Objective: To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury (CIRI) rats.Methods: Sixty healthy male Sprague-Dawley (SD) rats were routinely reared for 1 week. Ten rats were randomly selected as the sham operation group and 10 rats as the blank control group, while the remaining 40 rats were subjected to preparing the middle cerebral artery occlusion (MCAO) model by modified filament occlusion method. The 40 MCAO rats were further randomly divided into a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, with 10 rats in each group. Rats in the sham operation group, the blank control group and the model group did not accept treatment except binding; rats in the acupuncture group received acupuncture treatment; rats in the mild hypothermia group received mild hypothermia treatment; rats in the acupuncture plus mild hypothermia group received acupuncture and mild hypothermia treatment. 72 h after the treatment, neurological function impairment score was performed; the infarct area ratio was determined by 2,3,5-tripheyl tetrazolium chloride (TTC) staining; apoptosis of brain cells was observed by TUNEL method; the expressions of Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry.Results: Compared with the blank control group and the sham operation group, the neurological function impairment score, cerebral infarct area ratio, apoptosis, and the expressions of Bax and Caspase-3 in the model group were significantly increased, while the expression of Bcl-2 was significantly decreased, and there were significant between-group differences (allP<0.05). After the treatment, there were statistically significant differences among the treatment groups in the neurological function impairment score, cerebral infarct area ratio and apoptosis in the ischemic side of rats, as well as the expressions of Bcl-2, Bax and Caspase-3 (allP<0.05), and from the figures, tables and statistical analysis, it was found that a better tendency in the acupuncture plus mild hypothermia group than the acupuncture group or mild hypothermia group.Conclusion: Acupuncture plus mild hypothermia can protect the brain cells by improving neurological function impairment, decreasing cerebral infarct area ratio, reducing the number of apoptotic cells in the ischemic area and regulating the expressions of apoptosis related proteins to inhibit apoptosis.

Objective:To observe effect of acupuncture combined with hypothermia therapy on MAPK/ERK pathway and apoptosis related factorsin rats suffered cerebral ischemia reperfusion and to explore underlying mechanisms.Methods:Middle cerebral artery ischemia model were established.Ninety SD rats were randomly assigned into a blank group,a control group,a model group,an acupuncture group,a mild hypothermia group,and an acupuncture with hypothermia group.After 72 h treatment,nervefunction defect scores were observed,and infarction area percent was detected by 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) staining;expressions of Bcl-2 and Bax were examined by immunohistochemistry;apoptotic cells were detected by TUNEL assay;and expression levels of phospho-mitogen-activated protein kinase(p-MEK2) and phospho-extracellular signal regulated kinase 1/2 (p-ERK1/2) in the rats' hippocampus ischemic side were determined by Western blot.Results:In the rats of the model group,the neural function defect scores,the infarction area percent,the expression level of Bax,and apoptotic cells increased,while the level of Bcl-2 decreased significantly.The level of p-MEK2 and p-ERK1/2 increased obviously compared with the blank and control groups (P＜0.05 or P＜0.01).After treatment with acupuncture and hypothermia,the neural function defect scores,infarction area percent,and the level ofBax,apoptotic cells and the levels of p-MEK2 and p-ERK1/2 were significantly decreased,while the level of Bcl-2 in the treatment group was significantly elevated (P＜0.05 or P＜0.01) compared with the model group.Compared with the acupuncture group or the hypothermia group,the neural function defect scores and the levels of p-MEK2 and p-ERK1/2 in the acupuncture combined with hypothermia group were significantly reduced (P＜0.05 or P＜0.01).Conclusion:Acupuncture and hypothermia therapy can improve cerebral function,and reduce the cerebral injury through down-regulation of Bax level,and up-regulation of Bcl-2 level,which is related to reducing the levels of p-MEK2 and p-ERK1/2.The therapeutic effects on cerebral ischemia reperfusion injury for combination of acupuncture with hypothermia are better than those with single application of acupuncture or hypothermia.

