In the present study, a new compound named 17-(6-cinnamamido-hexylamino-)-17-demethoxygeldanamycin (CDG) was obtained by introducing the cinnamic acid (CA) group into the 17-site of geldanamycin (GDM). The anti-cancer effects of CDG in vitro and in vivo were evaluated. MTT assay was used to examine the inhibitory effect of CDG on the proliferation of MCF-7, HepG2, H460 and SW1990 cells. Immunofluorescent staining flow cytometry combined with Annexin V-FITC/PI staining were used to detect apoptotic cells. Transwell assay was used to analyze the effect of CDG on cell invasion and migration ability. Western blotting was used to detect the expression levels of RAF-1, EGFR, AKT, CDK4 and HER-2 of MCF-7, HepG2 and H460 cells. The toxicities of CDG and GDM were evaluated in mice. Using the subcutaneously transplanted MCF-7 xenograft in nude mice, inhibitory effect was evaluated in vivo. The results showed that CDG inhibited the proliferation of cancer cells (IC50: 13.6-67.4 microg.mL-1). After exposure to CDG for 48 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation or shrunken nucleus. The rates of apoptosis of MCF-7, HepG2, H460 and SW1990 cells incubated with 10 microg.mL-1 CDG were 23.16%, 27.55%, 22.21%, 20.47%, respectively. A dose-dependent reduction of migration of four cell lines was found after exposure to CDG. The decreased levels of RAF-1, EGFR, AKT, CDK4 and HER-2 showed that CDG possessed HSP90 inhibitory effect. The result of animal toxicity test on the mice suggested that CDG had lower toxicity than GDM. Meanwhile, CDG inhibited the growth of MCF-7 xenografts of athymic mice.

Objective To observe the changes in levels of plasma leukocyte cell-derived chemnotaxin 2 (LECT2) in patients with newly diagnosed type 2 diabetes (T2DM) complicated with non-alcoholic fatty liver disease (NAFLD),and to investigate the clinical significance.Methods A total of 137 subjects were enrolled in the study,including 50 patients with newly diagnosed T2DM complicated with NAFLD,47 patients with newly diagnosed T2DM with non-NAFLD and 40 healthy subjects.The level of plasma LECT2 was determined by ELISA.Anthropometric data and other biochemical indicators were measured in three groups.The body mass index (BMI) and waist-to-hip ratio (WHR) were calculated.Insulin resistance and pancreas β-cell function were determined by homeostasis model assessment (HOMA-IR,HOMA-％β).Results Plasma concentration of LECT2 in patients with newly diagnosed T2DM complicated with non-NAFLD was higher than that in type 2 diabetic patients with non-NAFLD [(32.95 ± 10.11 vs 29.08 ± 7.54) ng/mL,P ＜ 0.01].Plasma LECT2 levels in both groups were significantly higher than that in normal control group [(22.38 ± 4.40) ng/mL,P ＜ 0.01].Plasma LECT2 level was positively correlated with BMI,FPG,FINS,C peptide,HbA1c,GGT,TG and HOMA-IR,while negatively with HDL-C and HOMA-％ β (all P ＜ 0.01).Multivariate regression analysis showed that levels of BMI,FPG and HDL-C were important factors affecting plasma LECT2 level.Conclusions Plasma LECT2 concentration significantly elevates in patients with newly diagnosed T2DM complicated with non-NAFLD.Plasma level of LECT2 is closely correlated with insulin resistance and glycolipid metabolism.LECT2 may play an important role in the patho genesis of insulin resistance and T2DM.

AIMTo study the mechanisms of sodium ferulate (SF) on inhibition of collagen synthesis in hepatic stellate cells.

METHODSCollagen synthesis was analyzed by measuring 3H-proline incorporation. ELISA method was used to study the effect of SF on transforming growth factor beta1 (TGFbeta1) level in cultured HSC-T6 cell. The effect of SFon the TGFbeta1 activity in the supernatant of culture was analyzed by mink lung epithelial cell (Mv1Lu) proliferation inhibition by MTT assay.

RESULTSSF inhibited collagen synthesis in hepatic stellate cells stimulated with TGFbeta1. SF was shown to decrease TGFbeta1 level in the supernatant of HSC-T6 increased by oxidative stress. TGFbeta1 activity was intervened by SF.

