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OBJECTIVE@#To investigate the effect of acute myeloid leukemia cells in leukemia-microenvironment on proliferation and apoptosis of bone marrow-derived mesenchymal stromal cells (BM-MSC).@*METHODS@#Acute myeloid leukemia (AML) murine models overexpressing MLL-AF9 were established. The number of BM-MSC of wild type (WT) and AML-derived mice were analyzed by flow cytometry. Morphology and growth differences between WT and AML-derived BM-MSC were analyzed by inverted fluorescence microscope. Proliferation and apoptosis of BM-MSC between these two groups were detected by Brdu and Annexin V/PI.@*RESULTS@#Compared with WT-derived BM-MSC, the number and proliferation rate of AML-derived BM-MSC significantly increased (P<0.01, P<0.001), while apoptosis rate decreased (P<0.05). When cultured in vitro, BM-MSC grew faster under conditional medium.@*CONCLUSION@#AML cells can promote proliferation and inhibit apoptosis of BM-MSC.
Subject(s)
Animals , Humans , Mice , Apoptosis , Bone Marrow , Bone Marrow Cells , Cell Proliferation , Leukemia, Myeloid, Acute , Mesenchymal Stem Cells , Tumor MicroenvironmentABSTRACT
@#A 21-year-old male student was admitted to the emergency department of our hospital due to chest distress, dyspnea for 1.5 hours, and loss of consciousness for one minute. Before admission, the patient had been advised rest for two months because of left ankle sprain, leading to less activity. At admission, the patient was unconscious, with facial cyanosis, and his limbs were cold.
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Localized delivery, comparing to systemic drug administration, offers a unique alternative to enhance efficacy, lower dosage, and minimize systemic tissue toxicity by releasing therapeutics locally and specifically to the site of interests. Herein, a localized drug delivery platform ("plum‒pudding" structure) with controlled release and long-acting features is developed through an injectable hydrogel ("pudding") crosslinked
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Melanoma is a malignant tumor with a high degree of malignancy. The incidence of melanoma keeps increasing annually. In this study, a melanoma targeted hyaluronic acid (HA) nanogel was synthesized via crosslinking of thiolated HA with terminally functionalized F127-TPGS mixed micelles. Its stability in vitro was evaluated by the average particle size, and the cytotoxicity of the nanogel was investigated by in vitro cell based assays. Next, cell uptake studies were performed to quantitatively and qualitatively investigate the uptake of the nanogels in B16F10 cells. A small sized nanogel with a diameter of 30 nm was synthesized, which was proven to be minimally cytotoxic against both 3T3 or B16F10 cells. Compared with 3T3 cells with low levels of CD44, B16F10 cells with high levels of CD44 showed significantly higher cell uptake efficiency (P<0.05).
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Objective To detect the diatom population diversity in Dianchi by constructing a 18S rDNA clone library. Methods DNA from diatoms in 6 water samples of Dianchi was amplified with diatom 18S rDNA specific primer.The 18S rDNA clone library was constructed, and clones were randomly selected for sequence. Sequence alignment was performed by BLAST. The diatom population distribution in Dianchi was analyzed and the phylogenetic tree of diatom 18S rDNA in Dianchi waters was established with the MEGA v7.0.14 software. Results Two hundred and forty clones were sequenced, with 167 diatom sequences obtained, including 11 diatom species such as Stephanodiscus, Diatoma, and Melosira. There were certain differences in diatom population distribution among the 6 samples. Conclusion The population distribution of diatom species in Dianchi shows unique features and the sequence analysis of diatom 18S rDNA has a certain reference value to the inference of forensic drowning sites.
Subject(s)
Humans , China , DNA, Ribosomal/genetics , Diatoms/classification , Drowning , Forensic Sciences , Phylogeny , RNA, Ribosomal, 18S/geneticsABSTRACT
As the common pathway of chronic renal diseases leading to end-stage renal failure, renal tubulointerstitial fibrosis is characterized by the deposition of extracellular matrix and scar hardening. Our study aimed to construct an in vitro cell culture platform to explore the impact of matrix stiffness on cell morphology and function of renal tubular epithelial cells. Photopolymerized polyacrylamide gels (PAA gel) with varying stiffnesses as model substrates was selected to simulate the matrix stiffness of normal and fibrotic renal tissues with elastic moduli ranging from 1 to 40 kPa. The human renal tubular epithelial cells (HK-2) were seeded on the surface of PAA gels. The impact of matrix stiffness on the morphology of HK-2 were investigated via immunofluorescence staining and confocal microscopy. The expression levels of glucose transporter 1 (GLUT1), glucose transporter 2 (GLUT2), glucose transporter 5 (GLUT5) were semi-quantitatively analyzed. With increasing matrix stiffness, both the levels of GLUT1 and GLUT5 in HK-2 cells were significantly decreased, whereas the expression level and the distribution pattern of GLUT2 in HK-2 remained unchanged with stiffness variation.
