This study aimed to explore Semaphrin4D (Sema4D) expression and clinical significance in non-small cell lung cancer (NSCLC), and to define the roles and mechanisms of Sema4D in regulating the malignant behaviors of A549 cells by small interfering RNA (siRNA). Firstly, immunohistochemistry revealed that Sema4D was more frequently expressed in NSCLC than in lung benign lesion (P<0.05) and its overexprssion was associated with low differentiation (P<0.05), poor pTNM staging (P<0.05) and occurrence of lymph node (LN) metastasis (P<0.05). Endogenous Sema4D expression was suppressed by Sema4D siRNA in A549 cells overexpressing Sema4D. Protein levels of Sema4D, total Akt and p-Akt were examined by Western blotting. Cell proliferation, migration and invasion abilities were measured by MTT assay and Transwell assay respectively. Results showed that Sema4D siRNA significantly suppressed phosphorylation of AKT in A549 cells, but it did not alter total AKT expression. In addition, efficient down-regulation of SemaD significantly inhibit cell proliferation (P<0.05), migration (P<0.05) and invasion (P<0.05) in A549 cells. These findings suggest that Sema4D might serve as a reliable tool for early prediction of NSCLC poor prognosis. Sema4D could play an important role in promoting tumor proliferation, migration and metastasis in the NSCLC, by influencing the Akt protein phosphorylation. Inhibition of Sema4D may be a useful approach for the treatment of NSCLC.

Objective Discuss the clinical efficacy of caffeine citrate and aminophylline in treatment of primary apnea of prematurity.Methods 80 cases of primary apnea of prematurity were selected.They were divided into two groups randomly.The control group (40 cases) received aminophylline treatment and the observation group (40 cases) received caffeine citrate therapy.The efficacy of caffeine citrate and aminophylline in treatment of primary apnea ofprematudty was evaluated by efficacy,primary apnea episodes and disappeared time,adverse reaction during treatment.Results The effective rate of observation group was 85％,the effective rate of control group was 75％.The observation group had higher efficiency (P ＜ 0.05).The frequency of apnea of observation group was less than that of the control group.The disappearance time of observation group was shorter than that of the control group (P ＜ 0.05).During the treatment,the tachycardia,feeding intolerance,bronchopulmonary dysplasia and other adverse reactions rate of observation group was lower than that of the control group (P ＜ 0.05).Conclusion Compared with aminophylline,caffeine citrate had a good therapeutic effect of primary apnea of prematurity.It can reduce apnea frequency,eliminate clinical symptoms with high safety.It was an ideal drug for treatment of primary apnea.

Objective To investigate the effect of cocaine-amphetamine-regulated transcript peptide (CART) on the content of 4-hydroxy-2-noneral (HNE) and infarct volume after cerebral ischemia/reperfusion in mice.Methods A total of 96 healthy male mice were randomly divided into four groups:ischemia/reperfusion (n =27),CART (n =27),normal saline control (n =27) and sham operation (n =15) groups.A middle cerebral artery occlusion (MCAO) model was induced.Two hours after MCAO,CART 55-102 and equivalent normal saline were injected respectively via the tail veins of mice in the CART group and the normal saline control group,and then they were injected every other 24 hour.The neurological scores,infarct volume and the HNE content of lipid metabolism of oxidative stress were performed and detected respectively at 12,24,48 and 72hours after reperfusion.Results CART could significantly improve the neurological deficit scores (all P ＜0.05) and reduce infarct volume (all P＜0.05) at different time points after ischemia/reperfusion.The content of HNE was upregulated (all P＜0.05) at different points after referfusion.CART could significantly down-regulate the increased HNE levd in brain after ischemia (all P＜0.05).Conclusions CART may protect ischemic brain injury in mice by inhibiting lipid peroxidation.

Objective To assess the safety of early subcutaneous injection of a low-dose low molecular weight heparin (LMWH) nadroparin for prevention of deep vein thrombosis (DVT) in patients with spontaneous intracerebral hemorrhage (sICH).Methods The patients with sICH who early using nadroparin or lower limb intermittent pneumatic compression (IPC) for prevention of DVT were enrolled.A nadroparin group continuously injected nadroparin 0.4 ml/d subcutaneously for 10 days at day 4 after admission and an IPC group used lower limb IPC.Head CT was reexamined and hematoma volume changes were evaluated at day 3,5,and 14 after admission.The hemorrhagic events during the course of treatment were documented,and the lower limb DVT was examined by color Doppler sonography.Results A total of 94 patients with acute sICH (n =41 in the nadroparin group,n =53 in the IPC group) who early use of nadroparin or IPC for prevention of DVT were enrolled.Fourteen patients had lower limb DVT,5 (12.2％) of them were in the nadroparin group and 9 (17.0％) of them were in the IPC group.However,there was no significant difference in the incidence of DVT between the two groups (x2 =0.418; P =0.518).During the treatment,no patient experienced increased intracranial hematoma and rebleeding.Conclusion Early subcutaneous injection of low-dose nadroparin for the prevention of DVT in patients with sICH is safe.

