Objective To study the effects of helicase-like transcription factor (HLTF)transfection on DNA repair protein level in radiation-induced apoptotic cells.Methods Human lung carcinoma A549 cells were cultured and transfected with FLAG-tagged wild type HLTF (wild type HLTF transfection group),RING structure domain (ubiquitin conjugating region) mutatation HLTF expressing plasmid (mutant transfection group),empty plasmid (congtrol group) respectively.And the other cells were used as mock transfection group.All cells were irradiated with 15 Gy of 60Co γ-rays to induce apoptosis.Western blotting was used to detect the protein levels of the DNA repair proteins HRAD17 and HRAD52 in the transfected cells.Results The levels of HRAD17 and HRAD52 in the wild type HLTF transfection group was significantly lower than that of the control group.There was no significant difference in HRAD17 and HRAD52 levels between the mock transfection group and ubiquity in conjugating region mutation group.complexes of HLTF and HRAD17 and HRAD52 could be found in the irradiation-induced cells.Conclusions HLTF mediates the degradation of HRAD17 and HRAD52 in the irradiation-induced apoptotic cells possibly by the interaction of the protein complex causing ubiquitination of the repair proteins.

OBJECTIVESevere Epstein-Barr (EB) virus infection is potentially a devastating process that often leads to death encountered in pediatrics recently. Inappropriate control of EB virus replication may cause severe infection resulting in multiple organ dysfunction. However, little information is available on pulmonary complications associated with EB virus infection. The aim of the present study was to investigate severe EB virus (EBV) infection complicated with lung injury in pediatric intensive care unit (PICU), including clinical characteristics, laboratory or imaging feature and outcomes.

METHODA total of 45 children with severe EBV infection seen in PICU of Shanghai Children's Hospital between January 2011 and December 2014 were retrospectively reviewed. According to clinical characteristics and imaging feature, 45 children were divided into non-lung injury group (n =27), lung injury without pulmonary fibrosis group(n = 12) and pulmonary fibrosis group (n = 6).

RESULTIn totally 45 cases of severe EBV infection, 21 (46.7%) were male and 24 (53. 3%) were female, mean age was 2. 4 years; 18 cases were complicated with lung injury, including 8 male and 10 female, median age was 31. 2 months. All of 18 cases presented with fever and cough, 15 of them exhibited dyspnea,12 cases were complicated with gasping, and 6 cases with ARDS. Eight cases accepted mechanical ventilation for acute respiratory distress; 6 cases who developed pulmonary fibrosis had tachypnea, refractory hypoxemia and hypercapnia, severe pulmonary air leak. The average EBV-DNA level in peripheral blood was 4. 42 x 10(6) copies/ml (range: 3. 25 x 10(3) - 6.59 x 10(7) copies/ml). Anti-EBV antibodies were positive in 41 cases, 18 cases were positive (+) for VCA-IgM, 15 cases were VCA-IgG and EA-IgG (+), 8 cases VCA-IgM and VCA-IgG (+). The radiographic findings revealed pulmonary interstitial infiltrates in all 18 cases with lung injury, 4 cases with segmental consolidation and 2 cases showed pleural effusions. HRCT scanning found EBV associated fibrosis including multifocal patches and diffuse ground-glass attenuation in both lungs, reticular opacities and honeycombing changes were observed 4 weeks after illness onset. There were significant differences in respiratory failure, PICU stay (days), Pediatric risk of mortality III (PRISM III) and pediatric clinical illness score(PCIS), serum TNF-α, EBV-DNA levels, percentage of NK cells and CD4+/CD8+ T cell ratio among non-lung injury group, lung injury without pulmonary fibrosis group and pulmonary fibrosis group (X2 =27. 12, F = 85. 23, 78. 23, 88. 68, 323. 80, 7. 35, χ2 = 6. 71, 12. 15; all P < 0. 05). COX regression analysis revealed that EBV-DNA and serum TNF-α levels were correlated with pulmonary fibrosis significantly (OR = 3. 92, P = 0. 04; OR = 5. 95, P = 0. 01). The patients with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) had higher incidence of pulmonary fibrosis compared with non-EB-HLH (70% vs. 13%, χ2 = 4. 82, P = 0. 03). Of 18 cases, 8 cases died, including 3 cases with pulmonary fibrosis. The surviving cases with pulmonary fibrosis needed longer additional oxygen. Chest HRCT imaging of 3 cases with pulmonary fibrosis was improved at 6 months and oxygen therapy was discontinued at 18 months after discharge.