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Severe acute pancreatitis (SAP) is characterized by both local and systemic inflammatory responses. This study was designed to develop a site-specific delivery strategy for SAP therapy using celastrol (CLT). First, murine RAW264.7 cells were used as a model of macrophage cell line, cell membranes were obtained by emptying intracellular contents via hypotonic lysing, mechanical membrane disruption, and differential centrifugation. Poly(ethylene glycol) methyl ether-block-poly(lactic-co-glycolic acid) (PEG-PLGA) nanoparticles (NPs) were then prepared by sonication. With the collected membrane materials, macrophage membrane coated PEG-PLGA NPs (RNPs) were then prepared by extrusion through a 400 nm polycarbonate membrane. Biodistribution study in rats with SAP showed RNPs selectively accumulated in the inflamed pancreatic tissues. Compared with CLT loaded NPs, CLT loaded RNPs were proven to effectively attenuate local pancreatic inflammation and systemic inflammation in rats with SAP.
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In the past 20 years, tumor immunotherapy has made a significant progress in tumor inhibition effects in both laboratory studies and clinical trails. In the immune response to tumor, effective antitumor immunity is induced during the tumor progress; long-term monitoring of the tumor would be achieved through the immune memory, reducing the possibility of tumor recurrence. In the immune treatment strategies, a focus is delivery of therapeutic immune regulators with nanocarriers. It has been demonstrated that due to the special physical and chemical properties, nanocarriers are easily internalized by immune cells, which regulate the immune responses and effectively induce anti-tumor immune cascade to achieve the tumor inhibition effect. In this paper, we will discuss the progress of nanocarrier-mediated antitumor immunotherapy in recent years.
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Fluorescent polystyrene nanospheres (PS) were used to explore the impact of substrate stiffness on cell uptake of nanoparticles in the breast cancer cells. Polyacrylamide (PAA) gels with varying stiffness were prepared by photopolymerization, and type I rat tail collagen was covalently conjugated on the surface of PAA gels to facilitate cell adhesion. Type I rat tail collagen was also used to fabricate collagen gels for 3D cell culture. Cells of human breast cancer cell line MCF-7 were incubated in the 2D culture on PAA gels and 3D culture within collagen gels. Next, nanospheres of 20 nm and 50 nm polystyrene were applied to MCF-7 cells in the 2D or 3D cultures. Cell morphology and uptake efficiency were observed with confocal microscopy. Our study demonstrates that substrate stiffness differentially regulated the cell morphology as well as the cell uptake behavior of polystyrene nanospheres in MCF-7 cells under 2D or 3D culture conditions.
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In recent years, a new type of organic cation transporter has been found to transport organic cation drugs like pyrilamine, diphenhydramine and oxycodone in brain capillary endothelial cells, Caco-2 cells and other cells. Its transport activity can not be inhibited by typical organic cation transporter substrates or inhibitors, and its transport characteristics are different from those reported for the organic cation transporters, such as organic cation transporters (OCTs), organic cation/carnitine transporters (OCTNs), multidrug and toxin extrusion transporters (MATE) and the plasma membrane monoamine transporter (PMAT). It is a novel organic cation transporter, called pyrilamine-sensitive H+/OC antiporter. This review will present a comprehensive summary to elaborate the transport characteristics, structure of the substrates, tissue expression and the differences with other organic cation transporters.
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In the present report, we review the technical guidelines and principles on impurity research and control for antibiotics established by various agencies, including the International Conference of Harmonization (ICH), the US Food and Drug Administration (FDA), the European Medicines Agency (EMA) and the China Food and Drug Administration (CFDA). Progresses with the US Pharmacopoeia (USP), the European Pharmacopoeia (EP) and the Chinese Pharmacopoeia (ChP) to control impurities in antibiotics are also presented. Next, our discussion is focused on analyzing the CFDA's requirements on impurity research and control for antibiotics, and the implementation of ICH, FDA and other technical guidelines for generic drugs impurity control in China. Existing problems are further reviewed, in order to improve the overall process for the control of antibiotic purity.
