Yun-Long Tseng

Sterically stabilized anti‐idiotype immunoliposomes improve the therapeutic efficacy of doxorubicin in a murine B‐cell lymphoma model

A liposome containing diverse synthetic lipid derivatives of polyethylene glycol (PEG) results in smaller distribution volume and longer circulation time in blood and, thus, may improve drug targeting. The characteristics and therapeutic efficacy of immunoliposomes with similar liposomal formulation have never been studied in lymphoma models. We have developed immunoliposomes conjugated with S5A8 monoclonal antibody, an anti‐idiotype antibody to 38C13 murine B‐cell lymphoma, and loaded them with doxorubicin using an ammonium sulfate gradient. Purified antibodies were covalently coupled to the termini of PEG on the surface of small unilamellar liposomes. Cell binding and internalization ability of these immunoliposomes were estimated by a fluorescence assay using a pH‐sensitive fluorescent dye (HPTS). The in vitro cytotoxicity of doxorubicin encapsulated in immunoliposomes was greater for idiotype‐positive 38C13 cells than for the idiotype‐negative variant of this cell line. In syngeneic C3H/HeN mice, doxorubicin encapsulated in immunoliposomes exhibited a long circulation time and was more effective at prolonging survival of mice bearing 38C13 tumor than non‐targeted liposomal doxorubicin or free doxorubicin plus empty immunoliposomes. Our results demonstrate the superiority of targeted therapy with these immunoliposomes and its potential in lymphoma treatment. Int. J. Cancer 80:723–730, 1999.

Diagnostic and therapeutic evaluation of 111In-vinorelbine-liposomes in a human colorectal carcinoma HT-29/luc-bearing animal model

Colorectal carcinoma is a highly prevalent and common cause of cancer in Taiwan. There is still no available cure for this malignant disease. To address this issue, we applied the multimodality of molecular imaging to explore the efficacy of diagnostic and therapeutic nanoradiopharmaceuticals in an animal model of human colorectal adenocarcinoma [colorectal cancer (CRC)] that stably expresses luciferase (luc) as a reporter. In this study, an in vivo therapeutic efficacy evaluation of dual-nanoliposome (100 nm in diameter) encaged vinorelbine (VNB) and (111)In-oxine on HT-29/luc mouse xenografts was carried out. HT-29/luc tumor cells were transplanted subcutaneously into male SCID mice. Multimodality of molecular imaging approaches including bioluminescence imaging (BLI), gamma scintigraphy, whole-body autoradiography (WBAR) and in vivo tumor growth tracing, histopathology and biochemistry/hematology analyses were applied on xenografted SCID mice to study the treatments with 6% polyethylene glycol (PEG) of (111)In-NanoX/VNB-liposomes. In vivo tumor growth tracing and BLI showed that tumor volume could be completely inhibited by the combination therapy with (111)In-VNB-liposomes and by chemotherapy with NanoX/VNB-liposomes (i.e., without Indium-111) (P<.01). The nuclear medicine images of gamma scintigraphy and WBAR also revealed the conspicuous inhibition of tumor growth by the combination therapy with (111)In-VNB-liposomes. Animal body weights, histopathology and biochemistry/hematology analyses were used to confirm the safety and feasibility of radiopharmaceuticals. A synergistic therapeutic effect on CRC xenografted SCID mice was proven by combining an Auger electron-emitting radioisotope (Indium-111) with an anticancer drug (VNB). This study further demonstrates the beneficial potential applications of multimodality molecular imaging as part of the diagnostic and therapeutic approaches available for the evaluation of new drugs and other strategic approaches to disease treatment.