Objective To investigate the assessment methods and mechanisms of nonsteroidal antiinflammatory drugs (NSAID)-induced injury in rat small intestinal epithelial barrier,and to explore the protective effects of mucosal protective agents and antacids on it.Methods A total of 96 rats were evenly divided into the morphologic observation group and the mechanism research group,and 48 in each group.Then each group was evenly divided into eight subgroups:the healthy control group,the model group (model established with indomethacin),the teprenone prevention group,the rabeprazole prevention group,the treatment control group,the teprenone treatment group,the rabeprazole treatment group and the teprenone and rabeprazole combined group (combined group),six in each group.Exfoliated cells gap density of small intestine of each subgroup was determined by confocal laser endomicroscopy.Serum level of tumor necrosis factor-α (TNF-α)was measured with enzyme-linked immunosorbent assay (ELISA). The expression of nuclear factor-κB (NF-κB),caspase-3,zonula occludens-1 (ZO-1 )and occludin at protein level was detected by Western blotting.The LSD-t test or Hamhane′s T2 test was performed for statistical analysis.Results The exfoliated cells gap densities of the teprenone prevention group and the rabeprazole prevention group were (57.43 ± 24.55 )/1 000 and (59.80 ± 21 .14 )/1 000,respectively, which were both lower than that of the model group ((110.93±50.58)/1 000),and the differences were statistically significant (t= 53.50 and 54.13,both P < 0.01 ).The exfoliated cells gap density of the combined group was (40.53 ±15 .39)/1 000,which was lower than that of the treatment control group ((93.80±40.65 )/1 000 ),and the difference was statistically significant (t =44.27,P <0.01 ).The serum levels of TNF-α of the teprenone prevention group and the rabeprazole prevention group were (25 .80±8.97)ng/L and (22.74 ±7.15 )ng/L,repsectively,which were both lower than that of the model group ((44.48 ± 7.42 )ng/L),and the differences were statistically significant (t = 18.68 and 21 .74,both P <0.01 ).The serum level of TNF-αof the combined group ((13.66 ±4.98)ng/L)was lower than that of the treatment control group ((24.67±6.70)ng/L),and the difference was statistically significant (t = 9.02,P < 0.01 ).The caspase-3 levels of teprenone prevention group and rabeprazole prevention group were 1 .47 ±0.35 and 1 .58 ±0.34,and the NF-κB levels of these two groups were 1 .27±0.14 and 1 .21 ± 0.10,respectively.Compared with those of model group (2.44 ± 0.45 and 1 .69±0.13),the differences were statistically significant (t =0.97,0.86,0.42 and 0.48,respectively, all P <0.01 ).The levels of caspase-3 and NF-κB of the combined group were 0.66±0.06 and 0.44 ± 0.21 ,respectively,which were lower than those of the treatment control group,and the differences were statistically significant (t=0.34 and 0.56,both P <0.01).The expressions of occludin at protein level of the teprenone prevention group and the rabeprazole prevention group were 0.69 ±0.16 and 0.74 ±0.11 , and the levels of ZO-1 were 0.81 ± 0.08 and 0.84 ± 0.12.Compared with those of the model group (0.45 ±0.22 and 0.64±0.07 ),the differences were statistically significant (t =0.24,0.29,0.17 and 0.21 ,respectively,all P <0.01 ).The levels of occludin and ZO-1 of the combined group were 2.50 ± 0.46 and 1 .76±0.18,which were higher than those of the treatment control group,and the differences were statistically significant (t =1 .50 and 0.76,both P <0.01 ).Conclusions The exfoliated cells gap density may be a valuable indicator to predict the degree of inflammation response and permeability of epithelial barrier as well as to evaluate efficacy of medication.Teprenone and rabeprazole have prevention and treatment effects on NSAID-induced injury in rat small intestine.

