To reveal the mechanism of Cu enrichment of alfalfa, in-situ micro-X-ray fluorescence spectrometry (μ-XRF) and fractional extraction were used to explore in-situ Cu distribution information in alfalfa seedlings and different combination forms of Cu in organs.The results showed that alfalfa roots were enriched with Cu up to 12.06 mg/g, which was 8 times of stem and 4.9 times of leaves.The in-situ μ-XRF result showed that the root of alfalfa was the main site of enrichment of Cu, and there was a barrier of Cu at the rhizome junction to alleviate the toxic effect of excess Cu on the shoots.Excess Cu (more than 50 μmol/L) also inhibited the uptake of Zn and Ca in alfalfa, enhanced the uptake of Fe in alfalfa, but had no obvious effect on the uptake of K and Mn.Plant fractional extraction showed that the Cu in root cells was mainly fixed in the form of insoluble residues (41%) and cell wall chelate (20%), while in the stem, Cu existed as four forms including hydrophobic protein binding, cell wall binding state, residual state and water-soluble state, which further reduce the excess transport of Cu to the leaves.In the leaves, excess Cu in the leaf cells was mainly in the vacuole and insoluble residue to achieve tolerance and detoxification to Cu.

Moss has high tolerance and accumulating capacity to heavy metal.In this study, the distribution of heavy metal elements in moss sampled from lead-zinc mine was analyzed by X-ray fluorescence spectrometry.The speciation of lead was analyzed by X-ray absorption near-edge spectroscopy.Research showed that the contents of Pb, Zn, Cd and As in the moss of the mining area were extremely high, and their maximum concentration were 1.06 mg/g , 1.23 mg/g, 30.5 μg/g, 13.2 μg/g, respectively.Besides, the shoots especially the new tissue of the moss were the major sites for accumulation and storage of heavy metals.The micro-distribution characteristics varied among Hypnum plumaeforme and Brachytheciumprocumbens, indicating the difference of different species of moss in absorption pathway, accumulation and tolerance mechanisms for heavy metal.Linear combination fitting results indicated that the main lead speciation in moss was lead phosphate (78%) and lead oxide (22%), which suggested that the precipitation of lead phosphate might be the main mechanism of tolerance for moss.

Objectives To study the effects of pU- vascular endothelial growth factor (VEGF)- short interfering RNA (siRNA) on the formation and apoptosis of malignant melanoma in models of nude mice and its mechanism. Methods The siRNA eukaryotic expression vector for VEGF was constructed, then transfected into A375 (a human malignant melanoma cell line) by electroporation. The nude mice models of malignant melanoma were constructed. The protein expression of VEGF and factor Ⅷ related antigen (FⅧRAg) specific for vascular endothelial cells was detected by immunohistochemical technique and morphological quantitative analysis. Microvessel density (MVD) was counted based on the endothelial cells positively stained with anti-FⅧRAg antibody. The apoptosis of the neoplasms in nude mice was quantitatively determined by TdT mediated dUTP nick end labeling (TUNEL). Results Both the number and the growth speed of the neoplasm formation were lower in the experimental group than those in the controls (both P

Objective:To evaluate the cytotoxicity of ibuprofen and its 6 kinds of impurities.Methods:Different concentrations of ibuprofen and the impurities were used to act on mouse fibroblasts (L929) for 72 h,and the cytotoxicity was observed under a microscope.Results:In ibuprofen raw material,the cytotoxicity of impurity B was the weakest with slight toxicity,the cytotoxicity of impurity N,D,J and V was moderate,and that of impurity E was severe.Conclusion:At the same concentration,the toxicity of impurity E is the strongest,and its content in ibuprofen preparations should be strictly controlled.

Objectives To construct a small interfering RNA (siRNA) targeting vascular endothelial growth factor (VEGF) in eukaryotic expression vector, and to assess effects of this vector on proliferation and apoptosis of malignant melanoma cell line A375. Methods pU-VEGF-siRNA plasmid was transfected into A375 cells by electroporation. VEGF mRNA and protein were detected by reverse transcription polymerse chain reaction (RT-PCR) and ELISA, respectively, in A375 cells after gene transfer. Proliferation of A375 cells was assessed by cell counting. Human umbilical vein endothelial cells (ECV-304) were cultured in medium containing supernatants of treated and untreated A375 cells, and their viability was tested by cell counting. Apoptosis of A375 cells was observed by flow cytometry (FCM). Results VEGF mRNA and protein were significantly decreased in the experimental group, compared to those in the controls (P

