HCMV-infected human THP-1 cells induce expression of HLA-G and its receptors

AIM: To investigate the differential expression of human leukocyte antigen-G (HLA-G) isoforms and its receptors in human monocyte line THP-1 after human cytomegalovirus (HCMV) infection for exploring the role of HLA-G in HCMV escaping the immune response of the organism.METHODS: THP-1 cells were infected with HCMV Towne strain. The expression of HLA-G isoforms at mRNA and protein levels was determined by RT-PCR and Western blot, respectively. The surface expression of HLA-G and its receptors ILT2/ILT4 and the cell viability were analyzed by flow cytometry. The levels of soluble HLA-G (sHLA-G) and IL-10 were measured by ELISA.RESULTS: After infection of the THP-1 cells with HCMV, no obvious apoptosis in the cells was observed, and the viability of the cells was high. A significant up-regulation of HLA-G1, -G3, -G4 and -G5 at mRNA expression level 1 d after infection was found, while the protein expression of HLA-G1 and HLA-G5 isoforms was mainly detected. The expression of HLA-G/ILT2/ILT4 was evidently up-regulated 1 d after infection. The level of sHLA-G was significantly increased 1 d after infection as compared with control group (P<0.01). The expression of IL-10 was obviously up-regulated 1 d post-infection as compared with control group.CONCLUSION: The differential expression of HLA-G isoforms and secretion of the receptors ILT2/ILT4 and IL-10 in the THP-1 cells are induced after HCMV infection. This study provides experimental evidence for evaluating the immune mechanism of HCMV infection.     

Eeffect of HMGB1 on expression of NF-κB in BV-2 cells stimulated with Aβ25-35

AIM: To investigate the effect of high mobility group box-1 protein (HMGB1) on the expression of nuclear factor-κB (NF-κB) in BV-2 cells stimulated with amyloid β-protein (Aβ)_25-35. METHODS: Cultured BV-2 cells in logarithmic growth phase were divided into 4 groups:normal cell group (without any treatment), model group (treated with Aβ_25-35 at 40 μmol/L), RNA interference (RNAi) group (conducted with HMGB1-siRNA followed by Aβ_25-35 stimulation) and solvent control group (treated with 0.1% DMSO). After treatment with Aβ_25-35 for 24 h, the protein levels of HMGB1 and NF-κB in BV-2 cells were determined by Western blot. RESULTS: Aβ_25-35 at 40 μmol/L was used to stimulate BV-2 cells. The GFP fluorescence-tagged HMGB1-siRNA (30 nmol/L) was used to transfect BV-2 cells and its transfection efficiency was about 80%~90%. The results of Western blot showed that the protein level of HMGB1 was significantly decreased after the interference of siRNA fragment (P<0.05). The protein levels of HMGB1 and nucleic NF-κB p65 were dramatically increased in BV-2 cells stimulated with Aβ_25-35 (P<0.05). After RNA interference with HMGB1, the expression of HMGB1 and nucleic NF-κB p65 were significantly decreased in BV-2 cells stimulated with Aβ_25-35 (P<0.05). CONCLUSION: RNA interference with HMGB1 reduces the expression of nucleic NF-κB in BV-2 cells stimulated with Aβ_25-35.     

县/市尺度农用地整治新增耕地潜力评价——以浙江省温州市为例

[目的]探索提高县(市)尺度农用地整治新增耕地潜力评价效率、精度,降低评价成本的方法,为土地利用总体规划和土地整治专项规划提供参考。[方法]以GIS全域空间分析方法为主的农用地整治新增耕地潜力评价技术方法体系,即基于区域"二调"更新、土地利用总体规划、农用地整治经验等国土数据成果,在现阶段县/市尺度农用地整治新增耕地潜力评价技术思想及技术原则下,先确定待整治农用地规模,再测算新增耕地面积,最后计算新增耕地系数,进而进行新增耕地潜力类型区划分。[结果]温州市农用地整治新增耕地潜力测算结果合理,且具有较强的实用性。[结论]该技术方法体系实现了从基础图件数据输入整理、到潜力测算评价、再到成果图表输出的程序化操作,不仅有效提高县/市尺度农用地整治新增耕地潜力评价效率,降低评价成本,而且紧密衔接于当前土地管理工作实际,具有较高的推广应用价值。     

Cyanidin attenuates pressure overload-induced cardiac remodeling in mice via mitochondrial fusion enhancement

