The animal study was approved by the Animal Care and Use Committee of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. Twenty 6–8 weeks-old female NOD-PrkdcscidIl2rgem1/Smoc mice were anesthetized with 1%–1.5% isoflurane (R510-22-10, RWD Life science, Shenzhen, China). Subsequently, patient-derived xenografts (PDXs) were engrafted subcutaneously on the dorsum. After inoculation, mice weights and tumor sizes were measured twice weekly. When the tumors reached 50–100 mm3, mice were randomized into four subgroups. Niraparib was dosed (20 mg/kg, intragastric administration) once daily for 5 days per week while DAHP was dosed (60 mg/ml, intraperitoneal administration) once daily. Tumor volume based on caliper measurements will be calculated using the modified ellipsoidal formula: tumor volume = 1/2 length × width2. At experimental endpoints, the mice will be sacrificed after anesthesia. Tissue samples were taken for routine hematoxylin and eosin (HE) staining and immunohistochemistry.
R510 22 10
Manufactured by RWD Life Science
Sourced in China
The R510-22-10 is a laboratory equipment product. It is designed to perform specific functions within a laboratory setting. No further details about its core function or intended use are provided, as an unbiased and factual description while maintaining conciseness cannot be reliably given without the risk of interpretation or extrapolation.
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7 protocols using r510 22 10
1
Niraparib and DAHP for PDX Tumor Suppression
2
Cecal Ligation and Puncture Model in Rats
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The Sprague Dawley (SD) rats were purchased from Shanghai Jihui Laboratory Animal Care Co., Ltd. (SCXK (Hu) 2017-0012). All procedures on animals followed the Public Health Service Policy on Humane Care and Use of Laboratory Animals. After 7 days of adaptive feeding, the 200–230 g SD rats were divided into three groups for CLP model construction: the sham group, the CLP group, and the CLP + BBR group. Before the CLP surgery, SD rats were injected intraperitoneally with BBR (berberine hydrochloride, dose: 50 mg/kg; B21449, Yuanye, Shanghai, China) or PBS (as a solvent control) one time a day for 5 days. The CLP rats were anesthetized with isoflurane (R510-22-10, RWD, San Diego, CA, USA) and then cut from the middle of the abdominal wall. The cecum was gently pulled out to ligate at one-third from the end of the cecum with sterile No. 4 thread. Then, a 1 ml injection needle (21 G) was used to puncture the perforation in the middle between the ligature site and the top of the cecum about 3–4 times. The rats were sutured and resuscitated with 5 ml/100 g of normal saline. The sham rats underwent surgery similar to that performed on CLP rats, except for cecal ligation and puncture.
3
Cervical Lymph Node Removal in Rats
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The rat was anesthetized using oxygen and isoflurane (5%) (R510-22-10, RWD Life Science Co., Ltd., China) with a flow rate of 1.0 L/s in an induction chamber. To verify that the rat was asleep, its tail was pinched and rolled. If the rat did not react, the surgery was performed. Cervical lymph nodes (CLNs) were removed. Ointment was applied on the rat eyes before surgery to avoid eye dryness. A very small incision (approximately 5 mm) in the neck was made with sharp scissors. The incision was stretched with 2 forceps to see the LNs. The incision depth could reach 10 mm. The superficial CLN appeared grayish or darker than the surrounding fat. The fascia (the thin membrane covering the fat and tissue) on top of the LN was pinched with one set of forceps and pulled lightly without breaking the surrounding tissue. The second set of forceps was placed as far as possible underneath the LN. With the set of first forceps, the fascia was broken, and the superficial CLN was removed. The absence of hairs in the wound was verified.
4
Ultrastructural Analysis of Rat Tissues
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Rats were first anesthetized by inhalation with 3% isoflurane (R510-22-10, RWD Life Science) for induction and then 2% isoflurane (2–3 L/min) for maintenance. Next, rats were transcardially perfused with 0.1 M sodium carbonate in 2% paraformaldehyde and 2% glutaraldehyde, and then fixed in 0.1 M sodium dimethyl arsonate with 2% osmium tetroxide and 1.6% potassium ferrocyanide. Tissue samples were cut into sections (70–90 nm). The sections were dehydrated in ethanol and embedded in ebonite. Sections were then deposited onto a copper channel grid. Finally, 2% uranyl acetate (SPI-02624, HEAD) and lead citrate (HD17810, HEAD) were used to stain the sections. Transmission electron microscopic images were obtained by a transmission electron microscope (Tecnai F20, FEI, Hillsboro, Oregon, USA).
5
Low-Intensity Pulsed Ultrasound Stimulation for CKD
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The LIPUS device consists of an arbitrary waveform function generator (SDG1022X, Siglent, Shenzhen, China), an oscilloscope (TDS2022B, Tektronix, Shanghai, China) and ultrasound transducers (Φ = 3 cm). Among them, the ultrasound transducers were specially manufactured and tested by the Academy of Electronics and Engineering (the University of Electronic Science and Technology of China, Chengdu, China) for rat in vitro experiments. Before LIPUS stimulation, hair on the back of each rat was removed, the positions of bilateral kidneys were determined and marked by ultrasound imaging. Then, the ultrasound transducers were placed over the back marker with an ultrasonic gel pad (Mibo, Mp-A, Shenzhen, China) in between. Rats in the CKD + LIPUS group were stimulated by LIPUS for 20 min every 24 h, treatment duration was four weeks. During this period, isoflurane (R510-22-10, RWD, Shenzhen, China) was produced by the gas anaesthesia machine (MSSVAP01, MSS, Keighley, England) to keep rats under anaesthesia. According to previous studies [10 (link),11 (link)] and our preliminary experimental results, the parameters of LIPUS are set as follows: intensity of 60 mW/cm2 with a 50% duty cycle, and pulse frequency of 1.0 MHz with 1 kHz repetition rate. As for the Control group and the CKD group, which received the same treatments except LIPUS stimulation.
6
Cardiac Function Assessment in Rats
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At the end of the drug intervention period, the rats were anaesthetized with 2.5% isoflurane (RWD Life Science, R510-22-10), followed by an anaesthetic respiratory system (1.5% isoflurane to maintain anaesthesia), a M-mode ultrasound machine (GE Versana Premier) and L8-18i-RS ultrahigh-frequency line-array probe were performed to measure heart rate (HR), left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) from the left parasternal axis using two-dimensional echocardiography for five consecutive cardiac cycles.
7
FDG PET/CT Imaging of Tumor-Bearing Mice
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FDG was produced routinely in GE TracerLab with a commercially available module following the standard protocols with quality control and radiochemical purity reaching 99%. Six hours before the imaging session, the mice were fasted, and water was given ad libitum. Approximately 200 μCi 18F-FDG was injected into tumor-bearing mice via the tail vein. A total of 15 min static scanning was performed at 1 h after injection with mice under isoflurane (R510-22-10, RWD, Shenzhen, China) anesthesia using a SuperArgus small-animal PET/CT scanner (Sedecal, Madrid, Spain). Images were reconstructed and regions of interest (ROI) were manually drawn around tumors.
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