苄基化甘露糖原酸酯的开环研究

以甘露糖为原料合成了苄基化甘露糖原酸酯,并对其在酸催化下的开环反应进行了研究,采用1HNMR对各阶段产物进行了表征。结果表明,冰乙酸的催化效果最好,产率可达90%,同时对冰乙酸催化苄基化甘露糖原酸酯的开环机理进行了探讨,合理地解释了用80%或50%乙酸代替冰乙酸时产率下降的原因,主要是水与亲核试剂乙酸根形成了竞争,生成了副产物。

1,2-糖原酸酯的一锅法合成

以未保护的还原吡喃糖为原料,在甲基磺酸的催化下,与醋酐反应生成全乙酰化吡喃糖,随后经33%HBr/冰醋酸溶液溴代反应为溴代糖,最后与醇反应,一锅法得到目标化合物。整个过程无需分离对光、热不稳定的溴代糖中间体,为高效经济合成目标化合物提供了一种新方法。

氟化钾催化合成糖原酸酯

以葡萄糖,甘露糖和鼠李糖为原料,经羟基保护和溴代制得溴代糖(2a^2e);2在氟化钾催化下与醇反应合成了一系列新颖的单糖原酸酯(5a^5c),6-O-Ms-单糖原酸酯(5d)和6-O-Ts-单糖原酸酯(5e)。2与由半乳糖制得的糖基受体反应合成了糖-糖原酸酯(5a-Ⅵ,5b-Ⅵ)。5的结构经1HNMR,13C NMR和MS表征。

雷公藤甲素通过丝氨酸-苏氨酸蛋白激酶/糖原合成酶激酶-3β通路对MGC803细胞增殖、侵袭与迁移的影响

目的 探讨雷公藤甲素通过丝氨酸-苏氨酸蛋白激酶/糖原合成酶激酶-3β(AKT/GSK-3β)通路对MGC803细胞增殖、侵袭与迁移的影响。方法 体外培养MGC803细胞,确定雷公藤甲素的干预剂量和干预时间。将细胞分为阴性对照组(A组,不进行处理),阳性对照组(B组,10μmol/L顺铂),雷公藤甲素高、中、低剂量组(C组、D组、E组)。干预48 h,采用Transwell法检测细胞侵袭能力,采用划痕试验法检测迁移能力,采用免疫印迹(Western blot)法检测检测相关蛋白的表达。结果 随着雷公藤甲素浓度的增加,MGC803细胞增殖率降低(P <0.05);48 h时半数抑制浓度(IC50)最小,选择干预时间为48 h,C组、D组、E组的干预剂量分别为80,40,20 nmol/L。与B组比较,C组、D组、E组侵袭细胞数、细胞迁移距离及波形蛋白(vimentin),snail,p-AKT/AKT,p-GSK-3β/GSK-3β表达水平均显著升高(P <0.05),上皮钙黏素(E-cadherin)、β-联蛋白(β-catenin)表达水平均显著降低(P <0.05),且呈剂量依赖关系。结论 雷公藤甲素可通过抑制AKT/GSK-3β信号通路而抑制MGC803细胞增殖、侵袭与迁移。

糖基原酸酯的电子轰击质谱研究

本文报道了糖基原酸酯在电子轰击质谱中可能的裂解途径，并经同位素标记和亚稳离子扫描证实了所提出的裂解过程。

Physiological Responses of Yixing Lily Leaf and Bulb to Drought Stress

[Objective]To explore an effective method for evaluating drought resistance of Yixing lily and provide a reference for selecting drought-resistant Yixing lily varieties. [Method]Through artificial simulation of drought stress environment,the changes of physiological indexes,such as content of proline,malonaldehyde,and reducing sugar in Yixing lily leaf and bulb were observed,when soil water content changed. [Result]The response to drought stress of the Yixing lily leaf was greater than that of the bulb,and the content of the proline and malonaldehyde was higher than that of the reducing sugar. [Conclusion]The leaves as an appropriate material for evaluation and analysis on drought resistance can better reflect the physiological responses of Yixing lily to drought stress. And the content of the malonaldehyde and that of proline are preferable indexes.