Objective: To construct the insulin-like growth factor binding protein 7 (IGFBP7) expression plasmid (pEGFC1-IGFBP7) and to investigate the effect of IGFBP7 on the apoptosis of SK-MEL-28 (human malignant melanoma cell line) cells. Methods: The pEGFC1-IGFBP7 plasmid was constructed; pEGFC1-IGFBP7 and empty plasmids were transfected into SK-MEL-28 cells separately. The transfection efficiency was observed under fluorescence microscope. Apoptosis of SK-MEL-28 cells after transfection was detected by Annexin-FITC/PI staining. Results: The pEGFC1-IGFBP7 plasmid was successfully constructed and was effectively transfected into SK-MEL-28 cells by Effectene reagent, with the transfection rate being 61%. The results of flow cytometry showed that pEGFC1-IGFBP7 significantly induced apoptosis of SK-MEL-28 cells, with the apoptotic rates of pEGFC1-IGFBP7, empty vector, and non-transfected plasmid groups being (28.4±2.57)%, (5.8±0.44)%, and (6.4±0.71)% 24 h after transfection, respectively (F=406.138, P<0.05). Conclusion: pEGFC1-IGFBP7 can effectively induce apoptosis of malignant melanoma SK-MEL-28 cells, which provides an experimental basis for IGFBP7 gene-based therapy of malignant melanoma.

Objective To study the effect of Jade -Screen Powder (JSP)on regulating expression of 5 microRNAs associated with helper T cells in asthmatic mouse model.Methods Forty Balb /c mice were randomly di-vided into 4 groups,1 0 mice for each group,namely normal control,asthma model,JSP treatment and Dexamethasone treatment.The mouse models of allergic inflammation on both upper and lower airways were established by ovalbumin sensitization and challenge.Interleukin(IL)-1 3 and IL -1 7 expressions were detected from lung homogenates by ELISA.Hematoxylin and eosin staining was also performed to observe the pathological changes in the lung tissue.The expressions of miR -1 46a,miR -1 46b,miR -21 0,miR -1 26 and miR -21 a were detected by quantitative real time PCR from splenocytes.Results The lower levels of IL -1 3 [(6.382 ±1 .690)μg/L]and IL -1 7 [(24.21 2 ± 1 .250)μg/L]were found in JSP treatment group compared with those in the asthma model group [(20.1 54 ±7.960)μg/L;(50.31 2 ±5.770)μg/L,rseparately],there was significant difference in IL -1 3 between JSP group and the asthma model group,as well as IL -1 7 (t =3.785,P =0.005;t =9.891 ,P =0.000).Same findings were found in Dexamethasone treated group as well [IL -1 3:(9.366 ±3.460)μg/L,IL -1 7:(29.1 32 ±4.960)μg/L;t =2.779, P =0.024;t =6.225,P =0.000].However,upregulation of miR -21 0 was observed in JSP treatment group (2.052 ± 0.871 )compared with that in the asthma model group (4.034 ±1 .379)(3.95 folds,t =2.71 8,P =0.026).Mean-time,the expression of miR -1 26 in JSP group (4.920 ±0.924)and Dexamethasone group (3.862 ±1 .51 0)in-creased compared with asthma model group (6.024 ±0.447)(2.1 5 folds,t =2.405,P =0.043,and 4.48 folds,t =-3.069,P =0.01 5).Conclusions Th2 and Th1 7 T cells participate in the pathogenesis of asthma and the asthmatic process can be inhibited by JSP.JSP may affect the helper T cells by regulating miR -21 0 and miR -1 26.

Fenofibrate is a common lipid-lowing drug activating peroxisome proliferator-activated receptor alpha (PPRAα).Recent studies have found that fenofibrate possesses anticancer properties.Its specific mechanisms include inhibiting cell metabolism and formation of new tumor vessels,arresting cell circle,weakening cell motility,and inducing cell apoptosis.Those properties are partly independent of PPRAα.Due to its low side-effects,it's hopefully to be used as an anticancer adjuvant drug.

In order to investigate the expression of endothelin receptor B (ETR-B) in human malignant melanoma (MM) cells A375 and SK-mel-1 and the proliferative effects of endothelin 3 (ET3) on A375 cells, RT-PCR was applied to detect the expression of ETR-B gene in human MM cells A375 and SK-mel-1. MTT method was used to evaluate the growth enhancing effects of ET3 on A375 cell line in vitro. The results showed that ETR-B gene was expressed in both MM A375 and SK-mel-1 cells. ET3 had stronger ability to enhance the proliferation of A375 cells in vitro in a concentration-dependent manner. It was suggested that ET3/ETR-B might play an important proliferative role in MM.
Cell Line, Tumor ; Cell Proliferation/*drug effects ; Endothelin-3/*pharmacology ; Melanoma/*metabolism ; Melanoma/*pathology ; Receptor, Endothelin B/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Authors of the paper analyzed the motivation of building the healthy town,and interrelated the scientific aspects of such a town. It was found that this town of diversified resources and advantages had become small but beautiful,special and strong clustering and convergent by means of collecting high-end factors, selecting of essential health businesses and constructing an ecosystem of health industry clusters. Based on a definition of the government role,the paper described the innovation of health policy in view of policy supply.