AIM To investigate the effect of cyanidin （Cyn） on pressure overload-induced cardiac remodeling and the underlying mechanism. METHODS Six-week-old male C57BL/6 mice （n=120） were divided into 4 groups： sham group （n=20）, sham+Cyn group （n=20）, transverse aortic constriction （TAC） group （n=40） and TAC+Cyn group （n=40）. The model of cardiac chronic pressure overload was induced by TAC, and the first day of TAC was defined as day 0. The animals in sham+Cyn group and TAC+Cyn group were treated with Cyn dissolved in DMSO and normal saline （5 mg·kg^-1·d^-1） for 5 d before TAC, while the animals in sham group and TAC group were treated with the same amount of DMSO and normal saline. Four weeks after TAC, the survival rate of the animals in each group was analyzed, the heart function of the mice was measured by ultrasound echocardiography, and the heart weight/body weight and lung weight/body weight were calculated. The cross-sectional area of the cardiomyocytes was measured by wheat germ agglutinin staining and hematoxylin-eosin staining. The degree of cardiac oxidative stress was evaluated by dihydroethidium staining and measurement of superoxide dismutase （SOD） and malondialdehyde （MDA） levels. The cardiomyocyte apoptosis was detected by TUNEL method. The mRNA expression levels of atrial natriuretic peptide （ANP）, brain natriuretic peptide （BNP） and β-myosin heavy chain （β-MHC） were detected by RT-qPCR, and the protein expression levels of Bax, Bcl-2, optic atrophy protein 1 （OPA1） and dynamin-related protein 1 （Drp1） were determined by Western blot. The mitochondrial morphological changes were observed by transmission electron microscopy. RESULTS Compared with TAC group, the survival rate of the mice in TAC+Cyn group was significantly increased （P<0.05）, the myocardial apoptosis, the cross-sectional area of myocardial cells, the heart weight/body weight, the lung weight/body weight, the level of reactive oxygen species and the MDA content were decreased （P<0.05）, and the SOD was activated （P<0.05）. M-mode ultrasound tests showed that Cyn treatment significantly increased left ventricular ejection fraction and left ventricular fractional shortening in the mice after TAC （P<0.05）, while left ventricular end-diastolic diameter and left ventricular posterior wall thickness in diastole were reduced （P<0.05）. Transmission electron microscopic observation showed that the number of myocardial mitochondria was increased and the mitochondrial area was decreased after TAC （P<0.05）, while treatment with Cyn increased the area of myocardial mitochondria and decreased the mitochondrial number （P<0.05）. Compared with sham group, the protein level of OPA1 in TAC group was significantly reduced （P<0.05）, while treatment with Cyn significantly increased the protein level of OPA1. CONCLUSION Cyanidin significantly increases the survival rate, improves the cardiac function and attenuates the cardiac remodeling of the mice after TAC. The mechanism may be related to the inhibition of myocardial mitochondrial OPA1 cleavage and the promotion of mitochondrial fusion.     

Effect of intermittent hypoxia on bladder detrusor cell apoptosis and regulatory mechanism of Alpiniae oxyphyllae Fructus

AIM To investigate the effect of intermittent hypoxia （IH） on bladder detrusor cells apoptosis and calcium channel, and to discuss the regulatory mechanism of Alpiniae oxyphyllae Fructus （AOF）. METHODS IH model of bladder detrusor cells was established by treating the cells with 6 cycles of 5% O₂ for 60 min and 20% O₂ for 30 min. Human bladder detrusor cells were cultured in vitro, randomly divided into 6 groups, each group had 8 holes. P₂X₃ receptor antagonist + IH （A） group, M₃ receptor antagonist + IH （B） group, β₃ receptor antagonist + IH （C） group, AOF + IH （D） group, saline + IH control （NC） group and air simulation control （AC） group were set up. The cells density and morphology were identified by the methods of counting chamber and immunofluorescence light microscopy （LM） after interventions. The apoptosis was analyzed by flow cytometry. Calcium channel expression was detected by patch clamp. RESULTS （1） Compared with the cells in AC group, the cells density and activity were significantly increased in NC group （P<0.05）; some cells appeared protrusions, turned round and blur in cell borders. （2） The results of immunofluorescence for detecting α-SMA protein expression showed that, compared with the cells in group AC, the mean absorbance （MA） in group NC was significantly increased （F=3.25, P<0.05）; compared with the cells in group NC, that in group A and group D was both decreased significantly （P<0.05）. （3） Compared with the cells in group AC, the apoptotic rate was significantly decreased in group NC （P<0.05）; Compared with the cells in group NC, the apoptotic rates in group A and group D were both significantly increased （P>0.05）. （4） Compared with the cells in group AC, calcium ion channel expression was significantly decreased （P<0.05）. Compared with the cells in group NC, calcium ion channel expression in AOF （100 mg/L） and AOF （50 mg/L） group was significantly increased （P<0.05）. CONCLUSION IH regulates bladder detrusor cells proliferation and apoptosis through P₂X₃ bladder nerve receptors, high or moderate dose of AOF may change calcium channel and play a protective role in IH induced cell damage.     

