Correlation between uncoupling protein 2 expression and myocardial mitochondrial injury in rats with sepsis induced by lipopolysaccharide / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the correlation between uncoupling protein 2 (UCP2) expression and myocardial mitochondria injury in rats with sepsis induced by lipopolysaccharide (LPS).</p><p><b>METHODS</b>The rat model of sepsis was established through an intraperitoneal injection of LPS. Forty male Sprague-Dawley rats were randomly and equally divided into control group (an intraperitoneal injection of normal saline), sepsis 6 h group (LPS-6 h group), sepsis 12 h group (LPS-12 h group), sepsis 24 h group (LPS-24 h group), and sepsis 48 h group (LPS-48 h group). The serum and heart tissues were harvested at corresponding time points and myocardial mitochondria was extracted. The microplate reader was applied to measure creatine kinase (CK), creatine kinase-MB (CK-MB), and reactive oxygen species (ROS). Flow cytometry was applied to measure the degree of mitochondrial swelling and mitochondrial membrane potential (MMP). Western blot was used to measure the expression level of UCP2. Electron microscopy was applied to observe the morphological changes in heart tissues and myocardial mitochondria.</p><p><b>RESULTS</b>Compared with the control group, the LPS groups had significantly increased serum levels of CK, CK-MB, and myocardial ROS, as well as a significantly increased degree of mitochondrial swelling (P<0.05), and these values reached their peaks at 24 hours after LPS injection. The LPS groups had a significant decrease in MMP (P<0.05), which reached the lowest level at 24 hours after LPS injection. Western blot showed that the LPS groups had a significant increase in the expression level of myocardial UCP2 compared with the control group (P<0.05), which reached its peak at 24 hours after LPS injection. The results of electron microscopy showed mitochondrial swelling, partial rupture of the mitochondrial membrane, and cavity formation in rats in the LPS groups. The most severe lesions occurred in the LPS-24 h group. In rats with LPS, the ROS level in the myocardial mitochondria and the degree of mitochondrial swelling were positively correlated with the expression level of UCP2 (r=0.796 and 0.893, respectively; P<0.05), while MMP was negatively correlated with the expression level of UCP2 (r=-0.903, P<0.05).</p><p><b>CONCLUSIONS</b>In the rat model of sepsis, the myocardium and myocardial mitochondria have obvious injuries, and the expression level of UCP2 is closely correlated with mitochondrial injury. Therefore, UCP2 might play an important role in myocardial mitochondrial injury in sepsis.</p>

Implementation of a Pediatric Emergency Triage System in Xiamen, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pediatric emergency rooms (PERs) in Chinese hospitals are perpetually full of sick and injured children because of the lack of sufficiently developed community hospitals and low access to family physicians. The aim of this study was to evaluate the clinical value of a new five-level Chinese pediatric emergency triage system (CPETS), modeled after the Canadian Triage System and Acuity Scale.</p><p><b>METHODS</b>In this study, we compared CPETS outcomes in our PER relative to those of the prior two-level system. Patients who visited our PER before (January 2013-June 2013) and after (January 2014-June 2014) the CPETS was implemented served as the control and experimental group, respectively. Patient flow, triage rates, triage accuracy, wait times (overall and for severe patients), and patient/family satisfaction were compared between the two groups.</p><p><b>RESULTS</b>Relative to the performance of the former system experienced by the control group, the CPETS experienced by the experimental group was associated with a reduced patient flow through the PER (Cox-Stuart test, t = 0, P < 0.05), a higher triage rate (93.40% vs. 90.75%; χ2 = 801.546, P < 0.001), better triage accuracy (96.32% vs. 85.09%; χ2 = 710.904, P < 0.001), shorter overall wait times (37.30 ± 13.80 min vs. 41.60 ± 15.40 min; t = 11.27, P < 0.001), markedly shorter wait times for severe patients (2.07 [0.65, 4.11] min vs. 3.23 [1.90,4.36] min; z = -2.057, P = 0.040), and higher family satisfaction rates (94.23% vs. 92.21%; χ2 = 321.528, P < 0.001).</p><p><b>CONCLUSIONS</b>Implementing the CPETS improved nurses' abilities to triage severe patients and, thus, to deliver the urgent treatments more quickly. The system shunted nonurgent patients to outpatient care effectively, resulting in improved efficiency of PER health-care delivery.</p>

