Relationship Between Acute Lung Injury and Pulmonary Hyaluronic Acid Deposition After Subarachnoid Hemorrhage
Conditions: Subarachnoid Haemorrhage (SAH)
Sponsor: Tianjin Medical University

ALI & HA deposition after SAH                                            Identifier: IRB2024-YX-425-01 
 
Tianjin Medical University General Hospital                                                         - 1 - 
 
 
 
 
 
 
 
 
 
 
 
 
 
Relationship between acute lung injury and 
pulmonary hyaluronic acid deposition after 
subarachnoid hemorrhage 
 
 
 
 
 
 
 
 
 
 
 
 
Version: 1.0 
 
Date: September 30th, 2024 
 
 
 
 
Ethics Committee 
 
Tianjin Medical University General hospital 

ALI & HA deposition after SAH                                            Identifier: IRB2024-YX-425-01 
 
Tianjin Medical University General Hospital                                                         - 2 - 
Protocol Abstract 
Title 
Relationship between acute lung injury and pulmonary hyaluronic acid 
deposition after subarachnoid hemorrhage 
Ethics 
Committee 
(Grant Number) 
The ethics committee of Tianjin Medical University General Hospital 
(IRB2024-YX-425-01) 
Project Executor 
Xintong Ge (M.D. & Ph.D.) 
Sponsor 
Tianjin Medical University General Hospital 
Participant 
Tianjin Huanhu Hospital 
Principal Investigator 
Ye Tian (Ph.D.) - Tianjin Medical University General Hospital 
Yadan Li (M.D.) - Tianjin Huanhu Hospital 
Fundings 
High-Level Talent Project of Tianjin Health Research Program (Grant 
No. TJWJ2024RC002) 
Execute Time 
10/01/2024-09/30/2025 
Recruiting Time 
10/01/2024-09/30/2025 
Objective 
Clarify the correlation between pulmonary inflammatory injury and 
hyaluoronic acid deposition after subarachnoid hemorrhage 
Study Type 
Observational 
Time Perspective 
Prospective 
Enrollment 
24 participants 
Biospecimen 
Description: bronchoalveolar lavage fluid and serum 
Retention: Samples without DNA 
Condition 
Subarachnoid Hemorrhage 
Inclusion Criteria 
1) Age between 18 and 80 years old with independent behavior ability 
or authorized legal representative. 
2) A documented diagnosis of SAH within 5 days. 
3) A Hunt-Hess scale of Ⅳ or Ⅴ. 
4) Absence of clinical and etiological evidence of pulmonary infection. 
Exclusion Criteria 
1) Pregnant or lactating women.  
2) Present history of traumatic brain injury or intracranial hemorrhage.  

ALI & HA deposition after SAH                                            Identifier: IRB2024-YX-425-01 
 
Tianjin Medical University General Hospital                                                         - 3 - 
3) Past history of neurological disorders, lung infection within the past 
six months, cancer, chronic cardiopulmonary diseases, hematological 
diseases or renal failure. 
4) Have participated in clinical trials in the past 4 weeks. 
5) The investigator considers that not appropriate for inclusion. 
Withdrawal Criteria 
1) Clinical or etiological evidence of pulmonary infection after 
recruitment.  
2) Failure to complete tracheoscopy treatments within the time frame 
specified in the study.  
3) Voluntary withdrawal by the subject.  
4) Withdrawal requested by the physician or researcher due to issues 
such as poor compliance. 
Outcome Measures 
Hyaluronic acid and inflammatory cytokines (IL-1β, TNF-α and IL-10) 
levels in bronchoalveolar lavage fluid and serum at the acute phase (1-5 
days post-onset) and the chronic phase (10-14 days post-onset) of 
patients with SAH. 
Reference standard 
Hyaluronic acid is associated with organ dysfunction in acute respiratory 
distress syndrome. Critical Care. 2017, 21(1): 304. 
Statistical Analysis 
All statistical analyses were performed using SPSS Statistics Version 
27.0 (IBM, Armonk, NY, USA). A two-tailed p value of less than 0.05 
was considered to be statistically significant. 

