AIM:To investigate the effect of pretreatment of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) on the proliferation and the differentiation of mesenchymal stem cells(MSCs) into cardiomyogenic cells.METHODS:The MSCs,isolated primarily from bone marrow,and purified by passage culture,were obtained from the adult rats of four groups:the rats were pretreated by 5 daily injections of SCF;the rats were pretreated with G-CSF;the rats were pretreated with SCF and G-CSF;the rats were treated without any intervention.The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co-culture.The neonatal rat cardiomyocytes cultured for 3 days were co-cultured with DAPI-MSCs.The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co-culture days was analyzed.The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were recorded respectively with digital microscope camera system and immunofluorescence technique.The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated.RESULTS:The percentage of MSCs in G0/G1 phase in SCF/G-CSF group was significantly lower than that in SCF group,G-CSF group and the control group.The percentage of MHC protein-positive MSCs in SCF/G-CSF group was markedly higher than that in SCF group,G-CSF group and the control group,and that in SCF group and G-CSF group was significantly higher than control group.The percentage of TnT protein-positive MSCs in SCF/G-CSF group,SCF group and G-CSF group was significantly higher than that in control group.CONCLUSION:SCF and G-CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes.The combination of using SCF and G-CSF is more effective than using only SCF or G-CSF.

Aim To investigate the method of establishing the atopic dermatitis mice model induced by calcipotriol ( MC903 ) . Methods Induction dose exploratory experiment: since d 0, 0.33,1,3 nmol MC903 was smeared on the right ear of BALB/c mice in each dose group respectively , for 7 consecutive days . The ear swelling degree of the mice was observed every day and the bilateral ears thicknesses were measured .The materials were drawn and analyzed in d 3 and d 7.Induction days exploratory experiment: 2 nmol MC903 was smeared on the right ear of BALB/c mice, for 14 consecutive days .The ear swelling degree of mice was observed every day and the bilateral ears thicknesses were measured .The histopathological examination of the right ear was conducted in 3 d, 7 d, 11 d and 15 d respectively .The tissue homogenate of the mice right ear was prepared .The ex-pressions of thymic stromal lymphopoietin (TSLP),IL-33, IL-4 and IFN-γin the homogenate , CD40 +,CD86 +,the DC surface markers and ILC2 contents in the peripheral lymph nodes were detected.Results ①1,3 nmol MC903 induced ear swelling in mice was significantly increased in d 3, the levels of TSLP and IL-4 were significantly increased .The level of IL-33 in 3 nmol dose group was increased significantly in d 7.② The right ear swelling of 2 nmol MC903 induced atopic dermatitis mice was significant , the ear thickness was increased gradually and reached the peak in d 14.The histopathological examination of the mice ear tissue showed in d 7, the right ear tissue of the mice was swelling and red , the capillary vessels were dilated and the infiltration of inflammatory cells was obvious .The ear in-flammatory symptoms maintained and gradually aggravated for 15 days.Compared with the normal mice , MC903 increased the TSLP level in the right ear tissue homogenate significantly in d 3 and then decreased gradually .The levels of IL-4 and IL-33 were increased significantly in d 7 and then decreased gradually .The levels of ILC2, CD40 +, CD86 + in the peripheral lymph nodes were increased in d 7 and d 15.Conclusion The atopic derma-titis mice model can be successfully established using 2 nmol MC903 induced mice for 7 days.Appropriate testing point of TSLP is d 3.Appropriate testing point of IL-33 and IL-4 is d 7.

Allergic diseases such as allergic asthma, allergic der-matitis, allergic rhinitis, are polygenic diseases, involving the interaction between the environment, genes and immunity. In the past few decades, the incidence rate of allergic diseases in-creased predominantly and influenced the quality of people's lives seriously, so looking for new targets for the prevention and treat-ment of allergic diseases and drugs with less adverse reaction be-comes a hot topic for researchers. MicroRNAs(miRNAs)are a class of endogenous non-coding small RNAs that mediate nega-tively posttranscriptional regulation of gene expression by targe-ting specific mRNA sequences to inhibit the translation of mR-NAs. They are widely involved in the biological processes of cell differentiation, immune response and tumor development. The study shows that miRNAs can control the occurrence and devel-opment of allergic diseases. Studying the regulatory role of miR-NAs in allergic diseases has important implications for exploring the immunopathological mechanisms and discovering new thera-peutic targets of drugs.

