Effects of ZIP14 on biological behaviors of hepatocellular carcinoma cell line BEL-7404

AIM: To study the expression of zinc transporter ZRT/IRT-like protein 14 (ZIP14) in the hepatocellular carcinoma (HCC) tissues, and to investigate the effects of ZIP14 over-expression on the biological behaviors of HCC cells. METHODS: The expression of ZIP14 at mRNA and protein levels in the HCC tissues and adjacent non-tumor tissues were detected by real-time PCR and immunohistochemical staining, respectively. The lentivirus expression system containing GV365-ZIP14 was constructed, and was used to infect the HCC cell line BEL-7404, which had relatively poor expression of ZIP14. The expression of ZIP14 at mRNA and protein levels in the transfected cells were detected by real-time PCR and Western blot, respectively. Under the conditions of zinc sulfate stimulation at different concentrations, the cell viability, the cell cycle, and the cell migration and invasion abilities were detected by MTT assay, DNA ploid detection, and Transwell assay, respectively. RESULTS: The mRNA expression level and the strong-positive rate of protein expression of ZIP14 in the HCC tissues were significantly lower than those in the adjacent non-tumor liver tissues (P<0.01). The expression of ZIP14 at mRNA and protein levels in the BEL7404 cells was significantly enhanced by infection of GV365-ZIP14 expression lentivirus. Compared with negative control group (transfected with negative control lentivirus), the cell viability, migration and invasion in ZIP14 over-expression group (transfected with GV365-ZIP14 expression lentivirus) were significantly reduced, and the percentage of the cells in G₂/M phase was significantly increased, all of which were more obvious with the elevation of zinc concentration in the culture medium. CONCLUSION: ZIP14 is low expressed in the HCC tissues. The ZIP14 over-expression has inhibitory effects on the viability, migration and invasion of HCC cells, and blocks the cell cycle in G₂/M phase, which might be closely related to the elevation of zinc concentration in cytoplasma of HCC cells due to enchanced zinc transport by ZIP14.     

Neuroprotective effect of novel Rho kinase inhibitor FSD-C10 on a mouse model of Alzheimer disease

AIM: To explore the neuroprotective effect of novel Rho kinase inhibitor FSD-C10 on Alzheimer disease (AD) model of APP/PS1 double transgenic (Tg) mice. METHODS: Male APP/PS1 Tg mice (n=20) at 8 months of age were randomly divided into 2 groups:model group and FSD-C10 treatment group. The mice were treated with normal saline or FSD-C10 (25 mg·kg^-1·d^-1) by intraperitoneal injection, once daily for 2 months. Age-and sex-matched wild-type (WT) mice without treatment were used as the control. The Morris water maze (MWM) test and SMART 3.0 behavioral record system were applied to examine and analyze the spatial cognitive function of the mice. The protein levels and distribution of Aβ, p-tau and synapse-associated proteins such as synaptophysin, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and postsynaptic density protein 95 (PSD-95) were determined by immunofluorescence staining. The protein levels of phosphorylated amyloid precursor protein at Thr668[p-APP(Thr668)], beta-site APP-cleaving enzyme 1 (BACE1) and synapse-associated proteins in the brain were analyzed by Western blot. RESULTS: Compared with model group, FSD-C10 treatment significantly improved the cognitive function of the APP/PS1 Tg mice, accompanied by reduced Aβ deposition and p-tau level, increased protein level of p-APP (Thr668) in the central nervous system, decreased expression of BACE1, and increased expression of synapse-associated proteins in the brain of the mice (P<0.05). CONCLUSION: FSD-C10 has neuroprotective potential in the APP/PS1 Tg mice. The mechanism may be related to enhancing the non-amyloidogenic APP cleavage pathway, reducing the production of Aβ oligomers, the deposition of senile plaques and the amount of tau protein, up-regulating synapse-associated proteins, and increasing synaptic plasticity.     

