Combination immunotherapy of glioblastoma with dendritic cell cancer vaccines, anti-PD-1 and poly I:C.

Glioblastoma (GBM) is a lethal cancer with limited therapeutic options. Dendritic cell (DC)-based cancer vaccines provide a promising approach for GBM treatment. Clinical studies suggest that other immunotherapeutic agents may be combined with DC vaccines to further enhance antitumor activity. Here, we report a GBM case with combination immunotherapy consisting of DC vaccines, anti-programmed death-1 (anti-PD-1) and poly I:C as well as the chemotherapeutic agent cyclophosphamide that was integrated with standard chemoradiation therapy, and the patient remained disease-free for 69 months. The patient received DC vaccines loaded with multiple forms of tumor antigens, including mRNA-tumor associated antigens (TAA), mRNA-neoantigens, and hypochlorous acid (HOCl)-oxidized tumor lysates. Furthermore, mRNA-TAAs were modified with a novel TriVac technology that fuses TAAs with a destabilization domain and inserts TAAs into full-length lysosomal associated membrane protein-1 to enhance major histocompatibility complex (MHC) class I and II antigen presentation. The treatment consisted of 42 DC cancer vaccine infusions, 26 anti-PD-1 antibody nivolumab administrations and 126 poly I:C injections for DC infusions. The patient also received 28 doses of cyclophosphamide for depletion of regulatory T cells. No immunotherapy-related adverse events were observed during the treatment. Robust antitumor CD4+ and CD8+ T-cell responses were detected. The patient remains free of disease progression. This is the first case report on the combination of the above three agents to treat glioblastoma patients. Our results suggest that integrated combination immunotherapy is safe and feasible for long-term treatment in this patient. A large-scale trial to validate these findings is warranted.

A series of host-guest coordination polymers containing viologens: syntheses, crystal structures, thermo/photochromism and factors influencing their thermo/photochromic behaviors.

Responsive molecular chromic materials have shown potential applications in molecular optical switches and sensor devices as they undergo reversible colour changes upon application of external stimuli. Herein, six host-guest coordination polymers 1-6 and one organic supramolecular compound 7 containing viologens have been synthesized under hydrothermal conditions. Compound 1 shows thermochromic behavior with a colour change from green to blue upon heating at 150 °C in air. Compounds 3, 4 and 7 display photochromic behaviors with a colour change from pale yellow to green under UV light, visible light or sunlight. The relationship between their structures and chromic behaviors has been discussed. Thermo/photochromic mechanisms have been investigated by infrared spectroscopy, powder X-ray diffraction analysis, UV-vis absorption spectroscopy, electron paramagnetic resonance spectroscopy, and X-ray photoelectron spectroscopy. The results demonstrate the generation of viologen radicals caused by thermo/photo-induced electron transfer. It is found that compound 4 can be used as an inkless and erasable printing medium. In addition, compounds 3, 4, and 7 can be used as anti-counterfeiting materials for QR codes. Our findings demonstrate the possibility of applying thermo/photochromic coordination polymers in molecular devices.

Treatment with angiotensin-(1-9) alleviates the cardiomyopathy in streptozotocin-induced diabetic rats.

Diabetic cardiomyopathy, a disorder of the heart muscle in diabetic patients, is one of the major causes of heart failure. We hypothesized that angiotensin-(1-9) [Ang-(1-9)] attenuates cardiomyopathy in streptozotocin (STZ)-induced diabetic rats. Rats were injected with a single intraperitoneal injection of STZ (55 mg/kg body weight) to induced diabetic cardiomyopathy. 4 weeks later, diabetic rats were treated with Ang-(1-9) (200 ng/kg/min), angiotensin type 2 receptor (AT2R) blocker PD123319 (100 ng/kg/min), or Mas antagonist A779 (100 ng/kg/min) for 4 weeks. Although Ang-(1-9) treatment did not affect blood glucose and insulin levels, it significantly attenuated cardiac hypertrophy, reduced cardiac fibrosis and improved ventricular function in STZ-induced diabetic rats. Ang-(1-9) treatment suppressed cardiac NADPH oxidase activity and reduced formation of reactive oxygen species. Ang-(1-9) suppressed NFκB activation and reduced myeloperoxidase (MPO) activity and mRNA levels of TNFα and IL-1ß in hearts of diabetic rats. In addition, Ang-(1-9) treatment suppressed activity of ACE and reduced angiotensin II (Ang II) formation in hearts of diabetic rats. The beneficial effect of Ang-(1-9) was blunted by coadministration of PD123319 but not by coadministration of A779. Finally, it was found that Ang-(1-9) treatment could alleviate STZ-induced cardiomyopathy in a dose-dependent manner. In conclusions, Ang-(1-9) attenuates cardiac dysfunction in STZ-induced diabetic rats. The Ang-(1-9)/AT2R axis should be investigated as a novel target for treatment of type 1 diabetic cardiomyopathy.

