The effect and mechanism of sodium butyrate on alleviating renal and intestinal injury after cardiopulmonary resuscitation / 中华急诊医学杂志

Objective:To investigate the effect of sodium butyrate (NaB) on renal and intestinal injury after cardiac arrest and cardiopulmonary resuscitation (CA-CPR) and its related mechanism.Methods:Twenty-four domestic healthy male swines were randomly divided into 3 groups: sham group ( n=6), CA-CPR group ( n=10) and NaB group ( n=8). The animals only underwent operational preparation in the sham group. The animal model of CA and CPR was established by 9 min of ventricular fibrillation induced by electrical stimulation in the ventricle and then 6 min of CPR in the CA-CPR and NaB groups. At 5 min after resuscitation, a dose of 75 mg/kg of NaB was intravenously infused for 1 h in the NaB group, and meanwhile the same volume of vehicle was intravenously infused in the sham and CA-CPR groups. At 1, 2, 4, and 24 h after resuscitation, blood samples were collected to detect the renal and intestinal injury biomarkers, such as creatinine (Cr), blood urea nitrogen (BUN), intestinal fatty acid binding protein (IFABP), and diamine oxidase (DAO). At 24 h after resuscitation, renal and intestinal tissue specimens were harvested to detect the protein markers of cell autophagy including microtubule-associated protein light chain 3 Ⅱ (LC3Ⅱ) and p62 expression, and also renal and intestinal apoptosis. Statistical analysis was performed by SPSS software, and continuous variables were compared with one-way analysis of variance among the groups. Results:After CA-CPR, the renal and intestinal injury biomarkers including Cr, BUN, IFABP, and DAO were significantly increased at all time points after resuscitation in the CA-CPR and NaB groups compared with the sham group (all P<0.05). The injury biomarkers mentioned-above were significantly lower at all time points after resuscitation in the NaB group than in the CA-CPR group [Cr (μmol/L): (90±5) vs. (127±9) at 1 h, (135±14) vs. (168±9) at 2 h, (174±10) vs. (211±12) at 4 h, (192±10) vs. (253±13) at 24 h; BUN (mmol/L): (10.5±1.0) vs. (12.3±1.0) at 1 h, (12.2±1.2) vs. (15.3±0.9) at 2 h, (13.6±1.3) vs. (18.3±1.2) at 4 h, (15.4±1.4) vs. (21.5±1.4) at 24 h; IFABP (pg/mL): (502±33) vs. (554±32) at 1 h, (574±52) vs. (644±41) at 2 h, (646±44) vs. (732±43) at 4 h, (711±42) vs. (828±42) at 24 h; DAO (U/mL): (8.6±1.0) vs. (10.5±0.9) at 1 h, (10.6±1.2) vs. (12.8±1.0) at 2 h, (12.1±1.0) vs. (15.0±1.0) at 4 h, (14.1±1.1) vs. (17.6±1.0) at 24 h, (all P<0.05)]. Renal and intestinal tissue detection indicated that cell autophagy and apoptosis were significantly increased after resuscitation in the CA-CPR and NaB groups compared with the sham group, which was indicated by significantly increased LC3Ⅱ and decreased p62 expression, and markedly elevated apoptosis index (all P<0.05). However, cell autophagy and apoptosis in the kidney and intestine were significantly milder after resuscitation in the NaB group than in the CA-CPR group [renal LC3 Ⅱ: (1.15±0.17) vs. (2.23±0.31), p62: (1.60±0.10) vs. (1.17±0.08), apoptosis index (%): (21.2±5.3) vs. (50.9±7.9); intestinal LC3 Ⅱ: (1.03±0.17) vs. (1.71±0.21), p62: (1.30±0.29) vs. (0.79±0.29), apoptosis index (%): (25.6±6.1) vs. (61.7±10.7), all P<0.05]. Conclusions:NaB could alleviate the severity of renal and intestinal damage after CA-CPR in swine, and its protective mechanism may be related to the inhibition of cell autophagy and apoptosis.