Objective To systematically evaluate the effectiveness of delayed cord clamping (DCC) in term infants. Methods The data of the Cochrane library, PubMed, EMBASE, CNKI , VIP, Wanfang from 1 January 1970 to 30 April 2013 were searched. Randomized controlled trials (RCT) of DCC in term infants were included.RevMan 5.1.0 was used in the statis-tical analysis. Results Ten studies involving 1623 participants were included. Meta-analysis based on included studies showed that:compared with immediate cord clamping (ICC), DCC improved the hemoglobin levels at birth (MD=2.19, 95%CI:0.36, 4.02) and increased the incidence of polycythaemia (RR=2.87, 95%CI:1.24, 6.62). Compared with ICC, DCC showed no signi-ficant difference in the phototherapy for hyperbilirubinemia (RR=2.46, 95%CI: 0.93, 6.52), the hemoglobin levels within 6 months (MD=0.29, 95%CI:-0.17, 0.75), and the incidence of anemia (RR=0.71, 95%CI:0.45, 1.12). Conclusions DCC can improve the hemoglobin level in term infants after birth. However, the appropriate time of cord clamping has not been deter-mined. It is necessary to undertake further studies with higher quality and larger scale to evaluate the optimal time of cord clam-ping.

Objective To evaluate the effects of delayed cord clamping (DCC) on preterm infants with gestational age ＜32 weeks.Methods Literatures from January 1,1990 to April 30,2013 in Cochrane library,PubMed,EMBASE,China Academic Journal Network Publishing Database,Wanfang Medical Database and VIP Database were searched.Randomized controlled trials (RCT) of DCC in preterm infants with gestational age ＜32 weeks were screened and evaluated.DCC was defined as cord clamping in 30-90 s after delivery,and early cord clamping (ECC) (＜30 s) was as the control.Rev Man 5.1.0 was used for statistical analysis.Mean difference (MD) and 95％CI were used for continuous data while OR and 95％CI were for categorical data.Results Nine studies (11 articles) involving 373 infants were included.Compared with ECC,DCC improved hematocrit (MD=4.19,95％CI:2.97-5.40,Z=6.74,P＜0.000 01),blood volume (MD=11.70,95％CI:6.02-17.38,Z=4.04,P＜0.0001) and mean arterial pressure of preterm infants with gestational age ＜32 weeks (MD=3.11,95 ％CI:1.30-4.92,Z=3.37,P=0.0008),decreased the usage of volume expansion for hypotension (OR=0.32,95％CI:0.11-0.98,Z=2.05,P=0.04) and the incidence of necrotizing enterocolitis (OR=0.48,95％CI:0.25-0.92,Z=2.22,P=0.03).Meanwhile,DCC had no influence on the peak bilirubin concentration,the incidence of sepsis,patent ductus arteriosus,retinopathy and intracranial hemorrhage,also no influence on neonatal mortality on dcscharge,mental developmental index and psychomotor developmental index at seven-month old.Conclusions DCC might be a safe procedure to improve prognosis of preterm infants less than 32 weeks' gestational age.However,due to small sample size and lack of data on follow up,it is necessary to launch clinical trials with higher quality and larger scale to further evaluate the effect and safety of DCC.

Objective To investigate the effect of alpha 1-antitrypsin (A1AT) concerning in reducing the injury of transplanted islets by pancreas exocrine cells and promoting proliferation of the pancreas B cells.Method The pancreases of mice were digested with collagenase,islets were isolated artificially,and pancreatic exocrine cells were collected.In purified islet group (n =6),100 islets were seeded into a 6 well culture plate.In experimental group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells,and 0.5 mg/mL A1AT was added into a 6-well culture plate.In control group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells.After 48 h,insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured.The islets were cultured in low sugar and high sugar 1640 medium,then glucose stimulated insulin secretion (GSIS) test was carried out.In vivo,8-9-week old male BALB/C mice were induced with STZ (190 mg/kg body weight,i.p) to establish the diabetic model and randomly divided into two groups.In experimental(n =10) and control(n =10) groups,250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule,resepctively.The experimental group was injected with A1AT (83 mg/kg,qd,i.p) for 28 days after operation,and the control group was injected with the same amount of normal saline (qd,i.p) for 28 days.Both two groups were given EDU (5 μg/g,qd,i.p) for 28 days.The blood glucose level was monitored continually.Nephrectomies were performed after 28 days.The expression of anti-amylase antibodies in the renal subcapsule was detected by immunohistochemical staining,and the proliferation of islet beta cells was examined using immunofluorescence staining.Result Insulin levels and insulin stimulation index in the control group were decreased as compared with those in the purified islet group; those in the experimental group were higher than in the control group,but lower than in the purified islet group.Trypsin concentration in the control group was increased as compared with the purified islet group,that in the experimental group was lower than the control group,but higher than in the purified islet group (all P＜0.01).After islets transplantation,the blood glucose levels in control and experimental groups were normal,but those in the control group recovered later than in the experimental group (P＜0.01).At 3rd day after nephrectomy,the blood glucose levels were ＞21 mmol/L in both two groups.A large number of anti-amylase antibody-positive cells were found in the renal subcapsule in the control group while little seen in the experimental group after 28 days.The immunofluorescence showed that the insulin +/EDU + B cells in the experimental group were more than those in the control group.Conclusion Conclusion Co-culture of islets and pancreatic exocrine cells with A1AT can prevent islet cells from damage caused by trypsin.A1AT could inhibit the secretion of pancreatic amylase from pancreatic acinar cells and promote proliferation of islet beta cells.

