It is a brief introduction of necessity of uniform guidelines for pediatric emergency and critical care.Uniform guidelines are very important for development of discipline,safety of patients,protection of medical staff and administration.Uniform guidelines should be enacted according to our national condition with a scientific attitude of rigorous and gravity.

Objective To study the molecular characteristics of the Staphylococcus aureus isolated from the intensive care units (ICUs) of children's hospital.Methods From January 2016 to December 2016,a total of 39 S.aureus strains were collected and identified from various clinical specimens that were obtained from patients who were confined in the neonatal and pediatric ICUs of Beijing Childreng Hospital.Methicillin-resistant S.aureus (MRSA) and methicillin-susceptible S.aureus (MSSA) were identified using the cefoxitin disc method and the detection of the mecA gene.Multilocus sequence typing (MLST) and spa typing were analyzed using the PCR,and the staphylococcal chromosomal cassette mec (SCCmec) type was analyzed for the MRSA isolates.Twenty-one superantigen genes and the Panton-Valentine leukocidin (pvl) gene were also detected by PCR.Their susceptibility to 12 antibiotics was evaluated using the E-test method.The differences in prevalence of virulence genes and antimicrobial resistance were compared between the MRSA and MSSA isolates by Fisherg exact test.Results All the S.aureus strains were isolated from secretion inside the airway of pneumonia (including severe pneumonia),the blood of patients with bacteremia,and exudate of skin and soft tissue infections.ST59-SCCmecⅣa-t437 (55.6％) and ST398-t571 (28.6％) were the most predominant clones of MRSA and MSSA,respectively.Of the 39 isolates,26 strains (66.7％) had at least one superantigen gene,and seb (38.5％),sek (30.8％),and seq (20.5％) were the most common genes;seb-sek-seq (18.0％) was the main virulence genotype.The pvl geneg positive rate was 25.6％,and no significant difference between MRSA and MSSA was observed (P ＞ 0.05).Notably,79.9％ of the S.aureus isolates were multidrug resistant,and 94.9％,53.8％,and 51.3％ of the isolates were resistant to erythromycin,clindamycin,and chloramphenicol,respectively.The tested isolates were susceptible to trimethoprim/sulfamethoxazole,rifampicin,and vancomycin.Conclusions The S.aureus strains from the ICUs of childreng hospital were isolated from the secretion inside the airway of pneumonia (including severe pneumonia),the blood of patients with bacteremia,and exudate of skin and soft tissue infections.ST59-SCCmecⅣa-t437 (55.6％) and ST398-t571 (28.6％) were the common clones of MRSA and MSSA,respectively.The prevalence of superantigen genes and the multidrug resistant rate were relatively high.

Objective To assess the value of prenatal ultrasonography on fetal forearm and crus malformations by studying the ultrasonographic characteristic in relation to methods of examination. Methods All fetus were evaluated by using a systematic continuous sequence approach (SCSA) with ultrasonography. A close attention was paid on shapes, structures and movement of fetal forearm and crus, and fetus specimens after induced labor were rescanned by ultrasonography under the condition of mimic intra-uterus. Results Thirty of 33 cases (90.9%) with fetal forearm and crus malformations (totally 48 of 54 limb anormalies, 88.9%) were correctly diagnosed by prenatal ultrasonography. Six limb malformations of 3 cases were missed. Conclusion Our study demonstrates that the malformations of forearm and crus detected with prenatal ultrasonography are highly accordant with the malformations revealed in fetus in vitro. It is important to abide by the SCSA in ultrasonographic diagnosis of fetal forearm and crus malformation.

