天红2号苹果花芽分化期枝条和叶片碳水化合物含量和C/N变化

以天红2号苹果为试材,研究不同砧木短枝和中枝在花芽分化期间叶片和枝条中碳水化合物含量和C/N变化情况。结果表明,稳产期天红2号苹果成花率不受砧木影响。整个花芽分化期,无论嫁接在SH40中间砧还是八棱海棠乔砧上,在同一枝类中,天红2号苹果叶片可溶性糖、蔗糖、果糖和氮含量高于枝条,淀粉含量和C/N低于枝条。嫁接在SH40中间砧上的天红2号苹果成花率高的短枝叶片淀粉和果糖含量低于成花率低的中枝叶片,可溶性糖、蔗糖含量和C/N于花芽分化初期至花瓣原基分化期高于中枝叶片,短枝枝条可溶性糖、果糖含量和C/N低于中枝枝条,短枝枝条淀粉含量于花瓣原基分化期前高于中枝枝条;嫁接在八棱海棠乔砧上的天红2号苹果成花率高的短枝叶片可溶性糖、淀粉、果糖含量和C/N低于成花率低的中枝叶片,短枝枝条可溶性糖、果糖含量和C/N低于中枝枝条,短枝枝条淀粉含量于转化期低于中枝枝条。说明碳水化合物含量和C/N的高低对苹果成花不起决定性作用。在整个花芽分化期,叶片中可溶性糖含量呈上升趋势,枝条中淀粉含量和C/N呈波动性上升趋势,说明天红2号苹果花芽形态分化需要碳水化合物和C/N的积累。

Ginsenoside-Rg_6, a Novel Triterpenoid Saponin from the Stem-Leaves of Panax ginseng C. A. Mey.

A novel dammarane-type triterpene oligoglycoside, named ginsenoside-Rg6 3, was isolated from the stem-leaves of Panax ginseng C. A. Mey., together with two known ones, 20(S)-ginsenoside-Rg2 1 and 20(R)-ginsenoside-Rg2 2. On the basis of chemical and physicochemical evidence , the structure of ginsenoside-Rg6 have been elucidated as 6-O-(-L-rhamnosyl-(1?2)-(-D-glucopyranosyl-dammarane-(E)-20(22), 24-diene-3(, 6(, 12(-triol.

高效液相色谱法测定莱芜鸡腿葱不同食用部位中维生素C含量的方法优化

本文采用高效液相色谱法测定莱芜鸡腿葱葱茎及葱叶中的维生素C(L-抗坏血酸)含量,并进行了方法优化研究。样品采用1.0%的草酸溶液作为提取液超声提取2次,经Kromasil C_(18)柱分离后,用紫外检测器在254 nm波长处检测,以保留时间定性,外标法定量。结果显示,维生素C质量浓度在2.0~100.0 mg·L^(-1)时的线性关系较好(R=0.9995),精密度试验的相对标准偏差为1.33%(n=6),加标回收率为94.9%~96.1%,方法检出限为2.0×10^(-3)g·kg^(-1)。该测试方法简便快速,能准确测定莱芜鸡腿葱葱茎及葱叶中维生素C含量。

Ultraviolet-attenuated cercariae ofSchistosoma japonicum fail to effectively induce a Th1 response in spite of up-regulating expression of cytotoxicity-related genes in C57BL/6 mice

