不同储藏时间笔架茶生物碱成分分析

本实验以三种储藏时间不同的笔架茶1#、2#和3#茶为原料,用热回流法和有机溶剂萃取法联用的方法提取其生物碱,分析了热回流时间(A)和料液比(B)对三种茶生物碱提取率的影响。用M arqu is等五种试剂对提取的生物碱进行鉴定分析,并用HPLC分析生物碱主要成分的含量。结果表明:笔架茶中生物碱的含量随储藏时间的延长而降低。A和B对储藏时间不同的三种笔架茶生物碱提取率的影响各不相同。沉淀及显色反应鉴定显示提取生物碱的主要成分为咖啡因,HPLC定量分析表明3#新茶生物碱中咖啡因含量最高(94.33%),1#陈茶咖啡因含量最低(75.76%)。     

莫诺苷对大鼠局灶性脑缺血再灌注损伤神经功能的影响

目的观察中药山茱萸的有效成分莫诺苷对大鼠局灶性脑缺血再灌注后神经功能的影响。方法Wistar雄性大鼠随机分为假手术组、模型组、莫诺苷小剂量组（30 mg/kg）、莫诺苷中剂量组（90 mg/kg）、莫诺苷大剂量组（270 mg/kg）、维生素E（VE）（35 mg/kg）,采用线栓法制作大鼠局灶性脑缺血再灌注模型,缺血30 min后再灌注3 d,应用Zea Longa法、爬网格、平行木、吊绳、Ludmila Belayev 12分评分法,观察莫诺苷对神经功能缺损的改善作用。结果与模型组比较,莫诺苷给药组（小、中、大剂量）Zea Longa法评分均差异极显著（P〈0.01）;与模型组比较,莫诺苷给药组（小、中、大剂量）吊绳法评分均差异极显著（P〈0.01）;Ludmila Belayev 12分评分法评分与模型组比较,莫诺苷大剂量组差异极显著（P〈0.001）,中剂量组差异极显著（P〈0.01）。结论莫诺苷对局灶性脑缺血再灌注模型大鼠有改善行为学评分的作用。     

Effect of isoflavone administration on age-related hepatocyte changes in old ovariectomized femal Wistar rats

Aging seems to be due to the accumulation of oxidative damage in cells and molecules. On the other hand, menopause and ovariectomy induce deleterious effects on different organs and systems that have been shown to be counteracted by estrogens and in a not so evident form also with phytoestrogens. The present study has investigated whether the administration of a commercial soy extract that contains approximately 10% isoflavones was able to modify some parameters related to oxidative stress and inflammation in hepatocytes isolated from old ovariectomized female Wistar rats. Eighteen 22-month-old animals that had been previously ovariectomized at 12 months of age were divided into four groups: ovariectomized control rats, estradiol-treated ovariectomized females and ovariectomized rats treated with isoflavones. Six intact female rats of 2 months of age were used as reference group. Hepatocytes were isolated and cultured, and carbon monoxide (CO) and nitric oxide (NO) release, as well as adenosyl triphosphate (ATP), cyclic guanosyl monophosphate (cGMP), phosphatidylcholine (PC) and lipid peroxide (LPO) content of cells were evaluated. Uterus was also removed and weighed. Hepatocytes isolated from old ovariectomized rats showed a decrease in ATP content as compared to young animals. Age also induced an increase in LPO cell content. NO, CO and cGMP were augmented with age, and PC synthesis showed a dramatic reduction. Treatment with either estradiol or isoflavones were able to improve all the mentioned parameters altered in hepatocytes isolated from old ovariectomized rats, and the magnitude of the improvement was similar for both treatments. Ovariectomy induced a significant reduction in uterine weight, which was significantly counteracted by estradiol treatment but not by isoflavone administration. In conclusion, the administration of a soy extract containing isoflavones seems to prevent oxidative changes in hepatocytes isolated from old ovariectomized female rats, without modifying uterus weight.     

