Nitric oxide is involved in methyl jasmonate-induced defense responses and secondary metabolism activities of Taxus cells

Methyl jasmonate (MeJA), a methyl ester of jasmonic acid (JA), is a well-established signal molecule in plant defense responses and an effective inducer of secondary metabolite accumulation in plant cell cultures such as the valuable anticancer diterpenoid taxol (paclitaxel) in Taxus spp. This work examines the involvement of nitric oxide (NO) in MeJA-induced plant defense responses and secondary metabolism in Taxus chinensis cell cultures. Exogenously supplied MeJA at 100 microM induced rapid production of NO in the Taxus cell cultures, reaching a maximum within 6 h of MeJA supply. Several other responses occurred concomitantly, including the production of hydrogen peroxide (H2O2), and the increases in intracellular malondialdehyde (MDA) content, lipoxygenase (LOX) and phenylalanine ammonium-lyase (PAL) activities. The MeJA-induced H2O2 production was suppressed by an NO donor, sodium nitroprusside (SNP), but enhanced by NO inhibitors, N (omega)-nitro-L-arginine (L-NNA) and 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (PTIO). In contrast, the MeJA-induced MDA, LOX and PAL were all enhanced by the NO donor but suppressed by the NO inhibitors. The NO inhibitors also suppressed MeJA-induced taxol accumulation. These results are suggestive of a role for NO as a signal element for activating the MeJA-induced defense responses and secondary metabolism activities of plant cells.     

一个用于转染细胞分选的双顺反子载体的构建及其应用

为简化转染细胞的分选过程,构建了一个含有细胞表面标志 CD34 基因的双顺反子载体 p3.1-IRES-CD34. 利用来源于脑心肌炎病毒 (EMCV) 的内部核糖体进入位点 (IRES) ,实现目的基因与 CD34 基因的共同表达 . 将绿色荧光蛋白 (EGFP) 作为目的基因插入载体的多克隆位点,然后转染 NIH-3T3 细胞,通过免疫磁珠分选 (MACS) 方法来分选细胞 . 结果表明：对于转染细胞,均可实现快速分选 ( 瞬时转染细胞约 48 h ,稳定转染 10~15 天 ) ,并且获得较高纯度 (95% 以上 ) 的表达目的基因细胞 .     

Biosynthesis of fusarochromanone and its monoacetyl derivative by Fusarium equiseti.

One fluorescent compound previously named TDP-2 was isolated and purified from a rice culture of Fusarium equiseti (Alaska 2-2). Mass spectral and nuclear magnetic resonance data indicated that it is a C-3'-N-acetyl derivative of fusarochromanone, a newly discovered mycotoxin. Time course studies of synthesis of these two compounds on autoclaved rice and Czapek-Dox medium enriched with soybean peptone indicated that fusarochromanone was converted to TDP-2 in the cultures. A high concentration of peptone in the liquid medium may stimulate both fusarochromanone synthesis and its conversion to TDP-2.     

Orai1 and STIM reconstitute store-operated calcium channel function

The two membrane proteins, STIM1 and Orai1, have each been shown to be essential for the activation of store-operated channels (SOC). Yet, how these proteins functionally interact is not known. Here, we reveal that STIM1 and Orai1 expressed together reconstitute functional SOCs. Expressed alone, Orai1 strongly reduces store-operated Ca(2+) entry (SOCE) in human embryonic kidney 293 cells and the Ca(2+) release-activated Ca(2+) current (I(CRAC)) in rat basophilic leukemia cells. However, expressed along with the store-sensing STIM1 protein, Orai1 causes a massive increase in SOCE, enhancing the rate of Ca(2+)entry by up to 103-fold. This entry is entirely store-dependent since the same coexpression causes no measurable store-independent Ca(2+) entry. The entry is completely blocked by the SOC blocker, 2-aminoethoxydiphenylborate. Orai1 and STIM1 coexpression also caused a large gain in CRAC channel function in rat basophilic leukemia cells. The close STIM1 homologue, STIM2, inhibited SOCE when expressed alone but coexpressed with Orai1 caused substantial constitutive (store-independent) Ca(2+) entry. STIM proteins are known to mediate Ca(2+) store-sensing and endoplasmic reticulum-plasma membrane coupling with no intrinsic channel properties. Our results revealing a powerful gain in SOC function dependent on the presence of both Orai1 and STIM1 strongly suggest that Orai1 contributes the PM channel component responsible for Ca(2+) entry. The suppression of SOC function by Orai1 overexpression likely reflects a required stoichiometry between STIM1 and Orai1.     

Endothelium oriented differentiation of bone marrow mesenchymal stem cells under chemical and mechanical stimulations

Bone marrow mesenchymal stem cells (MSCs) have multi-differentiation capability. Their endothelial cell (EC) oriented differentiation is the key to vasculogenesis, in which both mechanical and chemical stimulations play important roles. Most previous studies reported individual effects of VEGF or fluid shear stress (SS), when MSCs were subjected to shear stress of 10–15 dyn/cm² over 24 hr. In this paper, we investigated responses of MSCs from young Sprague Dawley rats to shear stress, VEGF and the combination of the two stimuli. Our study showed that the combined stimulation of shear stress and VEGF resulted in more profound EC oriented differentiation of MSCs in comparison to any individual stimulation. Furthermore, we subjected MSCs to prolonged period of fluid shear stimulation, i.e. 48 hr rather than 24 hr, and increased the magnitude of the shear stress from 10 dyn/cm² to 15, 20 and 25 dyn/cm². We found that without VEGF, the endothelium oriented differentiation of MSCs that was seen following 24 hr of shear stimulation was largely abolished if we extended the shear stimulation to 48 hr. A similar sharp decrease in MSC differentiation was also observed when the magnitude of the shear stress was increased from 10–15 dyn/cm² to 20–25 dyn/cm² in 24 hr shear stimulation studies. However, with combined VEGF and fluid shear stimulation, most of the endothelial differentiation was retained following an extended period, i.e. at 48 hr, of shear stimulation. Our study demonstrates that chemical and mechanical stimulations work together in determining MSC differentiation dynamics.     