Protective role and mechanism of tubastatin A on renal and intestinal injuries after cardiopulmonary resuscitation in swine / 中华危重病急救医学

OBJECTIVE@#To investigate the protective effect and potential mechanism of tubastatin A (TubA), a specific inhibitor of histone deacetylase 6 (HDAC6), on renal and intestinal injuries after cardiopulmonary resuscitation (CPR) in swine.@*METHODS@#Twenty-five healthy male white swine were divided into Sham group (n = 6), CPR model group (n = 10) and TubA intervention group (n = 9) using a random number table. The porcine model of CPR was reproduced by 9-minute cardiac arrest induced by electrical stimulation via right ventricle followed by 6-minute CPR. The animals in the Sham group only underwent the regular operation including endotracheal intubation, catheterization, and anesthetic monitoring. At 5 minutes after successful resuscitation, a dose of 4.5 mg/kg of TubA was infused via the femoral vein within 1 hour in the TubA intervention group. The same volume of normal saline was infused in the Sham and CPR model groups. Venous samples were collected before modeling and 1, 2, 4, 24 hours after resuscitation, and the levels of serum creatinine (SCr), blood urea nitrogen (BUN), intestinal fatty acid binding protein (I-FABP) and diamine oxidase (DAO) in serum were determined by enzyme-linked immunoadsordent assay (ELISA). At 24 hours after resuscitation, the upper pole of left kidney and terminal ileum were harvested to detect cell apoptosis by TdT-mediated dUTP-biotin nick end labeling (TUNEL), and the expression levels of receptor-interacting protein 3 (RIP3) and mixed lineage kinase domain-like protein (MLKL) were detected by Western blotting.@*RESULTS@#After resuscitation, renal dysfunction and intestinal mucous injury were observed in the CPR model and TubA intervention groups when compared with the Sham group, which was indicated by significantly increased levels of SCr, BUN, I-FABP and DAO in serum. However, the serum levels of SCr and DAO starting 1 hour after resuscitation, the serum levels of BUN starting 2 hours after resuscitation, and the serum levels of I-FABP starting 4 hours after resuscitation were significantly decreased in the TubA intervention group when compared with the CPR model group [1-hour SCr (μmol/L): 87±6 vs. 122±7, 1-hour DAO (kU/L): 8.1±1.2 vs. 10.3±0.8, 2-hour BUN (mmol/L): 12.3±1.2 vs. 14.7±1.3, 4-hour I-FABP (ng/L): 661±39 vs. 751±38, all P < 0.05]. The detection of tissue samples indicated that cell apoptosis and necroptosis in the kidney and intestine at 24 hours after resuscitation were significantly greater in the CPR model and TubA intervention groups when compared with the Sham group, which were indicated by significantly increased apoptotic index and markedly elevated expression levels of RIP3 and MLKL. Nevertheless, compared with the CPR model group, renal and intestinal apoptotic indexes at 24 hours after resuscitation in the TubA intervention group were significantly decreased [renal apoptosis index: (21.4±4.6)% vs. (55.2±9.5)%, intestinal apoptosis index: (21.3±4.5)% vs. (50.9±7.0)%, both P < 0.05], and the expression levels of RIP3 and MLKL were significantly reduced [renal tissue: RIP3 protein (RIP3/GAPDH) was 1.11±0.07 vs. 1.39±0.17, MLKL protein (MLKL/GAPDH) was 1.20±0.14 vs. 1.51±0.26; intestinal tissue: RIP3 protein (RIP3/GAPDH) was 1.24±0.18 vs. 1.69±0.28, MLKL protein (MLKL/GAPDH) was 1.38±0.15 vs. 1.80±0.26, all P < 0.05].@*CONCLUSIONS@#TubA has the protective effect on alleviating post-resuscitation renal dysfunction and intestinal mucous injury, and its mechanism may be related to inhibition of cell apoptosis and necroptosis.

Effect of sodium octanoate on renal-intestinal ischemia-reperfusion injury after resuscitation from traumatic cardiac arrest in pigs / 中华创伤杂志