Objective： To better understand the reason that Schistosoma japonicurn （S. japonicum） ultraviolet （UV）- radiated cercariae could not induce high level of protection in C57BL/6 mice. Methods： Microarray technology was performed to investigate the gene transcription profile in skin draining lymph nodes （sdLNs） at 1 w after exposure to attenuated cercariae （AC） or normal cercariae （NC） of S. japonicum in C57BL/6 mice. The expressions of some representative genes were further confirmed by real-time PCR. Subsequently, the expressions of Th1/Th2 cytokine genes, cytotoxicity-related genes, as well as co-stimulator genes in spleens from AC-vaccinated and NC- infected mice were analyzed by real-time PCR at w 3 and 6 post-exposure. Results： The gene expressions of Th1 cytokines, including interferon-y （IFN-γ）, interleukin （IL）-12 and tumor necrosis factor-α （TNF-α） in the sdLNs were significantly lower in AC-vaccinated mice than in NC-infected mice. Furthermore, the gene expressions of Th1- and Th2- cytokines, including IFN-γ, IL-12, TNF-α, IL-4 and IL-10, in the spleens from AC-vaccinated mice showed little changes at w 3 and 6 post-vaccination. In addition, cytotoxicity-related molecules including granzyme A, granzyme B, granzyme K, perforin 1 and Fas L were up-regulated from the early stage of vaccination, and peaked at the 3rd w after vaccination with UV-AC. Conclusion： UV-AC of S. japonicum could not ef- fectively induce a Thl response in C57BL/6 mice, which may be an explanation for the low protection against parasite challenge, and the role played by up-regulated expression of cytotoxicity-related genes in mice needs to be further investigated.

Polysaccharide extracts of Cirsium japonicum protect rat H9c2 myocardial cells from oxidative stress induced by hydrogen peroxide

Daji （Cirsium japonicum） has been applied against gastric disorders, lung diseases, and cardiovascular problems in thetraditional Chinese medicinal system. The present study was to investigate the protective effects of Daji （Cirsiumjaponicum） polysaccharide extracts （CJP） against hydrogen peroxide （H2O2） shock in rat H9c2 myocardial cells. First,CJP was isolated by hot water extraction and ethanol precipitation; it was then characterized by high performance liquidchromatography and infrared spectrum analysis. Rat H9c2 cells were subjected to H2O2 treatment to establish a cellinjury model. The 3- （4,5- dimethylthiazol- 2-yl）-2,5- diphenyltetrazolium bromide assay showed that CJP pretreatmentsignificantly ameliorated the H2O2 injury in a dose-dependent manner. Furthermore, the cell apoptosis induced by H2O2was markedly inhibited by CJP pretreatment, whereas the cleavage level of caspase-3, -8, and -9 was reduced. Inaddition, the p38 mitogen-activated protein kinase pathway might be involved in the protective effect of CJP onmyocardial cells. Therefore, we conclude that polysaccharide extracts of Daji （Cirsium japonicum） protect rat H9c2myocardial cells from oxidative stress induced by H2O2.

Exogenous bone morphogenetic protein-7 reduces hepatic fibrosis inSchistosoma japonicum-infected micevia transforming growth factor-β/Smad signaling