Therapeutic Efficacy Evaluation of 111In-Labeled PEGylated Liposomal Vinorelbine in Murine Colon Carcinoma with Multimodalities of Molecular Imaging

In our previous studies using combined radioisotopes with chemotherapeutic liposomal drugs (i.e., 111In-labeled polyethylene glycol (PEG)ylated liposomal vinorelbine) we have reported possible therapeutic efficiency in tumor growth suppression. Nevertheless, the challenge remains as to whether this chemotherapy has a therapeutic effect as good as that of combination therapy. The goal of this study was to investigate the real therapeutic effectiveness of 6 mol% PEG 111In-vinorelbine liposomes via the elevation of the radiation dosage and reduction in the concentration of chemotherapeutic agents. Methods: Murine colon carcinoma cells transfected with dual-reporter genes (CT-26/tk-luc) were xenografted into BALB/c mice. The biodistribution was estimated to determine the drug profile and targeting efficiency of 111In-vinorelbine liposomes. Bioluminescence imaging and 18F-FDG small-animal PET were applied to monitor the therapeutic response after drug administration. The survival in vivo was estimated and linked with the toxicologic and histopathologic analyses to determine the preclinical safety and feasibility of the nanomedicine. Results: Effective long-term circulation of radioactivity in the plasma was achieved by 6 mol% PEG 111In-vinorelbine liposomes, and this dose showed significantly lower uptake in the reticuloendothelial system than that of 0.9 mol% PEG 111In-vinorelbine liposomes. Selective tumor uptake was represented by cumulative deposition, and the maximum accumulation was at 48 h after injection. The combination therapy exhibited an additive effect for tumor growth suppression as tracked by caliper measurement, bioluminescence imaging, and small-animal PET. Furthermore, an improved survival rate and reduced tissue toxicity were closely correlated with the toxicologic and histopathologic results. Conclusion: The results demonstrated that the use of 6 mol% PEG 111In-vinorelbine liposomes for passively targeted tumor therapy displayed an additive effect with combined therapy, not only by prolonging the circulation rate because of a reduction in the phagocytic effect of the reticuloendothelial system but also by enhancing tumor uptake. Thus, this preclinical study suggests that 6 mol% PEG 111In-vinorelbine liposomes have the potential to increase the therapeutic index and reduce the toxicity of the passively nanotargeted chemoradiotherapies.

Simple and efficient liposomal encapsulation of topotecan by ammonium sulfate gradient: Stability, pharmacokinetic and therapeutic evaluation

Topotecan (TPT), a topoisomerase I inhibitor, is presently undergoing clinical evaluation worldwide. Previous studies have shown that entrapping TPT within multi-lamellar vesicle liposome can stabilize the lactone moiety, which is structurally important for biological activity. However, low drug:lipid ratios due to the amphipathic character and small entrapment volume in the unilamellar vesicle limits the development of pharmaceutically acceptable liposomal formulation. With an aim to improve on this drawback, we herein describe a method that utilizes the ammonium sulfate gradient to entrap TPT into liposomes. By this method, the encapsulation efficiency was over 90% and a drug:lipid molar ratio as high as 1:5.4 was reached. In comparison with free drug, liposome-encapsulated TPT is more stable in physiological conditions and shows higher in vitro cytotoxicity. Because of increased blood circulation time, the initial plasma concentration and area under the plasma concentration of liposomal drugs were 14 and 40 times, respectively, of those of free drug. Furthermore, liposome encapsulation enhanced the antitumor activity of TPT in syngeneic murine C-26 and human HTB-9 xenograft models in vivo. At a dose of 5 mg/kg, the tumor growth delay of liposomal formulation was significantly than that of free TPT. Based on these results, we believe that this liposomal TPT formulation is worthy of further clinical study.

Improved Pharmacokinetics and Efficacy of a Highly Stable Nanoliposomal Vinorelbine

