Objective To construct the endothelial cell model with overexpressed human protein kinase C ?2(PKC?2) after high glucose inducement in order to study the function of human PKC?2. Methods The PRKCB1 gene was amplified from pMD18-T-PRKCB1 plasmid and then directly cloned into shuttle plasmid pDC315 to construct shuttle plasmid. Then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHGlox△E1,3Cre were cotransfected into 293 cells to construct recombinant adenovirus Ad-PRKCB1. The virus titer was calculated by TCID50. The Ad-PRKCB1 was verified by immunocytochemistry and RT-PCR. Ad-PRKCB1 was transfected into human umbilical vein endothelial cells (HUVECs) followed by the treatment of high glucose (25 mmol/L) for 96 h, and then the cells were observed by laser scanning confocal microscopy (LSCM). Results Restrictive endonuclease digestion and PCR result showed that the target gene was correctly cloned into shuttle plasmid. Cytopathic effect (CPE) was observed under inverted microscope through homologous recombination. The virus titer was 7.9?109 IU/ml. Immunocytochemical staining and RT-PCR indicated the expression of human PKC?2 in the transfected HUVECs. The translocation was observed under LSCM. Conclusion A recombinant adenovirus vector with human PKC?2 and the endothelial cell model with human PKC?2 overexpression by high glucose are constructed successfully.

0.05).Again,compared with NG group,the protein expression of PKC? in HUVECs was up-regulated,the cytosol/nuclei ratio of PKC? was decreased,cell cycle was blocked in G0/G1 phase,the apoptosis increased significantly,and the protein content of p-FOXO1(S256) and P27kip1 increased(P

Objective To investigate the roles of protein kinase C(PKC)β_2 and PPARα in the mRNA expression of vascular endothelial growth factor(VEGF)and vascular cell adhesion molecule-1(VCAM-1)in human umbilical vein endothelial cells(HUVECs)exposed to high glucose,and to explore their relationship.Methods The HUVECs were divided into eight groups:normal glucose(NG,5 mmoL/L D-glucose)group,high glucose(HG,25 mmol/L D-ucose)group,osmotic control(L,NG+20 mmol/L L-glucose)group,normal glucose transfected with empty vector(NN,NG+AdS-null)group,high glucose transfected with PKCβ_2(HB,HG+Ad5-PKCβ_2)group,high glucose plus fenofibrate(HF,HG+40 μmol/L fenofibrate)group,and HB plus fenofibrate (HBF,HB+40μmol/L fenofibrate)group.HUVECs were incubated with fenofibrate for 20 minutes as HF20 group.All cells in various groups were cultured for 6 days.The expressions of VEGF and VCAM-1 mRNA were detected by RT-PCR.PPARα protein expression was tested by Western blot.The expression and traaslocation of PKCβ_2 protein were observed by confocal laser scanning microscopo.Results (1)HG increased VEGF and VCAM-1 mRNA expressions,with 1.91 and 1.56 folds of NG group,respectively(both P<0.05).VEGF and VCAM-1 mRNA expressions in HB group further increased,with 2.59 and 2.07 folds of NG group,respectively(both P<0.05).Fenofibrate significantly decreased VEGF and VCAM-1 mRNA expressions,with 68%and 74%of HG group,respectively(both P<0.05).There were no significant difierences in the expressiong of VEGF,VCAM-1 mRNA between HF20 and HG groups.(2)The protein expression of PPARα decreased by 20%in HG group compared with NG group,and further decreased in HB group,being 78%of HG group.Compared with HG group,PPARαexpression increased by 13%in HF group(P<0.05).(3)HG induced PKCβ_2 translocation from cytosol to nucleus and quantitative analysis showed the ratio of plasma/nuclear fluorescence intensity in HG group decreased by 37% compared with NG group(P<0.05).The PKCβ_2 translocation was more obvious in HB group than in HG group.Conclusion Hiigll glucose stimulates VEGF and VCAM-1 mRNA expressions in HUVECs via PKCβ_2 activationdependent PPARα pathway.