CD36/Src/ERK pathway is involved in process of monocyte differentiation into macrophages

AIM To investigate the role of fatty acid translocase (FAT/CD36) on differentiation of monocytes to macrophages. METHODS Human monocyte THP-1 cells were treated with phorbol 12-myristate 13-acetate (PMA) at 0, 100 and 200 μg /L. Small interfering RNA (siRNA) targeting CD36 (siCD36) was employed to knock down the expression of CD36 in THP-1 cells. The CD36 over-expression (CD36OE) cell line was constructed by transfection with a recombinant lentivirus containing CD36 cDNA. Optical microscopy and crystal violet staining were used to detect the monocyte morphological changes and adhesion ability. The protein expression of CD36 was measured by flow cytometry and Western blot. The mRNA levels of CD36, CD11b and CD80 were detected by real-time PCR. The protein levels of extracellular signal-regulated kinase (ERK） and Src tyrosine kinase were determined by Western blot. RESULTS The cellular adhesiveness of THP-1 cells was elevated in the process of monocytes differentiation, and the expression of CD36 was increased in this process as well (P<0.01). siCD36 was transfected into the THP-1 cells (CD36i group) and the silencing efficiency was approximately 80%. The cell surface area and cellular adhesiveness were significantly decreased in CD36i group compared with scrambled siRNA (NCi) group (P<0.01). The mRNA levels of CD11b and CD80 were decreased in CD36i group compared with NCi group (P<0.01). The cell surface area and cellular adhesiveness were increased in CD36OE group compared with empty vector (vector) group (P<0.05). The mRNA levels of CD11b and CD80 were increased in CD36OE group compared with vector group (P<0.01). The phosphorylation levels of ERK and Src were decreased in CD36i group compared with NCi group (P<0.05). CONCLUSION CD36 promotes the differentiation of human monocyte THP-1 cells to macrophages by increasing the phosphorylation of Src and further activating ERK.     

Effect of FGFR1 expression knock-down on biological characteristics of infantile hemangioma endothelial cells

AIM: To study the effect of fibroblast growth factor receptor 1 (FGFR1) expression knock-down on the viability, apoptosis, invasion and migration of infantile hemangioma endothelial cells (HemECs). METHODS: FGFR1 was down-regulated by FGFR1 small interfering RNA (si-FGFR1) transfection. The viability of the cells was measured by CCK-8 assay. The apoptotic rate was analyzed by flow cytometry and the invasion and migration abilities were determined by Transwell assay. The protein levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and phosphorylated AKT (p-AKT) were examined by Western blot. RESULTS: Transfection of si-FGFR1 into HemECs had significant effects on inhibiting cell viability (P<0.05), promoting apoptosis (P<0.05), and decreasing cell invasion and migration abilities (P<0.05). The results of Western blot showed that knockdown of FGFR1 gene expression in the cells reduced the protein levels of PI3K and p-AKT (P<0.05), and had no significant effect on AKT protein level. CONCLUSION: Knock-down of FGFR1 expression changes the biological characteristics of endothelial cells in infantile hemangiomas by regulating PI3K/AKT signaling pathway.     

非氧化性修复剂在恢复NF/RO膜出水水质的应用研究

随着膜系统运行至一定年限,加上膜系统长期超负荷运行和膜化学清洗的不及时、不规范,都将加快膜元件的老化,造成膜元件对离子、小分子有机物等物质的截留率有所下降。膜出水COD偏高,电导率升高、颜色偏黄已成为纳滤/反渗透(NF/RO)系统运行过程中出现较频繁的问题之一,严重影响着膜系统的出水水质和出水美观度。本试验选取长沙生活垃圾渗滤液处理厂出水水质差的反渗透/纳滤处理系统为分析对象,通过冲击性投加的方式将专用NR-MRA-1000型修复剂粘附于膜表面,并24 h长期运行。试验结果表明,膜系统添加NR-MRA-1000型修复剂且运行48 h后,呈现出膜出水色度明显降低,COD截留率上升以及出水电导率下降、膜运行压力上涨等产水特征,对膜产水通量没有显著影响;且膜修复剂试验运行60 d后,膜机组相关产水性能恢复至原来的标准。     

Role of ghrelin in cell protection by up-regulating HSP70 through ERK1/2 signaling pathway in PC12 cells