Objective:To investigate the effect of sodium octanoate on renal-intestinal ischemia- reperfusion injury (IRI) after resuscitation from traumatic cardiac arrest in pigs.Methods:Twenty-two miniature piglets with a body weight of (37.6±2.5)kg were divided into three groups according to the random-number table method: normal group ( n=7), IRI group ( n=7) and IRI-treated group ( n=8). A renal-intestinal IRI model of the pig was established by allowing femoral artery to bleed through blood pump at a rate of 2 ml·kg -1·min -1 until cardiac arrest, followed by whole blood transfusion through the femoral vein at a rate of 5 ml·kg -1·min -1 after observation for 6 minutes, and 50% of total blood loss was reinfused before resuscitation. Both the IRI group and IRI-treated group were with IRI model, while normal group was just monitored without induction of IRI. Besides, IRI-treated group was injected intravenously with sodium octanoate (30 mg/kg) for 1 hour at 5 minutes after restoration of spontaneous circulation (ROSC). (1) The rate of resuscitation success, survival rate at 4, 24 hours after resuscitation, blood loss when reaching cardiac arrest criteria and resuscitation time when reaching the ROSC criteria were compared in the three groups. (2) Levels of serum creatinine (SCr), urea nitrogen (BUN), intestinal fatty acid binding protein (iFABP) and diamine oxidase (DAO) were measured before resuscitation and at 1, 2, 4, 24 hours after resuscitation. (3) The animals were sacrificed at 24 hours post-resuscitation to harvest renal and intestinal tissues rapidly. TUNEL test was applied for the cellular apoptosis index. Prussian blue was used to detect the rate of iron deposition. Western blot analysis was used to measure levels of glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member4 (ACSL4). Results:In three groups, all pigs survived. There was no significant difference in blood loss or resuscitation time between IRI group and IRI-treated group (all P>0.05). There was no significant difference in levels of SCr, BUN, iFABP or DAO before resuscitation and at 1, 2, 4, 24 hours after resuscitation in normal group (all P>0.05). But their levels were gradually increased at 1, 2, 4, 24 hours after resuscitation from that before resuscitation in IRI group and IRI-treated group (all P<0.01). Among three groups, levels of SCr, BUN, iFABP and DAO had no significant difference before resuscitation (all P>0.05), but showed obvious increase in IRI group and the IRI-treated group at 1, 2, 4, 24 hours after resuscitation compared with normal group, especially in IRI group (all P<0.01). In normal group, IRI group and IRI-treated group after 24 hours for resuscitation, the cellular apoptosis index of renal tissues was (2.3±0.8)%, (44.0±5.4)% and (13.8±4.3)%; the cellular apoptosis index of intestinal tissues was (2.6±0.9)%, (61.3±10.4)% and (20.8±3.7)%; the rate of iron deposition of renal tissues was (0.6±0.1)%, (3.9±1.0)% and (1.7±0.3)%; the rate of iron deposition of intestinal tissues was (0.8±0.1)%, (4.9±0.9)% and (2.1±0.5)% (all P<0.01). The cellular apoptosis index and rate of iron deposition of both renal and intestinal tissues were the highest in IRI group. The renal-intestinal expression of GPX4 in IRI group and IRI-treated group was lower than that in normal group at 24 hours after resuscitation (all P<0.05), with the lowest in IRI group. The renal-intestinal expression of ACSL4 in IRI group and IRI-treated group was higher than that in normal group at 24 hours after resuscitation (all P< 0.01), with the highest in IRI group. Conclusion:Sodium octanoate can reduce renal-intestinal IRI after resuscitation from traumatic cardiac arrest in pigs, the mechanism for which is probably due to that sodium octanoate can inhibit cellular apoptosis and reduce ferroptosis by regulating the expression levels of GPX4 and ACSL4.

The role and mechanism of sodium valproate in alleviating cardiac and cerebral injuries after cardiopulmonary resuscitation in pigs / 中华急诊医学杂志