A preliminary study of long-term mitochondrial dysfunction in rat brain caused by lipopolysaccharide-induced sepsis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To preliminarily investigate the long-term structural and functional injuries of mitochondria in rat brain caused by sepsis.</p><p><b>METHODS</b>Wistar rats were randomly assigned into sepsis and control groups. A rat model of sepsis was prepared by an intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS) of gram-negative bacteria, and the survival assay was performed. Eight rats in the sepsis group were sacrificed at 12, 24, 48, or 72 hours after LPS injection, while rats in the control group were sacrificed after an intraperitoneal injection of an equal volume of normal saline. Mitochondria were extracted from rat brain tissue. Mitochondrial membrane potential (MMP) and mitochondrial swelling level were determined by flow cytometry, and the activities of electron transport chain complexes (I-V) were measured using enzyme assay kits. Hematoxylin-eosin (HE) staining and electron microscopy were used to observe morphological changes in brain tissue and mitochondria.</p><p><b>RESULTS</b>The sepsis group had a significantly lower survival rate than the control group (P<0.01). The MMP and activities of electron transport chain complexes (I-V) in the sepsis group, which were significantly lower than those in the control group (P<0.05), were reduced to the lowest levels at 48 hours and partially recovered at 72 hours. The mitochondrial swelling level in the sepsis group, which was significantly higher than that in the control group (P<0.05), increased to the peak level at 48 hours and partially recovered at 72 hours. Hematoxylin and Eosin staining revealed substantial damages in the structure of brain tissue, and electron microscopy showed mitochondrial swelling, and vacuolization in a few mitochondria.</p><p><b>CONCLUSIONS</b>In the rat model of LPS-induced sepsis, both structural and functional injuries are found in cerebral mitochondria, and achieve the peak levels probably at around 48 hours.</p>

Effect of UCP2-siRNA on inflammatory response of cardiomyocytes induced by septic serum / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of uncoupling protein 2 (UCP2)-siRNA on the inflammatory response of rat cardiomyocytes (H9C2) induced by septic serum and to investigate the possible role of UCP2 in the development of septic cardiomyopathy.</p><p><b>METHODS</b>Serum samples were separately collected from normal rats and septic rats. Cultured rat cardiac cells (H9C2) were randomly divided into blank control, normal serum, 10% septic serum, UCP2-siRNA+10% septic serum and negative siRNA+10% septic serum groups. Stimulation with 10% septic serum was performed for 12 hours in relevant groups. The mRNA expression of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) was measured by RT-PCR. The expression of phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) and nuclear factor-kappa B (NF-κB) was measured by Western blot.</p><p><b>RESULTS</b>The expression levels of p-p38 and NF-κB in the UCP2-siRNA+10% septic serum group were significantly higher than in the 10% septic serum group (P<0.05). The UCP2-siRNA+10% septic serum group had a significantly higher TNF-α mRNA expression than the 10% septic serum group (P<0.01), but IL-1β mRNA expression showed no significant difference between the two groups.</p><p><b>CONCLUSIONS</b>UCP2 plays a regulatory role in the activation of p38 MAPK and NF-κB and the expression of downstream inflammatory mediators in H9C2 cells stimulated with septic serum.</p>

Early physical training improves long-lasting prognosis following hypoxic-ischemic brain damage in neonatal rats / 中华神经医学杂志