CONCLUSIONSF could decrease collagen synthesis, with mechanism may be associated with that SF intervened TGFbeta1 activity, and reduced the level of TGFbeta1 increased by oxidative stress.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Coumaric Acids ; pharmacology ; Epithelial Cells ; cytology ; Hepatocytes ; cytology ; metabolism ; Lung ; cytology ; Mink ; Oxidative Stress ; Rats ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1

This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.
DNA Primers ; genetics ; Flavivirus ; classification ; genetics ; isolation & purification ; Flavivirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Nonstructural Proteins ; genetics

Adult ; Benzene ; toxicity ; Chromatography, High Pressure Liquid ; Humans ; Occupational Exposure ; Reference Values ; Sorbic Acid ; analogs & derivatives ; analysis

In the present study, a new compound named 17-(6-cinnamamido-hexylamino-)-17-demethoxygeldanamycin (CDG) was obtained by introducing the cinnamic acid (CA) group into the 17-site of geldanamycin (GDM). The anti-cancer effects of CDG in vitro and in vivo were evaluated. MTT assay was used to examine the inhibitory effect of CDG on the proliferation of MCF-7, HepG2, H460 and SW1990 cells. Immunofluorescent staining flow cytometry combined with Annexin V-FITC/PI staining were used to detect apoptotic cells. Transwell assay was used to analyze the effect of CDG on cell invasion and migration ability. Western blotting was used to detect the expression levels of RAF-1, EGFR, AKT, CDK4 and HER-2 of MCF-7, HepG2 and H460 cells. The toxicities of CDG and GDM were evaluated in mice. Using the subcutaneously transplanted MCF-7 xenograft in nude mice, inhibitory effect was evaluated in vivo. The results showed that CDG inhibited the proliferation of cancer cells (IC50: 13.6-67.4 microg.mL-1). After exposure to CDG for 48 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation or shrunken nucleus. The rates of apoptosis of MCF-7, HepG2, H460 and SW1990 cells incubated with 10 microg.mL-1 CDG were 23.16%, 27.55%, 22.21%, 20.47%, respectively. A dose-dependent reduction of migration of four cell lines was found after exposure to CDG. The decreased levels of RAF-1, EGFR, AKT, CDK4 and HER-2 showed that CDG possessed HSP90 inhibitory effect. The result of animal toxicity test on the mice suggested that CDG had lower toxicity than GDM. Meanwhile, CDG inhibited the growth of MCF-7 xenografts of athymic mice.
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Benzoquinones ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase 4 ; metabolism ; Female ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Lactams, Macrocyclic ; chemical synthesis ; chemistry ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Proto-Oncogene Proteins A-raf ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; metabolism ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

Adult ; Female ; Forearm ; surgery ; Free Tissue Flaps ; Humans ; Male ; Middle Aged ; Oral Surgical Procedures ; methods ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation

Objective To investigate the effects of AGEs-RAGE on the apoptosis of GLUTag cells and explore the possiblc mechanism.Methods GLUTag cells treated with 0、100、200、300μg/ml of AGEs for 24h were examined for gene and protein expression of RAGE using RT-PCR and western blotting,respectively.GLUTag cells were randomly divided into four groups:control,200μg/ml AGEs,AGEs+siRNA-RAGE and AGEs+apocynin.The protein expression of p22phox、p47phox 、Bcl-2、Bax in the cells were detected with western blotting.The reactive oxygen species (ROS) levels were examined using 2'7'-dichlorodihydroflur-rescein diacetate (DCFH-DA) and the apoptosis of L cells were tested by AnnexinV-FITC/PI.Results AGEs increased thc cxpression of RAGE in a dose dependent manner.Treatment with AGEs induced a significant increase in the expression of p22phox,p47phoxand the activity of ROS,caused up-regulation of Bax and down-regulation of Bcl-2,which enhanced the apoptosis of GLUTag cells.Apocynin,the inhibitor of NADPH oxidase,prevented those responses and the effects caused by AGEs were abolished by inhibition of RAGE activity with siRNA.Conclusion AGEs positively regulate the exprcssion of NADPH oxidase-derived ROS and its down-steam signaling pathway p53/Bax by targeting RAGE,leading to the apoptosis of GLUTag cells.

·Cataract refers to the degenerative changes in lens quality caused by various causes of reduced transparency or color change. Surgical treatment is the main treatment modality at present. Among them, phacoemulsification has become the main surgical treatment for cataract because of its advantages such as short operation time, small incision and quicker healing. Today, cataract surgery has gradually shift to refractive surgery, and is no longer simply as cataract surgery. But after cataract phacoemulsification, the symptoms and visual quality are different. The main causes include refractive error, postoperative dry eyes and postoperative corneal astigmatism. This article reviews the factors that influence the visual quality of cataract phacoemulsification and its future trends.

Objective To assess the impact of additional cycles of temozolomide on the survival of glio-blastoma(GBM)patients after 6 months of maintenance temozolomide(TMZ)following concurrent TMZ chemo-therapy and radiation therapy. Methods Data of 51 GBM patients from 2009 to 2015 were retrospectively studied and the therapeutic effect was assessed according to whether receiving long-term treatment with TMZ. Results Sev-enteen of fifty-one GBM patients received 8 or more cycles and prolonged treatment improved progression-free sur-vival(P=0.011)and overall survival(P=0.004). Conclusions Extended use of TMZ is safe to GBM patients , which may improve response OS and PFS compared to conventional regimen. Prospective studies in larger popula-tions are needed to better-define the population to whom it can be proposed and its optimal duration.