Adolescent ; C-Reactive Protein ; metabolism ; Cell Aggregation ; Child ; Child, Preschool ; Cytodiagnosis ; methods ; Female ; Humans ; Infant ; Leukocytes ; cytology ; Male ; Meningitis, Bacterial ; blood ; cerebrospinal fluid ; diagnosis ; Meningitis, Viral ; blood ; cerebrospinal fluid ; diagnosis ; Sensitivity and Specificity

Objective To discuss the clinical feature ,diagnosis and treatment of the occult cerebrospinal fluid rhinorrhea after tracheotomy in patients with severe traumatic brain injury .Methods The clinical data of 18 cases of the occult cerebrospinal fluid rhinorrhea after tracheotomy in patients with severe traumatic brain injury were retro -spectively analyzed .Results 15 cases showed involuntary swallowing movements ,frequent stimulus-likecough, abnormal increased secretions in the oral and nasal;3 cases performance of aspiration ,hypoxemia ,respiratory distress . After a three -dimensional thin skull CT , cisternography , nasal endoscopic examination can confirm the diagnosis . After the treatment with replacing the tracheostomy tube with a balloon ,continuous lumbar drainage ,endoscopic repair leak,the cerebrospinal fluid rhinorrhea were cured .Conclusion Patients with occult cerebrospinal fluid rhinorrhea performance the diversity and easily missed ,early detection and timely treatment can prevent cerebrospinal fluid rhi-norrhea delayed healing and intracranial infection and promote patient recovery .

BACKGROUND: With mechanical ventilation widely used in intensive care unit, the ventilator associated pneumonia (VAP) has become a common and serious complication in critically ill patients. Compared with adults, the incidence of VAP and the mortality are higher in children in pediatric intensive care unit (PICU) because of immune deficiency, severe basic diseases, and increased use of artificial airway or mechanical ventilation. Hence it is of significance to study the epidemiology and changes of antibacterial susceptibility in order to reduce the incidence and mortality of VAP in children. METHODS: From January 2008 to June 2010, 2758 children were treated in PICU of Wuhan Children's Hospital. Among them, 171 received mechanical ventilation over 48 hours in PICU, and 46 developed VAP. The distribution and drug-resistance pattern of the pathogenic bacteria isolated from lower respiratory tract aspirations were analyzed. RESULTS: A total of 119 pathogenic microbial strains were isolated. Gram-negative bacilli (G-) were the most (65.55％), followed by fungi (21.01％) and gram-positive cocci (G+, 13.45％). Among them, the most common pathogens were Acinetobacter baummannii, Escherichia coli, Klebsiella pneumoniae, candida albicans and coagulase-negative staphylococci. Antibiotic susceptibility tests indicated that the multiple drug-resistances of G- and G+ to antibiotics were serious. Most of G- was sensitive to ciprofloxacin, amikacin, imipenem, meropenem, cefoperazone-sulbactam and piperacillin-tazobactam. The susceptibility of G+ to vancomycin, teicoplanin and linezolid were 100％. Fungi were almost sensitive to all the antifungal agents. The primary pathogens of VAP were G-, and their multiple drug-resistances were serious. CONCLUSION: In clinical practice we should choose the most sensitive drug for VAP according to pathogenic test.

