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Objective To explore the role and efficacy of VEGF and HGF gene adenovirus vector in promoting angiogenesis in ischemic tissue.Methods 84 Kunming mice were randomly divided into sham group,control group,VEGF group,HGF group and VEGF+HGF group,and the left lower limb ischemia model was established.The blood supply of ischemic tissue was observed by rheometer,and the expression levels of VEGF and HGF in each group were detected by Western Blot and ELISA.Immunohistochemical staining was used to detect angiogenesis(CD31,SMA)in ischemic tissues.Safety was assessed by side effects during treatment in mice.Results After the successful modeling,the blood flow velocity of the left lower limb in each group decreased significantly.On the 7th day after operation,the blood flow of the left lower limb in each group was significantly better than that on the 0th day after operation(P<0.05),and the blood flow of the left lower limb in Ad-VEGF-HGF group was significantly better than that in other groups(P<0.05).On the 28th day after operation,the blood flow of the left lower limb in Ad-VEGF-HGF group gradually stabilized,the blood flow in Ad-VEGF-HGF group was significantly better than that in other groups,and both VEGF group and HGF group were significantly better than the control group(P<0.05).On the 7th,14th,and 28th days following surgery,HGF and VEGF protein levels in the Ad-HGF,Ad-VEGF,and Ad-VEGF-HGF groups were substantially greater than those in the control group(P<0.05).The expression level in the Ad-VEGF-HGF group peaked on the 14th day(all P<0.001)and subsequently declined to preoperative levels on the 28th day after operation.Conclusion Ad-VEGF-HGF gene injection can effectively boost VEGF and HGF protein expression and rapidly reach the relative peak level,encour-aging angiogenesis after lower limb ischemia,increasing blood flow,and improving lower limb circulation.
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Objective:To enhance understanding of mental retardation autosomal dominant 35 (MRD35) by analyzing the clinical and genetic characteristics of the disease.Methods:Clinical and genetic data of 1 case of MRD35 in Beijing Children′s Hospital in July 2018 were reported, and literature review was conducted.Results:The male proband, 1 year and 3 months old, was admitted with the clinical manifestations including mental retardation, low-grade fever, a large forehead, flat nose, open mouth, and hypomyotonia. The brain magnetic resonance imaging showed enlarged lateral ventricles, cavum septum, cavum verge and cavum velum interpositum cyst. The whole exome sequencing test showed that the proband carried a missense mutation c.1258 G>A, (p.E420K) in the PPP2R5D gene, and the mutation was de novo confirmed by Sanger sequencing. There were ten literatures reported, including a total number of 31 cases. Counting on this case, totally 32 cases were included. Among the 32 patients, 32 cases (100.0%) had mental retardation, 26 cases (81.3%) with motor retardation, 26 cases (81.3%) with macrocephaly, 8 cases (25.0%) with epilepsy. Facial dysmorphic features, ocular abnormalities, skeletal abnormalities, and cardiac malformations were also reported. All reported individuals had missense mutations of PPP2R5D gene and were autosomal dominantly inherited. Conclusions:The main clinical manifestations of MRD35 include growth retardation/mental retardation, severe speech impairment, macrocephaly, hypomyotonia, seizures and dysmorphic facial features. A novel missense mutation in the PPP2R5D gene is the cause of MRD35.
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Influenza is an acute respiratory infectious disease caused by influenza virus, which can promote the adhesion and colonization of Streptococcus pneumoniae in respiratory epithelial cells, increasing the incidence of pneumococcal infections.Therefore, children should take early precautions against pneumococcal diseases during or before the influenza season.The progress in research on the epidemiology of influenza and the prevention of pneumococcal diseases in children was reviewed in this paper.
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Bacterial meningitis infections can cause serious complications and sequelae in children. Streptococcus pneumoniae is a major pathogen of the disease.Early and rapid identification of the pathogen of infection is of great significance for precise treatment, as well as mortality and reduction of sequelae.In recent years, the rapid progress of molecular biology methods has enormously changed the clinical pathogen diagnosis pattern.In this article, the progress in the pathogenic diagnosis of pneumococcal meningitis was reviewed, in order to acquire a better and systematic understanding of the role that emerging technologies play in the accurate diagnosis of pneumococcal meningitis.