AIM: To investigate the antifibrotic effects of bone morphogenetic protein-7 (BMP-7) on Schistosoma japonicum (S. japonicum )-induced hepatic fibrosis in BALB/C mice. METHODS: Sixty BALB/C mice were randomly divided into three groups, including a control group (group A, n = 20), model group (group B, n = 20) and BMP-7 treated group (group C, n = 20). The mice in group B and group C were abdominally infected with S. japonicum cercariae to induce a schistosomal hepatic fibrosis model. The mice in group C were administered human recombinant BMP-7. Liver samples were extracted from mice sacrificed at 9 and 15 wk after modeling. Hepatic histopathological changes were assessed using Masson's staining. Transforming growth factor-beta 1 (TGF-β1), alpha-smooth muscle actin (α-SMA), phosphorylated Smad2/3 (pSmad2/3) and Smad7 protein levels and localization were measured by Western blotting and immunohistochemistry, respectively, and their mRNA expressions were detected by reverse transcriptionpolymerase chain reaction (RT-PCR). RESULTS: The schistosomal hepatic fibrosis mouse model was successfully established, as the livers of mice in group B and group C showed varying degrees of typical schistosomal hepatopathologic changes such as egg granuloma and collagen deposition. The degree of collagen deposition in group C was higher than that in group A (week 9: 22.95±6.66vs 2.02±0.76; week 15: 12.84±4.36 vs 1.74±0.80; P<0.05), but significantly lower than that in group B (week 9: 22.95±6.66 vs 34.43±6.96; week 15: 12.84±4.36 vs 18.90±5.07;P<0.05) at both time points. According to immunohistochemistry data, the expressions of α-SMA, TGF-β1 and pSmad2/3 protein in group C were higher than those in group A (α-SMA: week 9: 21.24±5.73 vs 0.33±0.20; week 15: 12.42±4.88 vs 0.34±0.27; TGF-β1: week 9: 37.00±13.74 vs 3.73±2.14; week 15: 16.71±9.80 vs 3.08±2.35; pSmad2/3: week 9: 12.92±4.81 vs 0.83±0.48; week 15: 7.87±4.09 vs 0.90±0.45; P<0.05), but significantly lower than those in group B (α-SMA: week 9: 21.24±5.73 vs 34.39±5.74; week 15: 12.42±4.88 vs 25.90±7.01; TGF-β1: week 9: 37.00±13.74 vs 55.66±14.88; week 15: 16.71±9.80 vs 37.10±12.51; pSmad2/3: week 9: 12.92±4.81 vs 19.41±6.87; week 15: 7.87±4.09vs 13.00±4.98;P<0.05) at both time points; the expression of Smad7 protein in group B was higher than that in group A and group C at week 9 (8.46±3.95 vs 1.00±0.40 and 8.46±3.95 vs 0.77±0.42; P<0.05), while there were no differences in Smad7 expression between the three groups at week 15 (1.09±0.38 vs 0.97±0.42 vs 0.89±0.39; P>0.05). Although minor discrepancies were observed, the results of RT-PCR and Western blotting were mainly consistentwith the immunohistochemical results. CONCLUSION: Exogenous BMP-7 significantly decreased the degree of hepatic fibrosis in both the acute and chronic stages of hepato-schistosomiasis, and the regulatory mechanism may involve the TGF-β/Smad signaling pathway.

Osteopontin expression is associated with hepatopathologic changes in Schistosoma japonicum infected mice

AIM:To investigate osteopontin expression and its association with hepatopathologic changes in BALB/C mice infected with Schistosoma japonicum.METHODS:The schistosomal hepatopathologic mouse model was established by abdominal infection with schistosomal cercaria.Liver samples were obtained from mice sacrif iced at 6,8,10,14,and 18 wk after in-fection.Liver histopathological changes were observed with hematoxylin-eosin and Masson trichrome staining.The expression of osteopontin was determined with im-munohistochemistry,reverse transcription-polymerase chain reaction,and Western blotting.The expressionof α-smooth muscle actin(α-SMA)and transforming growth factor-β1(TGF-β1)were determined by im-munohistochemistry.Correlations of osteopontin ex-pression with other variables(α-SMA,TGF-β1,hepato-pathologic features including granuloma formation and degree of liver f ibrosis)were analyzed.RESULTS:Typical schistosomal hepatopathologic changes were induced in the animals.Dynamic changes in the expression of osteopontin were observed at week 6.The expression increased,peaked at week 10(P<0.01),and then gradually decreased.Positive correla-tions between osteopontin expression and α-SMA(r=0.720,P<0.01),TGF-β1(r=0.905,P <0.01),granu-loma formation(r=0.875,P<0.01),and degree of liver f ibrosis(r=0.858,P<0.01)were also observed.CONCLUSION:Osteopontin may play an important role in schistosomal hepatopathology and may promote granuloma formation and liver fi brosis through an un-explored mechanism.