Effective liposomal formulations of vinorelbine (5′ nor-anhydro-vinblastine; VRL) have been elusive due to vinorelbines hydrophobic structure and resulting difficulty in stabilizing the drug inside the nanocarrier. Triethylammonium salts of several polyanionic trapping agents were used initially to prepare minimally pegylated nanoliposomal vinorelbine formulations with a wide range of drug release rates. Sulfate, poly(phosphate), and sucrose octasulfate were used to stabilize vinorelbine intraliposomally while in circulation, with varying degrees of effectiveness. The release rate of vinorelbine from the liposomal carrier was affected by both the chemical nature of the trapping agent and the resulting drug-to-lipid ratio, with liposomes prepared using sucrose octasulfate displaying the longest half-life in circulation (9.4 h) and in vivo retention in the nanoparticle (t1/2 = 27.2 h). Efficacy was considerably improved in both a human colon carcinoma (HT-29) and a murine (C-26) colon carcinoma model when vinorelbine was stably encapsulated in liposomes using triethylammonium sucrose octasulfate. Early difficulties in preparing highly pegylated formulations were later overcome by substituting a neutral distearoylglycerol anchor for the more commonly used anionic distearoylphosphatidylethanolamine anchor. The new pegylated nanoliposomal vinorelbine displayed high encapsulation efficiency and in vivo drug retention, and it was highly active against human breast and lung tumor xenografts. Acute toxicity of the drug in immunocompetent mice slightly decreased upon encapsulation in liposomes, with a maximum tolerated dose of 17.5 mg VRL/kg for free vinorelbine and 23.8 mg VRL/kg for nanoliposomal vinorelbine. Our results demonstrate that a highly active, stable, and long-circulating liposomal vinorelbine can be prepared and warrants further study in the treatment of cancer.

Evaluation of Pharmacokinetics of 111In-Labeled VNB-PEGylated Liposomes After Intraperitoneal and Intravenous Administration in a Tumor/Ascites Mouse Model

Nanoliposomes are important drug carriers that can passively target tumor sites by the enhanced permeability and retention (EPR) effect in neoplasm lesions. This study evaluated the biodistribution and pharmacokinetics of 111In-labeled vinorelbine (VNB)-encapsulated PEGylated liposomes (IVNBPL) after intraperitoneal (i.p.) and intravenous (i.v.) administration in a C26/tk-luc colon carcinoma ascites mouse model. IVNBPL was prepared by labeling VNB-encapsulated PEGylated liposomes with 111In-oxine. BALB/c mice were i.p. inoculated with 2 x 10(5) C26/tk-luc cells in 500 muL of phosphate-buffered saline. Peritoneal tumor lesions were confirmed by 124I-FIAU/micro-PET (positron emission tomography) and bioluminescence imaging. Ascites production was examined by ultrasound imaging on day 10 after tumor cell inoculation. The pharmacokinetics and biodistribution studies of IVNBPL in a tumor/ascites mouse model were conducted. The labeling efficiency was more than 90%. The in vitro stability in human plasma at 37 degrees C for 72 hours was 83% +/- 3.5%. For i.p. administration, the areas under curves (AUCs) of ascites and tumor were 6.78- and 1.70-fold higher, whereas the AUCs of normal tissues were lower than those via the i.v. route. This study demonstrates that i.p. administration is a better approach than i.v. injection for IVNBPL, when applied to the treatment of i.p. malignant disease in a tumor/ascites mouse model.

Tumor Burden Talks in Cancer Treatment with PEGylated Liposomal Drugs

Purpose PEGylated liposomes are important drug carriers that can passively target tumor by enhanced permeability and retention (EPR) effect in neoplasm lesions. This study demonstrated that tumor burden determines the tumor uptake, and also the tumor response, in cancer treatment with PEGylated liposomal drugs in a C26/tk-luc colon carcinoma-bearing mouse model. Methods Empty PEGylated liposomes (NanoX) and those encapsulated with VNB (NanoVNB) were labeled with In-111 to obtain InNanoX and InVNBL in high labeling yield and radiochemical purity (all >90%). BALB/c mice bearing either small (58.4±8.0 mm3) or large (102.4±22.0 mm3) C26/tk-luc tumors in the right dorsal flank were intravenously administered with NanoVNB, InNanoX, InVNBL, or NanoX as a control, every 7 days for 3 times. The therapeutic efficacy was evaluated by body weight loss, tumor growth inhibition (using calipers and bioluminescence imaging) and survival fraction. The scintigraphic imaging of tumor mouse was performed during and after treatment. Results The biodistribution study of InVNBL revealed a clear inverse correlation (r 2 = 0.9336) between the tumor uptake and the tumor mass ranged from 27.6 to 623.9 mg. All three liposomal drugs showed better therapeutic efficacy in small-tumor mice than in large-tumor mice. Tumor-bearing mice treated with InVNBL (a combination drug) showed the highest tumor growth inhibition rate and survival fraction compared to those treated with NanoVNB (chemodrug only) and InNanoX (radionuclide only). Specific tumor targeting and significantly increased tumor uptake after periodical treatment with InVNBL were evidenced by scintigraphic imaging, especially in mice bearing small tumors. Conclusion The significant differences in the outcomes of cancer treatment and molecular imaging between animals bearing small and large tumors revealed that tumor burden is a critical and discriminative factor in cancer therapy using PEGylated liposomal drugs.