CONCLUSIONEB virus infection complicated with lung injury had higher incidence of respiratory failure, pulmonary fibrosis with a fatal outcome. EBV-DNA and serum TNF-α level were found to be statistically significant indicators of pulmonary fibrosis. Pulmonary fibrosis associated with severe EB virus infection may be reversible.

Antibodies, Viral ; blood ; CD4-CD8 Ratio ; Child, Preschool ; China ; DNA, Viral ; blood ; Epstein-Barr Virus Infections ; pathology ; Female ; Herpesvirus 4, Human ; Humans ; Intensive Care Units, Pediatric ; Killer Cells, Natural ; Lung Injury ; virology ; Lymphohistiocytosis, Hemophagocytic ; pathology ; virology ; Male ; Pulmonary Fibrosis ; pathology ; virology ; Retrospective Studies ; Tumor Necrosis Factor-alpha ; blood

Objective To study the relationship between the genetic polymorphisms of carboxylesterase 1 gene (CESI) and the susceptibility to antituberculosis drug-induced hepatotoxicity (ATBDIH).Methods Genetic polymorphisms of CES1 in 473 tuberculosis patients with or without hepatotoxicity (200∶ 273) after antituberculosis chemotherapy were analyzed by PCR-MassArray.Results In4 tags of CES1 single nucleotide polymorphism (SNP),the frequency of the rs1968753 allele had statistical difference between the hepatotoxicity group and the no-hepatotoxicity group ( P =0.0236 ).The characteristics of anti-hepatotoxicity had been shown relationship with rs8192950 ( P =0.044,OR =0.649,95％ CI =0.426-0.989,AC/AA ) and rs1968753 ( P =0.048,OR =0.556,95％ CI =0.311-0.995,GG/AA).The diplotypes with ‘ CGC' haplotype exhibited significant protection against hepatotoxicity at one copy (P=0.048,OR=0.654,95％CI=0.430-0.996).Conclusions The genetic polymorphisms of CESI might have significant association with ATBDIH.SNP rs8192950 AC genotype and rs1968753 GG genotype might be the candidates for risk prediction of ATBDIH.

Objective To study critical hemophagocytic syndrome (HPS) or macrophage activation syndrome (MAS) presented with multiple organ dysfunction syndrome (MODS) in pediatric intensive care unit (PICU),including clinical features and outcomes In order to explore the effect of bedside continuous hemodialysis/hemofiltration (CBP) as adjuvant treatment for severe HPS/MAS.Methods A total of 19 children with HPS/MAS were hospitalized met the diagnostic criteria for HPS from January,2009 to December,2012.Twelve cases were treated with CBP by continuous venin-venin hemodialysis/hemofiltration (CVVHDF) or high-volume hemofiltration (HVHF) following conventional anti-inflammatory therapy.The replacement liquid dose was 50-75 ml/ (kg · h).The organs function were evaluated and laboratory biomarkers including blood 、electrolytes,ferritin changes were measured before and after CBP treatment.Results Ninteen cases of HPS were acute onset and developed to MODS rapidiy after admission to PICU.The main clinical features were the irregular fever or high fever,hepatosplenomegaly and significant liver damage,nervous system dysfunction and disseminated intravascular coagulation (DIC).Eight cases were death and mortality rate was 42.1％,and all death occurred in those aged less than 3 years old.The mortality rate were 25％ (3/12) and 71.4％ (5/7) in CBP group and non-CBP group respectively.After CBP for 6-24 hours,the fever returned to normal range and blood electrolytes improved.The serum ferritin,serum alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) reduced significantly.Serum creatinine (sCr),blood urea nitrogen (BUN) level improved.Four cases with acute respiratory distress syndrome (ARDS) improved and the ventilator parameters were downregulated.Conclusions Our findings indicate that HPS/MAS complicated with MODS is life threatening with high mortality rate.CBP therapy can lower the fever within a short time,correct electrolyte imbalance,stable circulatory function,improve the lung,liver,and brain function.It is suggested that CBP may be the potential effective therapy in severe HPS/MAS with MODS in children.