重组C因子检测银杏叶提取物注射液细菌内毒素的方法学研究

目的:建立银杏叶提取物注射液的重组C因子细菌内毒素检测方法。方法:使用重组C因子法进行细菌内毒素方法学研究,并用鲎试剂动态显色法、家兔法对银杏叶提取物注射液进行对比研究。结果:金纳多注射液稀释至400倍及以下浓度进行试验,回收率均在《中国药典》规定的50%~200%范围内,符合药典规定,并与动态显色法和热原法结果一致。结论:本研究为重组C因子检测中药注射液细菌内毒素的方法学和标准的建立提供了实验依据。

Functional foods effective for hepatitis C: Identification of oligomeric proanthocyanidin and its action mechanism

Hepatitis C virus(HCV)is a major cause of viral hepatitis and currently infects approximately 170 million people worldwide.An infection by HCV causes high rates of chronic hepatitis(】75%)and progresses to liver cirrhosis and hepatocellular carcinoma ultimately.HCV can be eliminated by a combination of pegylatedα-interferon and the broad-spectrum antiviral drug ribavirin;however,this treatment is still associated with poor efficacy and tolerability and is often accompanied by serious side-effects.While some novel direct-actingantivirals against HCV have been developed recently,high medical costs limit the access to the therapy in cost-sensitive countries.To search for new natural anti-HCV agents,we screened local agricultural products for their suppressive activities against HCV replication using the HCV replicon cell system in vitro.We found a potent inhibitor of HCV RNA expression in the extracts of blueberry leaves and then identified oligomeric proanthocyanidin as the active ingredient.Further investigations into the action mechanism of oligomeric proanthocyanidin suggested that it is an inhibitor of heterogeneous nuclear ribonucleoproteins(hn RNPs)such as hn RNP A2/B1.In this review,we presented an overview of functional foods and ingredients efficient for HCV infection,the chemical structural characteristics of oligomeric proanthocyanidin,and its action mechanism.

A New Bisxanthone from Hypericum japonicum Thunb.ex Murray

A new bisxanthone, named bijaponicaxanthone C, was isolated from the whole plant of Hypericum japonicum. The structure was elucidated as 6-[1’’,5’’,6’’-trihydroxy-2’’’-(β-hydroxy-β- methylethyl)-2’’’,3’’’-dihydrofuran(5’’’,4’’’,3’’,4’’)xanthone-3’’’-oxyl]-1,3,5-trihydroxy-4-isoprenylxant- hone (1) on the basis of the spectral and chemical evidences.

法桐树叶衍生Fe-N-C氧还原电催化剂的制备及性能研究

Fe-N-C材料被认为是最有潜力替代贵金属Pt催化剂用于催化燃料电池阴极氧还原反应(Oxygen Reduction Reaction,ORR)的廉价非贵金属催化剂之一。本文以碱式碳酸锌(BZC)、法桐树叶(FT)、氯化铁和碳酸钾分别为反应性硬模板、碳氮前驱体、金属前驱体和激活剂,通过固相研磨法和高温热处理制备了生物质基Fe-N-C材料(FT-Fe-N-C(BZC))并考察了其在0.1M KOH溶液中对ORR的电催化性能。结果显示,其起始电势(Eonset)、半波电势(E1/2)和极限电流密度分别达到0.93V、0.81V和4.02mA·cm^(-2),且展示出高效的4电子ORR过程选择性;另外,在制备过程中引入过硫酸铵(AP),获得的FT-Fe-N-C(BZC/AP)催化ORR在维持4电子反应途径同时,其E_(onset)和E_(1/2)都提升了30mV,与商业Pt/C催化剂相当。

C-6膜粘附水中日本血吸虫尾蚴技术的研究

在实验室和现场水面,应用C-6膜多点抽样粘取方法检测水中尾蚴。获蚴率达40％,高于现用的尼龙绢条粘取沉淀法约20倍,在现场用现有方法未查获尾蚴的水面,调查钉螺阳性率为零或灭螺后的水面,本法可在一些环境查获尾蚴。本法操作简便,当天可有检测结果,费用仅约为哨鼠法的1％,利于广泛应用。