Pharmacokinetics and Dosimetry of 111In/188Re-Labeled PEGylated Liposomal Drugs in Two Colon Carcinoma-Bearing Mouse Models

PEGylated liposomes are important drug carriers for nanomedicine cancer therapy. PEGylated liposomes can encapsulate radio- and chemo-drugs and passively target tumor sites via enhanced permeability and retention effect. This study estimated the pharmacokinetics and dosimetry after administration of radio-chemotherapeutics ((111)In-labeled vinorelbine [VNB]-encapsulated liposomes, InVNBL, and (188)Re-labeled doxorubicin [DXR]-encapsulated liposomes, ReDXRL) for radionuclide therapy in two colon carcinoma-bearing mouse models. A C26 colon carcinoma tumor/ascites mouse model and a subcutaneous solid tumor-bearing mouse model were employed. Biodistribution studies of InVNBL and ReDXRL after intraperitoneal administration in tumor/ascites-bearing mice (protocol A) and intravenous administration in subcutaneous solid tumor-bearing mice (protocol B) were performed. The radiation dose to normal tissues and tumors were calculated based on the results of distribution studies in mice, using the OLINDA/EXM program. The cumulated activities in most organs after administration of InVNBL in either the tumor/ascites-bearing mice (protocol A) or the subcutaneous solid tumor-bearing mice (protocol B) were higher than those of ReDXRL. Higher tumor-to-normal-tissues absorption dose ratios (T/NTs) were observed after administration of InVNBL than those of ReDXRL for protocol A. The T/NTs for the liver, spleen, and red marrow after injection of InVNBL for protocol B were similar to those of ReDXRL. The critical organ was found to be red marrow, and thus the red marrow absorption dose defined the recommended maximum administration activity of these liposomal drugs. Characterization of pharmacokinetics and dosimetry is needed to select the appropriate radiotherapeutics for specific tumor treatment applications. The results suggest that InVNBL is a promising therapeutic agent, which is as good as ReDXRL, in two mouse tumor models.

Synergistic effect of combination 188Re-liposome and Lipotecan in Huh-7 subcutaneous xenograft model

