In the antitumor study, TFU-LNS were successfully used in the treatment of H 22 -bearing Kunming mice after intraperitoneal administration. The inhibitory rates of tumor weight and tumor volume were consistent with those of 5-FU injection. The body weight of mice could be monitored as an index of systemic toxicity. From the varia- tion of body weight in mice, blank LNS had no significant toxicity compared with NS during the therapeutic procedure. One reason was that absolutely no organic solution was added during the preparation of LNS, and another reason was the security of phosphatidylcholine. Phosphatidylcholine is one of the few injectable surfactants that have been used in US Food and Drug Administration-approved pharmaceuti- cal products and have well-established safety profiles and toxicological data. 35
The authors aimed to develop a MNP drug delivery system for intratumoral administration. A single emulsion evaporation method with magnetic Fe 3O 4 cores and a shell of biocompatible polymeric PLGA [poly(D,L-lactic-co-glycolic acid)] was used to prepare the MNPs. The physicochemical properties of the DOX-loaded MNPs (DOX-MNPs) were characterized in terms of morphology, size distribution, and drug loading content. In vitro release profiles of DOX from DOX-MNPs were examined in both acidic and neutral environments. The in vitro anticancer activity of DOX-MNPs was determined using a Lewis lung carcinoma (LLC) cell line, and the apoptotic rate was analyzed by flow cytometry. The uptake of nanoparticles by the LLC cells was visual- ized using confocal microscopy. Finally, in vivo antitumor activity was assessed by a single intratumoral injection of DOX-MNPs into C57BL/6 mice bearing subcutaneously established LLC.
Abstract: Over the last decade, magnetic iron oxide nanoparticles (IONPs) have drawn much attention for their potential biomedical applications. However, serious in vitro and in vivo safety concerns continue to exist. In this study, the effects of uncoated, Fe m O n -SiO 2 composite flake-like, and SiO 2 -Fe m O n core-shell IONPs on cell viability, function, and mor- phology were tested 48 h postincubation in human umbilical vein endothelial cell culture. Cell viability and apoptosis/necrosis rate were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay and annexin V-phycoerythrin kit, respectively. Cell mor- phology was evaluated using bright-field microscopy and forward and lateral light scattering profiles obtained with flow cytometry analysis. All tested IONP types were used at three different doses, that is, 0.7, 7.0, and 70.0 µg. Dose-dependent changes in cell morphology, viability, and apoptosis rate were shown. At higher doses, all types of IONPs caused formation of binucle- ated cells suggesting impaired cytokinesis. Fe m O n -SiO 2 composite flake-like and SiO 2 -Fe m O n core-shell IONPs were characterized by similar profile of cytotoxicity, whereas bare IONPs were shown to be less toxic. The presence of either silica core or silica nanoflakes in composite IONPs can promote cytotoxic effects.
In recent years, there has been interest in perovskite type of metal oxides containing manganese. These materials show interesting magnetic and electric properties, such as magneto resistivity, nanoscale charge ordering and high oxygen ion conductivity, opening the way to potential applications [1–3]. Gas detection problem was overcome by modified metal oxide semiconducting material. Due to the different applications and inherent limitations of different gas sensing technologies, researchers have been working on different scenarios with enhanced gas sensor calibration. Since 1962 it has been known that absorption or desorption of a gas on the surface of a metal oxide changes the conductivity of the material, so it brings high value of nanocrystalline thick film technology. This phenomenon being first time reported by Sieyama et.al.  for zinc oxide thick film.
The MTT assay is an important method for evaluating the in vitro cytotoxicity of biomaterials. In the MTT assay, absorbance has a significant linear relationship with cell numbers. Corresponding optical images of cells are shown in Figure 15. In the current work, the MTT assay showed that Fe 3O 4 -PLGA-PEG 2000 -doxorubicin has dose-dependent and time-dependent cytotoxicity against the A549 lung can- cer cell line (IC 50 0.13–0.26 mg/mL). Also, the MTT assay showed that Fe 3O 4 -PLGA-PEG 3000 -doxorubicin has dose- dependent and time-dependent cytotoxicity against the A549 lung cancer cell line (IC 50 0.08 mg/mL), that Fe 3O 4 -PLGA- PEG 4000 -doxorubicin has no dose-dependent cytotoxicity but does have time-dependent cytotoxicity against the A549 lung cancer cell line (IC 50 0.17–0.48 mg/mL), and that pure doxorubicin has dose-dependent but not time-dependent cytotoxicity against this cell line (IC 50 0.15–0.16 mg/mL). Therefore, there is a need for further study of doxorubicin- loaded Fe 3O 4 magnetic nanoparticles modified with PLGA- PEG copolymers using the A549 lung cancer cell line in the future. However, the results of the current work demonstrate that the IC 50 values for Fe 3O 4 -PLGA-PEG 4000 -doxorubicin, Fe 3O 4 -PLGA–PEG 3000 -doxorubicin, Fe 3O 4 -PLGA-PEG 2000 - doxorubicin, and pure doxorubicin are about 0.18 mg/mL, 0.08 mg/ml , 0.13 mg/mL, and 0.15 mg/mL, respectively, in this cell line.