东方田鼠感染血吸虫前后血清补体C_3 C_4水平的动态

目的 检测东方田鼠感染血吸虫前后血清补体C3 、C4 水平 ,阐明其动态规律 ,为东方田鼠抗血吸虫感染的免疫机理提供证据。 方法 免疫比浊法检测血清补体C3 、C4 水平。 结果 未感染血吸虫的东方田鼠血清补体C3 、C4 平均含量显著高于昆明系小鼠 ,东方田鼠感染后血清中C3 、C4 平均水平在第 11d以前较低 ,第 2 1d升高 ,第 2 6d较前显著升高 ,昆明系小鼠感染后C3 、C4 平均水平逐渐上升 ,直至第 2 6d非常显著地高于第 2 1d之前者。 结论 东方田鼠血清补体水平较高 。

肉桂及其混伪品的HSGC-MS的实验比较研究

目的 比较肉桂及其混伪品的顶空气相指纹图谱 ,以此鉴别肉桂药材的真伪。方法 采用顶空气相色谱 -质谱 (HSGC- MS)计算机联用技术 ,分别测定肉桂、桂皮、柴桂的指纹图谱并加以比较。结果 肉桂的顶空气相指纹图谱与混伪品桂皮、柴桂有明显区别 ,其指纹特征峰位于 10 .6 0 ,15 .6 0 min处 ,主要成分为桂皮醛。结论 此方法能有效地鉴别肉桂及其混伪品 ,具有快速、准确、操作简单 ,不需对样品进行提取分离 。

紫外线致弱日本血吸虫尾蚴诱导BALB/c小鼠免疫保护作用研究

目的观察紫外线(UV)致弱日本血吸虫尾蚴免疫BALB/c小鼠免疫保护作用。方法辐照致弱日本血吸虫尾蚴,经腹部皮肤用UV致弱日本血吸虫尾蚴免疫诱导BALB/c小鼠,3周后进行攻击感染,同时设正常感染对照组。攻击感染6周后解剖小鼠,计算减虫率、减卵率,并观察虫体发育情况和肝组织细胞免疫应答情况。结果免疫组的减虫率和减卵率分别为37.90%和51.16%;免疫组小鼠体内虫体发育明显滞后于正常感染对照组内虫体;免疫组小鼠肝脏虫卵肉芽肿较正常组明显减少。结论UV致弱日本血吸虫尾蚴免疫BALB/c小鼠能诱导较好的免疫保护力。

不同方法测定三七花、茎叶中维生素C含量

目的:比较3种测定三七花、茎叶中维生素C含量的方法,为三七地上部分开发利用提供科学依据。方法:采用2,6-二氯靛酚滴定法、紫外分光光度法、高效液相色谱法3种不同检测方法对三七地上部分维生素C含量进行测定。结果:三七花和茎叶中,采用紫外分光光度法和高效液相色谱法测定维生素C含量,其方法学考察良好,且两种方法测定结果高于滴定法所得结果。三七花和茎叶中维生素C平均含量无明显差异,其中采用HPLC法测得的结果分别为11.76mg/100g和11.39mg/100g。结论:三七花和茎叶中含有丰富的维生素C,紫外分光光度法和HPLC法均适用于三七地上部分中维生素C的含量测定。

