The role of SUMOylation in the protective mechanisms of therapeutic hypothermia on neonatal hypoxic-ischemic encephalopathy / 中国新生儿科杂志

Objective:To study the role of SUMOylation in the process of therapeutic hypothermia on neural stem cells (NSCs) in neonatal hypoxic-ischemic encephalopathy.Methods:SUMOylation is an essential post-translational modification involving small ubiquitin-like modifiers (SUMOs). Primary-cultured NSCs from mice were assigned into four groups: control group, hypoxia group, hypothermia group and hypoxia+hypothermia group. Western Blot was used to detect the protein levels of SUMO2/3, hypoxia-inducible factor-1α (HIF-1α), peroxisome proliferator-activated receptor γ coactivator factor 1α (PGC-1α) and octamer binding transcription factor 4 (Oct4). The diameters of NSCs were compared. ELISA was used to detect lactate dehydrogenase (LDH) level. Apoptosis was examined using flow cytometry. Immunofluorescence method was used to measure the differentiation of NSCs into neuronal cells.Results:Compared with the control group, the levels of SUMO2/3, HIF-1αand PGC-1α in NSCs of the hypoxia group increased 33%, 126% and 140%, respectively ( P<0.05). Compared with the control group, the levels of SUMO2/3 and PGC-1α in NSCs of the hypothermia group increased 52% and 536%, respectively ( P<0.05). Compared with the hypoxia group, the levels of SUMO2/3, HIF-1α, PGC-1α and Oct4 in the hypoxia+hypothermia group increased 44%, 40%, 230% and 59%, respectively ( P<0.05). The diameters of NSCs in hypoxia group, hypothermia group and hypoxia+hypothermia group were smaller than control group, and hypoxia+hypothermia group smaller than hypoxia group ( P<0.05). No significant differences existed in LDH levels between hypothermia group and control group ( P>0.05). LDH level in hypoxia+hypothermia group were significantly lower than hypoxia group ( P<0.05). No significant differences existed in the cell death rates between hypothermia group and control group ( P>0.05). The cell death rate in hypoxia+hypothermia group was significantly lower than hypoxia group ( P<0.05). Compared with the control group, the expressions of Nestin in both hypoxia group and hypothermia group were increased, but neuron specific enolase (NSE) were decreased ( P<0.05). Compared with hypoxia group and hypothermia group, the level of Nestin in hypoxia+hypothermia group was further increased, while NSE was further decreased ( P<0.05). Conclusions:Therapeutic hypothermia may increase the tolerance of NSCs to hypoxia by enhancing SUMO modification of proteins, providing theoretical basis for the treatment of hypoxic-ischemic encephalopathy with therapeutic hypothermia.

Early predictors of brain injury in patients with acute carbon monoxide poisoning and the neuroprotection of mild hypothermia.

INTRODUCTION: Carbon monoxide (CO) poisoning can cause serious neurological sequelae. However, there is neither effective treatment strategy nor reliable indicators to determine the prognosis of patients with CO poisoning. The present study aimed to observe the changes of neurological function score, disease severity score, cerebral oxygen utilization (O2UCc), bispectral (BIS) index and neuron-specific enolase (NSE) concentration, and to elucidate the clinical significance of these potential indicators and the neuroprotective effect of mild hypothermia on brain injury in patients with severe acute CO poisoning. MATERIALS AND METHODS: A total of 277 patients with acute severe CO poisoning from 2013 to 2018 were enrolled in our hospital. Patients were divided into three groups according to their body temperature on the day of admission and their willingness to treat: a fever group (n = 78), a normal temperature group (NT group, n = 113), and a mild hypothermia group (MH group, n = 86). All patients were given hyperbaric oxygen therapy, while those in the MH group received additional mild hypothermia treatment. The severity of the disease, the neurobehavioral status, the incidence of delayed encephalopathy after acute carbon monoxide poisoning (DEACMP), and other indicators including BIS, O2UCc, NSE were further evaluated in all patients at given time-points. RESULTS: Mild hypothermia therapy improved the prognosis of patients with CO poisoning, significantly decreased the value of O2UCc and NSE, and up-regulated BIS. The incidence of DEACMP at 6 months was 27% in the fever group, 23% in the NT group, and 8% in the MH group. The values of Glasgow-Pittsburgh coma scale (G-P score), BIS index and NSE were closely related to the occurrence of DEACMP, the cutoff values were 12.41, 52.17 and 35.20 ng/mL, and the sensitivity and specificity were 79.3%, 77.6%, 79.3% and 67.6%, 89.5%, 88.6% in the receiver operating characteristic curve (ROC), respectively. CONCLUSIONS: Early mild hypothermia treatment could significantly reduce the severity of brain injury after CO poisoning, and might be further popularized in clinic. G-P scores, NSE and BIS index can be regarded as the prediction indicators in the occurrence and development of DEACMP. CLINICAL TRIAL REGISTRATION: The study protocol was granted from Qingdao University Research Ethics Committee (Clinical trial registry and ethical approval number: QD81571283).

