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Background@#Limited data are available on the mortality rates of patients receiving extracorporeal membrane oxygenation (ECMO) support for coronavirus disease 2019 (COVID-19). We aimed to analyze the relationship between COVID-19 and clinical outcomes for patients receiving ECMO. @*Methods@#We retrospectively investigated patients with COVID-19 pneumonia requiring ECMO in 19 hospitals across Korea from January 1, 2020 to August 31, 2021. The primary outcome was the 90-day mortality after ECMO initiation. We performed multivariate analysis using a logistic regression model to estimate the odds ratio (OR) of 90-day mortality. Survival differences were analyzed using the Kaplan–Meier (KM) method. @*Results@#Of 127 patients with COVID-19 pneumonia who received ECMO, 70 patients (55.1%) died within 90 days of ECMO initiation. The median age was 64 years, and 63% of patients were male. The incidence of ECMO was increased with age but was decreased after 70 years of age. However, the survival rate was decreased linearly with age. In multivariate analysis, age (OR, 1.048; 95% confidence interval [CI], 1.010–1.089; P = 0.014) and receipt of continuous renal replacement therapy (CRRT) (OR, 3.069; 95% CI, 1.312–7.180; P = 0.010) were significantly associated with an increased risk of 90-day mortality. KM curves showed significant differences in survival between groups according to age (65 years) (log-rank P = 0.021) and receipt of CRRT (log-rank P = 0.004). @*Conclusion@#Older age and receipt of CRRT were associated with higher mortality rates among patients with COVID-19 who received ECMO.
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Purpose@#Real-world experience with tocilizumab in combination with dexamethasone in patients with severe coronavirus disease (COVID-19) needs to be investigated. @*Materials and Methods@#A retrospective cohort study was conducted to evaluate the effect of severity-adjusted dosing of dexamethasone in combination with tocilizumab for severe COVID-19 from August 2020 to August 2021. The primary endpoint was 30-day clinical recovery, which was defined as no oxygen requirement or referral after recovery. @*Results@#A total of 66 patients were evaluated, including 33 patients in the dexamethasone (Dexa) group and 33 patients in the dexamethasone plus tocilizumab (DexaToci) group. The DexaToci group showed a statistically significant benefit in 30-day clinical recovery, compared to the Dexa group (p=0.024). In multivariable analyses, peak FiO2 within 3 days and tocilizumab combination were consistently significant for 30-day recovery (all p<0.05). The DexaToci group showed a significantly steeper decrease in FiO2 (-4.2±2.6) than the Dexa group (−2.7±2.6; p=0.021) by hospital day 15. The duration of oxygen requirement was significantly shorter in the DexaToci group than the Dexa group (median, 10.0 days vs. 17.0 days; p=0.006). Infectious complications and cellular and humoral immune responses against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the convalescence stage were not different between the two groups. @*Conclusion@#A combination of severity-adjusted dexamethasone and tocilizumab for the treatment of severe COVID-19 improved clinical recovery without increasing infectious complications or hindering the immune response against SARS-CoV-2.
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Purpose@#In the intensive care unit, a peripherally inserted central catheter (PICC) may be an alternative option to standard central venous catheters, particularly in patients with coagulopathies or at high risk of infection. The purpose of this research was to assess the feasibility of bedside ultrasound (US)-guided PICC placement by critical care fellows on intensive care units. @*Methods@#All bedside US-PICCs inserted by critical care fellows from July 2013 to September 2015 were retrospectively reviewed focusing on the rate of successful insertion, complications of insertion, or during maintenance. @*Results@#A total of 177 US-guided PICCs were inserted in 163 patients and included in the analysis. The median age was 62 years (IQR 50-70 years) and 104 cases (58.8%) were male. There were 172 cases (90.4%) of PICCs inserted in the upper arm. Anticoagulant therapy was used in 26 patients (14.7%) and 8 patients (5.2%) had severe coagulopathies. The median procedural time was 30 minutes (IQR 19-45 minutes). Insertion success rate was 93.2%, and there were no major complications during insertions except for malposition (12.1%). Catheters remained in place for a total of 3,878 days (median 16 days: IQR 8-31 days). There was only 1 case (0.6%) of catheter-related bloodstream infection, and 2 cases (1.2%) of symptomatic venous thromboembolism. @*Conclusion@#Bedside US-guided placement of PICCs by critical care fellows is safe and feasible. The success rate of the procedure was “acceptable,” and was not associated with significant risks of infectious and non-infectious complications, even in patients with coagulopathies.