Synthesis, crystal structure, spectra and quantum chemical study on 1-phenyl-3-(4-nitrophenyl)-5-(2-thienyl)-2-pyrazoline.

1-Phenyl-3-(4-nitrophenyl)-5-(2-thienyl)-2-pyrazoline was synthesized and characterized by elemental analysis, IR and X-ray single crystal diffraction. UV-Vis spectra and fluorescence spectra were measured. Density functional theory calculations on the structure of the title compound were performed at the B3LYP/6-311G** level of theory. NPA atomic charge distributions indicate that, although the S atom in the thienyl ring loses coordination capacity, the title compound still may be used as a potential multi-dentate ligand to coordinate with metallic ions. The calculation of the second order optical nonlinearity was carried out. Natural bond orbital analyses indicate that the electronic absorption bands are mainly derived from the contribution of n→π* and π→π* transitions. Fluorescence spectra determination shows that the title compound is a potential orange-light emitting material.

Human kallikrein 6 inhibitors with a para-amidobenzylanmine P1 group identified through virtual screening.

A series of hK6 inhibitors with a para-amidobenzylamine P1 group and a 2-hydroxybenzamide scaffold linker was discovered through virtual screening. The X-ray structure of hK6 complexed with compound 9b was determined to a resolution of 1.68Å. The tertiary folding of the hK6 complexed with the inhibitor is conserved relative to the structure of the apo-protein, whereas the interaction between hK6 and the inhibitor is consistent with both the SAR and the in silico model used in the virtual screening.

Virtual Screening and X-ray Crystallography for Human Kallikrein 6 Inhibitors with an Amidinothiophene P1 Group.

A series of compounds with an amidinothiophene P1 group and a pyrrolidinone-sulphonamide scaffold linker was identified as potent inhibitors of human kallikrein 6 by structure-based virtual screening based on the union accessible binding space of serine proteases. As the first series of potent nonmechanism-based hK6 inhibitors, they may be used as tool compounds for target validation. An X-ray structure of a representative compound complexed with hK6, resolved at a resolution of 1.88 Å, revealed that the amidinothiophene moiety bound in the S1 pocket and the pyrrolidinone-sulphonamide linker projected the aromatic tail into the S' pocket.

Dy2(SO3)2(SO4)(H2O)2: the first lanthanide mixed sulfate-sulfite inorganic compound.

The first lanthanide mixed sulfate-sulfite inorganic coordination polymer, poly[diaqua-µ(4)-sulfato-di-µ(4)-sulfito-didysprosium(III)], [Dy(2)(SO(3))(2)(SO(4))(H(2)O)(2)](n), has been obtained, in which both sulfate and sulfite groups originate from the disproportionation of S(2)O(3)(2-) under hydrothermal and weakly acidic conditions. The crystal structure of the title compound exhibits a three-dimensional framework. The Dy(III) ion is surrounded by eight O atoms from one water molecule and two sulfate and five sulfite groups. These DyO(8) polyhedra have two shared edges and form an infinite zigzag Dy-O chain. In the bc plane, neighbouring chains are integrated through SO(3) trigonal pyramids, forming a two-dimensional sheet. Along the a-axial direction, the sulfate group, with the central S atom lying on a twofold axis, links adjacent two-dimensional sheets via two S-O-Dy connections, thus generating the three-dimensional framework.