Establishment of the Agrobacterium-mediated Genetic Transformation System of Ginkgo biloba and the Construction of the Expression Vector of Gb-DXR

[Objective] The research aimed to provide reference for increasing the genetic transformation efficiency of Ginkgo biloba mediated by Agrobacterium.[Method] Taking the mature embryos of Ginkgo biloba seeds as explants,after 48 hours＇ pre-cultivation on MS medium in the absence of phytohormone,GUS gene was transmitted into embryos of Ginkgo biloba mediated by three kinds of Agrobacterium.Transient expression of GUS gene activity was observed through histochemical staining,and the influencing factors of the expression of GUS gene were analyzed.And the expression vector of 1-deoxy-D-xylulose-5-phosphate reductoisomerase in the biosynthesis approach of biobalide precursor of Ginkgo biloba was constructed.[Result] A more suitable genetic transformation scheme was obtained as follows：taking embryos of Ginkgo biloba as explants,using EHA105 Agrobacterium with pCAMBIA1304＋ for infection,co-culture for 3 days and GUS staining.The results showed that transient expression rate of GUS after transformation was higher.[Conclusion] The research provide a more effective method for further study on the transgene of Ginkgo biloba.

电项针对全脑缺血VD模型大鼠PI3K/AKT/GSK-3β信号通路的影响

目的研究电项针对全脑缺血血管性痴呆(vascular dementia,VD)模型大鼠磷脂酰肌醇-3-激酶/丝氨酸-苏氨酸蛋白激酶/糖原合成酶激酶-3β(Phosphatidylinositol-3 kinase/serine-threonine kinase/glycogen synthase kinase-3β,P13K/AKT/GSK-3β)信号通路的影响。方法采用四血管阻断方法制备VD模型大鼠,电项针组取双侧风池穴、供血穴,电针30 min/次,1次/d,治疗14d。采用Y迷宫评价大鼠学习记忆能力;荧光定量PCR(RT-PCR)、Western blot法检测大鼠海马组织中磷酸化蛋白激酶B(phosphorylatedproteinkinaseB,p-AKT)、磷酸化糖原合成酶激酶-3β(Phosphorylated GSK-3β,P-GSK-3β)mRNA和p-AKT、p-GSK-3β蛋白的表达。结果与模型组比较,电项针组可显著提高VD大鼠Y迷宫学习与记忆正确次数(P<0.01)。与模型组比较,电项针组大鼠海马组织中p-AKT、p-GSK-3βmRNA和p-AKT、p-GSK-3β蛋白表达均有不同程度的升高(P<0.01)。结论电项针能够改善VD模型大鼠学习记忆能力,具体机制可能是激活PI3K/AKT/GSK-3β信号通路,发挥抗凋亡作用,起到对缺血海马神经元的保护作用。

Distinct immune response induced by peptidoglycan derived from Lactobacillus sp

AIM： To analyze the distinct immune responses induced by Lactobacillus peptidoglycan （PG）. METHODS： BALB/c mice were intraperitoneally injected with PG once a day for three consecutive days, Peritoneal macrophage and splenocyte mRNA was extracted and the gene expression profile was studied using high-density oligonudeotide microarrays. Inhibitory effects of Lactobacillus PG on colon tumor tissue were studied in vitro and in vivo, RESULTS： The gene expression profiles revealed that the TLR-NF-kB and Jak-STAT signaling pathways were highly activated. An inflammatory phenotype was induced when peritoneal macrophages were initially exposed to Lactobacillus PG and switched to a more complex phenotype when BALB/c mice were treated with three doses of Lactobacillus PG. A protective physiological inflammatory response was induced after three consecutive days of PG treatment. It was tending toward Thl dominant immune response. Lactobacillus PG also appeared to induce a significant in vivo anti-colon tumor effect. CONCLUSION： Lactobacillus PG is responsible for certain immune responses induced by Lactobacilli. Anti-tumor effects of Lactobacilliare likely to attribute to the activation of macrophages by PG expressed on the bacterial cell surface.

