Objective To determine the expression of adapter protein p66Shc in mediating alveolar epithelial cells induced by hyperoxia and to explore their relationship.Methods A549 cells were cultured in vitro and divided randomly into a control group and a hyperoxia group.The hyperoxia group was exposed to a mixture of oxygen(O2,900 mL/L) and carbon dioxide(CO2,50 mL/L) for 10 min,then cultured in a closed environment.The changes in morphology were observed under inverted microscope after exposure to oxygen or air for 24 hours.The cell apoptosis was detected by flow cytometry (FC) after 24 hours.And the expression of p66Shc was detected by immunohistochemical method after 24 hours.The correlation of the changes in mitochondrial membrane potential and p66Shc protein expression was analyzed by using Bivariate correlation analysis.Results 1.Under inverted microscopy,A549 cells from the air group significantly increased,stuck to each other tightly and grew very quickly.Their adhesion was better,multy-angle oblate and many cells were in division phase.Compared with the control group,the changes in morphology of A549 were remarked and obvious than those in the hyperoxia group.The cells grew slowly,their counts decreased and the cell morphology changed from typical multi-angle oblate to round or ellipse.2.Compared with the control group,after 24 h,in hyperoxia group of A549 cells,red fluorescence decreased,and green fluorescence enhanced.3.Compared with the controls (0.057 664 88 ± 0.006 517 84),the expression of p66Shc (0.123 600 50 ± 0.004 227 23) was significantly increased in the hyperoxia group(t =-24.006,P ＜ 0.001).4.The decline of membrane potential was negatively correlated with the increased expression of p66Shc protein (R =-0.988,P ＜ 0.001).Conclusions The hyperoxia induction could significantly increase in injured mediating alveolar epithelial cells induced by hyperoxia,the expression of p66Shc increases,the membrane potential declined,and they exhibit a negative correlation.So p66Shc may be involved in the process of high oxygen damage to human alveolar epithelial cells.

Objective To investigate the lacrimal gland normal and abnormal diagnosed characteristics.Methods One hundred and six cases were examined by color Doppler flow imaging(CDFI).The results were compared with B mode scan.Results CDFI was better than B mode ultrascan in showing the whole lacrimal gland and inter echo at the same time observing the blood flow of it.Configuration and blood flow combining were a good choice for lacrimal gland disease in the diagnosis.Conclusions CDFI was the best way for lacrimal gland diseases.

Objective:This study aims to investigate the protein expression of CD133, CD117, and Ki-67 in human osteosarcoma tissues and explore their relationships with the clinico-pathological features and risk of osteosarcoma. Methods:Immunohistochemical method was used to examine the protein expression of CD133, CD117, and Ki-67 gene in the paraffin specimens of 55 and 20 cases of osteosarcoma and osteochondroma, respectively. SPSS17.0 statistical software was used to explore the relationships among the expressions of CD133, CD117, and Ki-67 gene and the biological behavior and prognosis of osteosarcoma. Test criterion:P<0.05 was considered statistically significant. Results:The positive expression rates of CD133, CD117, and Ki-67 were significantly higher in the osteosarcoma tissues than in the benign osteochondroma tissues, and the differences were significant (P=0.016, P=0.008, and P<0.001, respectively). The mean survival and metastasis time were shorter in the CD133 or Ki-67 positively expressed osteosarcoma patients than in those with CD133 or Ki-67 negatively expressed osteosarcoma. The differences were significant (P<0.05). The expression of CD133 and Ki-67, surgical staging, and distal metastasis were relevant to the prognosis of osteosarcoma patients. The expression of CD133 and distal metastasis are the independent risk factors that affect the prognosis of these patients. Conclusion:The positive expressions of CD133 and Ki-67 closely correlated with the occurrence and progression of osteosarcoma, and may be used as an indicator for prognosis of the cancer.

Objective To explore the mechanism for the increase in reactive oxygen species regulated by p47phox of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit in peripheral blood mononuclear cells (PBMCs) after oxygen therapy in premature infants.Methods According to different volume fractions of oxygen,premature infants less than 32 weeks were divided into 3 groups:fractional concentration of inspired oxygen (FiO2) ＜ 30％ was low concentration oxygen group,FiO2 between 30％ and 40％ as middle concentration oxygen group,and FiO2 ＞ 40％ as high concentration oxygen group.Premature infants less than 32 weeks without oxygen was control group.After 48 h,3 mL blood was collected via radial artery from each group,PBMCs and serum were separated.Then intracellular reactive oxygen species (ROS) by confocal laser scanning microscopy,malondialdehyde (MDA) within serum by thiobarbituric acid colorimetric,and the location and activation rate of p47phox through immunofluorescence.Results After premature infants were exposed to oxygen,as the oxygen volume fraction was increasing,ROS and MDA gradually rised.More PBMCs with p47phox translocated to membrane,then the translocation rate of p47phox also increased.Compared with the control group,ROS were significantly higher(q =4.48,6.5,16.22,all P ＜ 0.05) among the other 3 groups ; MDA significantly increased as well(q =5.08,8.22,12.76,all P ＜ 0.05) ; the activation rate of p47phox also had significant differences (x2 =134.008,P ＜ 0.05);compared with the middle concentration oxygen group,the high concentration oxygen group had higher ROS and MDA(q =15.03,4.53,all P ＜ 0.05) ; the activation rate of p47phox increased significantly(x2 =19.26,P ＜ 0.05).Conclusions After oxygen exposure,p47phox translocated to membrane may regulate the NADPH oxidase-derived ROS increase in extremely premature infants.

