Overexpression of SmZIP plays important roles in Cd accumulation and translocation, subcellular distribution, and chemical forms in transgenic tobacco under Cd stress.

Plant ZIP genes represent an important transporter family and may be involved in cadmium (Cd) accumulation and Cd resistance. In order to explore the function of SmZIP isolated from Salix matsudana, the roles of SmZIP in Cd tolerance, uptake, translocation, and distribution were determined in the present investigation. The transgenic SmZIP tobacco was found to respond to external Cd stress differently from WT tobacco by exhibiting a higher growth rate and more vigorous phenotype. The overexpression of SmZIP in tobacco resulted in the reduction of Cd stress-induced phytotoxic effects. Compared to WT tobacco, the Cd content of the root, stem, and leaf in the transgenic tobacco increased, and the zinc, iron, copper, and manganese contents also increased. The assimilation factor, translocation factor and bioconcentration factor of Cd were improved. The scanning electron microscopy and energy dispersive X-ray analysis results of the root maturation zone exposed to Cd for 24 h showed that Cd was transferred through the root epidermis, cortex, and vascular cylinder and migrated to the aboveground parts via the vascular cylinder, resulting in the transgenic tobacco accumulating more Cd than the WT plants. Based on the transverse section of the leaf main vein and leaf blade, Cd was transported through the vascular tissues to the leaves and accumulated more greatly in the leaf epidermis, but less in the leaf mesophyll cells, following the overexpression of SmZIP to reduce the photosynthetic toxicity. The overexpression of SmZIP resulted in the redistribution of Cd at the subcellular level, a decrease in the percentage of Cd in the cell wall, and an increase of the Cd in the soluble fraction in both the roots and leaves. It also changed the percentage composition of different Cd chemical forms by elevating the proportion of Cd extracted using 2% HAc and 0.6 mol/L HCl, but lowering that of the Cd extracted using 1 mol/L NaCl in both the leaves and roots under 10 and 100 µmol/L Cd stress for 28 d. The results implied that SmZIP played important roles in advancing Cd uptake, accumulation, and translocation, as well as in enhancing Cd resistance by altering the Cd subcellular distribution and chemical forms in the transgenic tobacco. The study will be useful for future phytoremediation applications to clean up Cd-contaminated soil.

[Protocol of Miao medical LIU's infant tuina genre "Tui Wu Jing" in western Hunan province for prevention of asthma recurrence].

The Miao medical LIU's (LIU Kaiyun) infant tuina genre in western Hunan Province is one of the most famous infant tuina genres in China. Based on physiological and pathological characteristics of infants, generation-inhibition theory of five-elements and Miao medical's promotion-inhibition theory of five-meridians, the tuina protocol of "Tui Wu Jing" was flexibly adjusted; according to different constitution types, including lung-deficiency type, spleen-deficiency type, kidney-deficiency type, qi-deficiency type, yin-deficiency type, yang-deficiency type, phlegm-wet type, phlegm-heat type, different protocols were adopted to prevent or reduce the asthma recurrence and reach the aim of regulating constitution and disease prevention.

Neuroprotective effect of quercetin on beta-amyloid-induced PC12 cells injury and potential related mechanism / 中国药理学与毒理学杂志

OBJECTIVE To explore the estrogen- like neuroprotective effects and the related mechanism of quercetin by using PC12 cells induced with Aβ25-35, provided thought and strategy for the drug therapy of AD. METHODS Cells were cultured with Aβ25- 35 for 24 h, 17β-estradiol (0.1 μmol·L- 1), genistein(50 μmol·L-1) and three different concentrations of quercetin (200 μmol·L-1, 300 μmol·L-1 and 400 μmol·L-1) were respectively added after 24h. The effects of quercetin on activity of AD model were tested by MTT. Immunohistochemical stain and Western blot were used to detect the expression of estrogen receptors alpha and beta, p-ERK1/2 and apoptosis related proteins.The mechanism of quercetin estrogen-like neuroprotective effects was detected using estrogen receptor antagonist ICI182,780 and MAPKK inhibitor U0126. RESULTS The results revealed thatthe toxic effects showed in a dose-dependent increase of Aβ25- 35 on PC12 cells.Comparing with the control group,cells injury was observed after cultured with 10 μmol·L-1 Aβ25-35 for 24 h(P<0.01). The MTT results showed that 17β-estradiol, genistein and three different concentrations of quercetin could significantly enhance the cell survival rate compared with the model group (P<0.05). Compared with model group,Immunofluorescence and Western blot results show that quercetin could improve the estrogen receptor alpha and p- ERK1/2 protein expression (P<0.05), and the expression of estrogen receptor beta protein is increased without significant difference. And in the Western blot experiments, the ratio of Bcl- 2 and Bax was increased and the expression of Caspase 3 was decreased( P<0.05).When estrogen receptor inhibition ICI182,780 were reduced,the expression of p- ERK1/2 was decreased (P<0.05) and the ratio of Bcl- 2 and Bax was decreased, Caspase 3 protein expression was increased (P<0.05). In addition,pretreatment of cells with U0126 would reduce Bcl-2/Bax ratio and increase Caspase 3 protein expression increased (P<0.05). CONCLUSION Quercetin protected PC12 cells, which suffered from Aβ25- 35-induced cytotoxicity and exert neuroprotective effects. The estrogen-like neuroprotective effect can reduce the apoptosis in the classic estrogen receptor pathway and MAPK pathway. And quercetin can also active MAPK signaling pathways by the mediation of estrogen receptor alpha.

Protective estrogen-like properties and mechanism of quercetin in rat cerebral cortex neurons / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of quercetin on primary cultured newborn rat cortex neuron cell which is estrogen depletion, and discuss the possible mechanism, to provide new ideas and strategies for developing a drug of neurodegenerative disease. METHODS Rat cortex neurons were isolated from one day old Sprague Dawley rats and treated with estrogen, quercetin and estrogen receptor antagonists (ICI182,780). Cell viability was determined by MTT assay, neurite outgrowth was measured by fluorescent microsope and estrogen receptors were determine by Western blot. RESULTS Quercetin functions like estrogen to increase cortex neuronal cell viability, the Que (50, 100 μmol·L-1) group compared with the control group could significantly improve the activity of the cortical neurons(P<0.05). It can also increase neurite out growth, the Que (50,100 μmol·L-1) group significantly promoted the formation of synapse, most of the neurons were full, and the synapses of neurons became thick, growth, and connect to a dense neural network. And in the Western blot experiments, Que (50, 100 μmol·L-1) group could obviously increase the expression of estrogen receptor alpha protein, in addition, the neural protective effect of quercetin can be inhibited by ICI182,780. CONCLUSION Quercetin like estrogen can protected cortex neuronal and the effect of quercetin on cortex neuronal cells was mediated by estrogen receptor alpha.

Two new bisindole alkaloids from the seeds of Strychnos nux-vomica.

The reinvestigation of Strychnos nux-vomica resulted in the isolation of two colorless monoquaternary bisindole alkaloids from the seeds. The structures of the two new compounds named 4-N-hydroxymethyl strychnidin-17-acetic acid (1) and 10, 11-dimethoxy-4-N-hydroxymethyl strychnidin-17-acetic acid (2), were defined by detailed spectral analyses, especially (1)H NMR, (13)C NMR, DEPT, HSQC, (1)H NMR-(1)H NMR, NOESY, HMBC and TOF-MS.