Animals Healthy female SCID mice aged about 4 weeks were purchased from Shanghai Experimental Animal Center of Chinese Academic of Sciences (Shanghai, China), housed in specific pathogen-free conditions, and treated in accordance with guidelines of the Committee on Animals of Changhai Hospital affiliated to the Second Military Medical University (Shanghai China). Pharmacokinetics and in vivodistribution analysis The pharmacokinetics (PK) and in vivo distribution analysis was done following Joseph
M. Tuscano’s study with minor revisions . Briefly, Daudi cells (1 × 107) were inoculated subcutaneously into the right flank of 6-week-old SCID mice. For PK assays, when tumors reached about 50 to 60 mm3 Selleck Gilteritinib in volume (approximately 14 days), mice were randomly administrated tail vein injection of free ADR, non-irrad or irrad ADR-containing immunoliposomes at a dosage of 5 mg ADR/kg (n = 3 mice per treatment). Then, 10 μL of blood were collected through tail vein nicking from each mouse at 5, 15, and 30 min and 1, 2, 4, 6, 8, 12, 24, and 48 h after treatment, respectively. Samples were immediately diluted into 250 μL of 0.5 mmol/L
EDTA-PBS, followed by a centrifugation AG-881 clinical trial (300 g × 5 min). Plasma was collected and ADR was extracted by acidified isopropanol (75 mmol/L hydrochloric acid in 90% isopropanol) at 4°C for 20 h. The ADR concentrations were measured by UV at 480 nm and expressed as micrograms per milliliter (ADR/blood plasma). The data were analyzed by the PK solver software . For biodistribution assays, tumor-bearing mice were randomly administrated tail vein injection of free ADR, PC-ADR-BSA, or PC-ADR-Fab at a dosage of 5 mg ADR/kg (n = 3 mice per treatment). Mice were sacrificed 24 h after treatment; part of tumor, heart, liver, spleen, kidneys, and lungs were removed, washed, and weighed; and single-cell suspensions were made. ADR
was extracted from cells by the abovementioned acidified isopropanol for 20 h at 4°C. The ADR concentrations were determined as described above and expressed as micrograms per gram (ADR/tissue). What’s more, part of the tumor tissues were collected and subjected to frozen PTK6 sections, which were detected by a confocal microscrope (Zeiss, Oberkochen, Germany). In vivoantitumor activity assessment in disseminated human NHL xeno-transplant models Six-week-old SCID mice were injected via the tail vein with 5 × 106 Daudi cells in 100 μL PBS. Then, the inoculated mice were randomly assigned to 4 groups with 10 each for the treatment of PBS, free ADR, PC-ADR-BSA, and PC-ADR-Fab (with an equivalent amount of 5 mg/kg ADR) via the tail vein weekly for 3 times after 48 h. QNZ clinical trial Post-operation monitoring was exercised at least once a day until natural death in a range of 120 days.