In order to examine the expression of caspase-3, bax, bcl-2, NF- κ B, and survivin, we next performed Western blot analysis on whole cell protein extracted from cells treated for 48 hours as described previously. In brief, total protein was isolated on ice and subjected to 10% sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels using modified radio immunoprecipitation assay buffer, and transferred to a polyvinylidene difluoride membrane (Bio-Rad). Western blotting was performed with a 1:1000–1200 dilutions of monoclonal antibodies against either anti-human caspase-3, bax, bcl-2, NF- κ B, surviving, or β -actin anti-body in 5% nonfat dry milk, and then with horseradish peroxidase-conjugated goat anti-rabbit (1:5000) as a secondary antibody. The band was detected by using an enhanced chemiluminescence detection system (Amersham, Buckinghamshire, UK).
All animals were kept in stress-free, hygienic, and animal- friendly conditions (relative humidity 60% ± 10%, room tem- perature 20 ° C ± 2 ° C, and 12-hour light/dark cycle). Food and water were available ad libitum. All animal-study protocols were approved by the Ningbo University Institutional Animal Care and Use Committee (AEWC-2016–151), and experi- ments followed the Chinese “Regulation on the management of laboratory animals”, adopted in 1988. Healthy male rats (12 weeks old) were selected and randomly divided into one control group and six experimental groups (5, 15, and 30 mg/kg Fe 3O 4 -TiO 2 NPs or TiO 2 NPs, respectively) with six rats in each group. The observation period was 30 days. A staged approach to dosing method was used for selection of the dose ranges. When determining chemical dosage, care was taken to prevent any severe toxicity in the animal that could affect absorption, metabolism, distribution, or excretion. 22 Following this principle, the highest treatment
Abstract: While the potential impact of magnetic nanoparticles (MNPs) has been widely explored in almost all medical fields, including cardiology, one question remains; that is whether MNPs interfere with cardiac physiological processes such as the expression and function of ion channels, especially in vivo. KCNQ 1 channels are richly expressed in cardiac myocytes and are critical to the repolarization of cardiac myocytes. In this study, we evaluated the effects of Fe 3O 4 -magnetic nanoparticles (MNPs-Fe 3O 4 ) on the expression of KCNQ 1 in cardiac muscle of mice at rest and at different times following a single bout of swimming (SBS). Firstly, we demonstrated that the expression levels of KCNQ 1 channels are significantly up-regulated in mice following a SBS by means of reverse transcription polymerase chain reaction (RT-PCR) and western-blot. After treating mice with normal saline or pure MNPs-Fe 3O 4 separately, we studied the potential effect of MNPs-Fe 3O 4 on the expression profile of KCNQ 1 in mouse cardiac muscle following a SBS. A SBS increased the transcription of KCNQ 1 at 3 hours post exercise (3PE) 164% ± 24% and at 12 hours post exercise (12PE) by 159% ± 23% (P 0.05), and up-regulated KCNQ 1 protein 161% ± 27% at 12PE (P 0.05) in saline mice. In MNPs-Fe 3O 4
overexpression of NF- κ B might help circumvent resistance to conventional therapies. Our results showed that either adriamycin or daunorubicin increased apoptosis of Raji cells and that this was accompanied by upregulation of p53 protein and downregulation of NF- κ B protein. Interestingly, in the present study, MNP-Fe 3O 4 combined with adriamycin or daunorubicin increased upregulation of p53 protein and downregulation of NF- κ B protein, but similar results were not found with dexamethasone. Our findings for NF- κ B were similar to those reported by Ottonello et al, inhibition of NF-kB activation decreased the production of XIAP, an antiapoptotic molecule, leading to uncontrolled activity of regulators of apoptosis (eg, caspase 3). 26 Dexamethasone-
pro-caspase 9, caspase 9, pro-caspase 3, and caspase 3. In brief, total protein was isolated on ice and subjected to 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis using modified radioimmunoprecipitation assay buffer, and then transferred to a polyvinylidene difluoride membrane (65421, Pall Corporation, Port Washington, NY). The membrane was blocked with buffer containing 10% fat-free dry milk and performed with 1:200 dilution of monoclonal antibodies against either antihuman PI3K, p-PI3K, Akt, p-Akt, Bad, p-Bad, cytochrome c, pro-caspase 9, caspase 9, pro-caspase 3, caspase 3, or β -actin antibody, and then sub- sequently incubated with horseradish peroxidase-conjugated goat antirabbit (1:5000) as a secondary antibody. The band was detected using an enhanced chemiluminescence detec- tion system (Amersham, Buckinghamshire, UK). Each value is presented as the relative density of protein bands normalized to β -actin.