Impairment of insulin action in non-obese, normal-glucose tolerant, first-degree relatives of Chinese type 2 diabetic patients.

AIMS: To investigate first-phase insulin release and peripheral insulin sensitivity of non-obese, normal-glucose tolerant, first-degree relatives of Chinese type 2 diabetic patients. METHODS: 12 euglycemic first-degree relatives of type 2 diabetic patients (ERDM), 12 newly diagnosed type 2 diabetic patients (DM-2) and 12 healthy individuals (control) participated in the study. All subjects were non-obese (BMI< 25 kg/m(2)). Intravenous glucose tolerance test and euglycemic hyperinsulinemic clamp test were performed to evaluate first-phase insulin release and quantify insulin sensitivity, respectively. RESULTS: the first-phase insulin release did not differ between the ERDM and control subjects (p=0.532), while the acute insulin response was absent in the DM-2 patients (p=0.001). Peripheral glucose deposit rate (GDR) was significantly lower in the ERDM (10.6 ± 2.1mg/kg·min, p=0.000) and DM-2 (9.6 ± 1.1mg/kg·min, p=0.000) groups than that in the control group (13.2 ± 1.2mg/kg·min). There was no statistical difference in GDR between the ERDM and DM-2 groups (p=0.110). Fasting FFA levels of the ERDM (p=0.007) and DM-2 (p=0.000) subjects were significantly higher than those of the controls. CONCLUSIONS: non-obese, first-degree relatives of type 2 diabetic patients with normal glucose tolerance (NGT) exhibit remarkable impairment of insulin sensitivity and increased FFA levels. Insulin resistance is independent of obesity and blood glucose level. Progression from NGT to type 2 diabetes may mainly be attributed to deterioration of early insulin secretion.

[Effect of inflammation factors on endothelial function in insulin resistance SD rats].

OBJECTIVE: To test the effect of inflammation factors on endothelial dysfunction in insulin resistance SD rats. METHODS: Male SD rats were divided randomly into three groups. The rats in the NC group, OB group and OB-AS group were fed with normal chow, high-fat chow, and high fat chow plus aspirin, respectively. The perifereal insulin sensitivity was detected with hyperinsulinemic euglycemic clamp. The endothelial dependent and independent vasodilatation were evaluated by measuring blood flow through femoral artery. Serum concentrations of high-sensitivity (hs) CRP and TNF-alpha were measured. Quantitative analysis of the expression of IKKbeta and NF-kappaB was performed. RESULTS: The rats in the OB group had significantly higher levels of serum hsCRP and TNF-alpha than the rats in the NC group (P < 0.05). There was a significant difference in endothelium-dependent (Ach-mediated dilation) vascular response and serum NO level between the OB group and the NC group (P < 0.05). Meanwhile, the rats in the OB group showed higher expression of NF-kappaBp65 (P < 0.05) and IKKbeta/NF-kappaBp65 (P < 0.05) than those in the NC group. Interestingly, compared with the rats in the OB group, the serum concentrations of TNF-alpha and CRP decreased significantly in the rats in the OB-AS group, to which high dose aspirin was given (P < 0.05). Better endothelium-dependent vascular response and higher serum NO level were observed in the rats in the OB-AS group than those in the NC group (P < 0.05). The rats in the OB-AS group also had significantly lower expression of NF-kappaBp65 (P < 0.05) and IKKbeta/NF-kappaBp65 (P < 0.05) than those in the OB group. CONCLUSION: Elevated inflammation factors are involved in the endothelial dysfunction by inactivating IKKbeta-NF-kappaB pathway in rats with insulin resistance.

2,7-Dibromo-9,9-bis-[(pyridin-1-ium-4-yl)meth-yl]fluorene dinitrate.

In the title compound, C(25)H(20)Br(2)N(2) (2+)·2NO(3) (-), the cation lies on a twofold rotation axis which imposes disorder of the dibromo-fluorene unit. In addition, the unique nitrate anion is disordered over two general sites of equal occupancy. The crystal structure is stabilized by inter-molecular N-H⋯O hydrogen bonds.

Synthesis, crystal structure and quantum chemical study on 3-phenylamino-4-phenyl-1,2,4-triazole-5-thione.