Objective To investigate the effect of early physical training on long-lasting neurological function,pathology changes,spatial learning and memory abilities,sensorimotor function and ultrastructure changes in neonatal rats with hypoxic-ischemic brain damage (HIBD).Methods Ninety 7-day-old sprague-dawley rats were randomly divided into three groups:a group that was subjected to left carotid ligation followed by 2 hours hypoxic stress (vehicle group),a group that received physical trainings (grabbing,rotation,walking and balance) 1 weeks after HIBD event (trained group) and a sham-operated group that was subjected to a sham-operation without ligation and hypoxic stress (n=30).Following four weeks of physical trainings,neurological scale was performed on all the rats; the number of neurons in the CA1 region of hippocampus and cortex forehead was measured by Nissl count; the expression levels of synaptophysin and c-fos were examined; Morris water maze tests and cortex sensorimotor tests were performed to detect the spatial learning and memory abilities and sensory functions; transmission electron microscope was employed to observe the synapses and neuronal ultrastructure.Results As compared with those in the vehicle group,the neurological scale scores were significantly increased in the trained group and the sham-operated group 14,21 and 28 days after trainings (P＜0.05).As compared with those in the vehicle group,the number of neurons in the left hippocampal CA1 area and cortex increased in the trained group and sham-operated group (P＜0.05).The expressions of synaptophysin and c-los in the trained group increased significantly as compared with those in the vehicle group (P＜0.05).As compared with those in the vehicle group,the spatial learning and memory abilities and sensorimotor functions in the trained group were significantly increased (P＜0.05).The ultrastructure of the left hippocampus and cortex was remarkably abnormal in the vehicle group by the transmission electron microscopy,while no obvious abnormality was observed in the trained group.Conclusion Early physical training can restrain neuron damage in the hippocampus and cortex and enhance synaptic plasticity to ameliorate the long-term neurological function after HIBD; the strong expression of synaptophysin and c-fos in the hippocampus and cortex caused by early physical training may be one of the improvement mechanisms.

Research progress of microRNA in mitochondrial dysfunction of children with sepsis / 中华实用儿科临床杂志

Sepsis is the systemic inflammatory respond syndrome and autoimmunity injury caused by pathogenic microorganism or any other immunogenicity.Mitochondria,an important organelle,which has an essential role in cellular growth,metabolism,occurrence and development of disease by generating ATP following the oxidative phosphorylation from glycolysis.There had been some progresses on the research of sepsis in several decades.A number of studies had shown that the degree of mitochondrial dysfunction was related to the eventual outcome of sepsis.microRNA are endogenous small noncoding RNAs that post-transcriptionally regulate gene and bio-protein expression,and then adjust mitochondrial oxidative stress,has important influence on the process of sepsis and prognosis.Here,a current review of how microRNA impinge on mitochondrial dysfunction in sepsis was performed.

Correlation of the serum hydrogen sulfide level and neuron-specific enolase level in children with febrile seizures / 中华实用儿科临床杂志

Objective To investigate the pathophysiological role of serum hydrogen sulfide (H2 S) and its correlation with serum neuron-specific enolase (NSE) level in the process of febrile seizures (FS).Methods Sixty-five children with FS and acute upper respiratory infection were selected as FS group,51 children with acute upper respiratory infection associated with fever were taken as upper respiratory infection group and 43 healthy children in Child Health Section for physical examination were respectively chosen as the healthy control group.All children came from Department of Pediatrics of Zhujiang Hospital,Southern Medical University from Mar.15 to Nov.10,2012.The serum H2S and NSE levels of all groups were measured by Microplate Reader.Results The serum H2S in FS group was significantly lower than that in upper respiratory infection group and healthy control group (all P ＜ 0.01) ; the NSE level in FS group was significantly higher than that in upper respiratory infection group and healthy control group (all P ＜0.01) ;the levels of H2S and NSE had no significant differences between upper respiratory infection group and healthy control group (all P ＞ 0.05).The serum H2S level and NSE level in FS group were negatively correlated (r =-0.279,P =0.024) ; There were significant differences of serum NSE level between seizure frequency less than twice group and seizure frequency greater than or equal to twice group(t =-2.955,P =0.004).The seizure frequency was negatively correlated with serum H2S level (r =-0.269,P =0.03),and positively correlated with serum NSE level (r =0.322,P =0.009).The seizure durations (≥ 5 min) was negatively correlated with serum H2S level (r =-0.532,P =0.019).Conclusion The serum H2S level is expected to be an objective index for eva-luating the seizure brain injury in the early period of FS,and also potentially the important occurrence factors of brain injury.