Forty ICR mice, half male and half female, were individually and randomly divided into four groups: control group (0.5 mL of sterile physiologic saline), low-dose group (300 mg/kg Fe 3O 4 - MNPs), medium-dose group (600 mg/kg Fe 3O 4 -MNPs), and high-dose group (1200 mg/kg Fe 3O 4 -MNPs). The mice in the exposed group were given different concentrations of Fe 3O 4 - MNPs, and those in the control group were given RPMI-1640 (Gibco Chemical Co, Carlsbad, CA) medium by single gastric perfusion and observed for the poisoning symptoms after administration of the drug; finally, they were all sacrificed under Avertin anesthesia by cervical dislocation after 14 days and
Abstract: Topical vitamin D 3 ointments are widely used to treat psoriasis, sometimes in combination with cyclosporine, phototherapy, and biologic agents. However, the risk factors for hypercalcemia resulting from these ointments, and interactions with underlying disorders and age are unclear. We performed a retrospective study of psoriasis patients at Nagoya City University Hospital between January 1, 2004, and December 31, 2009, treated with a vitamin D 3 -containing topical drug, either calcipotriol, maxacalcitol, or tacalcitol. Data from blood samples and clinical scores collected during routine visits throughout the study period were analyzed. We assessed changes in the serum calcium levels over time in association with age, liver dysfunction, renal dysfunction, concomitant medication, and concomitant therapy. Serum calcium levels were significantly lower in the calcipotriol group than in the maxacalcitol group (P , 0.05), regardless of other factors, at the observation period. Calcipotriol was associated with lower serum calcium levels than maxacalcitol in patients $ 65 years (P , 0.05), those with renal disease (P = 0.0362), and those with liver disease (P = 0.0255). Only three patients using calcipotriol developed hypercalcemia that did not seem to be related to the treatment. Hypercalcemia was observed in 10 patients using maxacalcitol, although serum calcium levels rapidly recovered when use was discontinued. Only one patient using tacalcitol developed hypercalcemia. Hypercalcemia tended to occur in patients with conditions in which the skin is more vulnerable, even at standard doses; patients taking oral etretinate; patients requiring concomitant systemic therapy, even if the Psoriasis Area and Severity Index score was not severe; and patients with renal or liver dysfunction. These findings highlight the importance of the awareness of the risk factors for hypercalcemia in patients treated with topical vitamin D 3 ointment, and point to calcipotriol as the first-choice treatment for those over 65 years old, or those with renal or liver disease.
Our present work focuses on enzyme-enriched, magnetic functionalized nanoparticles in signal amplification and separation for ultrasensitive detection of AFP. Magnetic composite Fe 3O 4 /Au nanoparticles were synthesized by assembling gold nanoparticles on SH-modified Fe 3O 4 and using them as nanocarriers for immobilization of HRP and a secondary antibody (Ab 2 ). Greatly amplified sensitivity is achieved using bioconjugates featuring HRP and Ab 2 linked to Fe 3O 4 /Au at a high HRP/Ab 2 ratio for replacement of singly labeled secondary antibodies. A modified glass carbon electrode with porous nanostructured gold film was used to attach the primary antibody (Ab 1 ). The analytical procedure consists of immunoreaction of the antigen (AFP) with Ab 1 , followed by binding of Fe 3O 4 /Au-HRP-Ab 2 . Electrochemical detection was performed in the presence of H 2 O 2 and hydroquinone (Figure 1). The results demon- strated that an immunosensor based on this amplification strategy has a good dynamic range from 0.005 to 10 ng/mL and a low detection limit of 3 pg/mL for AFP. This simple, cost-effective, and sensitive immunosensor could find wide potential application in clinical analysis.
