非经典信号通路IKKε和TBK1与恶性肿瘤密切相关，多种因素激活IKKε和TBK1通路，可引起NF-κB途径的激活， 导致肿瘤细胞的凋亡减少、细胞周期加快，促进肿瘤发生和发展。抑制IKKε和TBK1信号通路，可增加多种细胞凋亡因子的表 达，抑制肿瘤细胞增殖，促进肿瘤细胞凋亡，同时提高化疗和放疗的敏感性。因此，阻断IKKε和TBK1信号通路可有效治疗恶性 肿瘤，已有的实验证实有多种阻断IKKε和TBK1通路的药物均具有良好的抗肿瘤作用。

Objective To investigate effects and its mechanisms of hypertonic saline hydroxyethyl starch 200/0.5 solution on intracranial pressure and brain water content in rats with ischemic cerebral edema.Methods All experiments were conducted in the animal experimental center of Sun Yat-sen University.The 28 male Sprague-Dawle (SD) rats were randomly (random number) divided into hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group,control group and sham operation group,each n =7.Ischemic cerebral edema model was reproduced by middle cerebral artery occlusion (MCAO),followed by reperfusion after ischemia for 2 hours (If the moldel was not successful,other rats were operated to fill the missing models).Then reperfusion after ischemia 2 hours and received hypertonic saline hydroxyethyl starch and hydroxyethyl starch via tail vein at the beginning of reperfusion.The colloidal osmotic pressure (COP) and intracranial pressure (ICP) were evaluated on 0,2,6,12,18,24 hours after the surgery.The water content of the right hemisphere was measured on 24 h after the surgery.Results The ICP of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 2,6,12,18,24 h after the surgery.The ICP of hypertonic saline hydroxyethyl starch group was significantly lower than those of hydroxyethyl starch group and control group on 2,6,12,18 and 24 h.But there was no significant difference in ICP of the hydroxyethyl starch group compared with that of control group at all time points.The COP of hypertonic saline hydroxyethyl starch group and hydroxyethyl starch group were significantly higher than the control group and sham operation group at each time point; There was no significant difference in COP (mmHg) of the hydroxyethyl starch group compared with that of hypertonic saline hydroxyethyl starch group at all time points.The brain water content (BWC) of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 24 hours after the surgery [(81.24±0.36)％,(83.04±0.10)％,(83.14±0.41)％ vs.(78.37±0.37)％,all P=0.000],BWC of hypertonic saline hydroxyethyl starch group lower than these of hydroxyethyl starch group [(81.24±0.36)％ vs.(83.04 ±0.10) ％,P =0.000] and control group [(81.24 ±0.36)％ vs.(83.14 ±0.41) ％,P =0.000].There was no significant difference in BWC of the hydroxyethyl starch group compared with that of control group [(83.04 ± 0.10) ％ vs.(83.14 ± 0.41) ％,P =0.578].Conclusion Hypertonic saline hydroxyethyl starch solution could significantly ameliorate ischemic cerebral edema and reduce ICP,but the relationship between its elevated COP and reduced ICP has not been confirmed.

Objective To explore the effect of erlotinib in patients with non-small-cell lung cancer (NSCLC)in stage ⅢA before operation and the relation with the rate of resection,good operability and postoperative complications.Methods 31 patients with NSCLC in stage ⅢA in group A were treated with erlotinib before operation;34 patients with NSCLC in stage ⅢA in group B only were treated with operation.Results The condition of 64.5%patients were improved.The operability in group A was more than that in group B(P=0.008),the good operability in group A was more than that in group B(P=0.011),the postoperative complications do not have statistical significance (P=0.07).Conclusion The erlotinib can increase the rate of resection in patients with NSCLC in stage ⅢA,and increase good operability in patients with NSCLC in stage ⅢA,but not increase the postoperative complications.