Dynamics of fine root carbon in Amazonian tropical ecosystems and the contribution of roots to soil respiration

Radiocarbon (¹⁴C) provides a measure of the mean age of carbon (C) in roots, or the time elapsed since the C making up root tissues was fixed from the atmosphere. Radiocarbon signatures of live and dead fine (<2 mm diameter) roots in two mature Amazon tropical forests are consistent with average ages of 4–11 years (ranging from <1 to >40 years). Measurements of ¹⁴C in the structural tissues of roots known to have grown during 2002 demonstrate that new roots are constructed from recent (<2‐year‐old) photosynthetic products. High Δ¹⁴C values in live roots most likely indicate the mean lifetime of the root rather than the isotopic signature of inherited C or C taken up from the soil. Estimates of the mean residence time of C in forest fine roots (inventory divided by loss rate) are substantially shorter (1–3 years) than the age of standing fine root C stocks obtained from radiocarbon (4–11 years). By assuming positively skewed distributions for root ages, we can effectively decouple the mean age of C in live fine roots (measured using ¹⁴C) from the rate of C flow through the live root pool, and resolve these apparently disparate estimates of root C dynamics. Explaining the ¹⁴C values in soil pore space CO₂, in addition, requires that a portion of the decomposing roots be cycled through soil organic matter pools with decadal turnover time.     

贵州苏铁根的解剖学研究

对贵州苏铁（Cycas guizhouensis K.M.Lan et R.F.Zou）的根进行了解剖学研究,结果表明：（1）据外部形态和内部结构不同,贵州苏铁的根可分为正常根、珊瑚状根和肉质根三种类型。（2）正常根与肉质根的木栓形成层起源于表皮内方第1～2层皮层薄壁细胞,珊瑚状根的木栓形成层则起源于外方皮层的第2～3层细胞,而非中柱鞘。（3）首次发现肉质根一种新的增粗方式,是由周皮内方的皮层薄壁细胞恢复分裂能力,持续或周期性产生新的皮层薄壁细胞,引起根的增粗。     

Real-Time PCR for the Detection and Quantification of Geodermatophilaceae from Stone Samples and Identification of New Members of the Genus Blastococcus

Real-time PCR (RT-PCR) technology was used for the specific detection and quantification of members of the family Geodermatophilaceae in stone samples. Differences in the nucleotide sequences of the 16S rRNA gene region were used to design a pair of family-specific primers that were used to detect and quantify by RT-PCR DNA from members of this family in stone samples from different geographical origins in Spain. These primers were applied later to identify by PCR-specific amplification new members of the family Geodermatophilaceae isolated from the same stone samples. The diversity and taxonomic position of the wild-type strains identified from ribosomal sequence analysis suggest the presence of a new lineage within the genus Blastococcus.     

Predation by invertebrate predators on the colonial rotifer Sinantherina socialis

Abstract. Colonies of the freshwater colonial rotifer Sinantherina socialis (Monogononta, Flosculariidae) have been shown to be unpalatable to a variety of small-mouthed, zooplanktivorous fishes. To test whether invertebrate predators ingest the rotifer S. socialis , we conducted two types of experiments: (1) Microcosm experiments—in separate experiments, four invertebrate predators (i.e., dragonfly nymphs, damselfly nymphs, notonectids, and Hydra ) were offered prey either singly or in combination. Prey were comprised of S. socialis; Epiphanes senta , a solitary, free-swimming rotifer; and Daphnia magna , a microcrustacean. In each experiment, the percent of prey surviving after 12, 18, and 24 h was recorded. (2) Paired-feeding experiments—in separate experiments, predators were offered prey in a pairwise fashion, in which members of D. magna were alternated with a rotifer, either S. socialis or E. senta. The results of the microcosm experiments showed that, after 24 h, 60–100% prey items of S. socialis survived the predators, but significantly fewer individuals of E. senta (6–89%) and D. magna (<25%) survived. When offered rotifers and individuals of D. magna simultaneously, predators tested consistently consumed more specimens of Daphnia. However, predators significantly reduced percent survival in E. senta but not in S. socialis. Predators, given a choice between the two rotifer species, all consumed significantly more specimens of E. senta than S. socialis after 24 h. In the paired-feeding experiments, three of the four predators captured members of S. socialis , but these colonies were frequently released rather than ingested, although in some cases colony structure was seriously disrupted. Our results suggest that the unpalatable nature of members of S. socialis to certain fishes extends to several invertebrate predators, but the nature of the putative factor(s) responsible for this remains unknown.     