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Purpose@#In the intensive care unit, a peripherally inserted central catheter (PICC) may be an alternative option to standard central venous catheters, particularly in patients with coagulopathies or at high risk of infection. The purpose of this research was to assess the feasibility of bedside ultrasound (US)-guided PICC placement by critical care fellows on intensive care units. @*Methods@#All bedside US-PICCs inserted by critical care fellows from July 2013 to September 2015 were retrospectively reviewed focusing on the rate of successful insertion, complications of insertion, or during maintenance. @*Results@#A total of 177 US-guided PICCs were inserted in 163 patients and included in the analysis. The median age was 62 years (IQR 50-70 years) and 104 cases (58.8%) were male. There were 172 cases (90.4%) of PICCs inserted in the upper arm. Anticoagulant therapy was used in 26 patients (14.7%) and 8 patients (5.2%) had severe coagulopathies. The median procedural time was 30 minutes (IQR 19-45 minutes). Insertion success rate was 93.2%, and there were no major complications during insertions except for malposition (12.1%). Catheters remained in place for a total of 3,878 days (median 16 days: IQR 8-31 days). There was only 1 case (0.6%) of catheter-related bloodstream infection, and 2 cases (1.2%) of symptomatic venous thromboembolism. @*Conclusion@#Bedside US-guided placement of PICCs by critical care fellows is safe and feasible. The success rate of the procedure was “acceptable,” and was not associated with significant risks of infectious and non-infectious complications, even in patients with coagulopathies.
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Purpose@#The aim of this study was to analyze the temporal change of resuscitation fluid use based on all fluids administered in a surgical intensive care unit (ICU). @*Methods@#The administration of resuscitation fluid to all patients admitted to a surgical ICU of a tertiary referral hospital was investigated from 2008 to 2015. The types and volumes of fluid, and laboratory data taken within 7 days after ICU admission were evaluated. Resuscitation fluids were defined as fluids infused according to stat orders, rather than routine orders. @*Results@#There were a total of 8,885 admissions to the ICU for 7,886 patients. The volumetric proportion of crystalloid to total resuscitation fluids increased significantly over the study period (p < 0.001; 79.6% in 2008; 93.7% in 2015). Although the proportion of 0.9% saline to crystalloids decreased, that of balanced solutions increased (p < 0.001; 29.5% in 2008; 55.6% in 2015). The use of colloids decreased from 20.4% in 2008, to 6.3% in 2015 (p < 0.001). Proportions calculated using the number of individual fluids administered revealed trends similar to those calculated using volumetric data. The amount of infused 0.9% saline was weakly correlated with the lowest blood pH and the highest serum chloride levels (ρ = -0.26 and 0.19, respectively). @*Conclusion@#Changes in the trends of fluid resuscitation practice were noted in a single surgical ICU over the 8-year study period. Crystalloid use increased owing to a rise in the utilization of balanced solutions with a downward trend in colloid use.
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Purpose@#The aim of this study was to analyze the temporal change of resuscitation fluid use based on all fluids administered in a surgical intensive care unit (ICU). @*Methods@#The administration of resuscitation fluid to all patients admitted to a surgical ICU of a tertiary referral hospital was investigated from 2008 to 2015. The types and volumes of fluid, and laboratory data taken within 7 days after ICU admission were evaluated. Resuscitation fluids were defined as fluids infused according to stat orders, rather than routine orders. @*Results@#There were a total of 8,885 admissions to the ICU for 7,886 patients. The volumetric proportion of crystalloid to total resuscitation fluids increased significantly over the study period (p < 0.001; 79.6% in 2008; 93.7% in 2015). Although the proportion of 0.9% saline to crystalloids decreased, that of balanced solutions increased (p < 0.001; 29.5% in 2008; 55.6% in 2015). The use of colloids decreased from 20.4% in 2008, to 6.3% in 2015 (p < 0.001). Proportions calculated using the number of individual fluids administered revealed trends similar to those calculated using volumetric data. The amount of infused 0.9% saline was weakly correlated with the lowest blood pH and the highest serum chloride levels (ρ = -0.26 and 0.19, respectively). @*Conclusion@#Changes in the trends of fluid resuscitation practice were noted in a single surgical ICU over the 8-year study period. Crystalloid use increased owing to a rise in the utilization of balanced solutions with a downward trend in colloid use.
