The 2012 version of international guidelines for management of severe sepsis and septic shock got re-revised on the basis of the 2008 version.For pediatric severe sepsis,the 2012 version enforces initial resuscitation;develops detailed ant-infection measures;stresses isotonic crystalloid fluid resuscitation and the use of albumin,hetastarch is no longer recommended ;reduces the value of early use of dopamine,and clears the use of corticosteroids indications and the use of blood products;mechanical ventilation is recommended to use lung protective ventilation strategy; standardizes sedation and analgesia applications; clears the level of blood glucose control;others include blood purification,nutrition,stress ulcer prevention recommendations.

Objective　To study the effects of systemic irradiation and conglutinant drug W11-a12 on the number and some functions of wound nentrophils (Neu). Methods　Wound Neu was collected from sponges which were implanted in rat's dorsum incision. The number of Neu, as well as the phagocytic function and motility of wound Neu were measured. Results　After 4,6,8 Gy systemic irradiation, the number of white blood cells and Neu in wound, as well as the phagocytic function and chemotactic motility of wound Neu, were significantly decreased at 24 h, 48 h after wounding. W11-a12 markedly increased the number of wound Neu, improved the phagocytic function and chemotactic motility of wound Neu at 24 h, 48 h after wounding despite the rats were radiated or not. Conclusion　The results indicated that the decreased number and function of wound Neu in the early stage of wound healing contributed to the impairment of repair after systemic irradiation. W11-a12 accelerated normal and irradiation-impaired wound healing partly by increasing the number of wound Neu and improving the Neu function.

OBJECTIVE:To study the bacterial endotoxin test for lomefloxacin hydrochloride and glucose injection.METHODS:According to the assigned method and guiding principle for bacterial endotoxin test in CHINESE PHARMACOPOEIA 2000,the interference test was performed.RESULTS:Lomefloxacin and glucose injection could interfere with limulus test,however,the interference could be eliminated after adjusting the pH of injection to 6.8～7.0.The proper concentration of limulus test agent of bacterial endotoxin test was 0.25EU/ml.CONCLUSION:Bacterial endotoxin test can be used for quality control of lomefloxacin and glucose injection.

OBJECTIVE:To establish the method for determining the content of Lomefloxacin in hollow type suppository of Lomefloxacin hydrochloride.METHODS:RP-HPLC was used with Eclipse XDB-C 18 column(4.6mm?250mm)and the mobile phase consisting of citric acid(0.05mol/L)-acetonitrile(82∶18)the pH of which was adjusted to4.0with triethy?lamine.The flow rate was1.0ml/min and the detection wavelength was287nm.RESULTS:The average recovery of Lome?floxacin was99.8%and RSD was0.6%.CONCLUSION:The method is rapid and accurate and can be used for determination of the content of Lomefloxacin in hollow type suppository.

Objective To construct the eukaryotic vector that expressing hepatitis B virus (HBV) S and the single fragment of variety chain (ScFv) of monoclonal antiboy against cytotoxic Tlymphocyte-associated antigen-4 (CTLA-4) and to analyze the immunological activity of recombinant S-ScFv protein.Methods The oringially constructed pSect2/ScFv4F10 and pSect2/S were double enzyme digested by Sfi I and Hind Ⅲ,respectively.Then the HBV S gene was cloned into the pSect2/ScFv4F10 vector.The pSect2/ScFv4F10 and pSect2/S-ScFv4F10 were expressed in Chinese hamster ovary (CHO) cells,and the expressed proteins were verified through sodium dodecyl sulfatepolyacrylamide gelelectrophoresis(SDS-PAGE)andWesternblotting.Afterultrafiltration concentration and affinity chromatography,the biological affinity of the expressed ScFv4F10 and S-ScFv4F10 proteins were examined by competitive enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) technology.The comparison between groups was done by One-way ANOVA.ResultsThe eukaryotic expression vector of pSect2/S-ScFv4F10was successfully constructed,and relative molecular mass of the expressed protein of S-ScFv4 F10 was about 52 000 that analyzed by SDS-PAGE and Western blotting.With the fixed concentration of 4F10-mAb against CTLA-4,the A570 value of the mixed reaction with purified CTLA-4 antigen gradually increased with the decrease of ScFc fusion protein proportion; when the molar ratio of ScFv,S-ScFv4F10∶4F10=2∶1,the competitiveinhibitionratesagainst 4F10conjugatedantigenwere72.6％and64.5％,respectively.The affinity constants of association kinetics for CTLA-4 mAb,ScFv4F10 and S-ScFv4F10 with CTLA-4 antigen were 7.29 × 108 mol/L,9.52 × 106 mol/L and 2.04 × 106 mol/L,respectively,and the dissociation constants of KD were 1.40 × 10-9 mol/L,1.05 × 10-7 mol/L and 4.91 × 10-7 mol/L,respectively.ConclusionsThe eukaryotic expression vector of pSect2/S-ScFv4F10is successfully constructed,and the recombinant protein of S-ScFv4 F10 has a fairly high affinity with CTLA-4 antigen.