BACKGROUND: With beta-lactam drugs and immunosuppressants widely used, the infection caused byAcinetobacter baumannii (Ab) has become more and more serious with multidrug resistant Acinetobacter baumannii (MDRAb) emerging and worsening rapidly. Compared with other patients, the incidence and multidrug resistance of MDRAb are higher in children in pediatric intensive care unit (PICU) because of immune deficiency, severe basic diseases, prolonged hospitalization and invasive operations. Hence it is significant to study the epidemiology and changes of antibacterial susceptibility in order to reduce the incidence of MDRAb in children. METHODS: A total 115 patients with MDRAb pneumonia and 45 patients with negative MDRAb (NMDRAb) pneumonia who had been treated from January 2009 to August 2011 were studied retrospectively at the PICU of Wuhan Children's Hospital. Clinical data were analyzed with univariate and multivariate Logistic regression. RESULTS: In 176 clinical strains ofAcinetobacter baumannii isolated, there were 128 strains of MDRAb, accounting for 72.73％. Drug susceptibility tests showed that the resistance rates of β-lactam antibiotics were more than 70％ except for cefoperazone sulbactam. The rates to carbapenems were higher than 90％. They were significantly higher than those of NMDRAb. Amikacin, levofloxacin, ciprofloxacin and minocycline had the lowest drug-resistance rates (<20％). Multivariate Logistic regression revealed that ICU stay, the time of mechanical ventilation, anemia, hypoproteinemia and the use of carbapenems were independent risk factors for MDRAb pneumonia. CONCLUSIONS: MDRAb is an important opportunistic pathogen to pneumonia in PICU, and its drug-resistance is severe. It increases significantly the mortality of patients. It is important to take the effective prevention measures for controlling it.

Objective ， To investigate the role of serum chemokines and oxidative and antioxidant biomarkers in occupational （ silicosis） Methods silicosis hereinafter referred to as . A total of 58 patients with stage Ⅰ silicosis were selected as the - （ ）， research subjects using convenient sampling method. The serum levels of nuclear factor erythroid 2 related factor 2 Nrf2 -（ - ） - （ - - ） - heme oxygenase 1 HO 1 and 8 isoprstaglandin F2α 8 iso PGF2α were determined by enzyme linked immunesorbent assay. （ ） （ - ） The serum levels of lipid peroxide LPO and total antioxidant capacity TAOC were determined by chemistry colorimetric method. - - （ - ）， Luminex flow fluorescence technology was used to detect the serum levels of interferon γ inducible protein10 IP10 macrophage （ ）- ， - - （ ） inflammatory protein MIP 1α MIP1β and macrophagederived chemokine MDC . The above indicators were analyzed by factor Results - analysis. The information extraction rate of the original indicators of the nine biomarkers was 58.5%96.5%. Four common ， ， （ ） ， factors were extracted including Nrf2 antioxidant signaling pathway helper T cell Th 1 dominant chemotaxis the total ， ， ， ， ， oxidation/antioxidant balance and Th2 dominant chemotaxis whose variance contribution rates were 32.2% 19.1% 16.4% ， ， Conclusion - and 11.8% respectively and the cumulative variance contribution rate was 79.5%. Both the oxidant antioxidant ， disturbance and the dominance chemotaxis are involved in the occurrence and development of silicosis and the Nrf2 antioxidant signaling pathway plays the most critical role.

OBJECTIVEStreptococcus pneumoniae necrotizing pneumonia (SPNP) was reported elsewhere but not in China yet. Inappropriate treatment due to poor recognition of this disease could influence its prognosis. This paper presents the clinical characteristics, diagnosis and treatment of SPNP hoping to elevate pediatrician's recognition level for this disease.

METHODClinical manifestations, radiological findings, treatment and prognosis of 20 patients (9 boys, 11 girls) who had been hospitalized with SPNP in Beijing Children's Hospital from 2004-2011 were retrospectively analyzed.

