Objective To evaluate the performances of glucose oxidase immobilized on hydrophilic-hydrophobic silica gel, which may be employed to prepare glucose sensor for the determination of glucose in body fluids. Methods The silica gel was prepared from precursors ?-aminopropyltrimethoxysilane (APTMOS) and methyltrimethoxysilane (MTMOS) by sol-gel technique. Glucose oxidase (GOD) was covalently attached to the silica gel via carbodiimide coupling reaction between a carboxylic acid group on enzyme and an amine group of the silica gel under the participation of the linking reagents 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide. The performances of immobilized GOD were explored. Results The optimum conditions were obtained as follows: volume fraction of APTMOS 70%, enzyme content given 16 800 U, the temperature of 35 ℃ and buffer pH 5.5. The decrement in the activity of immobilized GOD for the first 2 weeks was less than 10% of its original activity, and the activity of immobilized GOD retained more than 75% of its original activity after 1 month of testing. Six independently prepared immobilized GOD on the silica gel resulted in an average bioactivity of 1 290.9 ?mol?min -1?g -1 with an R.S.D. of 3.4%. The Michaelis constant (K m) of immobilized GOD was 9.1 mmol?L -1. Conclusion Immobilizing GOD on the silica gel via the formation of peptide bonds is an outstanding enzyme immobilization method.

Objective To analyze the amount and allocation of pediatric healthcare resources in Chengdu,and to recommend on local pediatric healthcare resources shortages.Methods Pediatric healthcare resources data of Chengdu came from pediatric relevant data reported regularly at yearend by the counties and districts to healthcare administration of the city.Data from such reports were subject to statistics with various indexes and descriptive analysis.Analyzed in focus was the distribution in 2015 of pediatric healthcare resources of the city among medical institutions of various types,levels and properties,as well as causes for such shortage.Results Tertiary hospitals hold 62.3% of the pediatric beds and 64.2% of pediatricians,and provide around 70% of the medical workload for pediatric outpatients and inpatients,upon the majority of pediatricians with master degree and above,and senior ones of/above associate chief physician titles;Tertiary hospitals have 58.7% of the pediatric beds and 49.7% of pediatricians,yet the outpatients served by specialized hospitals were 5% above tertiary hospitals.Conclusions The imbalance and shortage in total of pediatric resources in Chengdu result from stable source of manpower supply,high professional risk exposure,low income,and long training duration among others.Such measures as a better pediatrician development system,greater incentives for pediatric development,and enhanced development pediatric service consortiums,as well as greater support for private specialized pediatric hospitals.Those measures combined can effectively alleviate the shortage of pediatric resources.

Objective To investigate the effects of interventions on psychological stress factors of nurses in the emergency department of general hospital. Methods Symptom checklist 90(SCL-90)was used for investigation. The corresponding interventions were conducted based on the found problems. Results The mental health of the nurses in general hospital emergency department was worse than that of the general population. Each factor score, the total average score and the number of items after intervention had significant difference than that before intervention respectively. The incidence rate of top 10 symptoms was significantly decreased after the intervention than that before. Conclusions Implementation of comprehensive intervention can significantly improve the mental health of the nurses in emergency department of general hospital.

Objective To investigate the role of autophagy in the lung injury in the septic mice.Methods Thirty-six male C57BL/6 mice, aged 6 weeks, weighing 20-25 g, were randomly divided into 3 groups (n=12 each) using a random number table: sham operation group (group S);cecal ligation and puncture (CLP) group;CLP + autophagy inhibitor 3-methyladenine (3-MA) group (group CLP+3-MA).Sepsis was produced by CLP.In group CLP+3-MA, 3-MA 10 mg/kg was injected intraperitoneal at 1 h after operation.Arterial blood samples were taken at 24 h after operation for blood gas analysis, and the oxygenation index was calculated.The lungs were removed for microscopic examination of pathologic changes which were scored, and for determination of wet/dry lung weight ratio (W/D ratio) , myeloperoxidase (MPO) activity (using colorimetric method) and the expression of autophagy protein microtubule-associated protein 1 light chain 3 Ⅱ] (LC3 Ⅱ), Beclin-1 and lysosomes-associated protein Rab7 and lysosome-associated membrane protein-2 (LAMP2) (by Western blot).The lung was lavaged, and broncho-alveolar lavage fluid (BALF) was collected for determination of the total cell count and polymorphonuclear leukocyte (PMN) count.Results Compared with group S, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, and oxygenation index was decreased in CLP and CLP +3-MA groups, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was up-regulated in group CLP (P＜0.05).Compared with group CLP, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, the oxygenation index was decreased, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was down-regulated in group CLP+ 3-MA (P＜0.05).Conclusion Autophagy is involved in the endogenous protective mechanism of acute lung injury in the septic mice.

