Quercetin activates mitochondrial autophagy via PINK1/parkin pathway to alleviate cerebral ischemia-reperfusion injury in rats

AIM To analyze the regulatory effect of quercetin （QUE） on PTEN-induced putative kinase 1 （PINK1）/parkin mitochondrial autophagy pathway, and to explore the mechanism of quercetin in relieving cerebral ischemia/reperfusion （I/R） injury. METHODS Sixty SD male rats were randomly divided into sham operation group, model group （I/R group）, QUE group,3-methyladenine （3-MA） group and QUE+3-MA group. Administration started in each group 3 days before modeling, once a day, at 30 min after the last administration,except sham group, the other groups used 4-vessel blockage method to establish the whole brain I/R model. On the day after modeling, the neural function was evaluated by neuropathy disability score （NDS）. The volume of cerebral infarction was measured by 2,3,5-triphenyltetrazolium chloride （TTC） staining. The morphological changes of mitochondria in hippocampus were observed by transmission electron microscopy. The contents of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in hippocampus were measured by ELISA. The activity of superoxide dismutase （SOD） and contents of malondialdehyde （MDA） in hippocampus were detected by xanthine oxidase method, thiobarbituric acid condensation method. Western blot was used to detect the proteinex pression of PINK1, parkin and LC3-II in brain tissue. RESULTS Compared with sham group, the hippocampus of the rats in I/R group and QUE+3-MA group showed swelling of mitochondria, destruction or disappearance of internal crista and other pathological damage,also the volume of cerebral infarction, the contents of IL-6, TNF-α and MDA, the protein expression levels of PINK1, parkin and LC3-II were increased （P<0.05）, while NDS score and activity of SOD were decreased （P<0.05）. Compared with I/R group and QUE+3-MA group, the pathological damage degree of hippocampus in QUE group was reduced, the volume of cerebral infarction, the contents of IL-6, TNF-α and MDA were decreased （P<0.05）, the proteinexpression levels of PINK1, parkin and LC3-II, and NDS score and activity of SOD were increased （P<0.05）.The above indexes in 3-MA group were opposite to QUE group. No significant difference in the above indexes between I/R group and QUE+3-MA group was observed （P>0.05）. CONCLUSION Quercetin activates mitochondrial autophagy and reduces cerebral I/R by regulating the expression of PINK1/parkin pathway proteins.     

Progress in RNA-binding protein HuR and its roles in development of hepatocellular carcinoma

RNA-binding proteins （RBP） are molecules with a variety of biological functions discovered in recent years. Among them, HuR is an important RBP, widely expressed in various tissues of the body, and is a member of the Hu/embryonic lethal abnormal vision （ELAV） protein family. It mainly affects the expression levels of target genes in the cells by regulating the stability and/or translation efficiency of the mRNA of the genes, thus participating in the regulation of cell life activities. In recent years, more and more studies on HuR have revealed its important role in inflammation and cancer. This review summarizes the functions of HuR and probes into the regulation of HuR functions. In addition, the roles of HuR in the occurrence and development of hepatocellular carcinoma （HCC） are also discussed, which provides important information for exploring the pathogenesis, biomarkers and therapeutic targets of HCC.     

Correlation between progression of atherosclerosis accelerated by emotional stimulation and imbalance of SDF-1/CXCR4 in mice

AIM To observe effects of emotional stimulation on expression of stromal cell-derived factor-1（SDF-1） and CXC chemokine receptor 4 （CXCR4） in plasma, platelets, aortas, hippocampus and bone marrow of apolipoprotein E gene knockout （ApoE^-/-） mice, and to reveal the possible mechanism of the aggravated atherosclerotic plaque vulnerability by emotional stimulation. METHODS Thirty 8-week-old male ApoE^-/- mice were randomly divided into normal control group, high fat group, and emotional stimulation group. Ten 8-week-old inbred C57BL/6J mice served as blank control group. After 12 weeks of intervention, the serum levels of SDF-1 and CXCR4 were investigated by ELISA. The protein levels of SDF-1 and CXCR4 in platelets, aortas, hippocampus and bone marrow were determined by Western blot. The pathological damage of aortas was observed by oil red O staining. RESULTS Compared with blank control group, normal control group and high fat group, the mice subjected to emotional stimulation showed more serious atherosclerosis in aortas detected by oil red O staining, and increased levels of SDF-1 and CXCR4 in the plasma and aortas were also observed （P<0.05）. The results of Western blot showed that the protein levels of SDF-1 and CXCR4 in platelets, aortas and hippocampus were increased in the mice subjected emotional stimulation, but the expression of SDF-1 and CXCR4 in the bone marrow was decreased （P<0.05）. CONCLUSION Imbalance of SDF-1/CXCR4 may be the key target by which emotional stimulation accelerates the progression of atherosclerosis.     