Subject(s)
Humans , Anti-Bacterial Agents , Reference Standards , China , Drug Contamination , Drug and Narcotic Control , Drugs, Generic , Europe , Pharmaceutical Preparations , Reference Standards , Pharmacopoeias as Topic , Quality Control , Research , United States , United States Food and Drug AdministrationABSTRACT
A rapid, sensitive and specific reversed-phase high-performance liquid chromatographic method was developed for the determination of 3-n-butylphthalide, a drug currently being developed for treatment of stroke, in mice tissue. Ultraviolet detection were monitored at 228 nm for quantification of 3-nbutylphthalide. Ibuprofen was used as internal standard. The peak area ratio vs concentration in tissue was linear over the range of 25.625–1025.000 ng/mL and the limit of quantification was 25.625 ng/mL. The method was successfully applied to pharmacokinetic investigation in mice. The mean distribution half-life was 5.471 ± 4.736 min and elimination half-life was 58.459 ± 34.370 min. Finally, a preclinical biodistribution research of 3-n-butylphthalide in mice following intravenous administration had been studied. Brain targeting of 3-n-butylphthalide was strong and metabolism in the liver and blood was rapid.
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This study is to report the preparation of complexes of Ad5 and anionic liposomes (AL-Ad5), the amplification of adenoviruses with enhanced green fluorescent protein (eGFP) reporter gene performed by HEK 293 cells, the adenoviral vectors purified by cesium chloride gradient centrifugation, and the titer of adenovirus determined by cytopathic effect (CPE) method, hexon capsid immunoassay and quantitative-PCR (Q-PCR), separately. The prescription and experiment conditions were optimized by central composite design (CCD). The complexes of Ad5 and AL-Ad5 were formulated by the calcium-induced phase change method. The morpholopy, particle size and zeta potential were detected by dynamic light scattering (DLS) and transmission electron microscopy (TEM), respectively. Additionally, the bicolourable fluoresce-labeled complexes (F(labeled)-AL-Ad5) were prepared and their intracellular location in MDCK cells was detected by confocal laser scanning microscopy (CLSM). The results indicate that the complexes of AL-Ad5 exhibited a uniform distribution with a particle size of 211 +/- 10 nm and a zeta potential of -41.2 +/- 2.2 mV. The result of CLSM demonstrates that the intracellular location of red fluoresce-labeled adenovirus was consistent with that of green fluoresce-labeled liposomes suggesting that the naked adenovirus was well encapsulated by the anionic liposomes in complexes of AL-Ad5.
Subject(s)
Animals , Dogs , Humans , Adenoviridae , Genetics , Anions , Cytopathogenic Effect, Viral , Drug Compounding , Methods , Genetic Vectors , Green Fluorescent Proteins , Chemistry , HEK293 Cells , Liposomes , Chemistry , Pharmacokinetics , Madin Darby Canine Kidney Cells , Microscopy, Confocal , Microscopy, Electron, Transmission , Particle Size , Polymerase Chain Reaction , Methods , Recombinant Fusion ProteinsABSTRACT
<p><b>OBJECTIVE</b>To explore effects of acrylamide on synaptic plasticity of rat neuron and its mechanisms.</p><p><b>METHODS</b>24 Wistar rats were divided into control and test groups randomly, 12 rats in each group. The ratio of male and female in each group was 1:1. Acrylamide (30 mg/kg) was administered to rats by intraperitoneal injection in test group and normal saline (5 g/kg) was given to rats in control group. The neurobehavioral and pathologic changes of heart, liver, spleen, lung and kidney were observed. Changes of parameters in synapse were recorded by electron microscope. As an important target of synapse, change of Synapsin I was measured by immunohistochemical method.</p><p><b>RESULTS</b>Compared with the control group (male: 1.00 ± 0.00; female: 1.00 ± 0.00), the gait score was increased significantly in ACR treated group (male: 2.50 ± 0.55, t = -7.24, P < 0.01; female: 3.17 ± 0.41, t = -12.19, P < 0.01). No obvious pathological changes of heart, liver, spleen, lung and kidney were found in all rats. Compared with the control group (male: (0.41 ± 0.09) µm; female: (0.40 ± 0.06) µm), the length of active zone of synapse was decreased significantly in ACR treated group (male: (0.15 ± 0.05) µm, t = 6.59, P < 0.05; female: (0.14 ± 0.07) µm, t = 7.26, P < 0.05). The width and postsynaptic density of synapse in ACR treated group had no significant difference with control group. The location of Synapsin I of control group and ACR treated group was both in gray matter of spinal dorsal horn. Compared with the control group (male: 195.40 ± 12.30; female: 195.19 ± 6.71), the concentration of Synapsin I was decreased significantly in ACR treated group (male: 60.90 ± 29.19, t = 10.40, P < 0.05; female: 67.56 ± 20.23, t = 15.65, P < 0.05).</p><p><b>CONCLUSION</b>Neuronal synaptic plasticity was found in damage of nervous system induced by acrylamide in rats, which might be associated with the expression of Synapsin I.</p>
Subject(s)
Animals , Female , Male , Rats , Acrylamide , Toxicity , Neuronal Plasticity , Neurons , Rats, Wistar , SynapsesABSTRACT
The in vitro release behavior, in vivo biodistribution and antitumor activity of N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer-5-fluorouracil conjugates (P-FU) were studied. The in vitro release behavior was evaluated by determining the amount of 5-fluorouracil (5-FU) released from P-FU in mice plasma at 37 degrees C. The in vivo biodistribution and therapeutic evaluation were investigated with Kunming mice bearing hepatoma 22 (H22). The in vitro half-life (t1/2) of P-FU in mice plasma was 32.4 h. It appeared that the circulation life time of the conjugates were 166 times longer than that of 5-FU. The AUC and t1/2 of P-FU in tumor were 3.3 times and 2.3 times more than those of 5-FU, respectively. Therapeutic evaluation also demonstrated that the treatment with P-FU displayed stronger inhibition of the tumor growth when compared with that of 5-FU (P < 0.05). HPMA copolymer is a potential carrier for 5-FU for effective treatment of cancer.