Gene cloning and immunogenicity analysis of the structural proteins VP1-VP4 of enterovirus 71 / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone the genes encoding the structural proteins VP1-VP4 of enterovirus 71 and investigate the immunogenicity of the expressed recombinant proteins.</p><p><b>METHODS</b>The VP1-VP4 cDNAs were amplified by RT-PCR from the extracted viral RNA and cloned into pMD19-T vector. The cloned VP1-VP4 genes were then inserted into the multi-cloning sites of plasmid pQE30a and expressed in E. coli M15 with IPTG induction. After washing with 8 mol/L urea and purification with Ni-affinity chromatography, the recombinant proteins obtained were tested for immunogenicity by Western blotting and ELISA using rabbit antisera against enterovirus 71 and Coxsackie Virus A16.</p><p><b>RESULTS</b>The recombinant VP1-VP4 proteins were highly expressed in E. coli M15 and the purified proteins could be specifically recognized by the rabbit sera against enterovirus 71.</p><p><b>CONCLUSION</b>The expressed enterovirus 71 structural proteins show good immunogenicity and can be used for developing enterovirus 71 vaccine and detection kits.</p>

Adrenal mitochondrial injury in early stage of sepsis in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the pathogenesis of adrenal mitochondrial dysfunction in septic rats.</p><p><b>METHODS</b>Thirty SPF rats were randomized into 3 groups, including a normal control group and 2 sepsis groups receiving intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS) and observed at 6 and 24 h after the injection. The adrenal mitochondria were extracted from the rats at the corresponding time points for observation by electronic microscopy. The membrane potential of the mitochondria was detected by flow cytometry. The oxidative stress levels in the mitochondria (activities of NOS and levels of MDA and NO) were assessed.</p><p><b>RESULTS</b>With the progression of sepsis, the serum levels of corticosterone in LPS groups increased significantly as compared with that in the control group. Ultrastructural observation of the adrenal mitochondria showed mild mitochondrial injury in LPS groups in comparison with the control group. The mitochondrial membrane potential was lowered in the LPS groups, but all these changes appeared to be reversible. The activities of NOS and the levels of MDA and NO showed no significant difference between the sepsis groups and the control group.</p><p><b>CONCLUSIONS</b>No obvious adrenal dysfunction occurs in the early stage of sepsis in rats. Mitochondrial injury, which is reversible, occurs in early sepsis without obvious evidence of oxidative stress injury in the adrenal mitochondria, suggesting a strong resistance and capacity of self-repair of the adrenal gland and the mitochondria against sepsis-induced injury.</p>