Objective:To investigate the role and mechanism of sodium valproate (VPA) in cardiac and cerebral injuries after cardiopulmonary resuscitation (CPR) in pigs.Methods:Twenty-five healthy male domestic pigs, weighing (37±3) kg, were randomly divided into the sham group ( n=6), CPR group ( n=10), and CPR+VPA group ( n=9). Cardiac arrest was induced by alternating current delivered via a pacing catheter in the right ventricle and untreated for 9 min, and then CPR was performed for 6 min, in which this procedure was used to establish the animal model of cardiac arrest and CPR. At 5 min after resuscitation, a dose of 150 mg/kg of VPA was infused with a pump via the femoral vein in 1 h in the CPR+VAP group. At 1 h, 2 h, 4 h and 24 h after resuscitation, blood samples were drawn from the femoral vein, and then used to measure the serum concentrations of cardiac troponin I (cTnI), creatine kinase MB (CKMB), neuron specific enolase (NSE), and S100B protein (S100B) by ELISA. At 24 h after resuscitation, the animals were euthanized, and then tissue specimens in the left myocardium and brain cortex were rapidly harvested to detect the expression levels of C/EBP homologous protein (CHOP), caspase 12, and caspase 3 by Western blot, and the rate of apoptotic cells was detected by TUNEL. Continuous variables were compared with one way analysis of variance among the three groups. Results:(1) After resuscitation, cardiac and cerebral injury biomarkers including cTnI, CKMB, NSE, and S100B in serum were significantly increased in the CPR and CPR+VPA groups compared with the Sham group (all P<0.05). The serum concentrations of cTnI and NSE starting 1 h after resuscitation and the serum concentrations of CKMB and S100B starting 2 h after resuscitation were significantly decreased in the CPR+VPA group compared to the CPR group (all P<0.05). (2) Those proteins related to cell apoptosis mediated by endoplasmic reticulum stress, including CHOP, caspase 12, and caspase 3, were significantly increased, and meanwhile apoptosis index was markedly elevated after resuscitation in the CPR and CPR+VPA groups compared with the Sham group (all P<0.05). Nevertheless, the expression levels of CHOP, caspase 12, and caspase 3 were significantly decreased, and cell apoptosis was markedly reduced in the heart and brain after resuscitation in the CPR+VPA group compared to the CPR group (all P<0.05). Conclusions:VPA can alleviate cardiac and cerebral injuries after CPR in pigs, and its mechanism may be possibly related to the inhibition of cell apoptosis mediated by endoplasmic reticulum stress.

Research progress in application of extracorporeal membrane oxygenation in patients with severe trauma / 中华创伤杂志

Severe trauma can result in severe respiratory and circulatory failure with a high mortality rate，which is quite difficult for clinical treatment. In recent years，the application of extracorporeal membrane oxygenation（ECMO）to circulatory and/or respiratory failure caused by severe trauma has received more and more attention. Systemic anticoagulation is often considered as a relative contraindication to ECMO therapy in patients with severe trauma who are at higher risk of bleeding. However，recent studies have shown that venovenous（VV）-ECMO is safe and feasible for treatment of severe acute respiratory failure after trauma，and venoarterial（VA）-ECMO is of great value for treatment of cardiogenic shock and traumatic cardiac arrest. The issues such as the timing of application，anticoagulation strategies，impact on survival and risk-benefit evaluation related to ECMO application to trauma care need to be investigated further. In this study，the authors summarize advances in application of ECMO，prevention and management of related complications in patients with severe trauma，so as to provide a reference for improving the application level of ECMO.

Optimizing the strategy of radiological examination reduces iatrogenic radiation exposure in major trauma patients / 中华创伤杂志

Objective To investigate the effect of optimized radiological examination strategy on iatrogenic radiation exposure in severe trauma patients so as to provide scientific basis for standardized application of radiological examination.Methods A controlled, three-stage intervention study from April 2010 to November 2011 was carried out.From April 2010 to July 2010, a pre-intervention study was conducted and enrolled 60 patients [43 males, 17 females;age (50 ± 14)years, age range 23-78 years].From August 2010 to March 2011, optimized strategies of radiological examination were implemented, including improving clinicians' knowledge to the standardization of radiological examination and iatrogenic radiation injury and limiting frequency of CT scans through the electronic medical record.From April 2011 to November 2011, post-intervention study was conducted and enrolled 100 patients (81 males, 19 females;age (47 ± 14) years, age range 18-79 years].During this period, major trauma patients were analyzed with respect to the clinical information, radiation examination frequency, ionizing radiation dose and influencing factors.Radiation examination frequency and radiation dose were compared before and after the intervention.Results Radiological examinations were mainly X-ray and CT before the implication of optimized strategies.Of the 60 patients, median frequency of X-rays and CT scan was 6.0(3.0-11.0) and 10.0(8.0-13.8).Median frequency of CT scan was positively correlated with the injury severity score (ISS) and ICU length of stay (r =0.369 and 0.523, P ＜ 0.05).Of the 100 patients, median frequency of CT scan was significantly reduced after the optimization of radiological examination (8.0 vs.10.0, P ＜ 0.05).Total frequency of radiological examination was significantly reduced as well (13.6 vs.17.8, P ＜0.01).There was no significant difference in the treatment success rate before and after the optimization of radiological examination (85.0％ vs.88.3％, P ＞ 0.05).When the frequency of head and chest CT scan was limited, the frequency of radiological examination, radiation exposure and radiological examination expenses were greatly reduced.Conclusions Too much X-ray,CT or other radiological examinations are noted in major trauma patients during the treatment period.Improved understanding of radiation-induced injury, optimizing radiological examination and controlling the repeated radiological examinations of the same site contribute to reducing iatrogenic radiology exposure without affecting the outcome.