Effects of mild hypothermia therapy combined with minimally invasive debridement in patients with hypertensive intracranial hemorrhage: a randomized controlled study.

OBJECTIVE: To investigate the clinical effect of mild hypothermia therapy (MHT) combined with minimally invasive debridement (MID) in patients with severe hypertensive intracranial hemorrhage (HICH). METHODS: A total of 120 patients with severe HICH who received clinical intervention in our hospital were enrolled as study subjects. In this randomized, controlled, double-blind trial, they were divided into a study group (SG, n=70) and a control group (CNG, n=50). The CNG was treated with MID, and the SG was treated with MID combined with MHT. The general surgical indices, short-term postoperative outcomes, postoperative neurological and recovery in activities of daily living, and complications were compared between the two groups. Patients' Glasgow prognosis (Glasgow Outcome Scale, GOS) scores at 1 year after surgery were analyzed. RESULTS: The operative time, intraoperative blood loss and intensive care unit (ICU) admission were shorter/lower in the SG than in the CNG (P<0.05). The SG had higher hematoma clearance rate at 1 d and 3 d postoperatively, and lower residual hematoma volume at 3 d and 7 d postoperatively than the CNG (P<0.05). Patients in the SG had higher Barthel scores and lower National Institutes of Health Stroke Scale (NIHSS) scores than the CNG at 1-12 months after intervention (P<0.05). The incidence of complications in the SG was lower than that in the CNG (P<0.05). The percentage of GOS grade IV and V was significantly higher in the SG than in the CNG 1 year after surgery (P<0.05). CONCLUSION: The combination of MID and MHT in patients with severe HICH has better clinical results in the short and long term, and improves the postoperative outcomes and quality of life. It can also reduce the incidence of perioperative complications.

The efficacy of systemic hypothermia therapy initiated at different times on neonatal hypoxic-ischemic encephalopathy / 中国新生儿科杂志

Objective:To study the clinical efficacy, safety and prognosis of systemic hypothermia therapy on neonatal hypoxic-ischemic encephalopathy (HIE)initiated at different times after birth.Method:From January 2013 to August 2018, term neonates (within 12 hours after birth) diagnosed with neonatal moderate to severe HIE and received systemic treatment in the neonatal intensive care unit of our hospital were retrospectively included. According to the starting time of hypothermia therapy, the neonates were assigned into three groups: within 6 h after birth (TH1 group), 6~12 h (TH2 group) and conventional treatment group (control group). Their clinical data during perinatal period, hospitalization period and follow-up at 6-month were reviewed. Their clinical and neurodevelopmental outcomes were compared using SPSS 25.0 statistical software.Result:A total of 147 neonates with moderate to severe HIE were enrolled. 111 received 72-hour hypothermia therapy, including 79 in the TH1 group, 32 in the TH2 group and 36 in the control group. The neurobehavioral test scores at 10-day of life in the TH1 group were significantly higher than the control group ( P<0.05). No significant differences existed among the TH2 group, the TH1 group and the control group ( P>0.05). The brain magnetic resonance imaging (MRI) showed injuries in the TH1 group and the TH2 group were significantly milder than the control group ( P<0.05). No significant differences of brain injuries existed between TH1 group and TH2 group ( P>0.05). 100 patients completed Bailey Infant Intelligence Development Scale at 6-month follow-up. 21 had abnormal scores. No statistically significant differences existed in the psychomotor development index (PDI) scores among the three groups ( P>0.05). TH1 and TH2 groups had significantly fewer cases with mental development index (MDI) <70 points than the control group ( P<0.05). No statistically significant differences existed of MDI scores between the TH1 group and the TH2 group ( P>0.05). No statistically significant differences existed of PDI scores among the 3 groups ( P>0.05). At 6-month, the mortality rate of the control group (32.1%, 9/28) was significantly higher than the TH1 group (6.6%, 4/61) ( P<0.05). No significant differences existed of mortality rate at 6-month among the TH2 group, the TH1 group and the control group ( P>0.05). Conclusion:Systemic hypothermia therapy for neonatal HIE is safe. Starting systemic hypothermia therapy at 6~12-hour after birth may also be effective in reducing mortality rate and improving neurodevelopmental outcome.