Subject(s)
Animals , Female , Mice , Acrylamides , Pharmacokinetics , Pharmacology , Antimetabolites, Antineoplastic , Pharmacokinetics , Pharmacology , Area Under Curve , Cell Line, Tumor , Drug Carriers , Drug Compounding , Fluorouracil , Pharmacokinetics , Pharmacology , Liver Neoplasms, Experimental , Metabolism , Pathology , Neoplasm Transplantation , Random Allocation , Tissue Distribution , Tumor BurdenABSTRACT
<p><b>OBJECTIVE</b>To screen and optimize the extraction of Dingxiangjiangqi granules.</p><p><b>METHOD</b>The extraction route was screened by using pharmacodynamic experiment and the extraction conditions were optimized by orthogonal design and taking extract yield, content of naringin and tetrahydropalmatine as indexes.</p><p><b>RESULT</b>The pharmacodynamic result showed that aqueous extract had the best effect to cure the esophagitis of rats and the optimized extraction technique was adding 12 times water, extracting 0. 5 hour for 3 times.</p><p><b>CONCLUSION</b>The optimum extraction was simple, reasonable, stable and useful for further development.</p>
Subject(s)
Animals , Male , Rats , Berberine Alkaloids , Drug Combinations , Drugs, Chinese Herbal , Chemistry , Pharmacology , Esophagitis, Peptic , Drug Therapy , Pathology , Esophagus , Pathology , Flavanones , Phytotherapy , Plants, Medicinal , Chemistry , Random Allocation , Rats, Sprague-Dawley , Syzygium , Chemistry , Technology, Pharmaceutical , MethodsABSTRACT
A novel transferrin modified non-viral gene delivery system Tf-PLPD was developed and the related characteristics was investigated. Blank procationic liposomes were prepared by film dispersion-filteration method. PLPD was prepared as follows by first mixing the plasmid DNA and protamine together, then the resulted polyplexes were incubated for 10 min at room temperature, followed by addition of preformed blank procationic liposomes. Transferrin was adsorbed at the surface of PLPD via electrostatic interactions to form Tf-PLPD. Central composite design (CCD) was employed to optimize the formulation. The HepG2 cells were transfected using lacZ as reporter gene and characteristics such as the morphology, the mean particle size, the zeta potential and the transfection efficiency in HepG2 cells were further investigated by different methods. The resulting PLPD had a regular spherical surface with an average size of (228. 9 +/- 8. 0) nm (polydispersity index, PDI = 0. 122 +/- 0. 02, n = 3) , a zeta potential of ( - 25. 08 +/-2. 50) mV (n = 3) and a transfection efficiency of (12. 18 +/- 3. 80) mU x mg(-1) (protein). The Tf-PLPD had an average size of (240 +/- 12) nm (polydispersity index, PDI = 0. 150 +/- 0. 03, n = 3), a zeta potential of ( - 24. 10 +/- 2. 50) mV ( n = 3) and a transfection efficiency of (24. 26 +/- 2. 60) mU x mg(-1) (protein) , 20 times greater than that of the naked plasmid DNA. The presence of serum didn' t affect the tansfection activity of PLPD or Tf-PLPD. Compared to one kind of cationic liposomes (liposome-protamine-DNA, LPD), the PLPD and Tf-PLPD had much less cytotoxicity to three hepatic cell lines (including HepG2, SMMC7721 and Chang' s normal hepatocyte). The results indicated that the Tf-PLPD is a perspective non-viral vector for gene delivery systems.