Objective To observe the relationship between expression of retinoic acid receptor-β (RAR-β) in esophageal squamous cell carcinoma (ESCC) and chemotherapy response. Methods Fifty-two cases advanced ESCC patients treated by DDP and 5-FU, DDP 80 mg/m2, divided into 5 days;5-FU 375 mg/m2, dl-5. Immunohistochemistry was used to exmine the expression of RAR-β in ESCC. Fifty cases normal esophageal tissue were used as controls. Results RAR-β immunoreactivity was recognizd in both cytoplasm and nucleus, RAR-β positive rate was lower in ESCC compared with normal tissue (61.5%vs 92% ,P ＜0. 05 ). The 52 cases ESCC patients were treated 228 chemotherapy cycles, the overall response rate (OR) was 71.2%. The OR in RAR-β positive patients was 84. 4% (27/32), significant higher than RAR-β negative patients 50. 0% ( 10/20 ) ( P ＜ 0. 05 ). The time-to-progression ( TTP ) for RAR-β positive patients was 5.9 months, the median survival period was 12. 1 months, 2 years survival rate was 56. 7%;whereas TTP for RAR-β negative patients was 2. 1 months, the median survival period was 5.8 months,2 years survival rate was 32. 9%. There was signifcant difference between the 2 groups ( P ＜ 0. 05 ) .Conclusion RAR-β protein expression by immunohistochemistry may be a useful indicator to predict the chemotherapy response and clinical outcome for ESCC, meanwhile it may be an avenue for target therapy.

Objectives To explore the role of PKCβ/p66Shc oxidative stress signaling pathway in hyperoxia-induced apoptosis of alveolar epithelial cells A549. Methods A549 cells were cultured in vitro and divided randomly into control (incubated with 5%CO2), hyperoxia group (exposed to a mixture of 900 ml/L O2 and 50 ml/L CO2 at speed of 3 L/min for 10 mins, then cultured in a closed environment) and LY333531 group (treated with 10μmol/L of PKCβinhibitor LY333531 for 24h then induced with hyperoxia for 10 mins). The cellular morphology was observed under inverted microscope at 12, 24 and 48 h of treatment. The cell apoptosis was detected by lfow cytometry. Expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were detected by immunohistochemistry after 24 h of treatment. Results Comparing to the control group, the cellular morphology of A549 in the hyperoxia group changed to spherical shapes and space between cells increased, the living cell count decreased and suspension cell increased. The living cell count in LY333531 group increased and suspension cell decreased than those in hyperoxia group but not reach the levels of the control group. The apoptosis rate of A549 cells and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 at 24 h were signiifcantly increased in the hyperoxia group than those in the control group, while the apoptosis rate and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were greatly decreased in the LY333531 group than those in the hyperoxia group (all P<0.01). Conclusions The expression of PKCβin A549 cells can be increased by the hyper-oxia induction but reduced by LY333531, and then the expressions of Pin1, p66Shc and p66Shc-Ser36 are reduced. Thus the re-duced apoptosis of A549 cells relieve the cell injury induced by hyperoxia.