Intravenous thrombolysis in combination with mild hypothermia therapy in the treatment of acute cerebral infarction.

OBJECTIVE: To investigate the efficacy of recombinant tissue plasminogen activator (rt-PA) intravenous thrombolysis in combination with mild hypothermia therapy in the treatment of acute cerebral infarction. METHODS: One hundred and thirty-two patients with acute cerebral infarction who were admitted to our hospital were selected and grouped into a control group and an observation group, 66 each group. Patients in the control group were given conventional treatment in combination with local mild hypothermia therapy, and patients in the observation group were given rt-PA intravenous thrombolysis on the basis of conventional treatment and local mild hypothermia therapy. National institute of health stroke scale (NIHSS) score and intracranial pressure (ICP) of the two groups before and after treatment was recorded. The efficacy of the two groups was evaluated. The modified Rankin scale (MRS) score was followed up for three months. The blood samples of the patients were collected before and after thrombolysis. Superoxide dismutase (SOD) and malondialdehyde (MDA) levels in the plasma were detected. RESULTS: The NIHSS score of the two groups decreased in the 1st, 3rd and 7th day after treatment compared to before treatment (p<0.05), but the NIHSS score of the two groups had no significant difference at different time points after treatment (p>0.05). The ICP of the two groups decreased in the 1st, 3rd and 7th day after treatment compared to before treatment (p<0.05), and the decrease of ICP of the observation group was more significant than that of the control group at the same time point (1st, 3rd and 7th day after treatment) (p<0.05). The clinical efficacy of the observation group was higher than that of the control group after treatment, and the difference was statistically significant (p<0.05). The MDA concentration of both groups decreased at different time points after treatment (p<0.05), but the SOD concentration increased (p<0.05). The MDA concentration of the observation group was lower than that of the control group at different time points after treatment (p<0.05), and the SOD concentration of the observation group was higher than that of the control group (p<0.05). CONCLUSION: rt-PA intravenous thrombolysis in combination with mild hypothermia therapy has significant efficacy in the treatment of acute cerebral infarction. It can effectively relieve neurological function. Its action mechanism may be realized by relieving oxidative stress response.

Therapeutic effect of mild hypothermia on the inflammatory response and outcome in perioperative patients with acute Stanford type A aortic dissection: A randomized controlled trial / 中国胸心血管外科临床杂志