Objective To investigate the exercise performance in maintenance dialysis patients,and analyze its correlative factors.Methods Maintenance dialysis patients admitted in Tongji Hospital of Shanghai from December 2014 to March 2015 were enrolled,with their baseline data and biochemical measurement being collected.The anthropometric indexes including arm circumference,triceps skinfold,waist circumference and hip circumference were detected.The exercise activity was assessed by hand grip test,timed up and go test (3mTUG) and five times sit-to-stand test (FTSST).Patients were divided into fast group (3mTUG≤12 s) and slow group (3mTUG ＞ 12 s).Univariate and multivariable analyses were used to evaluate the factors influencing exercise performance in maintenance dialysis.Results There were 121 patients enrolled:62 on peritoneal dialysis and 59 on hemodialysis,76 men and 45 women.Patients' average age was (61.6± 13.0) years and median dialysis age was 31.7(12.3,69.0) months.There was no statistical difference between fast group (n=80) and slow group (n=41) in gender,dialysis method,dialysis age,body mass index (BMI),arm muscle area (AMA),waist-hip ratio (WHR),hemoglobin (Hb) and total cholesterol (TC).Patients in fast group were younger,had higher serum albumin,prealbumin,serum phosphate and iPTH,and less prevalence of diabetes than those in slow group.In exercise activity,patients in fast group had better performance in handgrip,3mTUG and FTSST (all P ＜ 0.05).Univariate analysis showed that,handgrip was correlated with sex (male),AMA,BMI,age,diabetes,serum phosphorus and TC;scores in FTSST was correlated with age,BMI,diabetes,WHR,dialysis method,dialysis age,prealbumin and serum phosphorus;scores in 3mTUG was correlated with age,diabetes,WHR,dialysis method and dialysis age,prealbumin,serum phosphorus and iPTH (all P ＜ 0.05).Multiple stepwise regression analysis showed that sex (male),age,AMA and diabetes were independently correlated with handgrip in dialysis patients (all P ＜ 0.05);age,dialysis method,BMI and diabetes were independently correlated with scores in FTSST (all P ＜ 0.05);age,dialysis method,diabetes and WHR were independently correlated with scores in 3mTUG (all P ＜ 0.05).Conclusions The exercise performances of patients on maintenance dialysis are impaired.Age and diabetes are independent factor associated with the exercise performances of patients on maintenance dialysis.AMA is independently associated with upper limb movement,and dialysis method,BMI and WHR are independent factors associated with lower limb movement in dialysis patients.

Objective:To examine the plasma soluble interleukin 2 receptor (sIL-2R)and the expression of IFN-γR in helper T (Th)cells in peripheral blood of patients with recurrent oral ulcer(ROU).Methods:The peripheral blood was obtained from 21 pa-tients in active stage of ROU and 21 healthy controls.Flow cytometry was used to detect the quantity of IFN-γR in CD +4 T cells,ELISA was used to detect the level of sIL-2R.Results:The expression of IFN-γR in CD +4 T cells and sIL-2R content in plasma of the patients with ROU were significantly higher than those in the healthy controls(P <0.05).Conclusion:Over expression of IFN-γR in CD +4 T cells and sIL-2R in peripheral blood may be related to the genesis of ROU.