Objective To evaluate the effect of Budesonide combined with Poractant Alfa(pulmonary surfactants,PS) on preventing the bronchopulmonary dysplasia (BPD) in preterm infants.Methods One hundred and twenty preterm infants 6 hours after birth(gestational ages≤32 weeks and birth weights ≤1500 g)admitted to the Department of Newborn Medicine,the Affiliated Hospital of Southeast Medical University from October 2013 to February 2015 were randomly divided into 4 group(30 cases in each group).Group A was a control group,group B was neonatal respiratory distress syndrome(NRDS) group,group C was NRDS with PS group,and group D was NRDS with PS and Budesonide group.Thirty-two-week preterms without other diseases and without uptaking oxygen within 48 h were assigned as group A.Group B were treated by continuous uptaking oxygen with continuous positive airway pressure(CPAP) (oxygen uptaken lasting more than 48 h and oxygen concentrations more than 40％).Group C were treated with 100 mg/kg PS within 48 h on the basis of group B.Group D were treated with 0.25 mg/kg Budesonide suspension for inhalation on the basis of group C.The pH value,partial pressure of oxygen [pa(O2)],partial pressure of carbon dioxide [pa(CO2)] in the blood gas analysis were all detected in all groups before treatment,1,6,12,24 and 48 hours after using drug,respectively.All groups were also observed for whether to use respirator assisted ventilation,the duration of high oxygen use,the total time of uptaking oxygen,the rate of using PS again,the rate of BPD,the total hospitalization days and the adverse effects.The adverse effects included high blood pressure,high blood sugar,necrotizing enterocolitis and the incidence of nosocomial infection.Results Compared with group B,the pH value at 1 and 6 hours after using drugs,the pa(O2) and pa(CO2) at 1,6 and 12 hours after using drugs were improved obviously in the group C,and the differences were statistically significant (all P＜0.01).Compared with group B,the above indicators were improved more obvious in group D,and the differences were statistically significant (all P＜0.01).Moreover,compared with the group B,the oxygen inhalation duration,the rate of having a respirator assisted ventilation and using PS again,and the incidence of BPD were obviously decreased in other groups,the differences were statistically significant (all P＜0.05).The incidence of BPD in group D was less than that of group C,and the differences were statistically significant (3.33％ vs 10.00％,x2=4.00,P=0.046).The total oxygen time and the rate of adverse effects of each group were similar.The differences were not statistically significant (all P＞0.05).Conclusions Budesonide combined with Poractant Alfa can prevent BPD in preterm infants.Its effect is better than the simple use of Poractant Alfa,and the rate of adverse effects are not increased significantly.

Objective To explore the optimization of intubations for severe brain injury patients in ICU. Methods Seventy-six severe brain injury patients were divided into research group and traditional group through the envelop method, with 38 patients in each group. Patients in research group was induced by dexmedetomidine, while patients in traditional group was induced by midazolam or propofol. Throat and intratrachea surface anesthesia was taken with 3 ml of 1%lidocaine on two groups before intubation, and they were intubated through direct laryngoscope. Patients were connected with breathing machines after intubation, and the arterial carbondioxide partial pressure was maintained at 30-35 mmHg (1 mmHg=0.133 kPa). Intubations time, one-time success rate, cardiovascular reaction index, myocardial damage and heart failure biomarkers, adverse reaction, patients comfort and intubationists satisfaction of the two groups were compared. Results The intubation time on research group was significantly shorter than that on traditional group [(27.1 ± 14.2) s vs. (42.2 ± 18.7) s], and there was statistical significance (P<0.01). The one-time success rates in two groups was not statistically significant (P>0.05). The heart rates and mean arterial pressure (MAP) at pre-intubation, post-intubation, 5 min after intubation and 10 min after intubation in research group were significantly lower than those in traditional group:heart rate:(77.8 ± 8.5) bpm vs. (85.1 ± 7.6) bpm, (85.3 ± 9.1) bpm vs. (106.4 ± 12.5) bpm, (84.4 ± 10.4) bpm vs. (96.4 ± 11.9) bpm, (80.5 ± 12.1) bpm vs. (89.0 ± 10.5) bpm;MAP: (82.6 ± 10.1) mmHg (1 mmHg = 0.133 kPa) vs. (93.2 ± 14.3) mmHg, (88.3 ± 13.2) mmHg vs. (113.7 ± 15.1) mmHg, (85.4 ± 14.0) mmHg vs. (101.3 ± 9.9) mmHg, (83.7 ± 10.7) mmHg vs. (90.3 ± 13.4) mmHg, and there were statistical significances (P<0.05). There was no statistical significance of SpO2 between groups at every time point (P>0.05). For 6 and 12 h after intubation, creatine kinase isoenzyme MB (CK-MB), cardiac troponin T (cTnT) and brain natriuretic peptide (BNP) levels in research group were significantly lower than those in traditional group:CK-MB:(30.5 ± 7.2) U/L vs. (35.2 ± 10.1) U/L, (25.7 ± 5.7) U/L vs. (27.7 ± 6.5) U/L;cTnT:(0.6 ± 0.2) μg/L vs. (0.8 ± 0.1) μg/L, (0.5 ± 0.2) μg/L vs. (0.6 ± 0.2) μg/L;BNP:(152.6 ± 13.7) pg/L vs. (189.4 ± 19.3) pg/L, (89.7 ± 27.5) pg/L vs. (111.8 ± 20.4) pg/L, and there were statistical significances (P<0.05). There were no significant adverse reactions in two groups after topical anesthesia. The comfort and intubationists satisfaction scores in research group were significantly higher than those in traditional group: (2.5 ± 0.4) scores vs. (1.8 ± 0.5) scores and (8.8 ± 1.1) scores vs. (7.1 ± 0.9) scores, and there were statistical significance (P<0.01). Conclusions Combining dexmedetomidine induction with topical anesthesia to intubate is safe and effective, which is the optimization of tracheal intubations to severe brain injury patients.