Objective To evaluate the synergistic effect of everolimus on cisplatin-mediated cytotoxicity against human cutaneous squamous cell carcinoma COLO-16 cells.Methods Cultured COLO-16 cells were divided into several groups to be treated with everolimus at different concentrations of 50,100 and 200 nmol/L or 25 μmol/L cisplatin for 12 and 24 hours.Acridine orange (AO)-labeled autophagic vesicles combined with lysomal enzyme inhibitors (E64d and pepstatin) were used to detect the levels of autophagy and autophagic flow.Western blot analysis was performed to track the conversion of the autophagosome marker microtubule-associated protein 1 light chain-3 (LC3)-Ⅰ to LC3-Ⅱ,as well as to detect cleavage levels of Caspase 3 and poly-ADP-ribose polymerase (PARP).Lactate dehydrogenase (LDH) assay was conducted to detect cell death,and Annexin V-EGFP staining to evaluate cell apoptosis.Results The LC3-Ⅱ / LC3-Ⅰ ratios (LC3-Ⅰ conversion to LC3-Ⅱ) after 12-and 24-hour treatment did not differ among the 50-,100-and 200-nmol/L everolimus groups (12 hours:3.52 ± 0.21 vs.4.03 ± 0.39 vs.5.05 ± 0.22,P ＞ 0.05;24 hours:3.38 ± 0.26 vs.3.29 ± 0.06 vs.6.57 ± 0.16,P ＞ 0.05),but were significantly higher in the three everolimus groups than in the control group receiving no treatment (12 hours:2.07 ± 0.05,P ＜ 0.05;24 hours:2.61 ± 0.16,P ＜ 0.05).After 12-hour treatment,no significant differences were observed in the ratio of LC3-Ⅱ to β-actin between the 50-nmol/L everolimus + E64d + pepstatin group (1.26 ± 0.40),100-nmol/L everolimus ± E64d + pepstatin group (1.16 ± 0.34),200-nmol/L everolimus + E64d + pepstatin group (1.21 ± 0.39) and E64d + pepstatin group (1.19 ± 0.27,P ＞ 0.05).Moreover,there was no significant difference in the percentages of autophagic vesicle-positive cells between the 100-nmol/L everolimus + E64d + pepstatin group and E64d + pepstatin group (2.06％ ± 0.61％ vs.1.68％ ± 0.62％,P ＞ 0.05).After 24-hour treatment,the everolimus + cisplatin group showed significantly increased rate of cell death compared with the cisplatin alone group (42.58％ ± 0.93％ vs.18.20％ ± 1.46％).However,no significant differences were observed in the cleavage levels of Caspase 3 and PARP,the number of annexin V-labelled cells and ratio of LC3-Ⅱ to β-actin between the everolimus + cisplatin group and the cisplatin-alone group (P ＞ 0.05).Conclusion Everolimus has a synergistic effect on the cisplatin-mediated COLO-16 cell death,and this effect does not depend on cell apoptosis or autophagy.

A pandemic influenza A (H1N1) virus strain was isolated from a pig farm in Korea in December 2009. The strain was propagated in and isolated from both the Madin-Darby canine kidney cell line and embryonated eggs. The partial and complete sequences of the strain were identical to those of A/California/04/2009, with >99% sequence similarity in the HA, NA, M, NS, NP, PA, PB1, and PB2 genes. The isolated strain was inactivated and used to prepare a swine influenza vaccine. This trial vaccine, containing the new isolate that has high sequence similarity with the pandemic influenza A (H1N1) virus, resulted in seroconversion in Guinea pigs and piglets. This strain could therefore be a potential vaccine candidate for swine influenza control in commercial farms.
Agriculture ; Animals ; Cell Line ; Eggs ; Guinea Pigs ; Influenza Vaccines ; Influenza, Human* ; Kidney ; Korea* ; Orthomyxoviridae* ; Ovum ; Pandemics* ; Seroconversion ; Swine

Objective To investigate molecular mechanisms underlying the synergistic damage to the human squamous cell carcinoma cell line COLO-16 by everolimus and cisplatin.Methods In the signaling pathway experiment,COLO-16 cells were divided into 4 groups:control group receiving no treatment,50,100 and 200 nmol/L everolimus groups treated with 50,100 and 200 nmol/L everolimus respectively.In the combined experiment,COLO-16 cells were divided into another 4 groups:control group,50 nmol/L everolimus group,25 mol/L cisplatin group,and 50 nmol/L everolimus + 25 mol/L cisplatin group.Western blot analysis was performed to analyze changes in mammalian target of rapamycin (mTOR) pathway,Akt pathway,DNA damage-related pathway and Csk homologous kinase (Chk) pathway.Results After the treatment with everolimus at different concentrations of 50,100 and 200 nmol/L for 12 and 24 hours,the phosphorylation levels of mTOR at ser2448 and ser2481 as well as Rictor at thr1 135 in COLO-16 cells were all decreased compared with the control group.However,there were no significant changes in the phosphorylation levels of downstream signals ULK1 at ser757,p70 S6 at thr389 and PKCα at thr638/64.The treatment with everolimus did not change the total protein level and phosphorylation of Akt.After the treatment with cisplatin for 12 and 24 hours,the phosphorylation levels of Rictor at thr1135 and Chk1 at ser345 were significantly increased,but the treatment with everolimus alone showed no such effects.After the combined treatment with everolimus and cisplatin for 12 and 24 hours,the upregulation of Chk1 and Rictor phosphorylation were significantly inhibited compared with the cisplatin alone group.Conclusions mTOR signaling is sensitive to everolimus in COLO-16 cells,but its targeted pathway is not regulated simultaneously to develop a cascade reaction.Everolimus may increase the cisplatin-induced death of COLO-16 cells by inhibiting the activation of Chk 1,but can not aggravate DNA damage induced by cisplatin.