Atest apparatus with electronic control system was devised to synthesis Carbon Nanoparticles (CNP) by submerged arc discharge between two pure graphite electrodes in pure liquid paraffin oil medium. Different grades of oil viscosity are used. It was found that the resultant oil from the arc process contains carbon nanoparticles. Transmission Electron Microscope (TEM) was used to characterize the output morphology of the resultant CNP. Physical and tribological properties of the resultant oil are assessed. It was found that presence of CNP in paraffin oil resulted in reduced coefficient of friction in four-ball testing machine, raising its flash point, but reduced slightly its viscosity. This system can be improved to be applied industrially in a continuous product line with added suitable dispersant. Further work is proceeding to optimize test conditions.Methods: Twenty four immature upper permanent incisors teeth, in 6 dogs of 6 month old, were selected and divided into 2 groups. Group (A), 12 teeth for dental pulp stem cells transplantation and group (B) 12 teeth for sealing with MTA paste only. Another 12 teeth, Group (C), for isolation of the dental pulp stem cells were grouped. Apical periodontitis were induced in groups (A) and (B). After disinfection, teeth were re-entered and mixture of antibiotic paste was applied for two weeks. Dental pulp stem cells were isolated from group (C) teeth of the same dog, prepared for transplantation, mixed with growth factors and transplanted in group (A) teeth while MTA was applied for group (B) teeth. Both groups (A) and (B) teeth were monitored radiographically for periradicular healing and root wall thickening and/or lengthening.W report on the effect of Au nanoparticle deposition on the colors of virus-based colorimetric sensors consisted of selfassembled M13 bacteriophage films decorated by Au nanoparticles. We observed the effect of the localized surface plasmons in the Au nanoparticles onto the virus-based self-assembled films. Recently, virus-based colorimetric sensors have demonstrated real-time detection of biochemicals with high sensitivity, and portability (Nature Communications, 5, 3043 (2014)). Therefore, it is desirable if we can obtain colored virus films in a reliable way. However, the M13 bacteriophage-based films show some variance in their colors after assembly. This can be a problem in the mass production process of virus-based colorimetric sensors for commercial use. In this study, we fabricated a biomimetic material-based colorimetric film using M13 bacteriophage, a filamentous virus that infects only bacteria, as the building block. Then, we deposited Au nanoparticles over the colorimetric film. We observed the effect of Au nanoparticle deposition on the color and morphology of the virus films by using optical and atomic force microscopy (AFM). From these studies, we provide a method for structural color enhancement of biomimetic material-based colorimetric films.B vanadate has become most promising material for photocatalytic applications from last decade due to its visible light driven reactions in the degradation of organic pollutants. However, it is well known that doping agents may enhance the catalytic activities. In this context, investigations on nano-sized BiVO4 oxides doped by suitable metal ions are conducted in the aim to enhance the absorption in the visible spectral range and increase the efficiency of photocatalytic responses. Thus, highly crystalline silver loaded BiVO4 are prepared by mechano-chemical reaction between bismuth oxide (Bi2O3) and vanadium oxide (V2O5) in a stoichiometric ratio of 1:1 and adding metallic silver particles (<10 mm) at different atomic percentages with respect to the host oxide material (2,4,6,8 and 10%). The suitable ball milling parameters are adjusted as well as the conditions of post-synthesis annealing in order to improve the crystalline features of the samples. Complementary characterization techniques were performed to analyze the main features of the samples. Thus, XRD analysis confirmed a major high crystalline monoclinic scheelite structure for all BiVO4:Ag samples. High resolution TEM image reveals Ag presence in nano-particles along with BiVO4. The morphology of the samples investigated by FE-SEM shows particles with 50 nm sized and the effective doping by Ag ions was indeed demonstrated by (EDAX) technique. Optical absorption measurements reveal an evolution from 2.55 to 2.33 eV of BiVO4 gap with Ag doping rates.S Enhanced Raman scattering (SERS) allows detection or state identification of molecules at sub-micromolar concentrations which are especially required for accurate analysis of physiological liquids. An enormous increase of Raman signal intensity from molecules adhered on metallic nanostructures is mostly connected with plasmonic properties of metals. By today solid SERS-active substrates have attracted a great attention due to enhancement factor (EF) reproducibility and simple use. On the other hand solid substrates have to be composed of highly ordered arrays of metallic nanostructures which are fabricated by complicated and expensive nanoengineering methods. In this report, results of a research of porous silicon (PS) as a template for the fabrication of SERS-active substrates are presented. PS was