3-Phenylamino-4-phenyl-1,2,4-triazole-5-thione was synthesized and characterized by elemental analysis, IR and X-ray single crystal diffraction. Density functional theory calculations of the structure, natural bond orbitals, atomic charge distributions and thermodynamic functions of the title compound were performed at B3LYP/ 6-311G** and PBE1PBE/6-311G**levels of theory, respectively. NPA atomic charge distributions indicate that the title compound can be used as a potential multi-dentate ligand to coordinate with various metallic ions. Calculation of the second order optical nonlinearity was also carried out. The thermodynamic properties of C(0)(p,m), S(0)(m) and H(0)(m) were calculated and correlative equations between the thermodynamic properties and temperatures were also obtained.

Teriflunomide reduces behavioral, electrophysiological, and histopathological deficits in the Dark Agouti rat model of experimental autoimmune encephalomyelitis.

Teriflunomide is an orally available anti-inflammatory drug that prevents T and B cell proliferation and function by inhibition of dihydroorotate dehydrogenase. It is currently being developed for the treatment of multiple sclerosis (MS). We report here for the first time the anti-inflammatory effects of teriflunomide in the Dark Agouti rat model of experimental autoimmune encephalomyelitis (EAE). Neurological evaluation demonstrated that prophylactic dosing of teriflunomide at 3 and 10 mg/kg delayed disease onset and reduced maximal and cumulative scores. Therapeutic administration of teriflunomide at doses of 3 or 10 mg/kg at disease onset significantly reduced maximal and cumulative disease scores as compared to vehicle treated rats. Dosing teriflunomide at disease remission, at 3 and 10 mg/kg, reduced the cumulative scores for the remaining course of the disease. Teriflunomide at 10 mg/kg significantly reduced inflammation, demyelination, and axonal loss when dosed prophylactically or therapeutically. In electrophysiological somatosensory evoked potential studies, therapeutic administration of teriflunomide, at the onset of disease, prevented both a decrease in waveform amplitude and an increase in the latency to waveform initiation in EAE animals compared to vehicle. Therapeutic dosing with teriflunomide at disease remission prevented a decrease in evoked potential amplitude, prevented an increase in latency, and enhanced recovery time within the CNS.

(1R,1'S)-1,1'-Dihydr-oxy-1,1'-biisobenzofuran-3,3'(1H,1'H)-dione.

In the title compound, C(16)H(10)O(6), the complete mol-ecule is generated by a crystallographic centre of symmetry. In the crystal, O-H⋯O hydrogen bonds link the mol-ecules into (100) sheets and C-H⋯O links also occur.

3-(4-Fluoro-phen-yl)-1-(4-methoxy-phen-yl)prop-2-en-1-one.

The title compound, C(16)H(13)FO(2), was prepared from 4-methoxy-hypnone and 4-fluoro-benzophenone by Claisen-Schmidt condensation. All the bond lengths and bond angles are in normal ranges. The dihedral angle formed by the two benzene rings is 33.49 (2)°. The crystal packing is stabilized by inter-molecular C-H⋯O hydrogen-bonding inter-actions.

Comparison of Suspension Array and EnzymE-Linked Immunosorbent Assay Methods for Detection of Three Kinds of Veterinary Drug Residues / 分析化学

A novel suspension array technology was established for the detection of three kinds of veterinary drug residues: chloramphenicol, clenbuterol and 17-β-estradiol. The three conjugates in which veterinary drugs coupled with BSA were immobilized on the solid carrier of the suspension microarray-polystyrene fluorescent microspheres/beads as detective probes. Indirect competitive technology was employed. Competitive reactions between the veterinary drugs in the aqueous phase and the veterinary drugs-BSA conjugates on the beads for coupling with their complimentary specific biotinylated monoclonal antibodies were carried out. And then, straptavidin-phycoerythrin was added for coupling and the fluorescent signals were captured. Afterwards the detective standard curves were plotted. The regular ELISA standard curves of the three veterinary drugs were also plotted. Comparison between suspension array and regular enzymE-linked immunosorbent assay(ELISA) was in the respects of the detective technology, the detection limits, the detective ranges, the samples detection and the multi-analysis. Suspension array technology is distinct advantageous except for specificity. There was well consistent performance between the two methods. The high-throughput suspension array provides a novel method for multi-analysis of veterinary drugs with simple operation, sensitive, rapid and low costing.