Shp2 regulates SRC family kinase activity and Ras/Erk activation by controlling Csk recruitment

The protein-tyrosine phosphatase Shp2 plays an essential role in growth factor and integrin signaling, and Shp2 mutations cause developmental defects and/or malignancy. Previous work has placed Shp2 upstream of Ras. However, the mechanism of Shp2 action and its substrate(s) are poorly defined. Additional Shp2 functions downstream of, or parallel to, Ras/Erk activation also are proposed. Here, we show that Shp2 promotes Src family kinase (SFK) activation by regulating the phosphorylation of the Csk regulator PAG/Cbp, thereby controlling Csk access to SFKs. In Shp2-deficient cells, SFK inhibitory C-terminal tyrosines are hyperphosphorylated, and the tyrosyl phosphorylation of multiple SFK substrates, including Plcgamma1, is decreased. Decreased Plcgamma1 phosphorylation leads to defective Ras activation on endomembranes, and may help account for impaired Erk activation in Shp2-deficient cells. Decreased phosphorylation/activation of other SFK substrates may explain additional consequences of Shp2 deficiency, including altered cell spreading, stress fibers, focal adhesions, and motility.     

Induction of heavy metal binding groups by dexamethasone and Zn in cultured Chinese hamster ovary cells

Cd binding capacity and pulse polarography were used to study the inducibility of sulfhydryl groups in cultured Chinese hamster ovary cells (wild type and a Cd-resistant mutant) in response to dexamethasone (dex) and Zn. Evidence is presented that both the wild type and the mutant responded to dex and Zn treatment by induction of sulfhydryl groups. In wild type for Zn and dex as well as in the mutant for dex, this induction seems to be in the form of sulfhydryls attached to particulate or membrane fractions in the cells. For Zn in the Cd-resistant mutant the induction was in the form of metallothionein.     

牛背梁自然保护区种子植物多样性研究

对牛背梁自然保护区的种子植物多样性进行了研究。据统计,种子植物950种,隶属105科433属,其中,裸子植物4科8属9种,被子植物101科425属941种;木本植物372种,草本植物578种;珍稀濒危保护植物17种,资源植物丰富,分为药用,观赏,食用等10大类型,生态系统多样,按大的类型可划分为森林,灌丛,草甸,裸岩和农田,其中森林面积15055hm^2,占总面积的91.7%。是生态系统的主体。在此基础上,本文还提出了保护植物多样性应采取的对策。     

The Morphology of Fruits and Starches in Bamboos,and Its Relation to Systematic Position

In this article, 30 speceis of bamboos, including 19 genera in 5 tribes, were collected and the morphology of fruits and starches of them was studied. The results are as follows. I. The morphology of fruits is important in studies of systematic position in bamboos. According to the systems of W. Munro and G. Bentham whether the pericarp is adhesive to or free from the seed coat may be taken as a basis of classification. It is also confirmed in this article. It is found in this work that all taxa with a binding pericarp and seed coat are of caryopsis that also has a ventral suture and hilum, while all others with a separated pericarp and seed coat are of bacca or nut, which has no ventral suture and hilum. The former has a hard and thin pericarp and rich endosperm, while the latter has a fleshy and thick pericarp and no endosperm. These characteristics form a basis of classification of major groups. II. In 1907, Brandis found that no any endosperm in matured fruit of Dinochloa, Melocalamus, Melocanna and Ochlandra. It has been proved by Stapf in at least one genus. We found that the baccae of Qiongzhuea, Melocanna, Ferrocalamus and Chimonobambusa Subg. Oerocalama were empty, with no endosperm. This may be a common character of the bacca. We believe, therefore, that the systematic position of Qiongzhuea, Ferrocalamus and Chimonobambusa Subg. Oreocalama is close to Melocanneae. III. Starch grains of bamboo fruits are complex in structure. They are round or ellipsoidal, consisting of 3-22 polyhedral or apple-like small grains. The morphology of starch grains is not so important as fruit in bamboo classification, but some characteristics are of a high value in the identification of genera and species, when they are combined with other features. In Cephalostachyum, the starch grain is very big, with 20-40 μm in diam, and the starch small grain is polyhedral or apple-like with 7.5-22.5 μm in diam, while in Dendrocalamus, the starch grain is small, with 10-28.9 μm in diam. and the starch small grain is only polyhedral, with 3-11.9 μm in diam. The morphology and size of the starch grain and starch small grain are also different in Melocanna and Chimonobambusa Subg. Oreocalama. IV. W. Munro’s system divided Bambuseae into three major groups according to the morphology of flower and fruit. Because the material was not sufficient at that time, the system wrongly put Cephalostachyum, Dendrocalamus into the group Bacciferea. Now it is found that both Cephalostachyum and Dendrocalamus have a nut. Later G. Bentham found this problem and divided the Bambuseae into four subtribes, treating Dendrocalamus as a separate subtribe, Dendrocalamae, and putting the bacca group into another subtribe, Melocannae. It is better, but it also has some shortcomings. Hackel, Gamble, E. G. Camus, A. Camus and Keng Pojie all accepted the view of Bentham, placing Dendrocalamus and Melocanna into different subtribes or tribes.