ABSTRACT:Objective To prepare polymeric nanomicelles capable of simultaneously loading doxorubicin (DOX) and Bcl‐2 small interfering RNA (Bcl‐2 siRNA ) , and to explore their in vitro cytotoxicity and cellular uptake in MCF‐7 human breast cancer cells .Methods Copolymer poly (ethylene glycol )‐g‐polyethylenimine‐g‐poly(γ‐benzyl‐L‐glutamate) was synthesized by the combination of reductive amination and carbodiimide methods , and its chemical structure was verified by 1 H NMR .Empty and drug‐loaded copolymeric nanomicelles were prepared by dialysis method and characterized by transmission electron microscope and dynamic light scattering .The ability of the nanomicelles to compress Bcl‐2 siRNA was measured by by agarose gel electrophoresis method . The release profiles of DOX and Bcl‐2 siRNA from the nanomicelles were explored by means of fluorescence spectrometry and dialysis method .The in vitro cytotoxicity and cellular uptake of DOX and Bcl‐2 siRNA co‐loaded nanomicelles in MCF‐7 human breast cancer cells were characterized by MTT assay and confocal laser scanning microscopy , respectively .Results The critical micelle concentration of the copolymer was about 4 mg/L ,and the sizes of self‐assembled empty and drug‐loaded nanomicelles were smaller than 200 nm .The drug‐loading efficiency and drug‐loading content of DOX in the nanomicelles were 88 .7% and 15 .1% ,respectively .The DOX‐loaded nanomicelles could efficiently compress Bcl‐2 siRNA when an N/P ratio was ≥64 .The zeta potential of DOX and Bcl‐2 siRNA co‐loaded nanomicelles was +30 mV .The release behavior of the cargoes from the nanomicells was pH‐sensitive , and the release of Bcl‐2 siRNA was more sensitive to acidic pH than that of DOX . The nanomicelles could simultaneously deliver DOX and Bcl‐2 siRNA into MCF‐7 cells , and the co‐delivered DOX and Bcl‐2 siRNA significantly increased the cytotoxicity of DOX (P<0 .05) .Conclusion The polymeric nanomicelles can co‐load DOX and Bcl‐2 siRNA and deliver them into MCF‐7 cells , and DOX in combination with Bcl‐2 siRNA can synergistically inhibit the growth of MCF‐7 cells and promote cell apoptosis ,suggesting that the nanomicells may be a promising carrier for the co‐delivery for chemotherapeutics and genes .

Objective To evaluate the effect of continuous renal replacement therapy (CRRT) on the remove of inflammatory mediators and the function of endothelial cells in patients with multiple organ dysfunction syndrome (MODS). Methods Thirty patients with MODS were enrolled in this study. All of the patients underwent CRRT for at least 24 hours. Peripheral blood levels of IL-1β, IL-4, IL-6, IL-10, TNF-α, E-selectin, sVCAM-1, slCAM-1 and PAF-AH were measured at the beginning and 3, 6, 12, 24 h after initiation of CRRT. Results Nineteen patients survived after 14 days and 17 patients survived after 28 days during therapy. The clinical oxygenation and bemodynamics were improved after 6 h of CRRT. Among inflammatory mediators, the levels of TNF-α, IL-6, IL-10 rose gradually from the beginning [(462.24±331.03) ng/L, (106.39±90.82) ng/L, (124.51±118.39) ng/L), and reached the peak at 12 h [(887.88±975.46) ng/L, (132.01±118.14) ng/L, (167.01±161.66) ng/L], and the levels of IL-1β, IL-4 decreased from initiation of CRRT. But there were no significant differences in the levels of above cytokines between at the beginning and at the end of CRRT. There were significant differences in the levels of cytokines between survival and death group. The level of IL-6 in death group [(145.45±14.28) ng/L] was significantly higher than that in survival group [(106.03±10.86) ng/L]. The level of IL-10 in death group [(94.93±16.09) ng/L] was significantly lower than that in survival group [(143.06± 12.24) ng/L]. Levels of E-select, sVCAM-1 and sICAM-1 elevated from the beginning and reached the peak at 12 h, but no significant differences were found between intiation and the end of CRRT. The level of PAF-AH increased after initiation, and there was a significant difference between beginning and the end of CRRT. Levels of cytokines for endothelial cell function were significantly different, such as E-selectin [(287.13±42.70) μg/L vs (266.26±65.26) μg/L], sVCAM-I [( 1697.25± 475.24) μg/L vs (1488.10±691.67) μg/L], sICAM-1 [(975.33±142.50) μg/L vs (835.40±332.41) μg/L], and PAF-AH [(9.07±6.38 ) μg/L vs (16.32±8.95)μg/L]. Conclusions Clinical oxygenation and hemodynamics can be improved, and endothelial cell function can be improved partly by CRRT. There were no significant differences of inflammatory mediator levels between initiation and the end of CRRT. IL-6 and IL-10 can be used as predicators for prognosis of MODS patients.

