Anti-inflammatory Effect of Therapeutic Hypothermia in Out-hospital Cardiac Arrest Patients With Cardiogenic Shock Via Interleukin-6 Trans-signaling
Phase: NA
Conditions: Cardiac Arrest
Sponsor: China Medical University Hospital

Study Protocol and Statistical Analysis Plan 
 
 
 
 
 
 
 
 
 
Official Title of the Study: 
Anti-inflammatory Effect of Therapeutic Hypothermia in 
Out-hospital Cardiac Arrest Patients with cardiogenic 
Shock via Interleukin-6 Trans-signaling 
 
 
NCT number: NCT02633358 
 
 
 
 
 
 
 
 
 
 
Da-Long Chen 
China Medical University Hospital, Taichung, Taiwan 
Date of the Document: August 25, 2018 
 
 

Study Design and Patients 
 
The clinical trial was designed to evaluate the effect of therapeutic hypothermia on IL- 
 
6 trans-signaling in patients with OHCA complicated with cardiogenic shock in 
comparison with placebo (ClinicalTrials.gov, NCT02633358). The study was 
conducted at the CMU hospital, Taichung, Taiwan from January 2015 to April 2018, 
and was approved by the institutional review board of the CMU hospital. All 
participants provided written informed consent through their surrogate decision-maker. 
Patients with OHCA who were aged more than 18 years old were enrolled after 
resuscitation. The clinical definition of cardiogenic shock used was patients with 
cardiac problems who require vasopressors to maintain a mean arterial pressure of more 
than 65 mmHg after adequate fluid hydration and an initial lactate level of more than 
18 mg/dL. The study 
design is outlined in the 
right figure. 

Exclusion criteria were recovery from OHCA without cardiogenic shock, 
pregnancy, metastatic cancer, and family disapproval of patient participation in the 
clinical trial. A CONSORT flow diagram is displayed in the below table. 
 
 
 
 
Therapeutic Hypothermia Protocol 
 
The therapeutic hypothermia protocol was in accordance with that proposed by Scirica 
et al. (2013), with minor modifications (12). The first stage was rapid immediate 
cooling. The second stage was maintenance of a core temperature of 33°C–35°C for at 
least 24 hours (TTM34oC). The third stage was slow rewarming. The fourth stage was 
maintenance of normothermia to prevent fever. The therapeutic hypothermia protocol 

is presented in the below figure. 
 
 
 
 
 
 
Several cooling devices were selected freely, including Arctic Sun (Medivance 
Inc., Louisville, CO, USA) and Thermogard XP (Zoll Medical, Chelmsford, MA, USA) 
automatic core temperature control devices in addition to classical ice blankets. 
 
 
Blood Sampling Protocol 
 
Peripheral venous blood was drawn into blood collection tubes containing 
ethylenediamine-tetra acetic acid to prevent coagulation at 6 and 24 hours after 
resuscitation. To obtain plasma, these tubes were immediately placed on melting ice 
and centrifuged within 30 minutes using Centrifuge 5810R (Eppendorf AG, Hamburg, 
Germany) set at 4°C, 450  g for 25 minutes. Immediately following centrifugation, 
plasma was stored at −70°C in a freezer until further analyses. 

IL-6 Trans-Signaling Immunoassay 
 
The sandwich enzyme-linked immunosorbent assay was performed to measure IL-6, 
sIL-6R, Il-6/sIL-6R complex, and soluble glycoprotein 130 (sgp130) by using a human 
kit and standard protocol (R&D systems Inc., MN, USA). The absorbance of each well 
was determined using a Synergy™ H4 microplate reader at a 450-nm wavelength 
(BioTek instruments, Inc., VT, USA). Results were calculated using a 4-parameter 
logistic standard curve. 
 
 
SOFA scores and Neurological Outcome 
 
To evaluate the severity of organ failure, we used a SOFA score. This score reflected 
the severity of organ failure in cardiovascular system (mean arterial pressure), 
respiratory system (arterial pressure of oxygen divided by the fraction of inspired 
oxygen), central nervous system (Glasgow coma scale), liver (bilirubin), kidneys 
(creatinine), and coagulation (platelet). 

Cerebral Performance Categories (CPC) scales of 1 (good recovery or slight 
disability) and 2 (moderate disability) were considered favorable neurological 
outcomes. CPC scales of 3 (severe disability) and 4 (comatose or persistent vegetative 
state) were considered poor neurological outcomes. A CPC scale of 5 (brain death) was 
considered as an indicator of mortality. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Statistical Analysis 
 
For assessing differences between the two groups, Student’s t-test was used. For two 
time points between something before and after, paired t-test was used. Interaction 
effect, differences between the two groups and over time was analyzed by using 
multivariate analysis of variance (MANOVA) model. For categorical data, Chi-square 
test was used. Kaplan–Meier survival curves were compared between the two groups 

by using the log-rank test. Risk ratios were reported as a measure of relative risk. A 
two-tailed p value of less than 0.05 was considered to indicate statistical significance. 
All statistical analyses were performed using SPSS Statistics version 22.0 (IBM Corp., 
Armonk, NY, USA) and SAS 9.4 (SAS Institute, Cary, NC, USA).