Development of a high-throughput suspension microarray technology for detection of three kinds of veterinary drug residues: chloramphenicol, clenbuterol and 17-beta-estradiol / 中华预防医学杂志

<p><b>OBJECTIVE</b>To establish a novel suspension microarray technology for the detection of three kinds of veterinary drug residues: chloramphenicol, clenbuterol and 17-beta-estradiol (CAP, CL and E2).</p><p><b>METHODS</b>The three conjugates that veterinary drug coupled with bovine serum albumin (BSA) were synthesized and identified by ultraviolet (UV) spectrophotometry and mass spectrum. The veterinary drug conjugates were immobilized on the polystyrene fluorescent microspheres/beads. There were competitive reactions between the veterinary drugs in the aqueous phase and that on the beads for combination with their specific biotinylated monoclonal antibodies. The optimum amount of the veterinary drug conjugates and the antibodies were optimized and selected. The detective standard curves were plotted. The specificity and the unknown samples were also determined by grouping according to different concentrations of the interferes and the samples. Meantime, the different microstructures of the surfaces of the beads were also observed by scanning electron microscope.</p><p><b>RESULTS</b>Couplings were completed between small molecular veterinary drugs and BSA. The amounts of the three conjugates and the antibodies were optimized. The detective standard curves of the suspension array and their corresponding coefficients of determination (R2) were good (R2 > 0.99). The detection ranges of the three veterinary drugs were (40.00 - 6.25) x 10(5) ng/L, (50.00-7.81) x 10(5) ng/L and 1.00 x 10(3) - 7.29 x 10(5) ng/L respectively. Simultaneously, the specific detection of the suspension microarray was excellent and did not indicate significant cross-reactions. Errors between the found and the real are in the range of 8.09% - 17.03%. It can be considered that the relative standard deviations were relatively small. Successful couplings were also directly confirmed by the observation for microstructures of the surfaces of the beads by scanning of electron microscope and laid good foundation for the following responses.</p><p><b>CONCLUSION</b>The high-throughput suspension microarray should provide a novel method for multi-analysis of the veterinary drugs and have a wide applicative prospects with simple operation, sensitive, rapid and low cost.</p>

Synthesis, IR spectra, crystal structure and DFT studies on 1-acetyl-3-(4-chlorophenyl)-5-(4-methylphenyl)-2-pyrazoline.

1-Acetyl-3-(4-chlorophenyl)-5-(4-methylphenyl)-2-pyrazoline has been synthesized and characterized by elemental analysis, IR and X-ray single crystal diffraction. Density functional (DFT) calculations have been carried out for the title compound by using the B3LYP method at the 6-311G** basis set level. The calculated results show that the predicted geometry can reproduce well the structural parameters. Predicted vibrational frequencies have been assigned and compared with experimental IR spectra and they are supported each other. On the basis of vibrational analyses, the thermodynamic properties of the title compound at different temperatures have been calculated, revealing the correlations between C(0)(p, m), S(0)(m), H(0)(m) and temperatures.

[Construction and expression of a prokaryotic vector of recombinant human adiponectin global domain].

OBJECTIVE: To construct and express the recombinant human adiponectin (gAd) global domain. METHODS: gAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG. RESULTS: The gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis. CONCLUSION: The recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.

A dinuclear CoII complex: bis(mu-dihydrogen phosphato-kappa2O:O')bis[(bipyridine-kappa2N,N')(dihydrogen phosphato-kappaO)cobalt(II)].

The title compound, [Co2(H2PO4)4(C10H8N2)2], is dinuclear, centred on a symmetry centre of the P1 space group. Each Co atom has a distorted square-pyramidal coordination involving two N atoms from a bipyridine molecule and three O atoms from two bridging and one terminal dihydrogen orthophosphate anion. The molecular structure and packing are stabilized by intermolecular hydrogen-bond interactions.