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Background@#Despite the increasing importance of rehabilitation for critically ill patients, there is little information regarding how rehabilitation therapy is utilized in clinical practice. Our objectives were to evaluate the implementation rate of rehabilitation therapy in the intensive care unit (ICU) survivors and to investigate the effects of rehabilitation therapy on outcomes. @*Methods@#A retrospective nationwide cohort study with including > 18 years of ages admitted to ICU between January 2008 and May 2015 (n = 1,465,776). The analyzed outcomes were readmission to ICU readmission and emergency room (ER) visit. @*Results@#During the study period, 249,918 (17.1%) patients received rehabilitation therapy. The percentage of patients receiving any rehabilitation therapy increased annually from 14% in 2008 to 20% in 2014, and the percentages for each type of therapy also increased over time. The most common type of rehabilitation was physical therapy (91.9%), followed by neuromuscular electrical stimulation (29.6%), occupational (28.6%), respiratory, (11.6%) and swallowing (10.3%) therapies. After adjusting for confounding variables, the risk of 30-day ICU readmission was lower in patients who received rehabilitation therapy than in those who did not (P < 0.001; hazard ratio [HR], 0.70; 95% confidence interval [CI], 0.65–0.75). And, the risk of 30-day ER visit was also lower in patients who received rehabilitation therapy (P < 0.001; HR, 0.83; 95% CI, 0.77–0.88). @*Conclusion@#In this nationwide cohort study in Korea, only 17% of all ICU patients received rehabilitation therapy. However, rehabilitation is associated with a significant reduction in the risk of 30-day ICU readmission and ER visit.
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BACKGROUND: Beyond its current function as a rescue therapy in acute respiratory distress syndrome (ARDS), extracorporeal membrane oxygenation (ECMO) may be applied in ARDS patients with less severe hypoxemia to facilitate lung protective ventilation. The purpose of this study was to evaluate the efficacy of extended ECMO use in ARDS patients. METHODS: This study reviewed 223 adult patients who had been admitted to the intensive care units of 11 hospitals in Korea and subsequently treated using ECMO. Among them, the 62 who required ECMO for ARDS were analyzed. The patients were divided into two groups according to pre-ECMO arterial blood gas: an extended group (n=14) and a conventional group (n=48). RESULTS: Baseline characteristics were not different between the groups. The median arterial carbon dioxide tension/fraction of inspired oxygen (FiO2) ratio was higher (97 vs. 61, p<0.001) while the median FiO2 was lower (0.8 vs. 1.0, p<0.001) in the extended compared to the conventional group. The 60-day mortality was 21% in the extended group and 54% in the conventional group (p=0.03). Multivariate analysis indicated that the extended use of ECMO was independently associated with reduced 60-day mortality (odds ratio, 0.10; 95% confidence interval, 0.02–0.64; p=0.02). Lower median peak inspiratory pressure and median dynamic driving pressure were observed in the extended group 24 hours after ECMO support. CONCLUSION: Extended indications of ECMO implementation coupled with protective ventilator settings may improve the clinical outcome of patients with ARDS.
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Adulto , Humanos , Hipoxia , Dióxido de Carbono , Oxigenación por Membrana Extracorpórea , Unidades de Cuidados Intensivos , Corea (Geográfico) , Pulmón , Mortalidad , Estudios Multicéntricos como Asunto , Análisis Multivariante , Oxígeno , Respiración Artificial , Síndrome de Dificultad Respiratoria , Estudios Retrospectivos , Ventilación , Ventiladores MecánicosRESUMEN
BACKGROUND@#Beyond its current function as a rescue therapy in acute respiratory distress syndrome (ARDS), extracorporeal membrane oxygenation (ECMO) may be applied in ARDS patients with less severe hypoxemia to facilitate lung protective ventilation. The purpose of this study was to evaluate the efficacy of extended ECMO use in ARDS patients.@*METHODS@#This study reviewed 223 adult patients who had been admitted to the intensive care units of 11 hospitals in Korea and subsequently treated using ECMO. Among them, the 62 who required ECMO for ARDS were analyzed. The patients were divided into two groups according to pre-ECMO arterial blood gas: an extended group (n=14) and a conventional group (n=48).@*RESULTS@#Baseline characteristics were not different between the groups. The median arterial carbon dioxide tension/fraction of inspired oxygen (FiO2) ratio was higher (97 vs. 61, p<0.001) while the median FiO2 was lower (0.8 vs. 1.0, p<0.001) in the extended compared to the conventional group. The 60-day mortality was 21% in the extended group and 54% in the conventional group (p=0.03). Multivariate analysis indicated that the extended use of ECMO was independently associated with reduced 60-day mortality (odds ratio, 0.10; 95% confidence interval, 0.02–0.64; p=0.02). Lower median peak inspiratory pressure and median dynamic driving pressure were observed in the extended group 24 hours after ECMO support.@*CONCLUSION@#Extended indications of ECMO implementation coupled with protective ventilator settings may improve the clinical outcome of patients with ARDS.
