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A tracheostomy was performed and a cannula (14 gauge) was inserted into the trachea.
An i.v. cannula (20 gauge) was inserted in the contralateral arm for venous blood sampling.
Next, an interspinous gauge was inserted through the cannula to determine proper implant size selection.
A flexible balloon-tipped rectal catheter (20 French gauge) was inserted to insufflate approximately 3 litres of CO2 gas into the colon, using an automated insufflator (ProtoCO2l, Bracco Diagnostics, Princeton, New York, USA).
Rats were anesthetized by injection of Avertin (Tribromoethanol, 200 mg/kg, ip) and placed in a stereotaxic frame (DAVIO Kopf Instruments, Tujunga, CA), and a stainless steel guide cannula (26 gauge) was inserted into the brain with the tip in the third ventricle (3v); 2.5 mm posterior from the bregma and 8mm below the skull).
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Two intravenous catheters (20 22 gauge) were inserted – one for the infusion of the study drug and the other for the withdrawal of blood for serum concentration assay.
For each subject, venous cannulae (17 gauge) were inserted into large subcutaneous veins of the ante-cubital fossae of both arms.
After adjusting the EA apparatus to 15 Hz/10 mA (Han's Healthronics Likon, Taipei, Taiwan), acupuncture needles (0.5 inch/32 gauge) were inserted 2 5 mm into the muscle layer of the selected acupoints.
Once the gauge is inserted in to the gap, tighten the adjusting screw gradually while sliding the gauge back and forth until there is a noticeable "drag".
A fine intrathecal catheter (32-gauge) was inserted from lumbar level L2/L3 and pushed up to T10 to deliver the Nogo-Ab from an osmotic minipump (Alzet© 2ML2; 5 μL/h, 3.1 μg/μL) for 2 weeks.
19 Venous cannulas (17-gauge) were inserted into large subcutaneous veins of the antecubital fossae of both arms.
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