Objective To evaluate the regulatory effects of lycopene on the key signaling receptors in human cutaneous squamous cell carcinoma cell line COLO16.Methods Cultured COLO16 cells were divided into 6 groups to be treated with lycopene at different concentrations of 0,5,10,15,20,and 25 μmol/L,respectively,for 24 hours (control group and 5,10,15,20,25 μmol/L lycopene groups),followed by estimation of the cell viability by lactate dehydrogenase (LDH) assay.Lycopene at a safe concentration was selected based on the LDH assay,and used for the determination of expression of signaling receptors,and Western blot analysis was performed to measure the expression of key signaling receptor proteins,including epidermal growth factor receptor (EGFR),glucocorticoid receptor (GR),retinoic acid receptor-alpha (RAR-α),retinoid X receptor-alpha (RXR-α),androgen receptor (AR) and progesterone receptor (PR).Statistical analysis was carried out by one-way analysis of variance (ANOVA),Tukey multiple comparison teat and Brown-Forsythe test with the SPSS software.Results After 24-hour treatment with lycopene at different concentrations,there were significant differences in the rate of cell death among these groups (F =13.116,P ＜ 0.05),and the rate of cell death in the 25 μmol/L lycopene group significantly differed from that in the control group (P ＜ 0.05).Therefor,lycopene at concentrations of 5,10 and 20 μmol/L were selected to treat COLO16 and HaCaT cells as well as human epidermal keratinocyte (HEK) for 24 hours in the following experiment.The treatment with lycopene significantly decreased the phosphorylation level of EGFR (P ＜ 0.05),but significantly increased the expression of GR protein (P ＜ 0.05),and showed no significant effects on the protein expression of RAR-α,RXR-α,AR,and PR in COLO16 cells.After 24-hour treatment with lycopene at concentrations of 5,10 and 20 μmol/L,there were no significant changes in the phosphorylation level of EGFR protein or the expression of GR protein in HaCaT cells and HEK (all P ＞ 0.05) compared with those without lycopene treatment.Conclusion Lycopene can decrease the viability of COLO16 cells,inhibit the activation of EGFR protein,and up-regulate the expression of GR,and these effects may be specific for tumor cells.

Objective To investigate the molecular characteristics,virulence genes and antimicrobial susceptibility to Staphylococcus aureus strains isolated from children with skin and soft tissue infections (SSTIs) in Beijing.Methods A total of 52 Staphylococcus aureus isolates were collected from children with SSTIs in Beijing Children's Hospital,Capital Medical University,and the clinical data were collected and analyzed.Methicillin-resistant Staphylococcus aureus(MRSA) and methicillin-susceptible Staphylococcus aureus(MSSA) were identified by using the cefoxitin disc method and the detection of mecA gene.Multilocus sequence typing (MLST) and staphylococcal protein A (spa) typing were analyzed by the PCR method,and the staphylococcal chromosomal cassette mec (SCCmec) type was analyzed for the MRSA isolates.The pvl,eta,etb,tsst-1 and hlg genes were also detected by PCR.The susceptibility strains to 16 antibiotics were evaluated by using the agar dilution method.Results A total of 52 Staphylococcus aureus SSTIs patients,30 with MRSA infections and 22 with MSSA infections were included in the study.There were 23 patients (44.2％) less than 1 year old.The most frequent infections were the newborn omphalitis (12/52 strains,23.1％)and abscess(11/52 strains,21.2％).ST59-MRSA-SCCmecⅣa-t437 was the most predominant clones of MRSA isolates.Among the MSSA isolates(14/30 strains,46.7％),no significant epidemic clone was found.Ten sequence types (STs) and 14 spa types were identified in MSSA,and the most common types were ST22(6/22 strains,27.3％)and t309 (5/22 strains,22.7％),respectively.Notably,the multidrug resistant rates of MRSA and MSSA isolates were all ＞ 85％.The percentages of the Staphylococcus aureus SSTIs strains resistant to Erythromycin,Penicillin,Chloramphenicol and Clindamycin were 100.0％,94.2％,69.2％ and 63.5％,respectively.The tested isolates were susceptible to Trimethoprim/Sulfamethoxazole,Mupirocin,Fusidic acid,Tigecycline,Linezolid and Vancomycin.The pvl gene's positive rate was 40.4％,and no significant difference between MRSA and MSSA was observed (P ＞ 0.05).Eta and etb genes were detected in 2 patients with staphylococcal scalded skin syndrome.Conclusions The Staphylococcus aureus SSTIs strains are most frequently isolated from newborn omphalitis and abscess in Beijing.The multidrug resistant rate is relatively high,so the erythromycin and clindamycin should not be preferred in empiric treatment of children with Staphylococcus aureus SSTIs.The prevalence of pvl gene is 40.1％.ST59-MRSA-SCCmecⅣa-t437 is the common clone of MRSA,while the MSSA isolates have a more diverse genetic background.