@#Objective To explore the therapeutic effect of mild hypothermia on the inflammatory response, organ function and outcome in perioperative patients with acute Stanford type A aortic dissection (AAAD). Methods From February 2017 to February 2018, 56 patients with AAAD admitted in our department were enrolled and randomly allocated into two groups including a control group and an experimental group. After deep hypothermia circulatory arrest during operation, in the control group (n=28), the patients were rewarmed to normal body temperatures (36 to 37 centigrade degree), and which would be maintained for 24 hours after operation. While in the experimental group (n=28), the patients were rewarmed to mild hypothermia (34 to 35 centigrade degree), and the rest steps were the same to the control group. The thoracic drainage volume and the incidence of shivering at the first 24 hours after operation, inflammatory indicators and organ function during perioperation, and outcomes were compared between the two groups. There were 20 males and 8 females at age of 51.5±8.7 years in the control group, 24 males and 4 females at age of 53.3±11.2 years in the experimental group. Results There was no obvious difference in the basic information and operation information in patients between the two groups. Compared to the control group, at the 24th hour after operation, the level of peripheral blood matrix metalloproteinases (MMPs) was lower than that in the experimental group (P=0.008). In the experimental group, after operation, the awakening time was much shorter (P=0.008), the incidence of bloodstream infection was much lower (P=0.019). While the incidence of delirium, acute kidney injury (AKI), hepatic insufficiency, mechanical ventilation duration, intensive care unit (ICU) stays, or hospital mortality rate showed no statistical difference. And at the first 24 hours after operation, there was no difference in the thoracic drainage volume between the two groups, and no patient suffered from shivering. Conclusion The mild hypothermia therapy is able to shorten the awakening time and reduce the incidence of bloodstream infection after operation in the patients with AAAD, and does not cause the increase of thoracic drainage volume or shivering.

The Profile of MMP-9, MMP-9 mRNA Expression, -1562 C/T Polymorphism and Outcome in High-risk Traumatic Brain Injury: The Effect of Therapeutic Mild Hypothermia.

The aim of this study was to investigate the effect of mild hypothermia therapy (34-36°C) and the alterations of matrix metalloproteinase-9 (MMP-9) in 20 patients with high-risk traumatic brain injury (TBI). The neurologic status and outcome were assessed using Full Outline of UnResponsiveness (FOUR) score and Glasgow Coma Scale (GCS). A prospective randomized control study involved patients with high-risk TBI (FOUR score ≤ 7). Patients were randomized into two groups, with and without mild hypothermia therapy which were investigated within 24 and 72 h. The MMP-9 level, MMP-9 mRNA expression and -1562 C/T polymorphism were estimated using enzyme-linked immune sorbent assay (ELISA), reversing transcription polymerase chain reaction (RT-PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). Different levels of these variables were compared in the two groups. In the hypothermia group, the expression of MMP-9 mRNA and the level of serum MMP-9 were significantly decreased (P < 0.05) within 72 h. There was a highly significant correlation between the expression of MMP-9 mRNA and the level of MMP-9 protein (R2 = 0.741, r = 0.861, P < 0.05). The study did not find in -1562 C/T polymorphism. The patients' outcome was improved significantly after mild hypothermia therapy (P < 0.05). The data obtained from this study show that mild hypothermia therapy down regulated the expression of MMP-9 mRNA, the MMP-9 protein level and increased the FOUR score and GCS in high-risk TBI patients within 72 h.

Interpretation of three important problems in clinical treatment from guidelines for the management of severe traumatic brain injury (fourth edition) issued by the United States in 2016 / 天津医药

Guidelines for the Management of Severe Traumatic Brain Injury (Fourth Edition) have received extensive attention both at home and abroad after releasing by the Brain Trauma Foundation of the United States in 2016. The interval between the published newest version of the Guidelines and the Third Edition has already approached 7 years. In accordance with more rigorous evidence-based medicine standards, the Fourth Edition includes 94 updated research findings as evidence, in combination with the proposition of more accurate treatment recommendations and problem solutions. Combining the domestic situation and problems in the treatment of severe traumatic brain injury at present, three most important clinical issues related to the Fourth Edition of the guidelines are interpreted and analyzed in the present study, including decompressive craniectomy, mild hypothermia therapy and intracranial pressure monitoring, so as to improve the level of treatment of traumatic brain injury in China.