AIM:To explore the effect of CXCL16 deficiency on streptozocin ( STZ)-induced diabetic nephrop-athy in mice.METHODS:CXCL16 knockout ( C16 KO) mice (8 years old) were used to build up diabetes model by treating with STZ.Age-and gender-matched wild-type ( WT) C57BL/6J mice treated with STZ were used as control.All mice were fed with chow diets for 12 weeks, and the development of diabetic nephropathy was evaluated.RESULTS:Compared with the WT mice treated with STZ, C16 KO mice treated with STZ presented lower fasting glucose levels and better glucose tolerance power.C16 KO mice treated with STZ also had lower urine protein levels and smaller areas of glo-merular injury as compared with WT mice treated with STZ.Furthermore, CXCL16 deficiency decreased the contents of re-nal reactive oxygen species ( ROS) , malondialdehyde ( MDA) and oxidized low-density lipoprotein ( ox-LDL) and the mR-NA expression of lectin-like oxidized low-density lipoprotein receptor 1 (Lox-1), and attenuated the expression of renal in-flammatory factors including tumor necrosis factor α( TNF-α) and interleukin 6 ( IL-6) , as well as chemokines including intercellular cell adhesion molecular 1 (ICAM-1) and chemokine C-X-C motif ligand 1 (CXCL1).CONCLUSION:CX-CL16 deficiency obviously inhibits the development of STZ-induced diabetic nephropathy in mice.

Objective To study the compatibility of TCM prescriptions of TCM practitioners of all dynasties of Alzheimer disease (AD). Methods Amnesia, forgetting, dementia, and idiot were set as search words to retrieve relevant literature in Encyclopadia of Traditional Chinese Medicine. Prescription information was screened and standardized to build database. Frequency analysis and association rules were used to mine TCM prescriptions and compatibility rules. Results Totally 449 AD related prescriptions were selected, involving 682 Chinese medicinal herbs. The individual herb with the highest frequency was Ginseng Radix Rhizoma (192);the herbal pair with the highest frequency was Ginseng Radix Rhizoma-Polyhalae Radix (182);the herbal combination with 3 Chinese medicinal herbs with the highest frequency was Poria with Hostwood-Ginseng Radix Rhizoma-Polyhalae Radix (79);the herbal combination with 4 Chinese medicinal herbs with the highest frequency was Polyhalae Radix-Ginseng Radix Rhizoma-Poria with Hostwood-Glycyrrhizae Radix et Rhizoma (37). The results of association rules showed that Ginseng Radix Rhizoma-Polyhalae Radix, Ginseng Radix Rhizoma-Glycyrrhizae Radix et Rhizoma, and Ginseng Radix Rhizoma-Poria with Hostwood were commonly used compatibilities in AD related prescriptions. Conclusion Treatment of TCM practitioners in all dynasties for AD mainly chooses Chinese medicinal herbs with the efficacy of tonifying qi and soothing nerves. The compatibilities and combinations are reasonable and with certain representativeness.