Monthly Changes of Glycogen, Lipid and Free Amino Acid of Oyster

Monthly difference of the chemical composition of oyster cultured along the eastern coast of Shandong Province was analyzed. The components analyzed included glycogen, fatty acid and free amino acid （FAA）. The content of glycogen was high in January and March （2.89 and 2.82 g（100 g） ^-1 on average, respectively） and low in October （2.07g（100g）^- 1 on average）. The low content of neutral lipids in October reflected a relatively poor nutritional value of oyster （1.42 g（ 100 g）^- 1 on average）. The main fatty acids of oyster were palmitic acid （16：0）, oleic acid （18： 1）, eicosapentaenoic acid （EPA, 20： 5（0 3） and docosahexaenoic acid （DHA, 22：6（0 3）. The major FAAs of oyster were Taurine, Glutamicacid, Glycin, Alanine, Arginine and Proline. Taurine was the most abundant FAA with its content ranging from 603 mg（100 g） ^-1 to 1 139 mg（ 100 g） ^-1. The high contents of glycogen, polyunsaturated fatty acid and FAA showed that oyster cultured along the eastern coast of Shandong Province was nutritionally good in January and March.

Synergistic effect of additives on electrodeposition of copper from cyanide-free electrolytes and its structural and morphological characteristics

Electrodeposition of acid copper plating on mild steel substrate is tedious due to the galvanic displacement reaction ofcopper on mild steel.This can be avoided by using a proper complexing agent,because the complexing agent tuned the potential ofnoble direction to less noble direction by complex formation.In this paper,environment friendly electrodeposition of copper fromnon-cyanide electrolyte using sodium gluconate as complexing agent was investigated in alkaline medium.The effects of additivessuch as1,2,3-benzotriazole,sodium lauryl sulphate,PEG8000and saccharin were studied.These additives are found to reduce thegrain size,grain boundaries and improve surface morphology of the copper deposits.Also they improve the throwing power of thedepositing electrolytes and hardness of deposits.The electrodeposited copper coatings were characterized by X-ray diffractiontechnique.XRD results indicate that the electrodeposited copper shows polycrystalline and face centered cubic structure.The crystalsize was calculated by XRD and AFM analysis.Among these additives studied,the mixture of benzotriazole and sodium laurylsulphate acts as the best additive.A uniform pore-free surface observed under SEM and AFM results reveal the grain refining broughtabout by the additives.

Jianpi Qingchang decoction alleviates ulcerative colitis by inhibiting nuclear factor-κB activation

To investigate the therapeutic effect of Jianpi Qingchang decoction (JPQCD) on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice.METHODSC57BL/c mice were injected intragastrically with 5% DSS instead of drinking water for 7 d, and their body weight, diarrhea severity and fecal bleeding were monitored, while the mice in the control group were treated with standard drinking water, without DSS. After 7 d, the DSS drinking water was changed to normal water and the DSS group continued with DSS water. The control and DSS groups were given normal saline by intragastric injection. The 5-aminosalicylic acid (5-ASA) group was treated orally with 5-ASA at a dose of 100 mg/kg daily. The JPQCD group was treated orally with JPQCD at a dose of 17.1 g/kg daily. On day 14, the colon length was measured, the colorectal histopathological damage score was assessed, and protein levels of interleukin (IL)-1β, IL-8 and tumor necrosis factor-alpha (TNF-α) in colon supernatants were measured by enzyme-linked immunosorbent assay. mRNA expression of IL-1β, IL-8, TNF-α and nuclear factor-kappa B (NF-κB) was detected by real-time quantitative polymerase chain reaction. Western blotting was used to detect the protein expression of NF-κB and inhibitor of kappa B.RESULTSAcute inflammation occurred in the mice administered DSS, including the symptoms of losing body weight, loose feces/watery diarrhea and presence of fecal blood; all these symptoms worsened at 7 d. The colons of mice treated with DSS were assessed by histological examination, and the results confirmed that acute inflammation had occurred, as evidenced by loss of colonic mucosa and chronic inflammatory cell infiltration, and these features extended into the deeper layer of the colon walls. The expression levels of IL-1β, IL-8 and TNF-α in the DSS group were higher than those in the control group (P < 0.05), and the expression levels of IL-1β, IL-8 and TNF-α in the JPQCD and 5-ASA groups were lower than those in the DSS group after treating with JPQCD and 5-ASA. Comparing with the DSS group, the mRNA level of IL-1β, IL-8, TNF-α and NF-κB was significantly reduced by 5-ASA and JPQCD. The difference between JPQCD and 5-ASA groups was not statistically significant (P > 0.05). Comparing with the DSS group, due to using JPQCD and 5-ASA, significant suppression of activation in DSS-induced NF-κB and increased phosphorylation of IκB in mice with experimental colitis occurred (P < 0.05). The difference between the JPQCD group and the 5-ASA group was not statistically significant (P > 0.05).CONCLUSIONActivation of the NF-κB signaling pathway is inhibited by JPQCD, which shows the potential mechanism by which JPQCD treats UC.