Objective: To investigate the effects of acupuncture on the number of associated phosphorylated proteins in brain tissues of middle cerebral artery occlusion (MCAO) rats, based on the protein microarray technique. Methods: The MCAO model was prepared according to the modified occlusion method using occlusion lines. Forty healthy Sprague-Dawley (SD) rats were randomly divided into 4 groups using the lottery method: a sham operation group, a model group, a control point group and an acupoint group, with 10 rats in each group. Rats in the sham operation group and the model group only received binding without acupuncture. Rats in the acupoint group received acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 25); rats in the control point group received acupuncture at non-acupoint control points. The needle was twisted once for 1 min after insertion and another time in the middle of the 30 min needle retaining. Acupuncture was conducted once every 12 h for 6 consecutive times. At the end of the experiment, the neurological impairment score was collected, and cells of the ischemic brain tissues were extracted. The protein phosphorylation of the related signaling was detected using the 720 phosphorylated antibody microarray technique, and the differentially expressed proteins between groups were screened. Results: The neurological impairment scores after 72 h of treatment: compared with the sham operation group, the scores of the model group, the control point group and the acupoint group were significantly increased (P<0.01); compared with the model group, the scores of the acupoint group and the control point group were significantly decreased (P<0.01,P<0.05); the score of the acupoint group was better than that of the control point group (P<0.05). The results of the protein microarray: compared with the sham operation group, 48 proteins showed up-regulated phosphorylation (≥1.5 times) in the model group and the down-regulated was 28; compared with the model group, 35 proteins showed up-regulated phosphorylation in the control point group, and the down-regulated was 24. There were 29 proteins showing up-regulated phosphorylation in the acupoint group and the down-regulated was 51. The numbers of proteins involved in the function and signal transduction pathways were also different. Conclusion: Acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 25) can effectively repair brain injury. The ischemic injury of brain tissue may be caused by imbalance of a variety of proteins, and acupuncture can promote brain tissue repair by multi-functional and multi-channel regulation of the protein disorders.

Objective:To observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury (CIRI), and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK1/2 in the mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases (ERK) pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke. Methods:Ninety Sprague-Dawley (SD) rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion (MCAO) models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride (TTC) staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay. Results: Compared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant (P<0.05 orP<0.01). Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant (P<0.05 orP<0.01). Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant (P<0.05 or P<0.01). Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant (P<0.01). Conclusion:Acupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels.

Objective To understand the clinical features of infant pulmonary hemorrhage of enterovirus 71 infection without skin rash, and to improve the diagnosis and treatment of this disease.Methods Six infants infected with enterovirus 71 and presented pulmonary hemorrhage but no skin rash between November 2007 and October 2008 were retrospectively reviewed. The clinical manifestations, clinical outcomes, treatments, laboratory data and chest imaging changes of the cases were analyzed. Results The 6 cases were all younger than 2 years old. The cases distributed throughout the whole year without peak season. Enterovirus 71 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and real time polymerase chain reaction from throat swabs and secretions of the respiratory tract. All the cases began with fever, and 4 of which were accompanied with vomit, and 2 accompanied with cough. After 1 to 3 days, they all got sudden deterioration, manifested with pale and cyanosis, and 1 had hyperspasmia. After intubation, they all had pink frothy fluid from the endotracheal tube. They all had obvious hyperglycaemia, 4 had tachycardia, and 2 had hypertension. All the 6 cases died, and 4 died within 6 h after deterioration. Conclusions Pulmonary hemorrhage of enterovirus 71 infection without skin rash is seen in infants. It is sporadic throughout the whole year. The disease is fulminant and the case often dies in short period of time.

Objective:Based on 108, 591 cases of pediatric emergency visits in a Level Ⅲ Grade A women and children′s hospital in Guangzhou area, we analyze the disease spectrum and epidemiological characteristics, and summarize the characteristics of patient flow changes.These investigations will provide an basis for scientific decision-making for manpower and material resource management of pediatric emergency and hospital workflow design.Methods:The children admitted to the Pediatric Emergency Department of the Zhujiang New Town District of Guangzhou Women and Children′s Medical Center from October 2016 to September 2018, including night emergency and inpatient observations, were analyzed according to the admission date, admission time, gender, age, initial diagnosis and etc.Results:There were more boys than girls in the emergency department, whose ratio was 1.46∶1 (64 480∶44 111 cases). The age of children ranged from 0 to 17 years old, with a median of 11 (23, 48) months.The age distribution was mainly under 5 years old, accounting for 84.14% (91 336/108 591). During the whole year, the number of children in July was the most, accounting for 10.53% (11 433/108 591), and the children in February were the least, accounting for 6.04% (6 555/108 591). The highest visit time of the whole day was 22-23 pm, accounting for 18.83% (20 443/108 591). The most of the diagnosis was respiratory disease, accounting for 53.83% (66 522/123 576). A total of 1 057 critically ill children were received, accounting for 0.97% (1 057/108 591). A total of 911 accidental injuries were received, accounting for 0.84% (911/108 591). Acute upper respiratory tract infection was the most among all diagnoses, accounting for 34.47% (42 541/123 576).Conclusion:Children in the pediatric emergency department of Guangzhou Women and Children′s Medical Center are mainly under 5 years old, and the number of children is the most in July of the year.The main disease is respiratory diseases.Medical staff can be trained according to the actual situation, and the disease spectrum can be updated in time to provide convenience for emergency rescue and improve service level.