Objective To explore the influence of inhaled glucocorticosteroid on ?-glutamylcysteine synthetase(?-GCS) in inflammatory cell of sputum in children with asthma.Methods Twenty-two asthmatic children were divided into 2 groups according to treatment.The children who were treated by inhaled budesonide combined with salbutamol were due to group A and the others inhaling salbutamol only were due to group B,the healthy children were acted as healthy control group(group C).The glutation(GSH),total GSH and the activity of ?-GCS in sputum were measured respectively;Expression of ?-GCS in inflammatory cell of sputum were detected by immunohistochemistry;the expression of ?-GCS heavy chain(?-GCS-h) mRNA were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Results 1.The total GSH[(1.08?0.14) ?mol/L] and oxidized glutathione(GSSG)[(0.37?0.09) ?mol/L] were decreased in sputum of group A of post-treatment compared with pre-treatment(Pa

flow cytometry.A strong linear relationship was observed in the standard curve of real-time PCR of each bacteria. Conclusion These three non-culture methods can be used in the quantitative analysis of oral microorganisms.Real-time PCR and laser scanning confocal microscopy are better than the traditional culture-based CFU count,and real-time PCR is the most sensitive method.

Objective To detect the differential gene expression between Streptococcus sobrinus(S.sobrinus) 6715 and its fluoride-resistant strains. Methods The fluoride-resistant strains of S.sobrinus 6715 was induced by increasing the concentration of fluoride step by step.Total RNA of both S.sobrinus 6715 and its fluoride-resistant strains was extracted,mRNA was separated and purificated,and then cDNA was obtained by reversed transcription.Suppression subtractive hybridization(SSH) technology was used to detect the differential gene expression between them.The differential gene expression fragments were cloned and compared with the GenBank by BLAST.Results After comparing with the GenBank by BLAST,it was identified that there were two differential gene expression fragments,fruA and SMU.438c. Conclusion The cDNA subtractive lib of differential gene expression between S.sobrinus 6715 and its fluoride-resistant strains was successfully constructed through SSH,which paves a way for the further study of fluoride-resistant mechanism.

Objective To evaluate the value of noninvasive monitoring of pulmonary arterial pressure in the children with severe pneumonia and respiratory failure.Methods A prospective study was adopted to investigate 69 patients who suffered from severe pneumonia and respiratory failure in Pediatric Intensive Care Unit in Shanghai Children's Hospital from June 2013 to December 2013 were involved in this study,except for heart disease.The pulmonary arterial pressure (PAP) and cardiac function were monitored by using bedside color doppler ultrasound cardiogram,such as PAP,cardiac index (CI),left ventricle ejection fraction(LEFT),and heart early diastolic filling velocity maximum/heart late diastolic filling velocity maximum (E/A ratio).They were divided into 2 groups according to PAP,one group as pulmonary arterial pressure normal group,the other group as pulmonary arterial hypertension(PAH) group,and the impact of the PAP on the prognosis and mechanical ventilation was assessed.Milrinone[0.5 μg/(kg · min)] were given the patients who were combined with pulmonary hypertension,and the PAP and cardiac function before using Milrinone and 24 h,48 h and 72 h after giving medicine was observed.Results Among 69 cases,40 cases were male and 29 cases were female,age ranging from 2 months to 12 years old,and the weight range was (14.3 ± 8.9) kg.The pediatric critical illness score(PICS) was 70.5 ± 9.6,and the pediatric risk of score m ortality Ⅲ was 13.5 ± 5.0.Among 69 cases,46 cases had pulmonary arterial hypertension,38 cases of them experienced mechanical ventilation,and 9 cases died.Among 23 cases who had no pulmonary arterial hypertension,only 8 cases experienced mechanical ventilation.There was a significant difference in the mechanical ventilation rate and mortality between two groups(x2 =15.78,P ＜0.0l ; x2 =5.18,P ＜ 0.05).The mechanical ventilation time was longer in pulmonary arterial hypertension group (t =3.89,P ＜0.01).PAP was (58.23 ±5.44) mmHg(1 mmHg =0.133 kPa),(49.10 ±4.69) mmHg,(42.53 ±4.54)mmHg and(35.63 ±4.78) mmHg respectively before and after using Milrinone 24 h,48 h and 72 h in 46 cases with pulmonary arterial hypertension,and the pressure decreased significantly after using medicine (F =67.11,P ＜ 0.01).There was no significant difference in CI,LVEF and E/A(all P ＞0.05).However,9 cases of them did not show any response to Milrinone,and in the end they couldn't live without mechanical ventilation,they died.Conclusions Noninvasive pulmonary arterial pressure monitoring could be beneficial in judging patient's condition and assessing prognosis of children with severe pneumonia and respiratory failure,and milrinone could decrease PAP.