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BACKGROUND: Admission blood glucose (BG) level is a predictor of mortality in critically ill patients with various conditions. However, limited data are available regarding this relationship in critically ill patients with cardiovascular diseases according to diabetic status. METHODS: A total of 1,780 patients (595 with diabetes) who were admitted to cardiac intensive care unit (CICU) were enrolled from a single center registry. Admission BG level was defined as maximal serum glucose level within 24 hours of admission. Patients were divided by admission BG level: group 1 (< 7.8 mmol/L), group 2 (7.8–10.9 mmol/L), group 3 (11.0–16.5 mmol/L), and group 4 (≥ 16.6 mmol/L). RESULTS: A total of 105 patients died in CICU (62 non-diabetic patients [5.2%] and 43 diabetic patients [7.9%]; P = 0.105). The CICU mortality rate increased with admission BG level (1.7%, 4.8%, 10.3%, and 18.8% from group 1 to group 4, respectively; P < 0.001). On multivariable analysis, hypertension, mechanical ventilator, continuous renal replacement therapy, acute physiology and chronic health evaluation II (APACHE II) score, and admission BG level significantly influenced CICU mortality in non-diabetic patients (group 1 vs. group 3: hazard ratio [HR], 3.31; 95% confidence interval [CI], 1.47–7.44; P = 0.004; group 1 vs. group 4: HR, 6.56; 95% CI, 2.76–15.58; P < 0.001). However, in diabetic patients, continuous renal replacement therapy and APACHE II score influenced CICU mortality but not admission BG level. CONCLUSION: Admission BG level was associated with increased CICU mortality in critically ill, non-diabetic patients admitted to CICU but not in diabetic patients.
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Humanos , APACHE , Glucemia , Enfermedades Cardiovasculares , Cuidados Críticos , Enfermedad Crítica , Diabetes Mellitus , Hipertensión , Unidades de Cuidados Intensivos , Mortalidad , Pronóstico , Terapia de Reemplazo Renal , Ventiladores MecánicosRESUMEN
This study aimed to investigate cognitive impairment and psychological distress of critically ill patients at discharge from intensive care unit (ICU). This study included 30 critically ill patients who had neither pre-existing dementia nor ongoing delirium. At ICU discharge, they performed a screening test for cognitive impairment (Mini-Cog test) and completed questionnaires for depression (Patient Health Questionnaire-2, PHQ-2) and for 4 stressful experiences during ICU stay including nightmares, severe anxiety or panic, severe pain, and trouble to breathe or feeling of suffocation (Post-Traumatic Stress Syndrome 14-Question Inventory, PTSS-14 Part A). Thirteen patients (43.3%) screened positive for cognitive impairment and 18 patients (60.0%) exhibited depressive symptoms. Twenty three patients (76.7%) recollected one or more stressful in-ICU experiences. Female patients (88.9%) was more likely to feel depressed at ICU discharge, compared to male patients (47.6%) (χ2=4.47, p=0.03). To the best of our knowledge, this is the first report on cognitive and psychological outcomes of ICU survivors in Korea. In this study, we observed that a considerable number of critically ill patients had experienced cognitive impairment or psychological distress at ICU discharge.