芥蓝二氢黄酮醇4–还原酶基因BoaDFR的克隆与功能鉴定

以白花芥蓝‘四季粗条’为材料克隆了Boa DFR基因,Gen Bank登录号为MT472647,CDS序列长为1158 bp,编码385个氨基酸,与甘蓝型油菜、甘蓝和白菜等具有高度同源性。Boa DFR表达水平在芥蓝不同发育时期、不同器官中存在显著差异,随植株发育总体呈现先下降后上升的趋势,在幼嫩种子中表达量最高。芥蓝原生质体的亚细胞定位结果显示Boa DFR定位在细胞核上。构建了农杆菌介导的芥蓝瞬时过表达体系,将Boa DFR转入白花芥蓝‘四季粗条’和黄花芥蓝‘福州黄花’中,发现Boa DFR在两种芥蓝中均高水平表达,转基因植株叶片颜色明显变紫,花青素苷显著积累,总含量最高达461.43μg·g-1 FW,而野生型和转空载体芥蓝叶片中几乎检测不到花青素苷。通过HPLC在转基因植株叶片中共检测到8种花青素苷,均为矢车菊花青素苷,其中矢车菊–3–阿魏酰–丙二酰–槐糖苷–5–葡萄糖苷占比最高。     

铁皮石斛人工繁育技术

以铁皮石斛商品瓶苗和盆栽苗茎段为试材,添加不同浓度6-BA、NAA,诱导茎段腋芽、原球茎和不定根,建立再生体系,快速繁殖铁皮石斛组培苗,以期为秦岭淮河一线以北地区人工繁殖铁皮石斛提供参考依据。结果表明:外植体宜采用70%酒精浸泡20 s,0.1%升汞浸泡8 min消毒;MS+6-BA 3.0~5.0 mg·L^-1+NAA 0.1~1.0 mg·L^-1适合腋芽分化和增殖,分化率92%~100%;1/2MS+6-BA 2.0 mg·L^-1+NAA 0.5 mg·L^-1+土豆泥15%适合原球茎诱导增殖,增殖倍数7.7;1/2MS+NAA 0.3 mg·L^-1生根率为100%;以松针土移栽,成活率83%。     

酒红球盖菇菌丝生长对不同培养基的响应

以酒红球盖菇R-4菌株为试材,采用微生物平板培养法,研究了12种氮源和8种碳源对酒红球盖菇生长的影响,以期为酒红球盖菇培养提供参考依据.结果 表明:酒红球盖菇菌丝生长对氮源和碳源的选择差异不显著(P＞0.05),均能利用试验所选的12种氮源和8种碳源,当酵母膏作为氮源和葡萄糖作为碳源时酒红球盖菇的菌落长势较优,菌丝生长速度较快.葡萄糖当作最优碳源时酒红球盖菇菌落生长速率、菌落直径、生长速度、菌丝生长指数分别是9.75 mm· d-1、50.22mm、1.41 mm· d-1和46.73,酵母膏当作最优氮源时酒红球盖菇菌落生长速率、菌落直径、生长速度、菌丝生长指数分别是14.12mm·d-1、52.54mm、2.10 mm·d-1和43.56,由此得出,酒红球盖菇生长的最优碳源是葡萄糖,最优氮源是酵母膏.     

