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25?g/kg, intraperitoneal injection) and then intubated with a tracheal cannula. The jugular vein was cannulated to administer histamine. The animal was placed within a body plethysmograph (a 45?cm��11?cm��15?cm plastic box) in supine position. A 10-min stabilization was allowed in order to record baseline values including tidal volume, flow rate BML-190 and intrathoracic pressure. An asthmatic response was evoked with histamine (15?��g/mL, 0.5?mL, intravenous injection) at 1?mL/min by syringe pump after inhalation of saline (3?mL in cup for 30?s). The changes in Raw (airway resistance) and Cdyn (dynamic compliance of lung) after the histamine challenge were expressed as percent of the baseline in either the R-salbutamol or racemic salbutamol group. The same animal was then allowed an additional 10?min for stabilization, after which a second histamine challenge was given (15?��g/mL 0.5?mL, intravenous injection) after the inhalation of either the aerosol of R-salbutamol (0.2?mg/mL, 30?s ) or racemic salbutamol (0.4?mg/mL, 30?s). The peak changes in Raw and Cdyn after histamine challenges were compared with the baseline values before the histamine challenge, and expressed as percent of the baseline (%). The percent changed from baseline (%) in Raw and Cdyn after being treated with saline (control), R-salbutamol or racemic salbutamol was then compared to determine the anti-asthmatic effects. All the samples from NGI Cyclopamine order and BRS2000 were analyzed quantitatively by HPLC according to the following equation: equation(1) m=(A+3.2442)9803.3��V��0.8299where m is the content of salbutamol for unknown samples (��g); A is the peak area of unknown samples; and BI 2536 concentration V is the volume of unknown samples (mL). Copley Inhaler Testing Data Analysis (CITDA, UK) software was used for analyzing the size distribution. Pulmonary function was characterized with two indexes: Raw and Cdyn, which were calculated from the parameters recorded using the following equations 26?and?27: equation(2) Raw=��PR/��VRaw=��PR/��V equation(3) Cdyn=VT/��PRCdyn=VT/��PRwhere ��PR is the intrathoracic pressure (cmH2O); ��V is the flow rate (derivation of tidal volume, cmH2O/s); and VT is the tidal volume (cmH2O). The percent changed from baseline (%) in Raw and Cdyn after treatment with saline (control), R-salbutamol or R,S-salbutamol was calculated using the following equations: equation(4) Raw%=(R1?R0)/R0��100%Raw%=(R1?R0)/R0��100% equation(5) Cdyn%=(C1?C0)/C0��100%Cdyn%=(C1?C0)/C0��100%where R1 is the maximum value of Raw after challenge; R0 is the mean of Raw before challenge; C1 is the minimum value of Cdyn after challenge; and C0 is the mean of Cdyn before challenge. Comparisons were made between groups using t-test. P