Subject(s)
Humans , Cations , Chemistry , Cell Line , Cell Line, Tumor , Cell Survival , DNA , Chemistry , Genetics , Hepatocytes , Cell Biology , Metabolism , Liposomes , Chemistry , Liver Neoplasms , Genetics , Pathology , Particle Size , Plasmids , Chemistry , Genetics , Protamines , Chemistry , Transfection , Methods , Transferrin , Chemistry , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study optimum inclusion conditions for volatile oil from three medicinal slices in Xiongdan Xiaoyan capsules.</p><p><b>METHOD</b>Saturated water solution method (agitation method), ultrasonic method and rubbing method were studied and compared. The preparation conditions of ultrasonic method were investigated by the orthogonal test.</p><p><b>RESULT</b>The optimum inclusion conditions of ultrasonic method were established.</p><p><b>CONCLUSION</b>The ultrasonic method can be used for production of inclusion complex in the factory with high inclusion rate and was effectiveness.</p>
Subject(s)
Angelica , Chemistry , Capsules , Drug Carriers , Drug Combinations , Drug Stability , Drugs, Chinese Herbal , Mentha , Chemistry , Oils, Volatile , Plants, Medicinal , Chemistry , Saururaceae , Chemistry , Technology, Pharmaceutical , Methods , Ultrasonics , beta-CyclodextrinsABSTRACT
<p><b>AIM</b>To prepare TK-gene nanoparticles and investigate its expression.</p><p><b>METHODS</b>Biodegradable and biocompatible polymer polylactic-co-glycolic acid (PLGA) was used to prepare recombinant plasmid pEGFP-AFP nanoparticles by double-emulsion evaporation technique. The characteristics of the nanopticicles including morphology, entrapment efficiency was investigated. The expression of TK gene was also investigated by MTT assay, which could determine the dying cells after the addition of gancyclovir (GCV). The enhanced green fluorescent protein (EGFP) expression in human hepatocarcinoma SMMC-7221 cells and human normal parenchymal Chang liver cells were assessed by flow cytometric analysis.</p><p><b>RESULTS</b>The resulting plasmid-nanoparticles had regular spherical surface and a narrow particle size with a mean diameter of (72 +/- 12) nm, The average entrapment efficiency was 91.25%, the enhanced transfection efficiency and ability protecting plasmid DNA from degraded by nuclease or sonication due to nanoparticles encapsulation.</p><p><b>CONCLUSION</b>DNA-nanoparticles need further study as gene delivery system.</p>
Subject(s)
Humans , Biocompatible Materials , Carcinoma, Hepatocellular , Metabolism , Cell Line, Tumor , Drug Delivery Systems , Ganciclovir , Pharmacology , Genes, Reporter , Green Fluorescent Proteins , Herpesvirus 1, Human , Lactic Acid , Liver , Cell Biology , Metabolism , Liver Neoplasms , Metabolism , Luminescent Proteins , Genetics , Metabolism , Nanotechnology , Particle Size , Plasmids , Polyglycolic Acid , Polymers , Recombinant Proteins , Genetics , Thymidine Kinase , Genetics , Metabolism , Transfection , alpha-Fetoproteins , Genetics , MetabolismABSTRACT
<p><b>AIM</b>To increase the accumulation of mitoxantrone in solid tumor by synthesis and characterization of N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer-mitoxantrone conjugate (p-DHAQ).</p><p><b>METHODS</b>HPMA copolymer-mitoxantrone conjugate was prepared by free radical precipitation copolymerization method. The in vitro stability of conjugate was investigated under different conditions. Biodistribution was examined in mice bearing Ehrlich solid tumor.</p><p><b>RESULTS</b>The p-DHAQ conjugate was characterized by UV, HPLC and size exclusion chromatography. The conjugate was stable in buffers of different pH and in mice plasma while the rate of drug liberation was faster in tumor. It appeared that the circulation lifetime of HPMA copolymer-bound mitoxantrone were three times more than that of the drug in free form. The AUC of p-DHAQ was three times more than the AUC of free drug. The p-DHAQ level in heart was five times lower than free drug. This reduces the possibility of toxicity to the heart.</p><p><b>CONCLUSION</b>HPMA copolymer-mitoxantrone conjugate was successfully synthesized and characterized. The biodistribution results showed the possibility of targeting anticancer drug-mitoxantrone with secondary amino residue to the tumor tissue by HPMA copolymer as carrier.</p>