Objective To compare a prospective ECG-gated high-pitch spiral technique (Flash) and conventional retrospective ECG-gated spiral technique for the image quality of coronary artery stent and radiation dose with a dual source CT.Methods One hundred and fifty five coronary stents in one hundred and twenty patients (mean age 64.9 ± 10.6 years,heart rates≤65 bpm) were examined using a dual source CT.All patients were divided in two groups,receiving either Flash or conventional coronary artery CT angiography separately.After images of coronary artery were reconstructed using both the smooth (B26) and sharp (B46) kernel,the coronary stent image quality and stent lumen were scored by two observers individually using four point scale (1 = excellent,4 = unvaluable) .The effective radiation dose of volume CT dose index (CTDIvol,mGy) and dose length product (DLP,mGy x cm) were also calculated for each patient.x2-test analysis of image quality and t-test analysis of radiation dose were used respectively for statistical difference between two groups.Results Interobserver agreement for stent image quality was good (Kappa =0.764,P＜0.001).The mean scores were 1.61 ±0.77 and 1.65 ±0.82 in Flash group and conventional group respectively.There was no significant difference in image quality between the two groups (x2 = 0.865,P = 0.834).The effective radiation dose in Flash group was significantly lower than that in conventional group.The mean values of CTDIvol were 3.24 ± 1.21 in Flash group and 31.26 ± 10.79 in conventional group (t = 19.83,P < 0.001) ,and the mean values of DLP in Flash group and conventional group were 54.61 ±19.88 and 468.30 ± 174.88,respectively (t = 18.06,P < 0.001).Conclusions Compared with the conventiaonal coronary artery CT angiography,the Flash coronary artery CT angiography technique has a similar coronary stent image quality,but at a lower radiation dose in patients with heart rates lower than 65 beats per minute.

Objective To investigate the characteristic changes in cerebral infarction and brain edema. Method A total of 122 Healthy adult male Spraque-Dawley rats were randomly (random number) divided into three groups: normal group ( n = 12), sham operated group (n=12) and cerebral ischemia group ( n = 98). Cerebral infarction and brain edema were induced by a permanent occlusion of right middle cerebral artery (POM-CA) with ligature. According to the duration of POMCA, the rats of cerebral ischemia group were further divided into seven sub-groups, 2 h, 4 h, 6 h, 12 h, 18 h, 24 h and 30 hours. The hemispheric ratio was detected by staining with 2% 2,3,5-triphenyltetrazolium chloride solution, and brain water content was assayed by dry/wet ratio 2 h, 4 h, 6 h, 12 h, 18 h, 24 h and hours after POMCA. Results There was a focal cerebral infarction in the rats of cerebral ischemia group 4 hours after POMCA. There was no significant difference in hemispheric ratio between 4 hours and 6 hours after POMCA by One-way ANOVA (P = 0.091). Compared with 6 h sub-group, the hemispheric ratio increased significantly in 12 h, 18 h, 24 h and 30 h sub-groups (P < 0.01), and the peak was in the 24 h sub-group. The brain water content began to increase 4 hours after POMCA and aggravated 6 hours later, and reached the peak 24 hours after POMCA. The brain water content of the non-ischemic hemisphere increased 18 h,24 h and 30 hours after POMCA. Furthermore, there was a significant correlation between the hemispheric ratio and brain water content ( r = 0.834, P < 0.01). Conclusions The critical point of cerebral infarction and brain edema aggravated is 6 hours after POMCA. Both brain edema and cerebral infarction reach the most serious degree 24 hours after POMCA. It is an important experimental evidence for evaluating the milieu conducive to the pathogenesis, and choosing the suitable time window for the treatment of cerebral infarction and brain edema.