A case with mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase deficiency(HMGCSD)was related and foreign and domestic reported cases were reviewed.The female proband was 7 months and 16 days old, and admitted to the hospital due to acute onset of " fever for 4 days, wheezing for 3 hours, dyspnea and moaning for 2 hours" . She was mainly manifested as encephalopathy, hepatomegaly, liver function damage, low ketone hypoglycemia, and hyperlipidemia.She died of respiratory and circulatory failure on the third day of hospitalization.Two compound he-terozygous variants in HMGCS2 gene were found by total exome sequencing, namely, c.1061+ 1 G> C and c. 476 G> T. HMGCSD could be diagnosed by gene detection in combination with clinical features of the patient. Thirteen literatures related to HMGCSD were collected, including 26 patients in total, with the age of onset ranging from 3 months to 6 years. The main cause of the disease was insufficient intake, mainly manifested as hypoglycemia accompanied by low ketone, hepatomegaly, liver damage, etc. A high level of urinary 4- hydroxy-6- methyl-2- pyrone might be a strong indicator of HMGCSD. Three died during the acute attack. Up to now, there were 32 mutations in HMGCS2 reported in 26 patients, and the main type was missense mutation. In this article, the second case of HMGCSD in China was identified, and 2 novel variants of HMGCS2 were found, which extended the clinical phenotype and mutation spectrum of HMGCSD.

Objective: To evaluate the effect of prophylactic nimodipine in vasospasm prevention and outcome improvement in children with subarachnoid hemorrhage (SAH). Methods: A prospective, randomized controlled clinical trial which enrolled children with SAH who were admitted to pediatric intensive care unit (PICU) of Beijing Children′s Hospital from January 2015 to October 2018 was conducted. A total of 43 patients were randomly divided into nimodipine group (24 patients) and control group (19 patients) according to random number table. Transcranial Doppler (TCD) was used to dynamically monitor blood flow velocity and spectrum monography of bilateral middle cerebral artery (MCA) for vasospasm evaluation. Pediatric cerebral performance category (PCPC) scale was used to evaluate patients′ brain function on 28^th day after discharge. Data were analyzed by t test, Mann-Whitney U test, χ² test. Results: Except heart rate ((157±26) vs. (137±34) beats/min, t=2.079, P=0.045), no significant differences existed between the two groups in basic demographic characteristics, primary diseases, and clinical manifestations (all P>0.05). The peak velocities of bilateral MCA on the 5^th day after admission were significantly lower in nimodipine group (left MCA (136±34) vs. (158±23) cm/s, t=-2.890, P=0.006; right MCA (129±34) vs. (176±27) cm/s, t=-3.717, P=0.001). Likewise, a lower peak velocity of left MCA was observed on the 7^th day after admission in nimodipine group ((127±45) vs. (152±13) cm/s, t=-2.903, P=0.007), but no significant difference existed in that of right MCA ((131±48) vs. (150±22) cm/s, t=-1.760, P=0.090). Eleven patients suffered from vasospasm, 25% (6/24) in nimodipine group and 26% (5/19) in control group (χ²=0.010, P=1.000), within whom 8 patients had complete remission after continuing nimodipine treatment, one died in hospital and the other two′s vasospasm still existed at the time of discharge. No significant differences were found between the two groups in mean length of hospitalization, proportion of mechanical ventilation, Glasgow coma scale at discharge, survival rate at discharge or survival rate on 28^th day after discharge (all P>0.05). However, nimodipine group had a higher proportion of favorable PCPC brain function (92% (22/24) vs. 63% (12/19), χ²=5.208, P=0.030). No side effects such as hypotension, rash or injection site erythema were observed. Conclusion Prophylactic nimodipine cannot reduce vasospasm incidence in children with SAH but may improve short-term brain function, without any significant safety issues.