ALI & HA deposition after SAH                                            Identifier: IRB2024-YX-425-01 
 
Tianjin Medical University General Hospital                                                         - 4 - 
1. Research Background and Objectives 
Acute lung injury is a common complication of subarachnoid hematoma (SAH), and a 
significant risk factor for death in patients with SAH. Unlike neurogenic pulmonary edema and 
pneumonia following brain injury, the clinical causes of pulmonary injury after SAH are not 
intracranial hypertension or pulmonary infection. Its occurrence is influenced by the release of 
catecholamines, the regulatory function of the hypothalamic-pituitary-adrenal (HPA) axis and 
systemic inflammatory response, but the specific mechanisms are still unclear. Therefore, delving into 
the pathological mechanisms of SAH-induced lung injury and developing therapeutic strategies based 
on the findings is of great importance to improve the prognosis of patients. 
Abnormal accumulation of hyaluronic acid in the lungs has been reported to be closely related to 
the pathological progression of various pulmonary injury diseases, such as chest trauma, pulmonary 
infection and chronic obstructive pulmonary disease. From this, the present research is aimed to 
explore the levels and dynamic changes of hyaluronic acid in the bronchoalveolar lavage fluid and 
blood of patients with acute lung injury following SAH, and to analyze its correlation with the 
prognosis of pulmonary complications, thereby providing assistance for the clinical diagnosis and 
treatment of SAH. 
2. Study design 
2.1 Overall design 
This research is an observational prospective cohort study. 
2.2 Estimated number of subjects 
Estimated by statistical analysis, a total of 24 subjects were included from patients admitted to 
the neurosurgical intensive care unit of Tianjin Medical University General Hospital and the 
intensive care unit of Tianjin Huanhu Hospital due to aneurysmal SAH. 
3. Inclusion, Exclusion and Withdrawal Criteria 
3.1 Inclusion Criteria 
1) Age between 18 and 80 years old with independent behavior ability or authorized legal 
representative. 
2) A documented diagnosis of SAH within 5 days. 
3) A Hunt-Hess scale of Ⅳ or Ⅴ. 
4) Absence of clinical and etiological evidence of pulmonary infection. 

ALI & HA deposition after SAH                                            Identifier: IRB2024-YX-425-01 
 
Tianjin Medical University General Hospital                                                         - 5 - 
3.2 Exclusion Criteria 
1) Pregnant or lactating women.  
2) Present history of traumatic brain injury or intracranial hemorrhage.  
3) Past history of neurological disorders, lung infection within the past six months, cancer, 
chronic cardiopulmonary diseases, hematological diseases or renal failure. 
4) Have participated in clinical trials in the past 4 weeks. 
5) The investigator considers that not appropriate for inclusion. 
3.3 Withdrawal criteria 
1) Clinical or etiological evidence of pulmonary infection after recruitment.  
2) Failure to complete tracheoscopy treatments within the time frame specified in the study.  
3) Voluntary withdrawal by the subject.  
4) Withdrawal requested by the physician or researcher due to issues such as poor compliance. 
4. Study procedure 
1) Researchers initiate the study according to the inclusion/exclusion criteria. After confirming 
the subject has signed the informed consent, doctors will record the basic information, GCS at 
admission, Hunt-Hess scale and past medical history. Besides, researchers will track the patient's 
surgical status during hospitalization and GOSE at discharge. 
2) The patient undergoes bronchoalveolar lavage treatment under bronchoscopy at the acute 
phase (1-5 days post-onset) and the chronic phase (10-14 days post-onset) of SAH. For each session, 
5 ml of bronchoalveolar lavage fluid is collected to quantify the levels of hyaluronic acid and 
inflammatory cytokines (IL-1β, TNF-α and IL-10). At the same timepoints, 5 ml of venous blood is 
drawn to assess serum hyaluronic acid and inflammatory factors (IL-1β, TNF-α and IL-10) levels. 
3) The patient undergoes a chest CT scan 14 days after the onset of SAH. 
5. Evaluation Parameters 
3.1 Primary outcome parameters 
Hyaluronic acid and inflammatory cytokines (IL-1β, TNF-α and IL-10) levels in 
bronchoalveolar lavage fluid at the acute phase (1-5 days post-onset) and the chronic phase (10-14 
days post-onset) of patients with SAH. 
3.2 Secondary outcome parameters 
Serum hyaluronic acid and inflammatory cytokines (IL-1β, TNF-α and IL-10) levels at the 
acute phase (1-5 days post-onset) and the chronic phase (10-14 days post-onset) of SAH patients. 