日本血吸虫中国大陆株TPI DNA疫苗诱导BALB/c小鼠保护性免疫力的研究

目的 研究日本血吸虫中国大陆株磷酸丙糖异构酶 (Sj CTPI) DNA疫苗诱导 BAL B/ c小鼠的保护性免疫作用。方法 将 39只 BAL B/ c小鼠随机分成 3组 :A组 (对照组 ) ,肌注 pc DNA3.1质粒 DNA10 0 μg/鼠 ;B组 (实验组 ) ,每鼠肌注 pc DNA3.1- Sj CTPI10 0 μg;C组 (加强组 ) ,每鼠肌注 pc DNA3.1- Sj CTPI10 0 μg及 pc DNA3.1- P35和 pc DNA3.1-P40的混合物 10 0 μg。每两周免疫 1次 ,共计 3次。末次免疫 4周后 ,每鼠用 45条尾蚴进行攻击 ,45 d后剖杀小鼠 ,计数成虫以及每鼠肝组织内虫卵数 ;于攻击前每组各取两只小鼠采用 51 Cr释放法测定 Sj CTPI介导的特异性细胞毒作用 ;EL ISA检测攻击前后的抗 TPI的抗体水平。 结果 用 EL ISA检测抗 TPI抗体的结果表明 ,攻击前 A组的 10份血清均为阴性 ,B组 5 / 10份血清出现弱阳性 ,C组也有 6 / 10份血清出现弱阳性反应。51 Cr释放法测定细胞毒活性结果表明 ,A、B、C组细胞毒活性分别为 9.1%、2 7.6 %和 5 4.4%。与对照组相比 ,实验组的减虫率、减卵率分别为 30 .2 %、5 2 .9% ;加强组的减虫率、减卵率分别为 32 .7%、47.0 %。 结论 进一步证实了 Sj CTPI DNA疫苗作为抗血吸虫病核酸疫苗的潜力。

血吸虫虫卵在BALB/c鼠各组织中的分布

目的研究血吸虫感染BALB/c鼠后,虫卵在其部分组织中特别是在肠壁中的分布状况。方法 10只BALB/c鼠腹部贴片感染血吸虫尾蚴(40±1条/只),感染42d后剖杀,分别取鼠的肝脏、脾脏、肠道(小肠上段、中段、下段、盲结肠、直肠)进行成虫计数、虫卵计数、毛蚴孵化、组织切片观察,并将得到的结果进行统计学分析。结果血吸虫虫卵在BALB/c鼠各组织的分布为:肝脏(59.64%)、脾脏(0.81%)、小肠上段(6.27%)、小肠中段(5.24%)、小肠下段(5.14%)、盲结肠(17.48%)、直肠(5.41%),肝脏的沉积虫卵数和孵化数与其他组织相比差异显著,肠组织中大肠段比小肠段的沉积虫卵数和孵化数多且差异显著。在孵化率的比较中肝脏的孵化率最高(1.09%)。病理切片观察,肝脏产生大量肉芽肿结节,肠壁出现不同程度的炎症坏死。结论 BALB/c鼠定量人工感染血吸虫后,肝脏中的沉积虫卵最多,其次是盲结肠,最少的是脾脏;且不同组织的沉积虫卵数目与病理损伤程度有关。

高速逆流色谱法分离纯化银杏叶中白果内酯和银杏内酯A、B、C

目的从银杏叶中分离纯化白果内酯和银杏内酯A、B、C。方法银杏叶提取液经醋酸乙酯萃取、D-101大孔吸附树脂柱和Al2O3柱纯化后得到总内酯提取物,提取物再经两次高速逆流色谱分离制备4种内酯单体。结果25%乙醇热提取、醋酸乙酯萃取、D-101柱、pH4.0的Al2O3柱对总内酯的提纯最终使总内酯质量分数达到44.98%。经高速逆流色谱制备得到的白果内酯和银杏内酯A、B、C最高质量分数分别为98.3%、98.9%、98.8%、98.4%。结论本法简便快速、回收率高。

HPLC法测定银杏叶粉针中总黄酮醇苷和萜类内酯

目的采用高效液相色谱法测定银杏叶粉针中总黄酮醇苷和总萜类内酯。方法黄酮醇苷水解后,采用HPLC-UV法测定;萜类内酯用醋酸乙酯萃取后采用HPLC-ELSD法测定。结果槲皮素、山柰素、异鼠李素分别在5.104～51.04、5.000～50.00、1.38～13.8μg/mL呈良好线性关系,平均回收率分别为99.34%、99.25%、101.2%(n=5);银杏内酯A、B、C及白果内酯分别在0.415～2.075、0.402～2.010、0.426～2.130、0.803～4.015mg/mL呈良好线性关系,平均回收率分别为99.28%、100.1%、101.0%、99.31%(n=5)。结论本实验采用的方法简便快速,结果准确,重现性好,可用于银杏叶粉针中黄酮和内酯类成分的质量控制。