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Objective@#To summarize the clinical characteristics of Listeria monocytogenes meningitis (LMM) with complications, and analyze the outcomes of next generation sequencing.@*Methods@#Clinical characteristics, laboratory findings, imaging features, antibiotics treatment, and next generation sequencing of cerebrospinal fluid were analyzed in 3 LMM patients who were hospitalized in the Department of Infectious Diseases of Beijing Children′s Hospital Affiliated to Capital Medical University from July 2015 to November 2017.@*Results@#The three patients were 1-year-old girl, 2-year-old girl, and 9-year-old boy, with normal immune function. They had eaten refrigerated food, milk or dairy products before onset. Symptoms included fever, headache, abdominal pain, diarrhea, vomiting, and convulsions, etc. The complications of two cases (case 2 and 3) were appendicitis and Meckel′s diverticulitis. The other one (case 1) was with sepsis and pneumonia. Leukocyte counts in cerebrospinal fluid were elevated in all the three cases, and cranial magnetic resonance imaging showed meningeal or periventricular involvement. All the children were diagnosed with LMM by positive CSF culture. CSF for next generation sequencing was sent after carbapenem antibiotics using, yet all the results were positive. The positive results were returned 2, 9, and 9 days earlier than culture results, respectively. The gene coverage was 5.00%, 7.00%, 0.04%, and the reads was 2 561, 1 011 and 8, respectively. All the three children had recurrent fever despite using cephalosporin. Levels of leukocytes in the blood and CSF further elevated. After using carbapenem antibiotics, patients improved eventually and were discharged from hospital.@*Conclusions@#LMM can occur in children with normal immune function and is usually associated with digestive system symptoms. Listeria monocytogenes can be detected quickly and accurately by the next generation sequencing technology, without being limited to sampling time and antibiotics application.
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Objective To explore the clinical value of 3D fast spin echo with an extended echo train acquistion (CUBE) T1WI enhancement sequence for purulent meningitis by comparing to contrast?enhanced T2WI?FLAIR sequence in the detection of lesions. Methods From August 2016 to July 2017, children with clinically suspected purulent meningitis underwent cranial magnetic resonance examination in our hospital. There were 35 children, 19 males and 16 females, aged 2 months to 3 years (median age 8 months) in total.A GE Discovery MR 750 3.0 T scanner was used to perform routine plain and enhanced scan in all children. After enhancement, the sequences of CUBE T1WI and FLAIR T2WI were applied randomly. Using the FLAIR T2WI enhancement sequence as the reference, we evaluated the detection rate of CUBE T1WI enhancement sequence for dura mater and leptomeningeal thickening. The number of enhanced lesions detected was tested by χ2 test. Results The enhanced FLAIR T2WI sequence showed 21 cases with dural enhancement, showing a rate of 60.0%. The enhanced CUBE T1WI sequence showed 31cases with dural enhancement, showing a rate of 88.6%. There were significant differences between the dural lesions detected(χ2＝6.058, P<0.01). The enhanced FLAIR T2WI sequence showed 16 cases of leptomeningeal enhancement,showing a rate of 45.7%. The enhanced CUBE T1WI sequence showed 19 cases of leptomeningeal enhancement,showing a rate of 54.3%.The enhanced CUBE T1WI sequence was not significantly higher than that of enhanced T2WI?FLAIR sequence in displaying leptomeningeal enhancement (χ2=0.229, P>0.05). Conclusion Enhancement of the CUBE T1WI sequence enables better visualization of meningeal thickening than FLAIR T2WI. It has great clinical value in the diagnosis of purulent meningitis in children.
