Uv spectrophotometer

Electronic micro-balance (Changsha Xiangping Science and Technology Development Co., Ltd., Changsha, China); ice maker (Sanyo Electric Co., Ltd., Moriguchi, Japan); −70°C ultra-low temperature freezer (Forma Scientific, Inc., Marietta, OH, USA); thermostat-controlled water-bath (Ningbo Xinzhi Biotechnology Co., Ltd., Ningbo, China); oscillation mixing device (Shanghai Huayun Analytical Instrument Co., Ltd., Shanghai, China); cryogenic supercentrifuge (DuPont, Wilmington, DE, USA); table concentrator (Barnstead International Co., Ltd., Boston, MA, USA); UV spectrophotometer (Hitachi, Tokyo, Japan); agarose gel electrophoresis apparatus (Beijing Liuyi Instrument Factory, Beijing, China); UV gel imaging system (Bio-Rad, Hercules, CA, USA); DNA Engine PCR instrument (University of Leicester, London, UK); clean bench (Beijing Semiconductor Equipment Factory, Beijing, China); CO₂ incubator (Thermo Electron Corp., Waltham, MA, USA).

The collected PCa cancer tissue and PCa adjacent tissues was performed with total RNA extraction by using TRIzol reagent, in strict accordance with the instructions for extraction. The concentration and purity of extracted RNA were detected by UV spectrophotometer (Hitachi, Tokyo, Japan). The OD value of the total RNA solution: A260/A280 ranged from 1.8–2.1, and if not, it was extracted again. The integrity of RNA was detected by 1% denaturing agarose gel electrophoresis. The total RNA was subjected to reaction system disposition and reverse transcription to cDNA by miRNA reverse transcription kit instructions (and stored at −20°C for later use). ABI StepOne Plus fluorescence quantitative PCR instrument was adopted, and the reaction system was according to the instructions. Reaction conditions: 95°C for 5 min; 95°C for 45 sec; 60°C for 60 sec; 72°C for 45 sec and 45 cycles. U6 was used as internal reference. The experiment was repeated 3 times, and the results were analyzed by using the 2^−ΔΔCq method (10 (link)).

Animals were injected with 2 g/l Evans blue dye (2 ml/kg) through the ear vein 1 h prior to sacrifice. Animals were perfused with 200–300 ml normal saline to remove Evans blue dye from the blood vessels. Brain tissues from the bilateral temporal lobes were carefully removed from the animals following sacrifice. A total of 3 ml formamide was added to 0.5 g of the brain tissue homogenate to dissolve the Evans blue dye. After incubation in a 37°C water bath for 48 h, the samples were centrifuged at 755 × g for 5 min. The supernatant was then collected before absorbance measurement at 632 nm was performed using a UV spectrophotometer (Hitachi, Ltd.). Evans blue content was calculated according to the standard curve.

All chemicals used in this study were of analytical grade and were obtained from
Merck (Darmstadt, Germany), while the nutrients and agar media were purchased from
HIMEDIA (Mumbai, India). Carbaryl was obtained from Dr. Ehrenstofer GmbH (Augsburg,
Germany). RP-C18 Bakerbond SPE 3 mL, 500 mg cartridges (J. T. Baker, Gross-Gerau,
Germany) were used to enrich samples, UV-Spectrophotometer and Dual wavelength flying
spot scanning densitometer were from Hitachi (Tokyo, Japan) and the solid phase
extraction (SPE) unit was from Supelco (Sigma-Aldrich, Missouri, USA).

Total RNA was extracted and the concentration was measured using a UV spectrophotometer (Hitachi, Ltd., Tokyo, Japan) and the integrity of RNA was examined by electrophoresis (A260/A280 ratio was 1.8–2). cDNA FastQuant was used for cDNA synthesis. The reaction conditions were: 95°C for 3 min and 42°C for 15 min, and then kept on ice. Primer sequences are presented in Table I.

A total of 10 µl CNP (100 mg/l) and CNPs@DOX (100 mg/l) solutions were dripped onto carbon-coated 400-mesh copper grids by pipette gun and air-dried for 24 h to prepare the sample for transmission electron microscopy (TEM; Hitachi, Ltd.) observation. The ζ potentials of aqueous CNPs and CNPs@DOX were measured using dynamic light scattering (Malvern Instruments, Ltd.). The ultraviolet (UV)-visible (vis) absorption and fluorescence spectra were measured using a UV spectrophotometer (Hitachi, Ltd.) and fluorescence spectrophotometer (Hitachi, Ltd.), respectively.