维泰醇对人骨肉瘤移植荷瘤裸鼠肿瘤生长的抑制作用及对Akt/GSK3信号通路的影响

目的:探讨维泰醇对人骨肉瘤移植荷瘤裸鼠肿瘤生长的抑制作用及对丝苏氨酸蛋白激酶(Akt)/糖原合酶激酶3(GSK3)信号通路的影响。方法:将30只BALB/c-nu品系裸鼠分为3组:正常对照组、环磷酰胺组(25 mg·kg^(-1))和维泰醇组(150 mg·kg^(-1)),每组10只。用培养的U-2OS细胞注射到裸鼠右胫骨骨髓腔内制备U-2OS荷瘤裸鼠模型,造模成功后环磷酰胺组和维泰醇组灌胃给予相应药物,正常对照组给予等量生理盐水,1次/d,给药周期为4周。末次给药次日,处死裸鼠,完整剥离出原位瘤块,计算抑瘤率,检测瘤块组织中相关蛋白表达水平及细胞凋亡情况。结果:与正常对照组比较,环磷酰胺组和维泰醇组瘤块重量、N-钙黏蛋白(N-cadherin)、波形蛋白(vimentin)、纤维连接蛋白(fibronectin)、p-Akt、磷酸化糖原合酶激酶3(p-GSK3)和锌指转录因子(Snail)蛋白表达水平降低,抑瘤率、上皮细胞钙黏蛋白(E-cadherin)蛋白表达水平和细胞凋亡率增加(P<0.05);与环磷酰胺组比较,维泰醇组瘤块重量、N-cadherin、vimentin、fibronectin、p-Akt、p-GSK3和Snail蛋白表达水平增加,抑瘤率、E-cadherin蛋白表达水平和细胞凋亡率降低(P<0.05);各组Akt和GSK3蛋白表达水平差异无统计学意义(P>0.05)。结论:维泰醇可能通过Akt/GSK3信号通路抑制上皮-间质转化(EMT)并降低骨肉瘤细胞的侵袭能力,促进骨肉瘤细胞凋亡。

Orexin-A stimulates the glucose output of porcine hepatocytes

Orexin-A （OxA） is a key neuropeptide involved in the central control of appetite and the maintenance of energy homeostasis, as well as in the regulation of glycemia. In the present study, OxA receptors, OXtR and OX2R were identified to be highly expressed in porcine hepatocytes. 100 nM OxA could rapidly stimulate the glucose output of porcine hepatocytes in 10min. Primary hepatocytes treated by 1 nM-500 nM OxA for 24 h significantly increase the released glucose and the concentration of albumin, total bile acids and triglyceride in the supernatant. The mRNA expression level of such gluconeogenesis and fat mobilization related genes as acetyl-coA carboxylase, glycogen phosphorylase, fatty acid translocase and phosphoenolpyrurate carboxykinase were also up-regulated accordingly. The results first demonstrated that OxA had direct effect on the hepatic glucose and lipid metabolism, which was an aspect for OxA to exert its maintenance function of energy homeostasis.