Objective To determine the likelihood of G-protein coupled receptor 56 (GPR56 ) induces axonal development and myelination in the corpus callosum of mouse brain.Methods A total of 64 Gpr56 +/-and Gpr56 -/-mice were selected and randomly divided into two groups:Gpr56 +/-group (n=32)and Gpr56 -/-group (n=32).According to number of days after birth,each group was further divided into 4 subgroups including P7d,P14d,P21d and P28d subgroups.Levels of neurofilament-200 (NF -200)and proteolipid protein (PLP ) of myelin basic protein in corpus callosum were measured with immunohistochemistry staining and Western blot in P7d、P14d、P21d、P28d Gpr56 +/- and Gpr56 -/-mice.Gpr56 +/-and Gpr56 -/-neurons were cultured using P1 d Gpr56 +/-and Gpr56 -/-mouse brain.The lengths of Gpr56 +/- and Gpr56 -/-neuronal axon were measured and compared with Image J software. Axonal myelination in the corpus callosum of mouse brain in each group was observed under electronic microscopy and the axonal diameters between subgroups were compared.Results The levels of NF-200 and PLP in the corpus callosum in P7d、P14d、P21d、P28d Gpr56 -/-mice decreased significantly compared with Gpr56 +/- mice.The length of Gpr56 -/-neuronal axon was shortened compared with Gpr56 +/-neuronal axon.The number of myelinated axons was obviously reduced in the corpus callosum in P28d Gpr56 -/-mice.The diameter of axon in the corpus callosum of P28d Gpr56 +/-mouse is longer than that of P28d Gpr56 -/-mouse. Conclusions GPR56 may be involved in axonal development and myelination in the corpus callosum of mouse brain.

Objective To study the effect of APETx2 on the expression of ASIC3 APETx2 in a rat model of acute gastric mucosal lesion(AGML). Methods Twenty-four Wistar rats were randomly assigned to three groups in equal number : normal control group, water immersion restraint stress (WIRS) group, APETx2 treatment group. AGML was induced by WIRS for 6 hours, and APETx2 (25 μg/kg) was injected intraperitoneally before the onset of stress. Intragastric pH and gastric histopathological changes were measured and the expression of ASIC3 mRNA in DRG neurons projecting to rat stomach was examined by real-time PCR. Immunohistochemistry was performed to detect the localization of ASIC3. Results Compared with the normal control group, the WIRS group showed obvious gastric injury with lower values of intragastric pH and extensive expression of ASIC3 in the DRG neurons (P < 0.05). The treatment with APETx2 before the onset of WIRS significantly alleviated the gastric mucosal injury, decreased gastric acidity and reduced ASIC3 expression in DRG neurons (P < 0.05). Conclusions ASIC3 expression in DRG neurons projecting to rat stomach is strongly associated with gastric mucosal lesion and acidosis in the WIRS model. APETx2 can improve gastric acidosis and prevent the occurrence of these lesions.

AIM: To observe the effects of Egr-1 gene knockout on the expression of inflammatory-related factors in pancreatic tissue in a mouse acute pancreatitis model.METHODS: The experimental pancreatitis was induced by high-dose of cearulein in wildtype mice and Egr-1 knockout mice.The pancreatitis indexes,such as serum amylase,pancreata edema,and myeloperoxidase(MPO) levels in pancreata and lungs were recorded.The mRNA levels of tissue factor(TF),plasminogen activator inhibitor(PAI-1),monocyte chemoattractant protein(MCP-1),Gro-1,IL-6 and ICAM-1 were measured by quantitative PCR.RESULTS: Contrary to wildtype mice,typical pancreatitis was not induced by high-dose cearulein in the Egr-1 knockout mice,not only markedly reduced edema in pancreata and lungs,but decreased MPO levels in lungs as well were found.Furthermore,the mRNA of TF,PAI,MCAP,ICAM-1 and IL-6 in pancreata were significantly decreased in Egr-1 knockout mice.CONCLUSION: The severity of pancreatitis and lung damage is ameliorated in Egr-1 knockout mice stimulated by high-dosage of cearulein,which was probably mediated by decreasing expression of inflammatory-related factors in pancreata,such as TF,PAI,MCP-1,ICAM-1 and IL-6.