Objective To investigate the clinical effect of mild hypothermia on neonatal bilirubin encephalopathy, and the value of 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)/CT and amplitude integrated electroencephalogram (aEEG) for diagnosis and evaluation of curative effect. Methods From May, 2013 to December, 2014, 29 newborns with bilirubin encephalopathy were divided into conventional group (n=15) and mild hypothermia group (n=14). The conventional group received conventional therapy, and the other group received mild hypothermia in addition. The aEEG and neuron-specific enolase (NSE) were measured before and after treatment, as well as the glucose metabolism rate with 18F-FDG PET/CT after treatment. Results The NSE was lower after treatment in both groups (t>9.670, P<0.001), and was lower in the mild hypothermia group than in the conventional group (F=46.146, P<0.001). After treatment, sleep-wake cycle (SWC), epileptiform activity and the degree of abnormality were obviously improved (P<0.05), and were better in the mild hypothermia group than in the conventional group (P<0.05). The cerebral glucose metabolism rate was significantly better in the mild hypo-thermia group than in the conventional group (t>2.943, P<0.01). The cerebral glucose metabolism rate was negatively correlated with aEEG and NSE (r>0.640, P<0.05). Conclusion Mild hypothermia therapy could further promote the energy metabolism of brain cells in neonatal bilirubin encephalopathy. 18F-FDG PET/CT and aEEG can be used for early diagnosis and therapeutic evaluation.

Objective To evaluate the efficacy and safety of inhaled corticosteroids for preventing chronic lung disease (CLD) in preterm infants. Methods PubMed, EMBASE, CENTRAL, the ISI Web of Knowledge databases, CBM, CNKI, VIP and Wanfang Data were searched for the period up to Oct. 2016. All randomized controlled trials (RCTs) about inhaled corticosteroids for preventing CLD in preterm infants were collected. The RCTs had been screened, data were extracted and assessed. The mata-analysis was performed by RevMan 5.3 software. Result A total of 12 RCTs were included (a total of 2051 preterm neonates). Compared with control group, in 28 day old group, the incidence of CLD was not significantly different between experimental and control groups (RR=0.87, 95%CI:0.74-1.03, P=0.11) and (RR=1.19, 95%CI:0.59-2.43, P=0.63) and no significant difference among subgroups budesonide (α), beclomethasone (β), fluticasone (γ) (RR=0.89, 95%CI:0.69-1.14, P=0.35), (RR=0.86, 95%CI:0.69-1.08, P=0.19) and (RR=0.91, 95%CI:0.60-1.38, P=0.19). In 36 wk postmenstrual age group,the incidence of CLD was decreased in experimental group and in subgroups inhalation (A), Intratracheal administration (B), α, γ (RR=0.70, 95%CI: 0.61-0.80, P<0.00001), (RR=0.74, 95%CI: 0.63-0.87, P=0.0003), (RR=0.57, 95%CI: 0.43-0.76, P=0.0002), (RR=0.67, 95%CI: 0.57-0.78, P<0.00001) and (RR=0.58, 95%CI: 0.36-0.94, P=0.03); but it is not significantly different in subgroup β(RR=0.98, 95%CI: 0.69-1.39, P=0.90); There was no difference in the motality in experimental and subgroups A ,B, α, β , γ (RR=1.07, 95%CI:0.86-1.33, P=0.55), (RR=1.24, 95%CI: 0.97-1.59, P=0.09), (RR=0.67, 95%CI: 0.43-1.03, P=0.07), (RR=1.04, 95%CI: 0.81-1.33, P=0.78), (RR=1.47, 95%CI: 0.79-2.74, P=0.22) and (RR=0.91, 95%CI: 0.47-1.74, P=0.77). No clinically significant adverse effects were observed during the study. Conclusions This updated review indicated that early administration of inhaled steroids to very low birth weight preterm neonates was effective in reducing the incidence of CLD. There was no statistically significant effect of inhaled steroids on motality, and there was no significant correlation between the mode of administration and the type of drug delivery, It is recommended to observe the 36 week gestational age as the outcome index. More and larger randomised placebo-controlled trials including long-term follow up are needed to establish the efficacy and safety of inhalation corticosteroids.