RESULTThe patients included in this study aged from 9 months to 6 years [(27.9 ± 15.8) m] and were healthy before admission. They were febrile for 8 to 50 days [(27.7 ± 13.5) d] and hospital day was 24 - 55 days [(36.5 ± 8.3) d]. The general condition of all subjects was relatively poor and they all had fever and cough. One child had moderate fever and nineteen children had high fever. Dyspnea was found in sixteen children. Fine rales were found on auscultation in 18 children, among whom diffuse wheeze appeared in 4 children, and wheezy phlegm was found in two children. Signs of pleural effusion were discovered in all cases by physical examination and chest X-ray. White blood cell (WBC) count was 16.2 - 60.95×10(9)/L and neutrophil was 70.5% - 80.2% in peripheral blood routine test. Erythrocyte sedimentation rate (ESR) was 44 - 109 mm/h [(69.6 ± 16) mm/h]and C-reactive protein (CRP) was 80 - > 160 mg/L. The pleural effusion biochemistry and routine test revealed a WBC count of 6400×10(6)/L-too much to count, polykaryocyte of 51% - 90%, glucose of 0.02 - 1.8 mmol/L, protein of 32 - 51 g/L and LDH of 5475 IU/L-or higher. Pleural effusion culture in all cases and blood culture in 2 cases was positive for Streptococcus pneumoniae. Chest X-ray or CT revealed high density and well-distributed lobar consolidation in one lung or two lungs initially. Single or multiple low density lesions in the area of lobar consolidation were found a week later, accompanied by multiple cystic shadow or cavity at the same time or afterwards. Bulla of lung appeared later. Pleural effusions were found in all patients. Seven cases complicated with hydropneumothorax, two with otitis media, one with heart failure, one with cardiac insufficiency. Seventeen patients were treated with vancomycin or teicoplanin or linezolid two with amoxicillin and clavulanate potassium. Other two patients had been treated with meropenem and cephalosporin antibiotics respectively before admission, and they had been at recovery stage when they were hospitalized. Thoracic close drainage and thoracoscopy were performed respectively in 18 cases and 3 cases, respectively. After a follow up of more than 6 months, chest CT showed that almost all lesions in lungs recovered during 4-6 months. No one received pneumonectomy.

CONCLUSIONSPNP has special manifestations. The incidence in infants is higher. Patients' general condition is poor and febrile course is relatively long. All patients manifested fever and cough, with a presence of dyspnea in most of them. WBC, neutrophil and CRP elevated apparently. The characteristic of pleural effusion indicates empyema. In early stage, the chest X-ray and CT showed high-density lobar lesions, followed by low-density lesions and cyst gradually. Bulla of lung and/or hydropneumothorax may appear at the late stage. But if diagnosed and treated promptly, the prognosis of SPNP was relatively good.

Anti-Bacterial Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Dyspnea ; diagnosis ; drug therapy ; epidemiology ; Female ; Fever ; diagnosis ; drug therapy ; epidemiology ; Humans ; Infant ; Leukocyte Count ; Lung ; diagnostic imaging ; pathology ; Male ; Methylprednisolone ; therapeutic use ; Pleural Effusion ; diagnosis ; drug therapy ; epidemiology ; Pneumonia, Pneumococcal ; complications ; diagnosis ; drug therapy ; Prognosis ; Retrospective Studies ; Streptococcus pneumoniae ; drug effects ; isolation & purification ; pathogenicity ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo investigate the effect of ketogenic diet (KD) on the clinical and electroencephalogram features in children with pharmacoresistant epileptic encephalopathy.

METHODThirty-one children (19 boys, 12 girls) aged 7 months to 7 years (mean 2 years 5 month) with epilepsy refractory to conventional antiepileptic drugs (AEDs) were included in this study. In addition to their original AED treatment, the children were assigned to different ketogenic diets based on their age. The prospective electro-clinical assessment was performed prior to the KD and then one week, one month and again 3 months after the initiation of therapy, respectively.

RESULTThe reduction of seizure frequency in 52%, 68% and 71% of all patients exceeded 50% one week, one month and three months after KD treatment respectively. KD is particularly effective in myoclonic astatic epilepsy (MAE; Doose Syndrome) and West syndrome with 100% and 81.25% of the patients having a greater than 50% seizure reduction, respectively. After 3 months of KD treatment, more than 2/3 patients experienced a reduction in interictal epileptiform discharges (IEDs) and improvement in EEG background.

CONCLUSIONThe clinical and electroencephalographic improvement confirms that KD is beneficial in children with refractory epilepsy.

Anticonvulsants ; therapeutic use ; Brain ; diagnostic imaging ; physiopathology ; Child ; Child, Preschool ; Diet, Ketogenic ; methods ; Dietary Fats ; administration & dosage ; Electroencephalography ; Epilepsy ; diagnosis ; diet therapy ; drug therapy ; Female ; Humans ; Infant ; Intellectual Disability ; diet therapy ; drug therapy ; Lennox Gastaut Syndrome ; Male ; Radiography ; Retrospective Studies ; Spasms, Infantile ; diet therapy ; drug therapy ; Syndrome ; Time Factors ; Treatment Outcome