S100β level and prognosis influence of mild hypothermia therapy assisted intracranial hematoma evacuation in treatment of patients with cerebral hemorrhage / 中国综合临床

Objective To investigate clinical effect of mild hypothermia therapy assisted intracranial hematoma evacuation in treatment of cerebral hemorrhage.Methods One hundred and ten patients with cerebral hemorrhage were selected in Affiliated Hospital of North China University of Science and Technology from December 2011 to December 2013,and were randomly divided into two groups.Fifty-five patients treated intracranial hematoma evacuation as control group.Another 55 patients treated mild hypothermia therapy assisted intracranial hematoma evacuation as observation group.Treatment effect was compared between two groups.Results Serum S100β,neuron specific enolization (NSE) enzyme,tumor necrosis factor α (TNF-α),creactive protein(CRP),cognitive function score,daily life ability score,neurological function defect score before and after treatment in control group were (0.82±0.12) μg/L and (0.53±0.09) μg/L,(19.42±2.30) μg/L and (10.36±1.07) μg/L,(3.62±0.57) mg/L and (1.54±0.30) mg/L,(29.43±4.36) g/L and (10.25± 1.07) g/L,(13.42± 1.58) points and (25.03± 1.19) points,(21.45± 3.27) points and (37.92 ± 5.83)points,(13.27± 1.35) points and (4.84 ± 1.08) points,the differences were significant (t =8.471,11.834,17.026,22.539,12.230,10.619,25.531,P ＜ 0.05).Serum S100β,NSE,TNF-α,CRP,cognitive function score,daily life ability score,neurological function defect score before and after treatment in observation group were (0.84±0.13)μg/L and (0.41±0.10) μg/L,(19.48±1.76) μg/L and (8.75±0.84) μg/L,(3.64± ±0.61) mg/Land (1.17±0.29) mg/L,(29.58±3.62) g/L and (6.02±1.18) g/L,(13.29±1.34) points and (27.58± 1.27) points,(21.68±4.02) points and (48.26±7.14) points,(13.46± 1.21) points and (3.57±0.85) points,the differences were significant(t=13.498,16.739,25.728,41.836,13.769,15.857,36.352,P＜0.05).Compared with serum S100β,NSE,TNF-α,CRP,cognitive function score,daily life ability score,neurological function defect score before treatment,there were no difference between two groups (P ＞0.05).Serum S100β,NSE,TNF-α,CRP,neurological function defect score after treatment in observation group were lower than control group(t =5.926,4.839,6.162,10.054,6.714,P＜0.05).Cognitive function score,daily life ability score after treatment in observation group were higher than control group (t =4.008,5.973,P ＜0.05).Postoperative Glasgow prognosis classification in observation group (14 cases of grade Ⅰ,27 cases of grade Ⅱ,11 cases of grade Ⅲ,2 cases of grade Ⅳ,1 case of grade Ⅴ) was better than control group(8 cases of grade Ⅰ,12 cases of grade Ⅱ,23 cases of grade Ⅲ,7 cases of grade Ⅳ,5 cases of grade Ⅴ),the differences were significant between the two groups (Z=17.085,P =0.002).Total effective rate in observation group 94.5％ (52/55) was higher than control group 78.2％ (43/55),the differences were significant between the two groups (Z =6.253,P=0.012).Conclusion Mild hypothermia therapy assisted intracranial hematoma evacuation in treatment of cerebral hemorrhage,can significantly reduce inflammatory factor and S100βlevel,improve neurological function,has significant effect and good prognosis.It is worthy of clinical use.

The clinical analysis of mild hypothermia therapy for severe traumatic brain injury in elderly patients / 中华行为医学与脑科学杂志

Objective To investigate the clinical therapeutic effect of mild hypothermia in elderly patients with severe traumatic brain injury.Methods 72 cases of elderly patients with severe traumatic brain injury(GCS ≤ 8) were divided into mild hypothermia therapy group(36 patients)and control group(36 patients) according to the random number table method.Mild hypothermia therapy group received mild hypothermia treatment while control group received normal treatment.The clinical prognosis was analyzed between the two groups.Results After 24h treatment,both mild hypothermia therapy group and control group intracranial pressure began to rise.But the intracranial pressure of the mild hypothermia therapy group(24 h:(13.0±4.5)mmHg,3 d:(16.6±4.0) mmHg,5 d:(19.9±3.9) mmHg,1 mmHg=0.133 kPa) were significantly lower than those of the control group (24 h:(16.6± 3.8) mmHg,3 d:(20.4±4.8) mmHg;5 d:(24.1 ± 6.2) mmHg),and the difference was statistically significant (t=2.225,2.260,2.192,P=0.035,0.033,0.039).The rate of good recovery to the control group and the mild hypothermia therapy group were 22.22％ and 47.22％ respectively while the mortality were 30.56％ and 13.89％ respectively,and the differences were statistically significant (x2 =4.936,5.675,P=0.047,0.035).Conclusion Mild hypothermia treatment can inhibit the increase of intracranial pressure and reduce disability rate and mortality in elderly patients with severe traumatic brain injury,which can increase the survival rate and improve the long-term prognosis.