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Objective To analyze the clinical characteristics and prognosis of disseminated cryptococcosis in children.Methods The data of disseminated cryptococcosis inpatients were reviewed retrospectively at Beijing Children's Hospital,Capital Medical University,from January 2002 to September 2014.The demographic data,clinical manifestations,laboratory findings,imaging,antifungal treatments and outcomes of all the patients were analyzed.Results Overall 25 children with disseminated cryptococcosis were enrolled including 17 boys(68.0％).The average age was 7 years old.Four cases (16.0％) had underlying conditions,among them 1 case had human immunodeficiency virus(HIV)-positive.The median time to diagnosis was 32 (23-47) days,44.0％ of the patients (11 cases) were misdiagnosed,and 8 cases misdiagnosed as tuberculosis.All patients had fever.Other common clinical manifestations included cough (16 cases),headache (10 cases),vomiting (10 cases),altered mental status (6 cases) and stomachache (6 cases).Respiratory system involvement was seen in all cases,central nervous system was involved in 18 cases,other organ involvement included lymph nodes,spleen,liver,kidney,skin,skeleton and costicartilage.Amphotericin B (AmB) + Fluconazole (Flu) ± 5-flucytosine (5-FC) was the most common therapy (15 cases),Flu ± 5-FC for 6 cases,AmB ± 5-FC for 2 cases,Voricanazole (VOR) for 1 case.Sixteen cases (66.7％) got recovery/improved on discharge,8 cases (33.3％) rejected to the advise and discharged with treatment failure,and the HIV-infected patient transferred to a special hospital.Fifteen patients (60.0％) were followed up,and 13 cases (87.0％) showed recovery,but 2 died in the long-term prognosis.Conclusions Disseminated cryptococcosis in children is more common in school-age boys.Most patients are without underlying conditions.Disseminated cryptococcosis can cause multiple organ damage.Patients with prolonged fever,cough,headache,with or without underlying diseases,can be suspected as disseminated cryptococcosis.Blood and cerebrospinal fluid culture for fungus,cryptococcal antigen should be performed for early diagnosis and treatment.
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Objective To investigate the clinical characteristics and antibiotic resistance of the bloodstream infections due to NDM-1 producing Klebsiella pneumoniae in children.Methods The nonduplicate carbapenem-resistant Klebsiella pneumoniae (CRKp) strains isolated from blood samples were collected in Beijing Children's Hospital from January 2011 to August 2014.Antimicrobial susceptibility was tested with broth microdilution method.PCR amplification and DNA sequencing were conducted targeting blaNDM-1 genes.Medical records were reviewed and analyzed.Results Of the 52 CRKp strains,blaNDM-1 gene was detected in 28 strains.All NDM-l-producing strains were multidrug-resistant.All the 28 isolates were resistant to penicillin,cephalosporins,piperacillin-tazobactam,and imipenem.More than 75.0％ of these NDM-1-producing strains were resistant to aztreonam,trimethoprim-sulfamethoxazole,gentamicin,and meropenem (92.9％,26/28).NDM-1-producing isolates had higher carbapenem MICs than non-NDM-1-producing isolates.Most (82.1％,23/28) of the NDM-1-producing isolates were isolated from hematology-oncology ward.The most common underlying disease was hematologic malignancy (78.6％,22/28).Febrile neutropenia was found in 20 (71.4％) patients.No difference was found between NDM-1-producing and non-NDM-1-producing CRKp infection in terms of repeated hospitalization (P=0.202),prior antibiotic use (P=0.615),underlying diseases (P=0.856),and deep venous catheter (P=0.099).After the susceptibility results were available,37 patients received carbapenembased combination regimen.The mortality did not show difference between NDM-1 producing CRKp infections and non-NDM-1-producing CRKp infections,7.1％ (2/28) vs.12.5％ (3/24),P=0.625.Conclusions The NDM-1 carbapenemase producing Klebsiella pneumoniae is emerging in this hospital.NDM-1-producing strains are resistant to multiple antimicrobial agents,associated with higher carbapenem MIC value.However,no difference was found in the clinical features between the bloodstream infections due to NDM-1-producing strain and those due to non-NDM-1-producing strains.
