HUVECs, and reduced the intercellular ROS level (P<0.05). CONCLUSION: Only Cordyceps sinensis crude extract effectively protects the HUVECs in high glucose-induced injury, which might be due to promoting more cells to enter to the cell cycle and down-regulating oxidative stress.     

Experimental study on effects of ATP and its derivatives on the proliferation of the immortalized human fibroblasts

AIM: To investigate the inhibitory effects of ATP on proliferation of immortalized human fibroblasts, and learn what subtypes of P-2 purinergic receptors are involved in ATP cytotoxicity.METHODS:The immortalized human fibroblast cell line KMST-6 was cultured with ATP and its derivatives including ADP, β,γ-methyleneadenosine5'-triphosphate(MeATP), 2'&3'-O-(4-benzoylbenzoyl) adenosine, triethylammonium salt (BzATP), adenosine 5'-O-(3-methiotriphosphate)(ATPγS), 2-methylthioadenosine 5'-triphosphate(2-MeATP) and UTP, and the cell cycle analysis was finished using flow-cytometer, the cell apoptosis analysis was made using Hoechst 33258 and examination of DNA ladder.RESULTS: The extent of cytotoxicity induced by these drugs was found to be in order of : ATP=ADP＞ATPγS＞MeATP=BzATP, neither 2-MeSATP nor UTP showed any cytotoxicity. No enhanced expression of P²¹ was observed and the cell cycle was held in G₁/S in the cells treated with 0.4mmol/L;no cell apoptosis was detected in the cells treated with 1mmol/L ATP.CONCLUSION: Connecting with P_2X or P_2Y purinergic receptors,ATP activated some intracellular signals to inhibit cell growth, the growth inhibition caused by ATP was not due to apoptosis or induction of cyclin/CDK kinase inhibitor,P²¹.