After treatment with drugs, total RNA was isolated using Trizol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) according to the manufacturer’s protocol. One microgram of total RNA was used to generate cDNA using SuperScript™ II reverse transcriptase (Invitrogen Life Tech- nologies). PCR primers were designed to amplify products within target and control sequences. (Primer sequences: P53(422bp):forward:5'-ACCCAGGTCCAGATGAAG-3'; reverse:5'-CACTCGGATAAGATGCTGA-3'.Caspase- 3(270bp):forward:5'-GCTATTGTGAGGCGGTTGT- 3',reverse:5'-TGTTTCCCTGAGGTTTGC-3'.GAPDH(205bp): forward:5'-CGGATTTGGTCGTATTG-3';reverse:5'- GAAGATGGTGATGGGATT-3') QPCR was performed by monitoring in real-time the increase in ﬂ uorescence of SYBR green I dye (Takara, Shiga, Japan) with Rotor-Gene 3000 (Corbett Research, Sydney, Australia). Each experi- ment was done in triplicate. The relative gene copy number was calculated by the concentration-CT standard curve method and normalized using the average expression of GAPDH. 19
in 6-well plates were treated with one of the following treatments: (a) DDP + MNPs-Fe 3O 4 , (b) DDP, (c) MNPs- Fe 3O 4 , (d) RPMI-1640 for 48 h, and harvested. Cells were dissolved in TRIzol reagent (Gibco BRL). Total RNA was extracted according to the manufacturer’s instructions. The RNA A 260 /A 280 ratios were between 1.6 and 1.8. The primers for human Bcl-2 (forward: 5’-GGGAGAACAGGGTAC GATAA-3’; reverse: 5’-CCACCGAACT CAAAGAAGG-3’), and β -actin (forward: 5’-TATGACTTAGTTGC GTTA- CACC-3’; reverse: 5’-CCTTCAC CGTTCCAGTTT-3’) were used. The amplified polymerase chain reaction (PCR) products were 452 bp and 155 bp, respectively. The copy number for each sample was calculated and all the data were normalized to β -actin. Briefly, cDNA was synthesized from 1 µ g of total cellular RNA using TaKaRa RNA PCR kit (AMV) (Ver. 3.0; Dalian, China). The newly synthesized cDNA was amplified by PCR (TaKaRa). The PCR conditions were 95 ° C for 3 min and 35 cycles of 95 ° C for 30 s, 56 ° C for 30 s, and 72 ° C for 1 min. Control amplifications were conducted either without reverse transcription (RT) or without RNA. Following PCR amplification, the reaction products were electrophoresed at 100 V on 1.5% agarose gels with 0.5 µ g/ml ethidium bromide (Sigma Aldrich) 16 and PCR fragments were visualized by UV
Researches on the nanofluids’ behaviors have almost commenced since the beginning of the twenty first century and annual publication of nanofluids has remarkably increased from 1999 to 2005 that has grown to more than 70% . In the previous decade, many studies have carried out about heat transfer of nanofluids in both experimental and numerical methods for different type and size of particles in various geometries. Although the thermal conductivity of nanofluids is considered as an encouraging feature for their use, a significant role is contributed to convection as it is a criterion of applicability and shows superiority of their heat transfer in comparison with other fluids. Wen and Ding  did an experimental study on nanofluids convective heat transfer in entrance region of a channel containing γ- Al 2 O3 particles
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich Co. (St Louis, MO, USA). RPMI 1640 medium was from Gibco/BRL (Thermo Fisher Scientific, Waltham, MA, USA). Newborn calf serum was from Hangzhou Sijiqing Biological Engineering Materials Co., Ltd. (Hangzhou, People’s Republic of China). Annexin V–Fluorescein Isothiocyanate Apoptosis Detection Kit and ECL (enhanced chemiluminescence) Western Blotting Detection Kit were from Nanjing KeyGen Biotech Co., Ltd. (Nanjing, People’s Republic of China). Monoclonal anti- bodies, including caspase-3, caspase-8, survivin, and Bcl-2, were supplied by Santa Cruz Biotechnology Inc. (Dallas, TX, USA).
After incubating with 0.1 (v/v) magnetic nanoparticles for 48 hours at 37 ° C, the cells were harvested and made into cell smears. Before observation by optical microscopy, the cells were stained with Prussian blue. The cells on the slide were continuously incubated for 30 minutes in 2% potassium fer- rocyanide and 6% hydrochloric acid, and then counterstained with nuclear fast red for 3 minutes. The smears were viewed under an optical microscope (B × 41M, Olympus, Tokyo, Japan) at 1000 × amplification.
indicated that the Fe 3O 4 -NPs are spherical with a diameter range between 3.21 and 2.22 nm. The VSM study demonstrated that the magnetic properties were enhanced with the decrease in the per- centage of honey. In vitro viability evaluation of Fe 3O 4 -NPs performed by using the MTT assay on the WEHI164 cells demonstrated no significant toxicity in higher concentration up to 140.0 ppm, which allows them to be used in some biological applications such as drug delivery.
approach for reducing the total cost of electricity generation [5-7]. It is well established that the use of suitable catalyst at the electrode of SOFC is the key issue for achieving the good SOFC cogeneration performance. Recently, several types of catalyst for OCM reaction have been studied and reported, from which the suitable catalysts for this reaction include SrF 2 /Nd 2 O3 , Mn/Na 2 WO 4 /SiO 2 , La 2 O3 , LaAlO 3 ,