肾气丸加减方对高糖高脂诱导的MIN6细胞保护作用及PI3K/AKT/GSK-3β信号通路的影响

目的利用MIN6细胞损伤模型探讨肾气丸加减方通过调控磷脂酰肌醇3-激酶/丝氨酸—苏氨酸激酶/糖原合成激酶3β(phosphoinositide3-kinase/threonine-protein kinase/glycogen synthase kinase-3β,PI3K/AKT/GSK-3β)信号通路保护胰岛β细胞功能的机制。方法将MIN6细胞分为空白组,模型组,肾气丸加减方低、中、高剂量组以及PI3K阻滞剂组(LY294002)。用CCK-8法检测各组细胞活性,胰岛素放射免疫分析试剂盒检测各组MIN6细胞上清液胰岛素含量,流式细胞术检测各组细胞凋亡率,免疫蛋白印迹法检测各组细胞中PI3K、磷酸化(phosphorylated,p)-PI3K、AKT、p-AKT、GSK-3β、p-GSK-3β及胰腺十二指肠同源盒-1(pancreatic duodenal homeobox-1,PDX-1)和肌腱膜纤维肉瘤癌基因同源物A(v-maf musculoaponeurotic fibrosarcoma oncogene homologue A,MafA)蛋白的表达。结果与空白组比较,模型组MIN6细胞活性明显下降,胰岛素分泌水平下降,细胞凋亡率明显升高(P<0.01);与模型组比较,肾气丸加减方低、中、高剂量组细胞活力升高,胰岛素分泌水平升高,细胞凋亡率均降低(P<0.05或P<0.01);各干预组间比较差异无统计学意义(P>0.05)。与空白组比较,模型组PI3K/p-PI3K、AKT/p-AKT、p-GSK-3β蛋白表达明显下调,GSK-3β蛋白表达上调(P<0.05),PDX-1和MafA蛋白表达明显下调(P<0.01);与模型组比较,肾气丸加减方中、高剂量组PI3K/p-PI3K、AKT/p-AKT、p-GSK-3β蛋白表达均明显上调(P<0.05或P<0.01),GSK-3β蛋白表达明显下调(P<0.01);PDX-1和MafA蛋白表达明显上调(P<0.01);加入通路阻滞剂LY294002后肾气丸加减方上述作用被抵消(P<0.05或P<0.01)。结论肾气丸加减方可提高糖脂毒性作用下MIN6细胞活力,减轻细胞凋亡,促进胰岛素分泌,以中、高剂量效果较佳,其机制可能与激活PI3K/AKT/GSK-3β信号通路、抑制GSK-3β的表达有关,进而升高PDX-1和MafA的蛋白表达,恢复胰岛β细胞功能。

Dietary supplementation of some antioxidants against hypoxia

The present study aims to clarify the protective effect of supplementation with some antioxidants,such as idebenone(200 mg/kg,ip),melatonin(10 mg/kg,ip) and arginine(200 mg/kg,ip) and their combination,on liver function(T.protein,albumin,alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase),energetic parameters(adenosine triphosphate,adenosine diphosphate,adenosine monophosphate,inorganic phosphate,total adenylate,adenylate energy charge and potential phosphate).The effect on glycolytic and glycogenolytic enzymes(glucose,glycogen,glycogen phosphorylase,pyruvate kinase and phosphofructokinase against hypoxia) was also studied.The drugs were administered 24 and 1 h prior sodium nitrite intoxication.All biochemical parameters were estimated 1 h after sodium nitrite injection.Injection of sodium nitrite(75 mg/kg,sc) produced a significant disturbance in all biochemical parameters of liver function,energetic parameters and glycolytic and glycogenolytic enzymes.Hepatic damage was confirmed by histopathological examination of the liver as compared to controls.The marked changes in hepatic cells induced by sodium nitrite were completely abolished by pretreatment with the drug combination,suggesting potential protection against sodium nitrite-induced hypoxia.It could be concluded that a combination of both idebenone and melatonin or idebenone and arginine provides potential protection against sodium nitrite-induced hypoxia by improving biochemical parameters and preserving liver histology.

葛根素激活AMPK/Akt/GSK-3β信号通路减轻高糖诱导的H9c2心肌细胞肥大作用研究

[目的]探讨葛根素对高糖诱导的H9c2心肌细胞的保护作用及机制。[方法]采用高糖损伤H9c2心肌细胞48 h,建立体外糖尿病心肌损伤细胞模型;罗丹明标记的鬼笔环肽标记细胞骨架,测定细胞表面积;蛋白免疫印迹法(Western Blot)检测细胞中心肌肥大相关蛋白β-肌球蛋白重链(β-MHC)的水平、AMP活化蛋白激酶α(AMPKα)、丝氨酸/苏氨酸激酶(Akt)和糖原合成激酶3β(GSK-3β)蛋白的磷酸化水平、以及1,4,5-三磷酸2型受体(IP3R2)和Sigma-1受体(Sig-1R)的蛋白水平。[结果]葛根素(20μmol/L)能显著抑制高糖诱导的心肌细胞表面积增大;Western Blot分析结果显示,高糖能上调β-MHC蛋白水平、抑制AMPK和Akt磷酸化,葛根素能下调β-MHC、IP3R2蛋白水平同时促进AMPK、Akt和GSK-3β的磷酸化。[结论]葛根素可能通过激活AMPK/Akt/GSK-3β信号通路减轻高糖诱导的心肌肥大。