formed by an electrochemical anodization of bulk silicon in HF-based solutions. Dimensions of pores and silicon crystallites in PS were varied from a couple of nanometers to micron by the anodization regimes and the type of bulk silicon. Metallic nanostructures were deposited on PSby “wet”methods. PS templates provided fabrication of different morphological forms of metallic structures (nanoparticles, dendrites, nanovoids, etc.). It was found that plasmonic properties of the metallic nanostructures strongly depend on the PS morphology and the metal deposition regimes. SERS-activity of the obtained substrates was studied using aqueous or alcoholic solutions of rhodamine6G and metallic porphyrins as analytes. It was estimated that EF of SERS-active substrate based on PS template can reach 109. Deviation of EF reproducibility from substrate to substrate was shown to be less than 20 %.W examined room temperature band-to-band tunneling in 2D InAs/3D GaSb heterostructures. Specifically, multisubband, gate-controlled negative differential resistance is observed in InAs/AlSb/GaSb junctions. Due to spatial confinement in the 10nm-thick InAs layer, tunneling contributions from two distinct subbands are observed as sharp steps in the current-voltage characteristics. It is shown that the relative position of the steps can be controlled via external gate bias. Additionally, the extracted separation in the subband energy agrees well with the calculated values. This is the first demonstration of a gate controlled tunneling diode with multiple subband contributions. By further improving the fabrication techniques, InAs/AlSb/GaSb FET is for the first time demonstrated. Device analysis indicates that Dit plays an important role on device performances and the experimental result clarifies the role of the tunneling junction width with respect to the gate width.H acid (HA) is a non-sulfated glycosaminoglycan organic polymer, found as structural component of the extracellular matrix (ECM) in mammalian bone marrow and loose connective tissues of our body. Hyaluronic acid also plays a major role in cell proliferation, motility, cell adhesion and gene expression. Cellular HA receptors such as CD44+ and RHAMM are over expressed in cancer which makes it a good candidate for tumor targeting purpose. Here we have developed a T1 gadolinium contrast agent based on Hyaluronic acid (HA) that target CD44+ receptor over expression in cirrhosis which also display higher relaxation property than primovist and better contrast–to–noise ratio (CNR) at 20 min and even at 2 hr time period. Superparamagnetic iron oxide nanoparticles (SPION) have emerged as an MRI contrast agent primarily for tumor imaging due to their efficacy and safety, proven by clinical application with a series of marketed SPION-based contrast agents. For the purpose of tumor diagnosis iron oxide nanoparticle coated with Hyaluronic acid (HA-SPION) as T2 contrast agent was synthesized with co-PEGylation strategy for increased bioavailabilty to tumor region. And finally, hyaluronic acid micelle that can encapsulate hydrophobic anti-cancer drug such as paclitaxel that specifically target CD44+ overexpressing tumor are synthesised which can act as promising drug delivery vehicle.P sulfide) (PPS) is well known for its heat resistance, high mechanical strength, excellent chemical resistance, and good electronic properties. It is used in many areas, especially in electronics, mechanical, and chemical engineering. In order to improve thermal and electrical properties of PPS as well as to expand its application, a series of PPSbased nanocomposite films containing multiwalled carbon nanotube (MWCNT) of 0.1~10.0 wt% as functional reinforcing nanofiller were manufactured by melt-mixing and -compression. The microstructures, thermal and electrical properties of the nanocomposite films were investigated as a function of the MWCNT content. FT-IR spectra and SEM images confirmed that PPS/MWCNT nanocomposite films supported that MWCNTs were randomly dispersed in the PPS matrix. DSC and TGA data indicated that the thermal transition temperatures as well as the degradation temperatures of the nanocomposite films were influenced by the presence of the MWCNT. The electrical resistivity of the nanocomposite films were dramatically changed with increasing the MWCNT by exhibiting a typical percolation threshold. Furthermore, the applications of PPS/MWCNT nanocomposite films as electric heating elements were demonstrated. The electric heating performance of the nanocomposite films with high MWCNT contents was systematically characterized in terms of temperature response rapidity, saturated maximum temperature, and electric power efficiency and operational stability at applied voltages up to 100V.G Quantum Dots (GQDs) are proving to be effective imaging paraphernalia for the comprehension of morphological alterations in the cellular membrane due to high absorption coefficients and quantum efficiency. Such quantum dots can be used in drug/delivery vehicles, biolabelling as well as in PCR. An upsurge of expanded interest in the field of magnetic nanotechnology has led us to allow indepth exploitation of magnetic nanoparticles in nanomedicine. Encapsulating the core made up of magnetic nanoparticles by Gold nanoshell leads to the