Objective To investigate whether timing of image acquisition influenced infarct size estimation using delayed CeMRI,and the association of left ventricular ejection fraction between magnetic resol3anee imaging and left ventrieulography Was also studied.Method From Junary 2005 to April 2006,27 first,onset AMI patients [23 male,mean age(54.3±10.5)years]were enrolledinthistudr.Allpatients receivedleft ventrictdographyas well as coronary angiography.The average checking time was(13.2±5.2)clays after the onset of AMI.MR imaging was performed with a 1.5-T magnet(SIMENS).After breath-hold eine images were acquired,patients re.ceived afI intravenous bolus of 0.05 mmol/kg Gd-DTPA at a rate of 5 ml/8.A first-pass perfusion scan was ac.qllired.Then a second bolus of 0.15 mmoVkg Gd-DTPA was give.at a rate of 2 mE/Is.After the hyperenhancement localized,the typical short axis slice with hyperenhancement WaS chosen to repeat imaging for IlleasuriIin.farct size every5minutesfrom5minutes after secondinjection ofcontrast until 20minutes.Results Twexty-seren patients showed hyperenhancement at the delayed CeMRI and hypoenhancement at the first pass enhancement(FPE).The average infarct size estimated by CeMRI WaS(17.9士9.8)%of LV nlass.Myocardial enhancement at a repesentative short-axis slice WIllS(7.2±6.2)%of LV Imss at 5 minutes,(8.5±7.4)%at 10 minutes,(7.3±6.3)%at 15 minutes and(6.9-t-6.4)%at 20 minutes respectively.There WltlS significant difference be-tween lmfninmes and 20-minutes enhancement size(P<0.05).Correlations of EF obtained by cineventriculo-grapIIy and MR irr,lg were significant(r=0.867,P<0.01).There were also correlations between infarction size and pe.k CK(r:O.819,P

ObjectiveTo investigate the noise level and influencing factors in metro platforms and station halls， thereby providing the scientific basis for the establishment of hygienic standards. MethodsDuring the morning peak（7：00‒9：30）and off-peak （9：30‒17：00） on weekdays， the noise levels were measured with noise meters at 39 monitoring points of 13 station platforms and 31 monitoring points of 6 station halls. The monitoring points arrangement and detection methods referred to the Examination methods for public places—Part 1： physical parameters（GB/T 18204.1‒2013）. ResultsThe measured noise level in the station ranged from 69.25 to 86.17 dB（A）， accounting for 44.74% below 75 dB（A）， 89.47% below 80 dB（A） and 97.37% below 85 dB（A）.The noise level of the platform ［（76.38±4.19） dB（A）］ was higher than that of the station hall ［（74.24±4.50） dB（A）］（P<0.01）. The noise level of the elevated platforms ［（80.01±2.25） dB（A）］ was higher than that of the underground platforms ［（75.73±4.13） dB（A）］（P<0.01）， and the noise level of the platforms without platform screen doors（PSD） ［（80.21±5.08） dB（A）］ was higher than that of platforms with PSD［（74.73±3.16） dB（A）］ （P<0.01）. No statistical significant differences were observed among the different areas of the platforms， monitoring periods， platform depth， exit mode and operation years （P>0.05）. ConclusionThe noise level in metro stations in the city does not fully meet the requirements of current relevant standards. It is suggested to take noise reduction measures to reduce the noise of metro stations.

Bacterial antitumor therapy has great application potential given its unique characteristics, including genetic manipulation, tumor targeting specificity and immune system modulation. However, the nonnegligible side effects and limited efficacy of clinical treatment limit their biomedical applications. Engineered bacteria for therapeutic applications ideally need to avoid their accumulation in normal organs and possess potent antitumor activity. Here, we show that macrophage-mediated tumor-targeted delivery of Salmonella typhimurium VNP20009 can effectively reduce the toxicity caused by administrating VNP20009 alone in a melanoma mouse model. This benefits from tumor-induced chemotaxis for macrophages combined with their slow release of loaded strains. Inspired by changes in the tumor microenvironment, including a decrease in intratumoral dysfunctional CD8⁺ T cells and an increase in PDL1 on the tumor cell surface, macrophages were loaded with the engineered strain VNP-PD1nb, which can express and secrete anti-PD1 nanoantibodies after they are released from macrophages. This novel triple-combined immunotherapy significantly inhibited melanoma tumors by reactivating the tumor microenvironment by increasing immune cell infiltration, inhibiting tumor cell proliferation, remodeling TAMs to an M1-like phenotype and prominently activating CD8⁺ T cells. These data suggest that novel combination immunotherapy is expected to be a breakthrough relative to single immunotherapy.