Application of fluorescent real-time polymerase chain reaction in analyzing the epidemic of influenza among children in Guangzhou area in 2006 / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of influenza virus infections in children in 2006 using the real-time PCR method.</p><p><b>METHODS</b>(1) Consulting the most conserved sequence NP gene of influenza virus, after comparing with the NP gene sequences of influenza virus in GenBank, one pair of specific primers and one TaqMan probe were designed for each subtype of influenza virus by the software Primer Express. The sensitivity of influenza was evaluated by testing known positive samples which had been two-fold diluted. The specificity of real-time PCR for influenza virus detection was assessed by cross testing 60 isolates of influenza A, 16 isolates of influenza B, and by testing a variety of other respiratory viruses positive samples; (2) 281 nasopharyngeal aspirate samples were detected by real-time PCR and virus isolation; (3) the 12 301 specimens from the patients of Guangzhou Children's Hospital were tested by using the real-time PCR method. Furthermore, the real-time PCR reagent was evaluated by comparing with the result of virus isolation.</p><p><b>RESULTS</b>(1) The sensitivity of real-time PCR developed in this study for influenza A detection was 1:2(22) and for influenza B was 1:2(20) in two-fold serially diluted way. (2) No positive results were found in cross testing of other viruses positive specimens. (3) Influenza virus was detected from 1687 cases (13.71%) out of the 12 301 cases, including 773 cases (45.8%) positive for subtype A and 914 cases (54.2%) positive for subtype B; 455 out of 525 (86.7%) of influenza B positive specimens and 70 out of 525 (13.3%) of influenza A (H1N1) positive specimens were from patients seen during January to April; 419 out of 1118 (37.5%) specimens positive for influenza B and 699 out of 1118 (62.5%) specimens positive for influenza A (H1N1) were from patients seen from May to August. Influenza virus could be identified from 1380 samples by the methods of virus isolation, accounting for 81.80% of the 1687 positive samples detected by real-time PCR. All the influenza virus subtype A was H1N1.</p><p><b>CONCLUSION</b>The real-time PCR method developed in this study was sensitive and specific for detecting influenza A and B in clinical specimens. During 2006, influenza A and influenza B co-circulated. The predominant virus was influenza B from January to April, peaking in April. Influenza A (H1N1) prevailed from May to August, with the peak in June.</p>

Clinical features and surgery in children with plastic bronchitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To review the clinical features and therapeutic experience in children with plastic bronchitis.</p><p><b>METHODS</b>Fourteen children with plastic bronchitis were reviewed retrospectively, 12 of which were under two years old. The clinical features are characterized by sudden onset, episodes of profound hypoxia and respiratory tract obstruction. SaO2 was between 0.70 and 0.80 even with mask oxygen inhalation. Eight cases were pyretic, 4 cases expectorated jel-like bronchial casts. The chest X-ray picture showed patchy consolidation or atelectasis unilaterally (10 cases) or bilaterally (2 cases). Pulmonary marking thickening and patchy shadow were observed in 2 cases. Twelve cases underwent rigid bronchoscopy and the bronchial casts were removed. Two cases underwent endotracheal intubation.</p><p><b>RESULTS</b>Eight cases of 12 children received therapeutic bronchoscopy were cured. Other 4 cases had second therapeutic bronchoscopy and bronchial casts were removed again in 3 cases, one died from pulmonary hemorrhage. Two cases who underwent endotracheal intubation died from the multiple organ failure (MOF). Pathologic results showed:the bronchial casts were composed mainly of mucus and fibrin, inflammatory cell infiltrate were observed in 6 cases (Type 1, inflammatory), no cellular infiltrate occurred in 8 cases (Type 2, acellular).</p><p><b>CONCLUSIONS</b>Plastic bronchitis is a severe and dangerous disease. The branching plastic casts may obstruct part or the entire tracheobronchial, causing respiratory failure. Bronchoscopy and pathologic examination are essential for it's diagnosis and treatment.</p>

Analysis of Respiratory Syncytial Virus Infection in Hospitalized Children with Pneumonia in Guangzhou Area from 2005 to 2007 / 实用儿科临床杂志

6-11 years old were 9.67%, 6.81%, 3.49% and 0.80%, respectively.Furthermore, the infection rates between each two age stages were significantly different(Pa0.05).4.Infection rates in 2005,2006 and 2007 were 4.0%, 8.92%, 8.85%,respectively.Infection rates between 2005 and 2006,2007 were significantly different(Pa

Clinical characteristics and prognosis of 33 children with severe acute respiratory syndrome in Guangzhou area / 中华儿科杂志

