This study aimed at investigating the inhibitory effects and the anti-tumor mechanisms of co-adminis-tration of fusion proteins mGM-CSF/βhCG ( GC ) and hVEGF121/βhCG ( VC ) on RM-1 prostatic cancer and B16 F10 melanoma in the C57 BL/6 J mouse model. Two recombinant stains containing pET-28 a-mGM-CSF-X10-βhCGCTP37 and pET-28 a-VEGF-M2-X10-βhCG-CTP37 were induced by lactose to express fusion proteins. The fusion proteins were separated and purified to prepare the anti-tumor protein vaccines ( VC protein vaccine and GC protein vaccine) , which were then mixed to prepare a combined protein vaccine named VGC protein vac-cine. The prostatic cancer and melanoma tumor-bearing mice C57 BL/6 J were immunized with described vac-cines, then the growth of each tumor was measured;splenocyte proliferation of immunized mice was detected;and the cytotoxic effects of the vaccine on tumor cells were tested. After that, the in vivo concentrations of IFN-γ and anti-hVEGF antibodies were investigated by ELISA. The difference between each experimental group and normal saline group ( NS) was statistically significant in both tumor-bearing mouse models ( P <0. 05) respectively. Besides, VGC group exhibited significantly better anti-tumor effect compared with the GC and VC groups with the anti-tumor rate ( 41. 7 ± 0. 83)% and ( 46. 4 ± 1. 27)% for prostatic cancer and melanoma tumor, respectively. The co-administration of the two proteins, VC and GC, could inhibit the growth of RM-1 prostatic tumor and B16F10 melanoma effectively via anti-tumor immunity and anti-tumor angiogenesis.

This study aimed at investigating the effects and mechanisms of 7-O-succinyl macrolactin A in inhibi-ting human lung cancer. After treatment of human lung cancer cell lines H460 with 7-O-succinyl macrolactin A, MTS assay was employed to determine cell proliferation;crystal violet staining was used to detect cell adhesion of H460;transwell chamber assay and wound healing assay were performed to evaluate cell invasion and migration;and flow cytometry assay was adopted to evaluate cell cycle. Western blotting and real-time PCR were also employed to determine the expression of β-catenin, c-Myc, Cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2 , Survivin and MMP-2/9. The phosphorylation of AKT and mTOR was determined as well. In vitro prolifera-tion of H460 was inhibited significantly by 7-O-succinyl macrolactin A. Cell adhesion, invasion and migration abilities were also attenuated. Western blot and real-time PCR showed that the expressions of β-catenin, c-Myc, cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2, Survivin and MMP-2/9 were down-regulated by 7-O-succinyl macrolactin A. It was also found that phosphorylation of AKT and mTOR was inhibited by 7-O-succinyl macrolactin A. 7-O-succinyl macrolactin A can inhibit the in vitro growth and invasion of human lung cancer cell lines H460.

Ubiquitous chromatin opening element ( UCOE ) , composed of the promoters of human housekeeping genes, prevents transgene from silencing and produces consistent, stable and high-level gene expression irrespec-tive of the chromosomal integration site. The research studied the influence of different UCOE element parts on antibody expression in CHO cells. UCOE 1. 5 kb from chromobox homolog 3 ( CBX3 ) , UCOE 2. 5 kb from the heterogeneous ribonucleoprotein A2/B1 ( HNRPA2 B1 ) and the whole UCOE 4. 0 kb were inserted into the anti-body light and heavy chain vectors, respectively, and transfected into CHO cells using antibiotics-Zeocin and Blasticidin for pressure selection. Four groups of monoclonal cells were harvested and antibody expression of each group was detected. The monoclonal cells with UCOE 1. 5 kb and UCOE 2. 5 kb increased 1. 5 to 2-fold in the level of antibody expression, wheareas, monoclonal cells with UCOE 4. 0 kb increased 3 to 4-fold. The enhance-ment of two housekeeping promoter genes on antibody expression could stack up.

In order to search for new antiplatelet agents with higher potency, a series of tetramethylpyrazine ( TMP) /chalcone hybrids ( 2-26) were synthesized and evaluated based on the principle of bioisostere and hybrid-ization. They exerted inhibitory activity against adenosine diphosphate ( ADP )-induced and arachidonic acid ( AA)-induced platelet aggregation to varied extent. Among them, compound 8 was the most potent with IC50 of 0. 14 mmol/L on ADP-induced platelet aggregation ( 9. 1 folds of TMP and 10. 5 folds of chalcone ) and 0. 09 mmol/L on AA-induced platelet aggregation ( 8. 8 folds of TMP and 10. 0 folds of chalcone) , which was superior to clinically used anti-platelet drug aspirin ( ASP, IC50 =0. 15 mmol/L) .