Objective To determine the vitamin D status in children with sepsis/severe sepsis in pediatric intensive care unit (PICU) in order to explor the association between vitamin D status and clinical outcomes,in turn to provide evidence for optimizing nutrition support.Methods It was a prospective,observational,multi-center study,carried out in patients with sepsis/severe sepsis from March 1,2013,to March 30,2014,in the PICUs of three tertiary-care children's hospitals.Total serum 25-hydroxy vitamin D [25 (OH) D] was measured by using an enzyme-linked immunosorbent assay at admission.The association of vitamin D status at admission with length of PICU length of stay,total hospital stay,in-hospital mortality,28-days mortality and costs were analyzed.Results A total of 194 patients includng 117 boys (60.3％)and 77 girls (39.7％) were enrolled.There were 96 patients with sepsis and 98 with severe sepsis.The mortality on discharge and 28 days were 6.7％ and 24.2％ respectively.The median vitamin D level was 9.79 ng/mL (5.32,18.46) at admission.Of them 77.8％ (151/194) had vitamin D deficiency and 50.5％ (98/194) had severe vitamin D deficiency.Patients with severe vitamin D deficiency,had higher mortality on discharge (P =0.011).Vitamin D status had no significant correlations with 28 days mortality,length of PICU stay,total hospital stay and costs.Conclusions More than three-quarters (77.8％) of children with sepsis/severe sepsis in PICUs had Vitamin D deficiency.Patients with severe vitamin D deficiency at admission had higher risk of mortality at discharge.

Objective:To optimize the clarification process of total flavonoids extract from Geum aleppicum with ZTC1+1-Ⅱtype clarifying agent. Methods:With the solid removal rate, retention rate of the total flavonoids and clearance rate of protein as the indi-ces, single factor tests were adopted to study the effects of extract concentration, amount of clarifying agent, reaction temperature and reaction time on the clarification results. Results:The optimal clarification process was as follows:the extract concentration was 0. 5 g ·ml-1 , the amount of clarifying agent was 4% component B and 2% component A, the reaction temperature was at 60℃, and the soaking time was 60 min. The retention rate of total flavonoids was 91. 32%, the clearance rate of protein was 35. 82%, and the solid removal rate was 8. 1%. Conclusion:ZTC1+1-Ⅱ type clarifying agent has good effect on the clarification of total flavonoids extract from Geum aleppicum with promising feasibility and stability.