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<p><b>OBJECTIVE</b>To analyze risk factors, clinical features, outcomes and antibiotic resistance of Escherichia coli(E.coli) causing bloodstream infections in children.</p><p><b>METHOD</b>All inpatients with E. coli positive blood culture in Beijing Children's Hospital from January 2012 to May 2014 were enrolled; 112 cases were included, 66 cases (58.9%) were male, and 46 cases(41.1%) were female. Age range was 2 days to 16 years. Among them, 43 cases (38.4%) were neonates, 19 cases (17.0%) aged from 1 month to 1 year, 14 cases (12.5%) were 1-3 years old, and 36 cases (32.1%) were over three years old. We analyzed the divisions to which the patients were admitted, source of infection, underlying diseases, clinical characteristics, antibiotic resistance, and treatment outcomes, etc.</p><p><b>RESULT</b>Forty-six cases (41.1%) were treated in division of hematology, 42 (37.5%) in neonatology, 9 (8.0%) in internal medicine, 8 (7.1%) in surgery, and 7 (6.3%) in pediatric intensive care unit. Sixty-five cases(58.0%) had underlying diseases. Fever was the most frequently presented symptom, as it was seen in 91 cases (81.3%); 52 cases(46.4%) had respiratory symptoms. Among these, 43 cases had pneumonia, 3 cases had respiratory failure, 3 cases were diagnosed as upper respiratory tract infection, 2 had pulmonary hemorrhage and 1 case had bronchitis. Twenty-six cases (23.2%)were diagnosed as severe sepsis and purulent meningitis separately, 14 cases(12.5%) had urinary tract infection. There were 73 (65.2%) strains inducing extended spectrum β-lactamases (ESBLs), of which 6 (8.2%) and 10 (13.7%) strains were resistant to amikacin and carbapenems respectively. Resistance rate against other antimicrobial agents varied from 64.6% to 100%.</p><p><b>OUTCOMES</b>92 (82.1%) cases were cured or had improvement while 20 patients (17.9%) died or could not be cured at the end of treatment. Positive ESBLs (χ(2) = 6.609, P = 0.010), being complicated with severe sepsis (χ(2) = 40.253, P = 0.000) and requiring mechanical ventilation (χ(2) = 34.441, P = 0.000) indicate poor prognosis.</p><p><b>CONCLUSION</b>Patients with underlying diseases and newborns are susceptible to E. coli bloodstream infection. ESBLs infection, severe sepsis and mechanical ventilation indicate poor prognosis in E. coli blood stream infection. Clinicians may use carbapenems as empirical treatment for ESBLs infection. There may be carbapenem-resistant enterobacteriaceae strains infection if patients receiving treatment with carbapenems have no response.</p>
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Adolescent , Enfant , Enfant d'âge préscolaire , Femelle , Humains , Nourrisson , Nouveau-né , Mâle , Bactériémie , Carbapénèmes , Utilisations thérapeutiques , Résistance bactérienne aux médicaments , Escherichia coli , Infections à Escherichia coli , Sang , Traitement médicamenteux , Méningite bactérienne , Tests de sensibilité microbienne , Pneumopathie infectieuse , Infections urinaires , bêta-LactamasesRÉSUMÉ
Objective To investigate the effects of trimebutine maleate (TM) on the expression of large conductance calcium-activated potassium channel (BKCa) and ryanodine receptors (RyR)channels at mRNA and protein level in colonic smooth muscle cell of cold restraint stress(CRS)induced rats.Methods A total of 24 Wistar rats were divided into CRS group,CRS with TM group and control group equally.The rats of CRS group were gavaged with 0.9％NaCl (6 ml/kg) daily; the rats of CRS with TM group were gavaged with 15 g/L TM (6 ml/kg) daily and activity was restricted in wire cage at 4 ℃ for two hours,continuously for five days.The rats of control group were gavaged with 0.9 ％ NaCl (6 ml/kg) once without CRS.The amount and characteristics of stool of rats in each group were observed.The colonic smooth muscle was isolated to detect the expression of BKCa and RyR at mRNA and protein level by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot.Results The median of rats defecation particles of CRS group was six,control group was one and CRS with TM group was five.Compared with control group,the defecation appearance of CRS group and CRS with TM group was looser and wetter observed by naked eyes.Compared with control group,there was no obvious pathological changes in CRS and CRS with TM group.There was no significant difference in the mRNA expression of BKCa and RyR channels between control group and CRS group.Compared with control group,the BKCa expression at mRNA level of CRS with TM group increased 1.45 fold.Compared with control group,the RyR2 expression at mRNA level of CRS with TM group increased 1.32 fold.Compared with control group,the BKCa expression at protein level of CRS with TM group increased 1.39 fold,and there was no RyR2 expression band at protein level.Conclusion TM might affect colonic smooth muscle contraction through the upregulation of BKCa expression at mRNA and protein level and RyR expression at mRNA level.