Objective To determine the effects of feeding donor human milk versus formula milk on very low birth weight infants (VLBWI) and extremely low birth weight infants (ELBWI).Methods The Cochrane library,PubMed,EMBASE,Wanfang,CNKI and VIP database were searched for the randomized controlled trials (RCT) that compare donor human milk with formula milk in VLBWI and ELBWI from the establishment of database up to February 2014.The quality of the included studies was assessed.Meta analysis was performed using RevMan 5.2.9 software.The results were expressed by mean difference (MD) and 95％CI for continuous variables,RR and 95％CI for categorical variables.Results Only five trials were included:in quality evaluation,two trials were graded B,and the other three were graded C.Meta-analysis showed that,compared with the formula milk,feeding of donor human milk could reduce the risk ofnecrotizing enterocolitis (RR=0.36,95％CI:0.18-0.73,P＜0.01),but not the risks of sepsis (RR=0.92,95％CI:0.50 1.72,P=0.80),retinopathy of prematurity (RR=1.21,95％CI:0.84-1.74,P=0.31) and in-hospital mortality (RR=0.66,95％CI:0.18-2.37,P=0.52).The significantly lower rates in weight gains in neonatal period (MD=-6.58,95％CI:-11.19 to-1.98,P＜0.01) and body length (MD=-0.30,95％CI:-0.41 to-0.20,P＜0.01)were found in donor human milk compared with formula milk.No significant difference in head circumference (MD=-0.16,95％CI:-0.33 to 0.01,P=0.13) was seen in comparison of donor human milk with formula milk.Conclusions Feeding with donor human milk can reduce the risk ofnecrotizing enterocolitis in VLBWI and ELBWI,but its effects on neonatal growth need to be further studied in large scale RCT.

Objective Toinvestigatetheclinicalsignificanceofplasmamatrixmetalloproteinase9 (MMP-9)intheformationofdelayedcerebraledemaafterintracerebralhemorrhage.Methods The clinical data of 107 patients with spontaneous intracerebral hemorrhage treated with conservative medical treatment were analyzed retrospectively. According to the clinical features and imaging changes,they were divided into either a delayed cerebral edema group (case group n=39)or a non-delayed cerebral edema group (control group n =68 ). The plasma MMP-9 level was detected with enzyme-linked immunosorbent assay within 24 h after onset. The patients performed head CT scan again at day 7 and 14 after admission. The changes of hematoma and edema volume were detected. All the possible factors associated with the formation of delayed cerebral edema were firstly analyzed by the univariate analysis. Univariate analysis showed that the variables with significant differences were enrolled into multiple logistic regression analysis. Results TheplasmaMMP-9levelofthedelayedbrainedemagroupwassignificantlyhigherthanthatof the control group,they were 189 ± 51 and 118 ± 27 mg/L respectively (P<0. 01). The result of univariate analysis showed that age,history of smoking,blood glucose level,baseline hematoma volume,and National Institute of Health stroke scale (NIHSS )score on admission might be associated with the formation of delayed cerebral edema after intracerebral hemorrhage. Logistic regression analysis showed that MMP-9 level (OR,9. 745,95%CI 6. 754-15. 466,P<0. 01),baseline hematoma volume (OR,2. 411,95%CI 1. 190-2. 728,P =0. 018),blood glucose level on admission (OR,1. 327,95%CI 1. 133 -1. 850,P =0.004),and NIHSS score (OR,1. 867,95%CI 1. 272-2. 364,P=0. 020)were the independent risk factorsfortheformationofdelayedcerebraledemaafterintracerebralhemorrhage.Conclusion Theamount of bleeding,NIHSS score,and hyperglycemia are the risk factors for the formation of delayed cerebral edema in patients with spontaneous intracerebral hemorrhage,while high MMP-9 level on admission indicated that the risk of the formation of delayed cerebral edema is high.