Effects of mild hypothermia therapy on the levels of glutathione in rabbit blood and cerebrospinal fluid after cardiopulmonary resuscitation.

OBJECTIVES: The aim of this study was to investigate the effects of mild hypothermia therapy on oxidative stress injury of rabbit brain tissue after cardiopulmonary resuscitation (CPR). MATERIALS AND METHODS: Rabbit models of cardiac arrest were established. After the restoration of spontaneous circulation, 50 rabbits were randomly divided into normothermia and hypothermia groups. The following five time points were selected: before CPR, immediately after CPR, 2 hr after CPR (hypothermia group reached the target temperature), 14 hr after CPR (hypothermia group before rewarming), and 24 hr after CPR (hypothermia group recovered to normal temperature). Glutathione (GSH) concentrations in both the blood and cerebrospinal fluid of the normothermia and hypothermia groups were measured. RESULTS: At 2, 14, and 24 hr after CPR, the GSH concentrations in both the blood and cerebrospinal fluid were significantly higher in the hypothermia group than in the nomorthermia group. CONCLUSION: Mild hypothermia therapy may increase GSH concentrations in rabbit blood and cerebrospinal fluid after CPR as well as promote the recovery of cerebral function.

Evaluation of mild hypothermia therapy for neonatal hypoxic-ischaemic encephalopathy on brain energy metabolism using 18F-fluorodeoxyglucose positron emission computed tomography.

It remains unclear whether mild hypothermia affects energy metabolism in the brain tissue of newborns with hypoxic-ischaemic encephalopathy (HIE). The current study aimed to investigate the effect of mild hypothermia on energy metabolism in neonatal HIE and assess brain energy metabolism using position emission tomography/computed tomography (PET/CT) scanning. The mean standardised uptake values of 18F-fluorodeoxyglucose (18F-FDG) were used to determine the glucose metabolic rate in various brain anatomical regions, including the thalamus, basal ganglia and the frontal, parietal and occipital lobes. The rate of glucose metabolism significantly improved following treatment with mild hypothermia therapy and conventional therapy (P<0.001). Prior to the treatment, no significant differences were identified between the groups (P>0.05). Following treatment, the rate of glucose metabolism was significantly improved in the mild hypothermia therapy group compared with that in the conventional therapy group (P<0.001). Thus, these results indicate that mild hypothermia therapy effectively promotes the recovery of patients with neonatal HIE. 18F-FDG PET/CT scanning may be used to provide reference values for the assessment of energetic metabolism in patients with neonatal HIE.

<b>Effect of Orengedokuto on Hyperthermia </b><b>after mild Hypothermia Therapy </b> / 日本東洋医学雑誌

We studied the effects of orengedokuto on central hyperthermia after mild hypothermia therapy for cardiac arrest. The subjects in this study were 7 patients who showed central hyperthermia (>38.3 °C) after mild hypothermia therapy. Orengedokuto 7.5-15 g/day was administered and central temperature was monitored. Maximum change in central temperature was 1.55 ± 0.71 °C(from 39.1 ± 0.7 °C to 37.6 ± 0.7 °C) (p < 0.05). Mean change was 0.35 ± 0.77 °C, (from 37.7 ± 0.6 °C to 37.5 ± 0.7 °C). Thus we conclude that orengedokuto is a drug with applications in the treatment of central hyperthermia after mild hypothermia therapy.