Risk factors and clinical effects of trauma induced coagulopathy in ICU patients with major trauma / 中华急诊医学杂志

Objective To investigate the risk factors of trauma induced coagulopathy and its effect on the outcome of ICU patients with severe trauma.Methods Totally 223 severe trauma patients admitted to emergency ICU within 24h after injuring between June,2008 and September,2009 were retrospectively analyzed.Injury severity score (ISS),APACHE Ⅱ score,coagulation function,routine blood test,biochemical test,and blood gas assay were completed for each patient. Hypoperfusion was defined as vasoactive agents usage,or base deficit (BD) ≥ 6 or shock index ≥ 1. Patients were divided into coagulopathy group and non-coagulopathy (control) group according to coagulation function.ISS,APACHE Ⅱ score,the occurrence of hypothermia and hypoperfusion were compared between the two groups.The risk factors of trauma induced coagulopathy were analyzed,and the multivariate logistic regression equation was formulated.Coagulation function and incidence of trauma induced coagulopathy were compared between nonsurvival and survival group.Results Fifty-two of 223 (23.3 ％ ) patients met the criteria of trauma induced coagulopathy.Mortality rate in this group was significantly higher than that in non-coagulopathy group (36.5％ vs 9.4％, P ＜ 0.01 ). Patients in both groups had the comparability in age,sex, injury mechanism and time after trauma.ISS,the incidence of hypothermia,hypoperfusion and severe traumatic brain injury in coagulopathy group were higher than those in non-coagulopathy group ( P ＜ 0.01 ).GCS,hemoglobin,hematocrit,and platelet counts in coagulopathy group were significantly lower than that in noncoagulopathy group (P＜ 0.01).Base deficit ≥6,GCS ≤ 8,and platelet counts were considered as the independent risk factors involved in trauma- induced coagulopathy according to logistic regression in this study.Coagulation function of non-survivors also remarkably attenuated when compared with survival group.Conclusions The incidence rate of trauma induced coagulopathy is high in severe trauma patients admitted to ICU within 24h. Trauma induced coagulopathy correlates well with ISS core,severe traumatic brain injury,shock and hypothermia,and results in high mortality.

Cox regression analysis of 6246 critically ill patients with prolonged stay in emergency room / 中华急诊医学杂志

Objective To explore factors associated with prolonged emergency room (ER) stay of critically ill patients admitted so as to accelerate throughput of emergency patients.Methods Data of critically ill patients admitted into the emergency room of a tertiary teaching hospital in 2010 were retrospectively studied.Stepwise Cox regression analysis was used to determine factors likely associated with prolonged stay in ER.Results ( 1 ) A total of 6246 critical illnesses were admitted into emergency room,the ER length of stay [M (Qr)] was 11 h (3 ～23 h).Of them,56.6％ patients stayed in ER more than 6 h and 21.6％ over 24 h.(2) Univariate analysis showed the major factors contributing to prolonged stay in ER were insufficient inpatient bed capacity,followed by poor family finances,complicated diseases needed care from multiple departments,emergency operation,lack of specialty wards,lack of department bearing main responsibility of critical care,age,gender and arrival time to ER.(3) Multivariate analysis showed that the main factors contributing to prolonged stay were insufficient inpatient beds,poor family finances,complicated diseases needed treatment from multiple departments,emergency operation,lack of specialty wards,lack of department bearing main responsibility of treatment,gender and arrival time to ER.Age was not an independent factor.Conclusions Plenty of critically ill patients admitted to this hospital had prolonged stay in emergency room with variety of factors.The possible factors contributing to this were insufficient inpatient bed capacity,poor family finances and complicated diseases needed care from multiple departments,and this investigation deserves a further study.