Objective To analyze the expression of the placenta Grb10 from women conceived by transferred thawed blastocyst,and to evaluate the security of blastocysts vitrification.Methods A cross-sectional study was performed in the Department of Obstetrics and Gynecology of Fujian Provincial Maternity and Children's Hospital from January 2012 to May 2014,50 women conceived by transferring thawing blastocyst and 50 natural pregnancy control women were enrolled in this study.The expression of Grb10 protein was detected by immunohistochemistry and Western blot,and the expression of Grb10 mRNA was detected by Realtime PCR method.Results Comparison of two cases of gestational age,gestational age,fetal sex,fetal body weight,body length,head circumference,abdominal circumference,there were no significant differences(P＞0.05),comparison of placental area,placental weight,the difference was statistically significant(P＜0.05).Real-time PCR and Western blot results showed that,there was no significant difference in the expression of Grb10 mRNA and protein between the two groups(P＞0.05).Conclusion Blastocysts vitrification may increase the area and quality of delivery of placenta,however,there was no significant change in the expression of Grb10 in placenta.

Objective To evaluate the effect of hydrogen ( H2 ) inhalation on mitochondrial bio?synthesis in lung tissues during acute lung injury in mice with sepsis. Methods One hundred and four male ICR mice, aged 6 weeks, weighing 20-25 g, were divided into 4 groups ( n=26 each) using a ran?dom number table: sham operation group ( group S) , sham operation + H2 group ( group S+H2 ) , sepsis group ( group Sep) and sepsis + H2 group ( group Sep+H2 ) . Sepsis was produced by cecal ligation and puncture. In S+H2 and Sep+H2 groups, the mice inhaled 2% H2 for 1 h starting from 1 and 6 h after opera?tion. Twenty mice in each group were selected, and the survival rates on postoperative days 1, 2, 3, 5 and 7 were recorded. On the postoperative day 1, 6 mice in each group were selected, and blood samples were collected from the common carotid artery for measurement of arterial oxygen partial pressure, and the oxygenation index was calculated. The pulmonary specimens were obtained for examination of the pathologi?cal changes which were scored and for determination of the expression of peroxisome proliferator?activated re?ceptor gamma coactivator?1α ( PGC?1α) in lung tissues by Western blot. The pulmonary mitochondria were isolated for determination of mitochndrial membrane potential ( MMP ) and ATP contents using spectropho?tometry and a bioluminescence technique, respectively. Results Compared with group S, the survival rate, oxygenation index and MMP and ATP content in lung tissues were significantly decreased, and the pathological scores and PGC?1α expression in lung tissues were significantly increased in Sep and Sep+H2 groups (P<0.05). Compared with group Sep, the survival rate, oxygenation index, and MMP, ATP content and PGC?1α expression in lung tissues were significantly increased, and the pathological scores were significantly decreased in group Sep+H2 ( P<0.05) . Conclusion H2 inhalation can ameliorate acute lung injury in mice with sepsis, and the mechanism is associated with the enhanced function of PGC?1αand promoted mitochondrial biosynthesis in lung tissues.