Effect of curcuma aromatica oil on learning,memory and hippocampal p-CaMKII expression in rats exposed to chronic hypoxia

AIM: To investigate the effects of different concentrations of curcuma aromatica oil on learning and memory in rats exposed to chronic hypoxia. METHODS: The rats were divided randomly into the control, chronic hypoxia and chronic hypoxia with low (LC), middle (MC) and high (HC) concentrations of curcuma aromatica oil groups. After 29 d, all animals were examined to obtain the scores of leaning and memory. The SOD activity and MDA content were determined in the serum and hippocampus, the ［Ca2+］_i in hippocampus was also detected. The staining and expression of p-calcium/calmodulin-dependent protein kinase II (p-CaMKII) in the hippocampus was observed and measured. RESULTS: ① In the chronic hypoxia group, the latency to find the hidden platform remarkably prolonged and the MDA content was obviously higher, but the SOD activity was significantly lower. Meanwhile, hippocampal ［Ca2+］_i was markedly increased. The immunostaining of p-CaMKII was much weaker in hippocampus as well as its expressions (P<0.01). ② The latency to find the hidden platform was remarkably shorter in groups with MC and HC (P<0.05). The MDA content was obviously lower among groups treated with curcuma aromatica, but SOD activity was significantly higher in groups with MC and HC. Meanwhile, hippocampal ［Ca2+］_i was markedly decreased in all groups treated with curcuma aromatica oil (P<0.01). The hippocampal immunostaining of p-CaMKII was much stronger in the MC and HC as well as its expression (P<0.05，P<0.01). Under the electron microscope, synaptic boundaries were not distinct, the edema of dendrite spine and axon was seen, synaptic vesicles and postsynaptic densities (PSD) were disappeared in the chronic hypoxia group. With rising of the concentration of curcuma aromatica oil, the edema of synapse and mitochondria was mitigated and the PSD was increased gradually. CONCLUSION: Curcuma aromatica oil might enhance learning and memory capacities of rats exposed to chronic hypoxia by cleaning up and antagonizing the production of the free radical and increasing the p-CaMKII expression in PSD. The effects are dose-dependent.     

Protective effect of Ligustrazini on gut barrier function

AIM:To study the protective effect of Ligustrazini(LGT) on gut barrier function after hemorrhagic shock-reperfusion. METHODS: Thirty white rabbits were divided randomly into 3 groups: control group (A),shock group (B) and LGT group (C). Malondialdehyde(MDA), tumor necrosis factor-α(TNF_α), interleukin-1β(IL-1β) and nitric oxide products(NO₂^-/NO₃^-) contents were measured in intestinal mucosa at 3 hours following reperfusion,culture of bacteria in blood from rabbits of 3 groups was carried out,the intestinal mucosa was examined under optical and electron microscope. RESULTS: MDA, TNF_α, IL-1β and NO₂^-/NO₃^- contents of intestinal mucosa remained unchanged in group C,but increased significantly in group B, compared with group A. Incidence of bacterial translocation in group B was markedly higher than that in group A at 30 min following reperfusion,there was not any difference between group A and group C. Under light and electronic microscope,in comparison with A and C groups,intestinal mucosa damage in B group became more severe. CONCLUSION: LGT can protect gut barrier from intestinal ischemia-reperfusion injury induced by hemorrhagic shock through reducing oxygen free radicals,raising nitric oxide and preventing inflammation.     

基于BP神经网络的杨梅大棚内气温预测模型研究

利用2009年12月-2010年5月塑料大棚内外观测的气象数据,构建了基于BP神经网络的杨梅生产大棚内的最高、最低气温预测模型,根据逐时转化系数计算出棚内相应的逐时气温,达到逐时预报大棚内气温的目的。通过模拟回代和对独立试验数据的验证,基于BP神经网络模型对大棚内日最低气温、日最高气温和逐时气温预测值与实际值的回归估计标准误差（RMSE）分别为0.8℃、1.4℃和0.7℃,精度明显高于同时利用逐步回归法建立的模型。该模型所需参数少,实用性强,模拟精度高,可为设施杨梅气象服务和环境调控提供依据。     

温度对真鲷Pagrus majoor(Temmincket Schlegel)蛋白酶(Protease)和淀粉酶(Amylase Activity)活力的影响

本文研究了不同温度条件下真鲷(Pagrus major)胃、肠、肝胰脏和幽门盲囊4个部位蛋白酶和淀粉酶活力的变化。结果表明:在5~50±0.5℃范围内(每隔5℃为一个梯度),胃蛋白酶的最适温度为25℃,胃淀粉酶的最适温度是30℃;肠蛋白酶的最适温度是35℃,而肠淀粉酶的最适温度是20℃;肝胰脏蛋白酶的最适温度是40℃,肝胰脏淀粉酶的最适温度是30℃;幽门盲囊蛋白酶的最适温度是30℃,幽门盲囊淀粉酶的最适温度在40℃。