Analysis on the characteristics and associated factors of shunt of critically ill patients in emergency room / 中华急诊医学杂志

Objective To investigate the characteristics of patient throughput in emergency room (ER),and the factors causing increase in ER length of stay in order to improve the quality of emergency service.Methods Data of critically ill patients presented to an emergency room in a tertiary teaching hospital in 2010 were retrospectively studied,and the characteristics of patient throughput including patients referred to different departments with different outcomes,and variation in number of patients round the clock in workdays and weekends were analyzed.Results ( 1 ) The median length of stay (LOS) of 7966 critical patients in emergency room (ER) was 11 h,and of them,56.6％ stayed in ER for more than 6 h,and 21.6％ over 24 h.(2) There were significant differences in LOS in ER among patients treated by different departments leading to no shorter length of time consumed for treatment by many departments other than the following specialties of emergency department,neurosurgery,neurology and general medicine department in sequence from less time required to the longest length of time consumed.( 3 ) There were significant differences in LOS in ER among patients with different courses after disposition leading to the longest length of time consumed by those discharged by patients 'own decision and admitted into the hospital,and the shortest time required in patients after emergency operation.(4) There were also significant differences in specialty,outcomes and time needed for throughput between workdays and weekends,and during different time intervals round the clock.Conclusions The situation of patient throughput of critical illness in emergency room in this hospital was not ideal.The factors associated with prolonged stay in ER included different specialties in charge of patients,different courses and outcomes after disposition,severity of illness,different time intervals round the clock,and this investigation deserves a further study.

Protective effect of raloxifene on lung function after acute lung injury in rats / 中华创伤杂志

Objective To evalhate the protective effect of oral raloxifene on lung function after acute lung injury (ALI) in rats. Methods Thirty male adult Sprague-Dawley rats were used and divided into three groups: LPS raloxifene hydrochloric acid. group before secondary impact ( Group A, n = 10 ), LPS raloxifene hydrochloric acid group after secondary impact ( Group B, n = 10) and control group ( n = 10). All the rats were injected intraperitoneally with 5 mg/kg LPS. Raloxifene (30 mg/kg) was orally administered one hour before LPS injection and 14 hours after LPS injection in Groups A and B. The con-trol group remained free. All the animals were anesthetized by intraperitoneal injection of pentobarbital so-dium at 40 mg/kg and the femoral artery was cannulated 16 hours after LPS injection to measure the mean arterial pressure (MAP). All the rats received a direct intratracheal injection of hydrochloric acid ( pH = 1.2, 0.5 ml/kg). Before injection of hydrochloric acid and at 0. 5,1.5 and 4 hours after injection of hy-drochloric acid, the blood gas was measured. Fifteen rats ( five from each group) underwent a micro posi-tron emission tomography ( [18F] FDG microPET) scan of the thorax four hours after hydrochloric acid in-stillation. Then, the lung tissue was collected for histopathological examination. Results The Group B showed better pulmonary gas exchange and more stable MAP compared to the control group. The [18F] fluorodeoxyglueose uptake and histological lung injury score were 9. 01 ± 1.58 and 12.6 ± 0.97 respec-tively in Group B, which were higher than 4. 67 ± 1.33 and 9. 01 ± 1.58 respectively in control group (P < 0. 01 ). Conclusions Raloxifene exerts significant protective effect on lung function after ALI. [18F] FDG microPET is a useful method to evaluate the inflammatory reaction during ALI.

Identification of EOLA1 gene promoter sequence / 第三军医大学学报

Objective To identify the promoter sequence of endothelial-overexpressed lipopolysaccharideassociated factor 1 ( EOLA1) gene and to elucidate the molecular mechanisms controlling EOLA1 expression. Methods A DNA fragment containing 1 723 bp 5' upstream of the EOLA1 gene and the transcription start site was generated by polymerase chain reaction and then cloned into a luciferase reporter gene vector,pGL3-basic. The relative luciferase activities driven by this 5'-upstream fragment and a series of deletion mutants were measured in transiently transfected human ECV304 cells,respectively. At last,the 1 723 bp upstream of the EOLA1 gene was analyzed online with Cluster Buster. Results A fragment 785 bp upstream of the EOLA1 coding region was sufficient to promote transcription. Further deletion analysis of the 785 bp fragment indicated that a 68 bp element from-738 to -676 was important for EOLA1 transcription in ECV304 cells. The 1 723 bp sequence contains binding sites for Sp1 and Myf. Conclusion We map the EOLA1 promoter by deletion analysis and reveal that the proximal region ( -738 to -676 bp) ,which contains binding sites for Sp1 and Myf,is essential for human EOLA1 promoter activity in ECV304 cells.