Purpose To investigate the clinical utility of DAB2IP (DOC-2/DAB2 interactive protein)and β-catenin expression in bladder urothelial carcinoma (BUC).Methods The expression of DAB2IP and β-catenin was detected in 104 BUC cases and 40 peritumorial tissues using EnVision two-step immunohistochemical method,and the association with BUC clinicopathological parameters was analyzed.Results The expression of DAB2IP in BUC was significantly less than that of peritumorial normal tissues,and the expression of β-catenin in BUS was significantly higher than that of peritumorial normal tissues (P ＜ 0.05).DAB2IP expression and histologic grading,clinical pathologic staging and 5 years survival rate had statistical significance (P ＜ 0.05).No statistical significance with gender,age,tumor diameter and in patients with incipient/recurrence.β-catenin expression and age,histologic grading,clinical pathologic staging,tumor diameter and 5 years survival rate have statistical significance (P ＜ 0.05).No statistical significance correlated with gender and in patients with incipient/ recurrence,DAB2IP and β-catenin expression in BUC are negatively correlated (P ＜ 0.05).Conclusion In bladder urothelial carcinoma,down-regulation of DAB2IP and up-regulation of β-catenin,are in a negative correlation.Abnormal expression of DAB2IP and β-catenin is correlated with histologic grading,clinical pathologic staging and prognosis.

Objective:To analyze and optimize the extraction technology of Geum aleppicum by Box-Behnken design and response surface methodology. Methods:With the content of total flavonoids as the index, the effects of three factors including the solvent-solid ratio, ethanol concentration and extraction time on the content of total flavonoids were studied by single factor experiment, and the ex-traction technology was optimized by Box-Behnken design and response surface methodology. Results: The optimal extraction condi-tions of Geum aleppicum were as follows:the solvent-solid ratio was 4. 2, the ethanol concentration was 50% and the extraction time was 80 min. Under the above conditions, the average yield was 12. 590 0 mg·g-1 . Conclusion: Optimizing extraction technology of the total flavonoids in Geum aleppicum by Box-Behnken design and response surface methodology is reasonable, and the mathematical mod-el is consistent with the experimental data, which has good predictability.

Objective To investigate the value of the interhospital transportation and the advantage of the cooperative network to the critically ill children.Methods The clinical data of 232 critically ill chil-dren transported from other hospitals in long distances to PICU of Fudan University Affiliated Children Hos-pital cooperated with Shanghai 120 interprovincial transport Department,between Oct 2010 and Oct 2013, were analyzed retrospectively.Results At otal of 232 critically ill childrne were transported from 31 hospi-tals of Eastern China region including five provinces and the city of Shanghai.Among thse e critically ill chil-dren 141 casse were male and 91 cases were femla e,with age from 29 days ot 13 years( median age was 12 months) and weight from 2.5 to 66 kg ( median weight was 10 kg ) .The average pediatric clinical illness score was 80.4 ±7.7,155 cases(66.8%) were single organ dysfunction,55 cases(23.7%) were multiple organs dysfunction,105 cases(45.3%) were mechanical ventilation required fort ransportait on.Referral radi-us was 50-1 000 km(median).Among these patients,87 cases(37.3%) weret ransported over 200 km radius and73 cases(36%) were transop rted 101-200 km,62 cases(26.7%) were transported 100 km or less.D ur-ing the transportation,0 case died,3 cases(1.3%) received cardiopulmonary resuscitation treatment,2 cases (0.9%) received electrical conversion treatment,2 cases(0.9%) were replaced endotracheal intuab tion,the success rate of transportation was 100%.The top 4 disae ses were respiratro y id seases(90 cases,38.8%), neural diseases(43 cases,18.5%) ,cardiovascular diseases(36 cases,15.5%) and acute pediatric trauma(28 cases,12.1%) .Al l 232 cir tically ill children were admitted to our PICU for further treatment through the Green Channel.In the end, 178 cases ( 76.7%) discharged, 24 cases ( 10.3%) gave up and 32 cases (13.8%) died.Among thse e 232 rc itically ill children,30 cases(12.9%) received special organ replacement treatment,4 casse ( 1.7%) of whom were treated with extracorporeal membrane oxyg enation, 26 cases (11.1%) received blood purification therapy.Conclusion Establishing the system of transportation between PICU of hospitals will be propitious to treat the critically ill children energetically and effectively.It is worth pro-moting to master the transportation indication of children with critical illness,which is safe and reliable.