AIM: To construct a recombinant lentivirus RNAi vector carrying cytochrome C oxidase gene to obtain the titer of the lentiviral stock for investigation of the expression in the eukaryotic cell and the affection of the COX gene silencing in the eukaryotic cells. METHODS: According to the DNA of the cytochrome C oxidase gene, we designed and synthesized complementary single-strand DNA oligos, annealed the single-stranded oligos to generate a ds oligo, cloned the ds oligo into pENTR/U6 to obtain an entry clone; An LR recombination reaction was performed between the pENTR/U6 entry construct and pLenti6/BLOCK-iT-Dest to generate expression construct, the 293FT cell line was cotransfected with pLenti6/BLOCK-iT expression construct, and the viral packaging mix, viral supernatant was harvested to determine the titer. RESULTS: The DNA sequence of interest clone to the vector was constructd to generate an entry clone and an expression clone successfully, which were proved by sequence determination. A vector producing cell line 293FT was established, and the titer for transfection was obtained. Western blotting analysis demonstrated that COX shRNA expression construction could suppress the expression of MTCOX-I. CONCLUSION: A lentivirus RNAi vector containing cytochrome C oxidase gene was successfully constructed.

Objective To investigate the clinical value of the ratio of plasma vascular endothelial growth factor level to platelet count (VEGF/PLT) in predicting 28-day prognosis in patients with sepsis.Methods A prospective cohort study was conducted.From September 2009 to March 2013,164 sepsis patients in Intensive Care Unit (ICU) of Guangdong General Hospital were included for study.Patients with age younger than 18 years old,the illness already reaching final stage of chronic diseases,suffering from two or more organs dysfunction within 3 days,acute pancreatitis without infection,or less than 28 days of expected survival time were excluded.Finally,135 patients were included in the further analysis.Peripheral blood samples were collected at admission.Routine blood tests were done,and then VEGF levels in plasma were measured by enzyme linked immunosorbent assay (ELISA).Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores were recorded every day for 7 days.Patients' prognosis was assessed during the following 28 days.The patients were divided into 28-day survival group and non-survival group.Comparison between two groups was done by single factor analysis.Spearman rank correlation was used to analyze the correlation between VEGF levels and PLT.Mutivariate logistic regression analysis was performed to identify the independent risk factor for 28-day prognosis.Receiver operating characteristic curve (ROC curve) was plotted,and the effect of related indexes on predicting 28-day survival was evaluated by area under ROC curve (AUC).Results There were no significant differences in VEGF (ng/L:471.73 ± 198.34 vs.383.49 ± 266.54,t=-1.918,P=0.057),PLT (× 109/L:220.40±127.60 vs.246.42± 100.72,t=1.275,P=0.204),leucocyte counts (× 109/L:12.48 ±4.62 vs.13.70 ±5.97,t=1.063,P=0.292),mean arterial pressure [mmHg (1 mmHg=0.133 kPa):86.50 ± 12.04 vs.91.03 t 13.10,t=1.557,P=0.123] and blood lactic acid (mmol/L:1.79 ± 1.30 vs.1.50 ± 0.60,t=-1.768,P=0.079) at admission between the non-survival group (n=42) and survival group (n=93).VEGF/PLT (2.59 ± 1.44 vs.1.73 ± 1.13,t=-3.756,P=0.000) as well as APACHE Ⅱ scores (15.50 ± 4.50 vs.13.28 ± 4.61,t =-2.022,P=0.045) of the non-survival group were significantly higher than those of survival group,and oxygenation index (PaO2/FiO2) of the non-survival group was significantly lower than that of survival group (kPa:32.38 ± 11.12 vs.37.04 ± 10.97,t=2.278,P=0.024).Correlation analysis showed that the concentration of VEGF was positively correlated with PLT (r=0.271,P=0.001).It was shown by multivariate logistic regression analysis that only VEGF/PLT was the independent risk factor in predicting 28-day prognosis in patients with sepsis [odds ratio (OR) was 1.591,95％ confidence interval (95％CI) 1.164-2.175,P=0.004].AUC of VEGF/PLT was 0.704 ± 0.047 (P=0.000,95％CI:0.611-0.797) for predicting 28-day survival.The optimal cut-off point was 1.32,and the sensitivity and specificity were 81.0％ and 48.4％,respectively.Conclusion VEGF/PLT can be used as one of the indicators to predict 28-day survival in patients with sepsis.