ALI & HA deposition after SAH                                            Identifier: IRB2024-YX-425-01 
 
Tianjin Medical University General Hospital                                                         - 6 - 
6. Withdrawal of the subject 
The subject has the right to withdraw from this trial at any time for any reason. The researcher 
should have a thorough understanding and a record of the reasons for withdrawal. Doctors and 
researchers also have the right to stop the subject from continuing to participate in the trial at any 
time during the research. Besides, subjects with early withdrawal should not be replaced by others. 
As too many subjects withdrawing from the trial will lead to unreliable results, unnecessary 
withdrawal should be avoided. The falling rate of the trial should be controlled within 10%. 
7. Statistical analysis 
1) The difference between hyaluronic acid and inflammatory cytokines (IL-1β, TNF-α and 
IL-10) levels in bronchoalveolar lavage fluid at the acute phase (1-5 days post-onset) and the 
chronic phase (10-14 days post-onset) was compared using one-way ANOVA followed by LSD 
post-hoc test. 
2) The difference between serum hyaluronic acid and inflammatory cytokines (IL-1β, TNF-α 
and IL-10) levels at the acute phase (1-5 days post-onset) and the chronic phase (10-14 days 
post-onset) was compared using one-way ANOVA followed by LSD post-hoc test. 
3) Pearson correlation coefficient was used to evaluate the correlation between hyaluronic acid 
levels in bronchoalveolar lavage fluid and serum, hyaluronic acid and inflammatory cytokines 
(IL-1β, TNF-α and IL-10) levels in bronchoalveolar lavage fluid, as well as serum hyaluronic acid 
and inflammatory cytokines (IL-1β, TNF-α and IL-10) levels. 
8. Trial Management 
8.1 Ethics 
To obtain the approval documents of the clinical trial, researchers should submit the trial 
protocol and a copy of the research documents including the informed consent to the ethics 
committee. 
The approval documents from the ethics committee should be accompanied by the name list of 
ethics committee members and their respective responsibilities. These documents will be delivered 
to the researchers in written form before the start of the study.  
Any safety-related issues must be promptly reported to the ethics committee, which includes 
revision on the trial protocol and modification on the subject information page. The end or early 
termination of the trial should also be reported. 

ALI & HA deposition after SAH                                            Identifier: IRB2024-YX-425-01 
 
Tianjin Medical University General Hospital                                                         - 7 - 
8.2 Informed consent and data protection agreement 
It is the responsibility of the researchers to explain the objectives, study procedures, benefits 
and potential risks of the trial for each subject. The researchers should receive the informed consent 
signed by the subject before starting the trial, and keep it properly. The subject should also permit 
researchers and the clinical trial management agency to check his or her original data. Thus, the 
reliability of the research findings could be ensured. 
The personal information of each subject, including name, gender, age, home address and 
telephone number, should be collected in detail. And the researcher/doctor should give the subject 
his or her contact information, so that the subject can contact with him or her when needed. This is 
also helpful for the research and medical care. 
8.3 Subjects privacy 
The researchers should protect the privacy of the subject. All research documents can only be 
identified using the subject's number instead of name or physical examination number. In addition, 
the grouped table that records the correspondence between the subject's name and number can only 
be kept by the researcher, and could not be submitted to any institution or individual. 
8.4 Modification of the trial protocol 
The trial protocol is approved by the ethics committee before starting the trial. During the 
research, any proposed modification on the trial protocol should be reported to the principal 
investigator, and then be submitted to the ethics committee for review. The clinical research can 
only be resumed after the re-approval of the ethics committee. 
8.5 Original records certification 
Researchers should ensure that the subject's privacy is protected when collecting and 
organizing data. In addition, the data manager is authorized to review the original records in order 
to confirm the accuracy of the original data. 
8.6 Documents on file 
According to relevant laws and regulations, researchers should keep the original records 
properly for at least 5 years from the end of the study. 
8.7 Quality control 
The clinical trial management agency has the authority to review the study progress, in order to 
ensure that the trial is carried out as predetermined and that the research data be veritably recorded.