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Femenino , Humanos , Masculino , Ansiedad , Asfixia , Trastornos del Conocimiento , Cuidados Críticos , Enfermedad Crítica , Delirio , Demencia , Depresión , Sueños , Unidades de Cuidados Intensivos , Corea (Geográfico) , Tamizaje Masivo , Pánico , SobrevivientesRESUMEN
No abstract available.
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Aconitum , Arritmias Cardíacas , Reanimación CardiopulmonarRESUMEN
Limited data are available on improved outcomes after initiation of neurointensivist co-management in neurosurgical intensive care units (NSICUs) in Korea. We evaluated the impact of a newly appointed neurointensivist on the outcomes of neurosurgical patients admitted to an intensive care unit (ICU). This retrospective observational study involved neurosurgical patients admitted to the NSICU at Samsung Medical Center between March 2013 and May 2016. Neurointensivist co-management was initiated in October 1 2014. We compared the outcomes of neurosurgical patients before and after neurointensivist co-management. The primary outcome was ICU mortality. A total of 571 patients were admitted to the NSICU during the study period, 291 prior to the initiation of neurointensivist co-management and 280 thereafter. Intracranial hemorrhage (29.6%) and traumatic brain injury (TBI) (26.6%) were the most frequent reasons for ICU admission. TBI was the most common cause of death (39.0%). There were no significant differences in mortality rates and length of ICU stay before and after co-management. However, the rates of ICU and 30-day mortality among the TBI patients were significantly lower after compared to before initiation of neurointensivist co-management (8.5% vs. 22.9%; P = 0.014 and 11.0% vs. 27.1%; P = 0.010, respectively). Although overall outcomes were not different after neurointensivist co-management, initiation of a strategy of routine involvement of a neurointensivist significantly reduced the ICU and 30-day mortality rates of TBI patients.
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Humanos , Lesiones Encefálicas , Causas de Muerte , Resultados de Cuidados Críticos , Cuidados Críticos , Unidades de Cuidados Intensivos , Hemorragias Intracraneales , Corea (Geográfico) , Mortalidad , Neurocirugia , Estudio Observacional , Estudios RetrospectivosRESUMEN
No abstract available.
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Aconitum , Arritmias Cardíacas , Reanimación CardiopulmonarRESUMEN
Invasive aspergillosis (IA) is most commonly seen in patients with risk factors, such as cytotoxic chemotherapy, prolonged neutropenia, corticosteroids, transplantation and acquired immune deficiency syndrome. IA commonly occurs in the respiratory tract. Extrapulmonary aspergillosis is usually a part of a disseminated infection, and primary invasive intestinal aspergillosis is very rare. Herein, we report a case of an immunocompetent 53-year-old male who suffered recurrent septic shock in the intensive care unit (ICU) and was finally diagnosed as invasive intestinal aspergillosis without dissemination. IA is rarely considered for patients who do not have an immune disorder. Thus, when such cases do occur, the diagnosis is delayed and the clinical outcome is often poor. However, there is a growing literature reporting IA cases in patients without an immune disorder, mostly among ICU patients. Primary intestinal aspergillosis should be considered for critically ill patients, especially with severe disrupted gastrointestinal mucosal barrier.
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Humanos , Masculino , Persona de Mediana Edad , Síndrome de Inmunodeficiencia Adquirida , Corticoesteroides , Aspergilosis , Enfermedad Crítica , Diagnóstico , Quimioterapia , Enfermedades Gastrointestinales , Enfermedades del Sistema Inmune , Huésped Inmunocomprometido , Unidades de Cuidados Intensivos , Neutropenia , Sistema Respiratorio , Factores de Riesgo , Choque SépticoRESUMEN
Invasive aspergillosis (IA) is most commonly seen in patients with risk factors, such as cytotoxic chemotherapy, prolonged neutropenia, corticosteroids, transplantation and acquired immune deficiency syndrome. IA commonly occurs in the respiratory tract. Extrapulmonary aspergillosis is usually a part of a disseminated infection, and primary invasive intestinal aspergillosis is very rare. Herein, we report a case of an immunocompetent 53-year-old male who suffered recurrent septic shock in the intensive care unit (ICU) and was finally diagnosed as invasive intestinal aspergillosis without dissemination. IA is rarely considered for patients who do not have an immune disorder. Thus, when such cases do occur, the diagnosis is delayed and the clinical outcome is often poor. However, there is a growing literature reporting IA cases in patients without an immune disorder, mostly among ICU patients. Primary intestinal aspergillosis should be considered for critically ill patients, especially with severe disrupted gastrointestinal mucosal barrier.
