the Change of R-spondin Proteins Plasma Concentration Level Caused by Mechanical Ventilation and Its Effect on Mechanical Ventilation Induced Lung Injury
Conditions: Acute Lung Injury
Sponsor: Xinhua Hospital, Shanghai Jiao Tong University School of Medicine

Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine 
 
 
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Research Protocol 
 
 
 
 
 
 
TITLE: the Change of R-spondin Proteins Plasma Concentration Level caused by 
Mechanical Ventilation and its Effect on Mechanical Ventilation Induced Lung Injury 
  
 
 
PROTOCOL ID: XH-17-015 
 
 
 
NCT number:  NCT03315702 
 
 
 
SPONSOR: Xinhua Hospital, Shanghai Jiao Tong University School of Medicine 
 
 
 
 
INVESTIGATOR: Lai Jiang 
 
 
 
Date： July 20, 2017 
 
 
 
 
 
 
 
 
 

Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine 
 
 
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Materials and methods 
1.Patients recruitment 
Research protocol was approved by Xinhua Hospital Ethics Committee Affiliated to 
Shanghai Jiao Tong University School of Medicine. With written informed consents 
given by patients undergoing elective surgery, individual information and blood 
samples were collected. 
Inclusion Criteria is shown as followed: patients undergo elective surgery with 
mechanical ventilation lasting for > 4 hours; classified as physical status I to III 
according to the American Society of Anesthesiologists Physical Status Classification 
System; Written informed consent is approved. 
Participating subjects with any one of criteria shown as followed were excluded: 
chronic lung disease; recent lung infection; recent anaesthetics or mechanical 
ventilation treatment; hemodilution with massive fluid supply during surgery; children；
women during pregnancy or lactation; being involved in other clinical subjects. 
2.Mechanical ventilation 
Mechanical ventilation protocol: tidal volume 6-8 ml/kg, positive end-expiratory 
pressure 5 cmH2O, oxygen concentration 40%; respiratory rate 10-15/min, 
inspiratory/expiratory ratio 1:1.5. 
3.Blood samples collection and detection 
With written informed consents given by patients undergoing elective surgery, collect 
2ml venous blood samples each time before endotracheal intubation and third hour after, 
thus mechanical ventilation for one patient lasts for four hours. Store samples in EDTA 
plus blood collection tubes. All samples were stored in 4 ℃ refrigerator and 
centrifugated in 1hours after being collected. Plasmas were separated from venous 
blood samples via centrifugation at 4 ℃ 1000rpm for 10 min, and stored in -80 ℃ before 
being analyzed with Elisa. 
4. Statistical analysis 
Analyze R-spondin proteins plasma centration levels of two samples from the same 
patient collected relatively before endotracheal intubation and fourth hour after as 
paired-samples. Quantitative data were expressed as mean and 95 % confidence 
interval (CI) for mean. Differences between paired-samples are assessed by paired t-
test, performed using the SPSS13.0 package. P < 0.05 is considered as statistically 
significant criteria.