development of a proficient biocompatible and stabilized drug/delivery system under physiological conditions. Further a nanocomposite was created by allowing conglomeration of AuFe3O4 core-shell with GQDs. This modular design enables Au-Fe3O4-GQDs to perform multiple functions simultaneously, such as in multimodal imaging, drug delivery and real-time monitoring, as well as combined therapeutic approaches. The ability of MNPs to enhance proton relaxation of specific tissues and serve as MR imaging contrast agents is one of the most promising applications of nanomedicine. In the present work, Au-Fe3O4 nanoparticles are used as able cargo for the docking of anti-cancer drug such as Doxorubicin (DOX) using cysteamine as a linker for the attachment. The attachment could be monitored using UV-visible spectroscopy. The stability of Au-Fe3O4 nanoparticles was scrutinized by measuring the flocculation parameter which was found to be in the range of 0-0.65. Further, zeta potential measurements confirmed the pH of 7.4 at which maximum drug attachment can take place. The amalgamation of the drug along with activated folic acid as a navigational molecule is the critical phase for targeted drug delivery. Attachments were verified using FTIR and NMR which confirmed the formation of non-covalent interactions. The drug loading capacity of the Au-Fe3O4 was found to be 76%. Drug-release was studied using the AC magnetic field generator and was found to be temperature dependent phenomena. GQDs were found to be effective players in tracking the drug-delivery vehicle around the miscreant cell and inside them. Au-Fe3O4-GQD-FA-DOX complex was found to be comparatively non-toxic for normal cells and considerably toxic for Hep-2 cells due to hyperthermal properties of SPIONS and targeted-mechanism of folic acid.S is severe chronic disabling brain disease affecting about 24 million people worldwide and 4 million people in India as WHO 2011 report. Asenapine maleate (ASN) is a novel psychopharmacologic agent recommended for treatment of Schizophrenia and Bipolar disorder. ASN suffers with the problems of poor aqueous solubility, very low oral bioavailability (1-2%) and high patient non-adherence. Therefore, objective of the present study was to prepare Nanostructured Lipid Carriers (NLCs) of Asenapine maleate to improve the bioavailability and enhance the uptake of ASN to the brain via intranasal route. Asenapine loaded NLCs (ASN-NLCs) were prepared by melt emulsification-high shear homogenization method. For preparation of NLCs, Glyceryl monostearate, Oleic acid and Tween 80 were used as solid lipid, liquid lipid and surfactant, respectively. ASN-NLCs were characterized for particle size, zeta potential, entrapment efficiency, in-vitro drug release study, XRD and DSC. Stability study was performed at 25°C/60% RH for three months. Further, pharmacodynamic studies (paw test and l-dopa induced locomotor activity test) were performed on rat model to evaluate the efficacy of formulation. ASNNLCs were successfully prepared and optimized with particle size below 200 nm, zeta potential -15.38±2.17 mV, EE 82±3.5% and more than 85% drug release in 24 h. The XRD and DSC analysis indicate that Asenapine was present in amorphous state in NLCs. The ASN-NLCs were stable over 3 month studies. In pharmacodynamic studies, significant increase (p<0.05) in antipsychotic potential was observed in ANS-NLCs as compared to pure drug. These results indicate that the NLCs are having a potential to deliver drug into the brain from the non-invasive intranasal route.P is extremely abundant in the earth’s crust and distributed very widely. Due to the particularity of it, pyrite is an important research subject in several different fields such as gold prospecting, metallogenic environment, metallogenic prediction, and even functional material. Studies have shown that different Fe/S atom ratio, impurity, and temperature, could all cause crystal defects in the crystal interior, which could lead to the occurrence of crystal lattice structure distortion, which then could affect the thermoelectricity of pyrite. Thus, there might exist a response relationship between the mineral crystal structure and thermoelectricity. However, presently, the research on the relation between the response researches is still rare. In this paper, the samples were obtained by thermal sulfidation during 320~420°C and studied using a SEM ,a XRD and thermoelectricity instrument respectively, to study the crystal morphology, structure and thermoelectricity. The research shows that:(1) Pyrite prepared by thermal sulfidation, are p-type with a small scope of thermoelectric coefficient and good stability. (2) The value of the thermoelectric coefficient was correlated with the growth degree of crystal planes, the higher degree of crystallinity, the higher thermoelectric coefficient. (3) There exists a significantly different influence from different directions on the thermoelectric coefficient; the thermoelectric coefficient of low-index surface (111), (200) and (210) was significantly greater than high-index surface (220) and (311). (4) In the pyrite crystal, the different reticular density caused different thermoelectric. The greater the reticular density, the greater is the influence on the thermoelectric coefficient.