26 ghamilton microsyringe

All
animal procedures were approved by the Ethical Committee on Animal
Experiments of the University Medical Center Utrecht and Utrecht University,
(DEC: 2012.I.11.116) and conducted in accordance with the guidelines
set by the European Community Council Directives 86/609/EEC. Experiments
are reported in compliance with the ARRIVE 2.0 guidelines (Animal
Research: Reporting in Vivo Experiments).
Male Wistar rats (300–330
g) (Harlan, Horst, The Netherlands) were endotracheally intubated
and mechanically ventilated with a mixture of 2% isoflurane in 70%
air/30% O₂ for anesthesia, and placed in a stereotaxic
apparatus (Kopf Instrument, Tujunga, CA, USA). The skull was exposed
between bregma and lambda. A burr hole was drilled using a dental
drill during continuous irrigation with 0.9% saline at room temperature
to prevent overheating of the underlying cortex.
Five microliters
of each hydrogel formulation was stereotaxically
injected over a 10 min period in the right striatum (stereotaxic coordinates
from bregma (mm): AP, 0.3; ML, 3.0; DV, 4.0 from dura) using a 26-G
Hamilton microsyringe (80330, Hamilton Company, Reno, NV). The following
hydrogels were used: LC Thermo (blank: n = 2; Galbumin-loaded: n = 3) and HC Thermo (blank: n = 3; Galbumin-loaded: n = 3). In each case, the needle was left in place after
injection for 7 min before being slowly withdrawn.

Experimental groups were assigned in a randomized and blinded manner. The rats were divided into five groups: (1) sham-operated rats housed in SE (Sham); (2) MCAO rats housed in SE (MCAO); (3) MCAO rats housed in EE (EE); (4) EE rats treated with anti-IL-17A mAb (EE + anti-IL-17A); (5) EE rats treated with mouse IgG1 isotype (EE + IgG1 isotype). In more detail, for group (4), anti-IL-17A mAb (2 μg/rat; Sigma-Aldrich, St Louis, MO, USA) was administered intracerebroventricularly (i.c.v.) into the left lateral ventricle (coordinates:+0.8 mm posteriorly, +1.5 mm lateral from the bregma, –3.5 mm depth) using a stereotaxic instrument (ZH-Lanxing B-type stereotaxic frame, Anhui Zhenghua Biological Equipment Co., Ltd., Huaibei, China) every 24 h for 1 week starting at 14 days post-ischemia (dpi). I.c.v. injection was performed with a sterile 26G Hamilton microsyringe (80330; Hamilton Company, Reno, NV, USA). Moreover, for group (5), 2 μg IgG1 isotype control anti-IL-17A mAb was administered i.c.v. into the left lateral ventricle of EE rats every 24 h for 1 week starting at 14 dpi. A schematic representation of the experimental timeline is presented in Figure 1A.

Forty-eight Sprague Dawley rats (male, 6–8 weeks, 240–280 g) were purchased from Vital River (Beijing, China).
The rats were randomly divided into six groups: control, control + esketamine, propofol, propofol + esketamine, propofol + esketamine + IgG and propofol + esketamine + anti-BDNF (n = 8/group). Rats in the propofol-treated groups were intraperitoneally injected with 50 mg/kg propofol (Sigma–Aldrich, Louis, MO, USA), followed by an injection of 50 mg/kg propofol again after 1 h when the rat righting reflex recovered [19 (link)]. Rats in the control group were administrated with 100 mg/kg intralipid [19 (link)]. Thirty minutes after propofol injection, rats in the esketamine-treated groups were intraperitoneally injected with 15 mg/kg esketamine (Cristalia, Brazil) [20 ]. Normal saline (NS) was used as a control for esketamine.
In some experiments, to examine the role of the BDNF/TrkB/PI3K signaling pathway, anti-BDNF was used for BDNF knockdown assays. Rats were intranasally administrated with anti-BDNF or control IgG (100 μg/kg) [21 (link)] using a sterile 26-G Hamilton microsyringe (Hamilton Company, Reno, NV, USA) 30 min before propofol treatment.