Arachidonic acid induces an increase of β-1,4-galactosyltransferase I expression in MDA-MB-231 breast cancer cells

Arachidonic acid (AA) is a common dietary n‐6 cis polyunsaturated fatty acid that under physiological conditions is present in an esterified form in cell membrane phospholipids, and it might be present in the extracellular microenvironment. AA and its metabolites are implicated in FAK activation and cell migration in MDA‐MB‐231 breast cancer cells, and an epithelial‐to‐mesenchymal‐like transition process in mammary non‐tumorigenic epithelial cells MCF10A. During malignant transformation is present an altered expression of glycosiltransferases, which promote changes on the glycosilation of cell‐surface proteins. The β‐1,4‐galactosyltransferase I (GalT I) is an enzyme that participates in a variety of biological functions including cell growth, migration, and spreading. However, the participation of AA in the regulation of GalT I expression and the role of this enzyme in the cell adhesion process in breast cancer cells remains to be investigated. In the present study, we demonstrate that AA induces an increase of GalT I expression through a PLA2α, Src, ERK1/2, and LOXs activities‐dependent pathway in MDA‐MB‐231 breast cancer cells. Moreover, MDA‐MB‐231 cells adhere to laminin via GalT I expression and pretreatment of cells with AA induces an increase of cell adhesion to laminin. In conclusion, our findings demonstrate, for the first time, that AA promotes an increase of GalT I expression through an AA metabolism, Src and ERK1/2 activities‐dependent pathway, and that GalT I plays a pivotal role in cell adhesion to laminin in MDA‐MB‐231 breast cancer cells. J. Cell. Biochem. 113: 3330–3341, 2012. © 2012 Wiley Periodicals, Inc.     

Allopregnanolone Potentiates the Glutamate-Mediated Seizures Induced by 4-Aminopyridine in Rat Hippocampus in vivo

Excitatory and inhibitory neurotransmission in the central nervous system can be modulated by neurosteroids. We previously found that in rat hippocampal slices allopregnanolone (3α-hydroxy-5α-pregnan-20-one), a positive GABA_A receptor modulator, suppresses the epileptic discharges induced by 4-aminopyridine (4-AP), a convulsant K⁺ channel blocker that stimulates glutamate release. Here, we tested the action of allopregnanolone on the epileptogenic and excitotoxic effects of the intrahippocampal administration of 4-AP in vivo. Drugs were perfused by a microdialysis cannula-electrode in the dorsal hippocampus and the EEG was recorded. Extracellular levels of aspartate, glutamate and GABA were analyzed by HPLC in the microdialysis fractions, and 24 h after the experiment the hippocampus was studied histologically. 4-AP induced intense epileptic discharges, increased the extracellular levels of aspartate, glutamate, and GABA by 383, 420, and 245%, respectively, and produced a notable neurodegeneration in CA1 and CA3 areas. Allopregnanolone administration alone did not affect the electrical activity, amino acids levels or cellular morphology, but when co-infused with 4-AP incremented 55–77% the duration of the epileptic discharges, and potentiated 32–49% the release of glutamate in comparison with 4-AP alone. The 4-AP-induced neurodegeneration was not modified by allopregnanolone. The NMDA receptor antagonist MK-801 protected against the epilepsy and neurodegeneration produced by 4-AP, and allopregnanolone did not affect this protection. We conclude that, differently from the observations in vitro, allopregnanolone potentiated the stimulatory effect of 4-AP on glutamate release and that this may explain the potentiation of the epileptogenic effect of 4-AP in vivo.     

Risk of immunodeficiency virus infection may increase with vaccine-induced immune response

To explore the efficacy of novel complementary prime-boost immunization regimens in a nonhuman primate model for HIV infection, rhesus monkeys primed by different DNA vaccines were boosted with virus-like particles (VLP) and then challenged by repeated low-dose rectal exposure to simian immunodeficiency virus (SIV). Characteristic of the cellular immune response after the VLP booster immunization were high numbers of SIV-specific, gamma interferon-secreting cells after stimulation with inactivated SIV particles, but not SIV peptides, and the absence of detectable levels of CD8(+) T cell responses. Antibodies specific to SIV Gag and SIV Env could be induced in all animals, but, consistent with a poor neutralizing activity at the time of challenge, vaccinated monkeys were not protected from acquisition of infection and did not control viremia. Surprisingly, vaccinees with high numbers of SIV-specific, gamma interferon-secreting cells were infected fastest during the repeated low-dose exposures and the numbers of these immune cells in vaccinated macaques correlated with susceptibility to infection. Thus, in the absence of protective antibodies or cytotoxic T cell responses, vaccine-induced immune responses may increase the susceptibility to acquisition of immunodeficiency virus infection. The results are consistent with the hypothesis that virus-specific T helper cells mediate this detrimental effect and contribute to the inefficacy of past HIV vaccination attempts (e.g., STEP study).