<p><b>OBJECTIVE</b>Since the outbreak of a highly contagious new pneumonia, atypical pneumonia or severe acute respiratory syndrome (SARS) occurred in Guangzhou area, 33 children with this syndrome were treated in the authors' hospital. The present study aimed to understand clinical characteristics and prognosis of pediatric SARS patients in Guangzhou area.</p><p><b>METHODS</b>Clinical manifestations, laboratory and radiologic findings, therapeutic approaches and prognosis of the 33 children with SARS in Guangzhou area were analyzed.</p><p><b>RESULTS</b>Of the 33 cases, 17 were males and 16 were females. The age was between 3 months to 13 years, and 3 - 12 years old patients accounted for 82%. Five (15%) cases had an evident history of contacting SARS patient before the symptoms occurred. Another 5 (15%) cases had a history that contacts of these patients (family members or friends) developed fever and/or cough later. The most common symptoms in this cohort were fever (100%) and cough (91%). Most of the cases had high fever, higher than 39 degrees C. Near half of the cases had nonproductive cough. The initial blood cells count showed that total white blood cell (WBC) count was (2.5 - 9.7) x 10(9)/L. In 22 (67%) cases the WBC count was < 5.0 x 10(9)/L, and in 10 (30%) WBC was (5.0 - 7.0) x 10(9)/L, in 18 cases most of the WBC were lymphocyte count. Chest radiograph showed patchy infiltrates, in 15 cases the changes were unilateral, and in 18 were bilateral. The radiologic changes developed fast, in some cases the changes progressed from one side to both sides. The opacity was absorbed slowly, significant absorption took in average two weeks. Elevated ALT was found in 3 cases and elevated CK-MB in 2 cases. Treatment included isolation, good ventilation of the ward, bed rest, supportive regimens, low volume oxygen inhalation, use of Chinese traditional medicine, antibiotics to prevent bacterial infection, and anti-inflammation therapy. All the patients recovered and discharged from hospital after a mean period of 10.0 +/- 3.8 days.</p><p><b>CONCLUSION</b>SARS in children may have its own characteristics. The main clinical manifestations were high fever and cough while no severe toxic symptoms, nor respiratory failure was seen; few symptoms or signs suggesting involvement of systems other than respiratory system were seen. Chest radiograph showed uni- or bilateral asymmetric air-space infiltrates which could worsen quickly and were absorbed slowly. Though there were severe changes in the lung, the patients might not have corresponding symptoms or signs. The total white blood cell count in peripheral blood did not increase. All the patients studied had a favorable outcome after the combined treatment.</p>

Clinical Application of Pediatric Risk of Mortality Score in Pediatric Intensive Care Unit / 实用儿科临床杂志

15, the mortality was38.1%(n = 8).There was significantly difference both in mortality (x2 = 4.14 P

Significance of Monitoring Serum Level of Gentamycin in Children with Serious Illness / 实用儿科临床杂志

Objective To explore the serum level of gentamycin for orally in children with serious illness.Methods The serum level of gentamycin in 41 children who were in serious illness [multiple organ dysfunction(MODS)group with 21 cases and non-MODS group with 20 cases ] were monitored and the patients were treated with select decontamination of the digestive tract(SDD) from October 2004 to April 2005.Dosage:10 mg/(kg?d),orally taken three times(every 8 hours) one day.The blood after taking the drug one hour later in the fourth day was selected and the serum level of getamycin was monitored.Results Thirty-six children of 41 cases serum level of gentamycin were negative and 5 children(4 in MODS group and 1 in non-MODS group) who had alimentary tract hemorrhage were masccline in serum after taking gentamycin one hour later in the forth day.The absorption of gentamycin from enteric after orally was not(rela)-ted to MODS.There were statistics value between the gestrintestinal tract ulcer and serum level of gentamycin.Conclusions The safety for treating the children in serious illness with gentamycin for SDD is obvious.But we suggest to monitor the serum level of gentamycin for who has severe alimentary tract hemorrhage together with insufficiency of liver and kindey.