白及愈伤组织与内生真菌共培养及次生代谢产物含量变化

以白及愈伤组织为试材,采用与内生真菌共培养的方法,研究了该过程中白及愈伤组织、内生菌生长速率及4种主要次生代谢含量的变化,以期为白及内生菌在药用成分的开发利用提供参考依据.结果 表明:7株受试真菌菌株与白及愈伤组织分别共培养3、6d时均表现出高于真菌单培养形成的菌面积,共培养9d时各组愈伤增值率显著低于对照(CK)组;1株受试内生真菌生长缓慢,在共培养与单独培养3、6、9d均具有相近菌面积.进一步对8株内生真菌共培养处理下的白及次生代谢产物含量检测发现,不同内生真菌对dactylorhin A、1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯、4-甲氧基-9,10-二氢菲-2,7-二醇、对羟基苯甲醇的积累趋势具有不同影响,主要呈现出3种累积趋势.     

海藻酸对萝卜生长及镉积累的影响

以"满身红"萝卜为试材,通过盆栽试验的方法,研究了叶面喷施不同倍数海藻酸对萝卜生长及镉积累的影响,以期为镉污染土壤生产安全蔬菜提供参考依据.结果表明:与对照相比,喷施不同倍数的海藻酸均不同程度地增加了萝卜根、块根与叶片的生物量,并且随着海藻酸浓度的增大,各器官生物量也随之增加,其中在600倍海藻酸处理中,萝卜根系、块根与叶片较对照分别显著增加98.59％、16.95％和39.13％.海藻酸处理还不同程度地提高萝卜光合色素(叶绿素a、叶绿素b、叶绿素总量和类胡萝卜素)含量和抗氧化酶活性.在镉含量方面,与对照相比,600倍和900倍海藻酸处理显著促进了萝卜根系对镉的吸收,600倍海藻酸显著增加了块根的镉含量,600、900倍和1 200倍海藻酸处理均显著增加了萝卜叶片和可食用部分的镉含量.因此,海藻酸处理促进萝卜的生长,但也促进了萝卜对镉的吸收,不适合用于镉污染土壤的萝卜栽培.     

绒毛白蜡组培快繁体系的建立

以绒毛白蜡带腋芽茎段为试材,采用组织培养法,并进行组培苗驯化移栽试验,研究了不同消毒时间对外植体、不同植物激素种类及浓度对腋芽增殖、壮苗培养和生根培养的影响,以期建立完善的绒毛白蜡组培快繁体系,为绒毛白蜡优良无性系繁育及工厂化生产提供参考依据.结果 表明:最佳外植体消毒处理为70％酒精消毒30 s+2％次氯酸钠消毒10 min,污染率仅4.44％,存活率最高为90.00％,死亡率为5.56％;最佳增殖培养基为MS+0.5 mg· L-1BA+1.0 mg·L-1 IAA,增殖系数达5.52;最佳壮苗培养基为MS+0.5 mg·L-1IAA,平均苗高5.83 cm;最佳生根培养基为1/2MS+0.5 mg· L-1NAA,生根率达98.89％,平均每株根条数5.62条;生根后自然光下驯化炼苗1周,移栽到草炭:蛭石:珍珠岩=1∶1∶1基质中,存活率达90％以上.     

海南西瓜蔓枯病病原菌的鉴定及8种杀菌剂对其室内毒力测定

对海南西瓜蔓枯病病原菌进行了分离鉴定及室内杀菌剂筛选,为西瓜蔓枯病的防控提供参考依据.采用PDA培养基,对西瓜病株病原菌进行分离、培养,经致病性测定,形态学分析及rDNA-ITS測序,鉴定其致病菌为Stagonosporopsis cucurbitacearum.选用8种不同类型杀菌剂对西瓜蔓枯病菌进行室内毒力测定,结果表明,8种杀菌剂均对西瓜蔓枯病菌有一定的抑制作用,其中24％双胍.吡唑酯可湿性粉剂(WP)和11％精甲.咯.嘧菌悬浮种衣剂(FSC)的毒力较强,EC50值分别为0.101 7、0.391 1 mg/L,均小于1 mg/L;其次是10％苯醚甲环唑水分散粒剂(WDG)和18.7％丙环.嘧菌酯悬浮剂(SC),EC50值分别为1.393 3、3.037 6 mg/L;27％春雷?溴菌腈WP和22.5％啶氧菌酯SC毒力一般,EC50值分别为10.753 0、20.982 6 mg/L;毒力效果较差的是46％氢氧化铜WDG和50％多菌灵WP,EC50值分别为60.703 9、151.477 0 mg/L.