Objective To explore the changes of renin-angiotensin-aldosterone system in pediatric severe sepsis treated with continuous blood purification(CBP).Methods Prospective study,35 cases of critically ill children diagnosed with severe sepsis and admitted to PICU of Shanghai Children's Hospital of Shanghai Jiaotong University from June 2012 to May 2014 served as reseach objective.Based on the monitoring of vital signs,including central venous pressure,arterial blood pressure,mean arterial pressure,patients were treated with conventional therapy,as antibiotics,fluid therapy,and CBP by mode of continuous veno-venous hemodiafiltration or high volume hemofiltration.Plasma levels of rennin activity,angiontensin Ⅱ and aldosterone were measured by radioimmunoassay before and 24 h after CBP.Twenty-five cases of blood samples taken from the children collected from health care for liver function examination were matched as control group.Results Plasmalevelsofrenninactivitywere(2.11 ±1.93) pg/(L·h),(1.27±1.56) μg/(L·h),(0.37 ± 0.22) μg/(L· h) before and 24 h after CBP and control group,respectively.The levels of angiontensin Ⅱ were (426.78 ±332.37) ng/L,(364.40 ± 325.51) ng/L,(41.70 ± 10.81) ng/L,respectively.And the levels of aldosterone were (255.12 ± 218.18) ng/L,(134.92 ± 104.13) ng/L,(106.88 ±43.18) ng/L,respectively.The plasma levels of rennin activity,angiontensin Ⅱ,and aldosterone were higher in sepsis cases than in control group,while decreased obviously after CBP treatment(P ＜0.01,P ＜0.05).Eleven cases died and mortality was 31.4％ (11/35).After 24 h of CBP,the mean arterial pressure improved in 26 cases with septic shock and dopamine dose reduced(P ＜ 0.01).Conclusion The reaction of renin-angiotensin-aldosterone system is increased significantly in pediatric severe sepsis.CBP can down-regulate the levels of rennin activity,angiotensin Ⅱand aldosterone,but not worsen the circulation function.

Objective To explore the relationship of expression and the effect of preoperative radiotherapy of endothelin-1 (ET-1) and pyruvate kinase M-2 (PKM2) in rectal carcinoma.Methods Immunohistochemical method was used to detect the expression of ET-1 and PKM2 proteins of rectal cancer tissues in 96 cases.The expressions of ET-1 and PKM2 were analyzed with the effect of preoperative radiotherapy in rectal cancer tissue.Results The high expression of ET-1 protein was 59 cases (61.46％).The high expression of PKM2 proteins was 54 cases (56.25％).The high expressions of ET-1 and PKM2 protein were worsen the effect of tumor regressive grade (TRG) than lower expressions of those after preoperative radiotherapy of rectal cancer tissue (P ＜ 0.05).The protein expression of ET-1 and PKM2 were positively correlated (P =0.006).Conclusions The high expressed ET-1 and PKM2 proteins in rectal cancer are closely related to preoperative radiotherapy resistance.ET-1 and PKM2 proteins are expected to become new targets of radiotherapy sensitivity and radiotherapy sensitization of rectal cancer.