A series of 5-amino-2-( benzylthio ) thiazole-4-carboxamide derivatives were designed and synthesized to discover novel compounds with anti-tumor activity. Compounds DDO-5401-DDO-5416 were synthesized using 2-amino-2-cyanoacetamide as the start material. The structures of the synthesized compounds were confirmed by IR, 1 H NMR and ESI-MS. The in vitro anti-tumor activities of the synthesized compounds were determined by MTT assay in HCT116 , HepG2 , A549 , MDA-MB-231 and MCF-7 cell lines. Target compounds showed good anti-tumor activity especially in A549 cell line. SAR study showed that electron donating groups were more favorable than electron absorption ones. Compound DDO-5413 exhibited noteworthy activity in MDA-MB-231 and MCF-7 cell lines with IC50 value lower than the positive reference dasatinib. It suggested that DDO-5413 might be the candidate for further investigation.

Arterial thrombosis (AT) is a common disease which usually causes acute myocardial infarction,ischemic stroke and other ischemic cardiovascular and cerebrovascular diseases,which shows high rates of morbidity and mortality,and has become a serious problem to human health.It is increasingly clear that the interactions among platelets,the endothelium and leukocytes are important throughout all stages of the atherothrombotic process.It is of great significance to search for therapeutic targets in the process of AT and developing the therapeutic drugs based on those newly discovered targets.This article reviews the research advances in the discovery of antithrombotic targets and the current situation of drug development based on those antithrombotic targets found in the recent 5 years,in order to provide some references or clues for the development of innovative drugs to prevent and treat AT.

A five-step synthetic method of ambrisentan starting from benzophenone was developed.The reaction methods of resolution and substitution were optimized.A stable,facilitating method of detection and efficient resolution reagent,L-phenylglycinamide,was used and helped to obtain highly optical pure intermediates.In the substitution reaction,NaOH was chosen as the base,which is safe and suitable for the industry.This method was mild,easily operated with a total yield up to 31.0％,and may have a good application prospect.

The aim of this study was to synthesize 1-(5-(hydroxymethyl)-4-methyl-3-oxo-3,4-dihydro-pyrazin-2-yl) guanidine (mucunaguanide),a pyrazinone alkaloid compound extracted from the seed of Stizotobium cochinchinensis.Starting from benzyloxy acetaldehyde,following four steps in reaction including condensation,cyclization,substitution and hydrogenation reaction,mucunaguanide was synthesized with total yield of 27.9％,and purity of more than 97.5％.Structures of all the intermediates and target compound were confirmed by IR,1H NMR and MS.This synthetic process was characterized with raw materials available,simple in operation with much milder reaction conditions,and was an ideal method of synthesis of this compound.

The purpose of this study was to synthesize and evaluate the necrosis target of MRI contrast agent based on rhein.The novel ligand 10-{ [6-(1,8-dihydroxyanthraquinone-3-carboxamido) hexyl] amino} acetyl-1,4,7,10-tetraazacyclododecan-1,4,7-triacetic acid (DO3A-rhein) was synthesized by two-step acylation and two-step deprotection.The paramagnetic contrast agent gadolinium 10-{ [6-(1,8-dihydroxyanthraquinone-3-carboxamido) hexyl] amino} acetyl-1,4,7,10-tetraazacyclododecan-1,4,7-triacetate (Gd-DO3A-rhein) was obtained by coordination of Gd3+ with the synthesized ligand.Its necrosis affinity was evaluated by liver infarction and muscular necrosis on rat models.The MRI was performed before administration of Gd-DO3A-rhein and during 0 h to 12 h after administration of Gd-DO3A-rhein (0.1 mmol/kg),respectively,and Gd-DOTA was used as control.After MRI scanning,rats were sacrificed and necrotic tissues were stained using triphenyltetrazolium chloride (TTC) and hematoxylin-eosin (HE).MRI images of liver infarction and muscular necrosis on rat models showed significantly enhanced signal intensity compared with normal tissues.The contrast ratios of necrotic liver/normal liver were 1.61 ±0.14 and 2.36 ±0.20 at 3 h and 12 h postinjection of Gd-DO3A-rhein (0.1 mmol/kg) respectively,demonstrating a significant difference compared with pre-administration of Gd-DO3A-rhein (1.16 ±0.10;P ＜ 0.05).The same results were obtained from necrotic muscles.These findings suggested that Gd-DO3A-rhein possessed the necrosis target and imaging capability of necrotic tissues.

5-(4-Hydroxyphenyl)-3H-1,2-dithiole-3-thione (ADT-OH) is a slowly releasing H2S donor with some neuroprotective effect.In order to study the structure-activity relationships,seventeen compounds (Y1-Y17) were synthesized by modification of ADT-OH at the aromatic ring position,and their structures were confirmed by 1H NMR,13C NMR and HR-MS.Among them,6 compounds (Y4,Y13-Y17) were novel compounds.Their effects had been evaluated on HT-22 hippocampal neuron cells damaged by glutamate with MTF method.The pharmacological results demonstrated that all the Y compounds had potent neuroprotection at most of the tested concentrations (1-100 μmol/L).Compounds Y1-Y9 and Y11 significantly improved the survival rates of the damaged cells at 1-100 μmol/L (P ＜0.01).Specially,compounds Y1,Y4,Y6-Y9,Yu are more potent than their parent compound ADT-OH at concentration of 1-10 μmol/L,which is worthy of further study.