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Humanos , Masculino , Persona de Mediana Edad , Síndrome de Inmunodeficiencia Adquirida , Corticoesteroides , Aspergilosis , Enfermedad Crítica , Diagnóstico , Quimioterapia , Enfermedades Gastrointestinales , Enfermedades del Sistema Inmune , Huésped Inmunocomprometido , Unidades de Cuidados Intensivos , Neutropenia , Sistema Respiratorio , Factores de Riesgo , Choque SépticoRESUMEN
Although shock in sepsis is usually managed successfully by conventional medical treatment, a subset of cases do not respond and may require salvage therapies such as veno-arterial extracorporeal membrane oxygenation (VA ECMO) support as well as an attempt to remove endotoxins. However, there are limited reports of attempts to remove endotoxins in patients with septic shock on VA ECMO support. We recently experienced a case of septic shock with severe myocardial injury whose hemodynamic improvement was unsatisfactory despite extracorporeal membrane oxygenation (ECMO) support. Since the cause of sepsis was acute pyelonephritis and blood cultures grew gram-negative bacilli, we additionally applied polymyxin B direct hemoperfusion (PMX-DHP) to the ECMO circuit and were able to successfully taper off vasopressors and wean off ECMO support. To the best of our knowledge, this is the first adult case in which PMX-DHP in addition to ECMO support was successfully utilized in a patient with septic shock. This case indicates that additional PMX-DHP therapy may be beneficial and technically feasible in patients with septic shock with severe myocardial injury refractory to ECMO support.
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Adulto , Humanos , Cardiomiopatías , Endotoxinas , Oxigenación por Membrana Extracorpórea , Hemodinámica , Hemoperfusión , Membranas , Oxígeno , Polimixina B , Polimixinas , Pielonefritis , Terapia Recuperativa , Sepsis , Choque , Choque SépticoRESUMEN
We found an error in this article. The Fig. 1A.
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BACKGROUND: Colistin (polymyxin E) is active against multidrug-resistant Gram-negative bacteria (MDR-GNB). However, the effectiveness of inhaled colistin is unclear. This study was designed to assess the effectiveness and safety of aerosolized colistin for the treatment of ventilator-associated pneumonia (VAP) caused by MDR-GNB. METHODS: In this retrospective longitudinal study, we evaluated the medical records of 63 patients who received aerosolized colistin treatment for VAP caused by MDR-GNB in the medical intensive care unit (MICU) from February 2012 to March 2014. RESULTS: A total of 25 patients with VAP caused by MDR-GNB were included in this study. The negative conversion rate was 84.6% after treatment, and acute kidney injury (AKI) occurred in 11 patients (44%, AKI group). The average length of MICU stay and colistin treatment- related factors, such as daily and total cumulative doses and administration period, were not significantly different between groups. In-hospital mortality tended to be higher in the AKI group (p = 0.07). Multivariate analysis showed that a body mass index less than 18 was an independent risk factor of mortality (odds ratio [OR] = 21.95, 95% confidence interval [CI] 1.59-302.23; p = 0.02). Notably, AKI occurrence was closely related to the administration of more than two nephrotoxic drugs combined with aerosolized colistin (OR = 15.03, 95% CI 1.40-161.76; p = 0.025) and septic shock (OR = 8.10, 95% CI 1.40-161.76; p = 0.04). CONCLUSIONS: The use of adjunctive aerosolized colistin treatment appears to be a relatively safe and effective option for the treatment of VAP caused by MDR-GNB. However, more research on the concomitant use of nephrotoxic drugs with aerosolized colistin will be necessary, as this can be an important risk factor of development of AKI.
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Humanos , Lesión Renal Aguda , Índice de Masa Corporal , Colistina , Farmacorresistencia Microbiana , Bacterias Gramnegativas , Mortalidad Hospitalaria , Unidades de Cuidados Intensivos , Estudios Longitudinales , Registros Médicos , Mortalidad , Análisis Multivariante , Neumonía , Neumonía Asociada al Ventilador , Estudios Retrospectivos , Factores de Riesgo , Choque Séptico , Resultado del TratamientoRESUMEN
We found an error in this article. The Fig. 1A.