AIM:To study the role of ghrelin in cell protection by up-regulating heat shock protein 70 (HSP70) and inhibiting apoptosis induced by oxidative stress through extracellular regulated protein kinases 1/2 (ERK1/2) signaling pathway in the PC12 cells. METHODS:Sodium nitoprusside (SNP) was used to induce oxidative stress injury in the PC12 cells. The cultured PC12 cells were divided into SNP-injured group (incubated with SNP at 0.5 mmol/L for 6, 12, 18 and 24 h), ghrelin pretreatment group (ghrelin at 100 nmol/L was given 30 min before adding SNP); HSP70 inhibitor group (quercetin at 10 μmol/L was added 60 min before ghrelin treatment), ERK inhibitor group (ERK 1/2 inhibitor PD98059 was added 60 min before ghrelin treatment) and control group (added same amount of culture medium only). The apoptotic rate was detected by flow cytometry. The protein expression was determined by Western blot and immunocytochemistry. RESULTS:Compared with control group, the apoptotic rate of PC12 cells in SNP-injured group was significantly increased (P<0.05). Compared with SNP-injured group, ghrelin (100 nmol/L) pretreatment significantly inhibited SNP-induced apoptosis of PC12 cells (P<0.05), and significantly up-regulated the protein expression of HSP70 (P<0.05). Time-effect analysis showed that ghrelin had the most significant effect at 18 h after SNP injury. Quercetin, an inhibitor of HSP 70, significantly reduced the anti-apoptotic effect of ghrelin (P<0.05). Ghrelin pretreatment promoted the phosphorylation of ERK1/2. ERK1/2 inhibitor PD98059 significantly inhibited the effects of ghrelin on up-regulation of HSP70 expression (P<0.05). CONCLUSION:Ghrelin upregulates the expression of HSP70 and inhibits the apoptosis in the PC12 cells induced by oxidative stress by promoting the phosphorylation of ERK1/2.     

基于B/S的农业气象自动化观测系统国家级平台设计与应用

农业气象自动化观测系统国家级平台是依托中国气象局组织的农业气象自动化观测试点工作而设计开发,平台的搭建采用B/S架构实现多级用户、跨平台登录,采用XML及JSON等数据传输格式实现国家级、省级、台站级用户信息交互,实现农业气象观测XML数据传输状态的监控、台站端设备状态的评估以及数据内容的实时查询。     

SPME/GC-MS法分析不同龙眼品种果实中的香气成分

采用 SPME/GC-MS法对4个龙眼品种果实香气成分进行了分析鉴定。结果表明，4个龙眼品种共检测出44种芳香物质，其中烷类11种，烯类18种，酯类10种，醇类3种，酮类1种，炔类1种，它们构成4种龙眼主要的香气成分。4个龙眼品种在香气组成和含量上有所差异，储良、东良、东丰、石硖中分别含有25种、24种、26种、21 种香气成分。其中，4种龙眼共有的香气成分有7种：分别是十七烷、罗勒烯(顺式)、罗勒烯(反式)、别罗勒烯、1,3,8-对-薄荷三烯、α-石竹烯、(E)-β-金合欢烯，但其相对含量都有所差异；此外，各品种也具有自己独特的香气成分，如储良特有的香气成分有9种：包括十六烷、2-甲基-4-亚甲基-5 -(2,2-二甲基环丙基)-1-戊烯、5-环丙基戊酸乙酯、二十酸乙酯、棕榈酸乙酯、9-十六碳烯酸乙酯、香叶基芳樟醇、芳樟醇、2,7-二甲基-3-辛烯-5-炔；东丰特有的3种：包括1-碘十一烷、反式，反式-法尼基酸甲酯、2-羟基十二烷酸甲酯；石硖特有的4种：包括十一烷、(-)-异丁香烯、1,3,3-三甲基-2-乙基环己烯、喇叭茶醇；东良没有特有的香气成分。     

Detection of cigarettes and other tobacco products by giant African pouched rats (Cricetomys gambianus)

If the illicit tobacco trade were eliminated, governments could gain at least $31.3 billion a year, and more than 164,000 premature deaths a year could be avoided after 2030 (Joossens, Merriman, Ross, and Raw, 2009). Dogs have been used successfully in tobacco control programs, and there is a good chance that rats could also play an important role. In the present experiment, giant African pouched rats were trained to respond to filters that had been stored together with cigarettes (i.e., soaked) and to not respond to filters that had been soaked with noncigarette items. Generalization to untrained types of tobacco was then tested. The sensitivity of 4 rats trained on filters soaked with 1 of 7 types of cigarettes ranged from 86% to 100% (mean, 95%). There was very little evidence of generalization when the rats were tested on tobacco leaves and snuff but good evidence of generalization when the rats were tested on cigarettes that had been soaked with strong-smelling additives. These findings suggest that rats may be a valuable asset in the global effort to control illicit cigarette trade.