Aim To investigate the effects of bradykinin on the expression of IL-1? in C6 glioma cells and provide an experimental basis for the mechanism of bradykinin opening the blood tumor barrier.Methods Semi-quantitive RT-PCR and radioimmunoassay were used to detect the expression of IL-1?mRNA in C6 glioma cells and the content of IL-1? in the culture media treated with bradykinin respectively.Results RT-PCR revealed that the expression of IL-1?mRNA was significantly elevated with bradykinin treated for 15min(P

Aim To investigate the effects of PB-19 on action potential (AP) of ventricular cells in vitro and ischemia and reperfusion induced arrhythmias in vivo in rats. Methods ① We established the model of isolated rat right ventricular wall and perfused the models with normal Tyrode’s solution and PB-19.Microelectrode was inserted into the cells to record the AP. ② We ligated the left anterior descending coronary artery of rats to study the effects of PB-19 on ischemia and reperfusion induced arrhythmias. 50 rats were divided into 5 groups randomly and were given iv. injection with normal saline and PB-19 of different concentrations respectively. We recorded Ⅱ ECG of the rats. Results ①In vitro models, the APD_ 50、APD_ 90 of PB-19 group were significantly longer than that of control group respectively (P

Aim To investigate the effects of bradykinin preconditioning on the damage produced by focal cerebral ischemia-reperfusion in rats.Methods Rats were scheduled to undergo middle cerebral artery ischemia-reperfusion by intraluminal suture.Prior to ischemia,bradykinin was pumped into the brain via extra carotid artery and control group was given the same amount of normal saline.The infarct volume、brain water content、permeability of blood brain barrier and histological neuronal changes were evaluated after 2 h ischemia and 24 h reperfusion.Results Compared with other groups, bradykinin preconditioning 15min before ischemia reduced infarct volume、brain edema、permeability of blood brain barrier and histological neuronal damage.Conclusion Bradykinin preconditioning may provide protective effects against cerebral ischemic injury.This protection may be due to the protection of cerebral vasculature and the decrease of infarct volume.

Aim To study the activation of transcription factor cAMP response element binding protein(CREB)in the rat C6 glioma cells induced by bradykinin,and discuss its possible significance.Methods Immunocytochemistry and western-blot method were used to determine the phosphorylation of CREB after stimulated by 1 ?mol? L-1 bradykinin at various time points(0,5,10,15,20,30min).Results Bradykinin at 1 ?mol? L-1 induced phosphorylation of CREB inthe glioma cells at the indicated time points(0～30min)(P

Objective To investigate the clinical efficacy of ambroxol intravenous and budesonide atomizing inhalation combined with cefodizime on infantile pneumonia and its effects of serum PCT, IL -6 levels and related immune factors.Methods 95 children with pneumonia from May 2013 to October 2015 in Sanya Hospital of Traditional Chinese Medicine were collected and randomly divided into control group(n=47) and experiment group (n=48), two groups were treated by clinical routine treatment, such as antipyretic, anti-inflammatory, strengthen nutrition in children, and control group were added with ambroxol, iv, qd; experiment group were added cefodizime on the basis of control group, the course was one week.Clinical efficacy,serum PCT, IL -6 levels, immune factors and adverse reactions were observed and compared.Results The serum PCT and IL -6 levels of experiment group were lower than control group post-treatment, and CD4 +, CD4 +/CD8 +levels were higher than control group, CD8 +level was lower than control group, the differences were all significant (P<0.05).The effective rate of experiment group was 93.75%, higher than 80.85% of control group(P<0.05).Incidence of adverse reactions between two groups had no statistical difference.Conclusion Ambroxol intravenous and budesonide atomizing inhalation combine with cefodizime in treatment of infantile pneumonia has better clinical efficacy, could effectively reduce the serum PCT and IL-6 levels, effectively improve the clinical symptoms.

Objective To explore the role of xCT in the metastasis and drug resistance of nasopharyngeal carcinoma (NPC),to provide new evidences for the mechanism of NPC metastasis and drug resistance,and to enhance effects of chemotherapy.Methods Fluorescence quantitative polymerase chain reaction (PCR) and Western blot were used to detect the xCT expression of 5-8F and 6-10B.The xCT eukaryotic expression vector was constructed to transient transfect 6-10B cell lines.The 5-8F cell lines were treated with xCT antisense oligodeoxynucleotide (ASO).Scratching assay and transwell method were used to detect the cell metastasis and invasion abilities.The expressions of matrix metalloproteinase 1 (MMP1) of all the cell lines were surveyed by Western blot.Results The results of fluorescence quantitative PCR and Western blot showed that the expression of xCT in 5-8F was higher than 6-10B in the levels of mRNA and protein.Exogenous overexpression of xCT enhanced metastasis and invasion abilities of 6-10B cells.Silencing and inhibition of xCT could decrease metastasis and invasion abilities of 5-8F cells.The expressions of MMP1 protein in 5-8F were higher than 6-10B,and they were positively correlated with the expression of xCT.Conclusions xCT is closely related to the metastasis and invasion abilities of nasopharyngeal carcinoma.xCT could enhance the metastasis and invasion abilities of NPC cells.xCT might mediate proliferation and matastasis/invasion of NPC cells through MMP1.

Insulin resistance in insulin sensitive organ results in metabolic disorder such as hyperglycemia, hyperinsulinemia and hyper triglyceridemia which are common features of type 2 diabetes.Insulin resistance in liver cells mainly causes impaired glycogen synthesis, failed to suppress glucose production which is the major contribution to hyperglycemia.FGF-21 as a new metabolic regulator can control fasting blood glucose.The mechanism of FGF-21 effects on regulating plasma glucose has little to known.In order to establish an in vitro insulin resistant model of liver cells and evaluate the effects and mechanism of FGF-21 on glucose metabolism in the cell model, HepG2 cells were incubated with 10-7 mol/L insulin for 24 h to build insulin-resistant cell model.To evaluate the cells for insulin resistance, the cells were stimulated with fresh insulin for 24 h and the glucose uptake by these cells was carried out.The insulin-resistant cells were treated with different concentrations of FGF-21 for 24 h and insulin-treated cells were used as a control.The glucose uptake by the cells was detected by the method of glucose oxidizes/peroxides(GOD-POD);the synergy between insulin and FGF-21 was evaluated.The mRNA expression of GLUT1 in the insulin-resistant cells was detected by the real-time PCR.Glycogen synthesis of the cells was examined by the anthrone method.The results showed that HepG2 cells treated with 10-7 mol/L insulin for 24 h became resistant to insulin and the insulin resistance status was maintained for 48 h without change of cell morphology.FGF-21 could stimulate glucose consumption of the insulin-resistant model in a dose-dependent manner.The glucose consumption and glycogen synthesis of the insulin-resistant model were significantly improved by FGF-21 treatment.FGF-21 showed strong synergy with insulin in glucose uptake and glycogen synthesis of the model cells.While the cells became resistant to insulin, FGF-21 could increase the mRNA expression of GLUT1.Thus, It is concluded that FGF-21 stimulates glucose uptake in insulin resistant HepG2 cells through GLUT1 expression, stimulates glycogen synthesis and improves the glucose metabolism in the insulin resistant liver cell model.

Aim To explore the effect of shikonin on stemness maintance of glioma stem cells ( GSCs ). Methods After the U87-MG cells were cultured and isolated, the sphere cells were identified by immuno-fluorescent staining. The alteration of stemness of GSCs by shikonin treatment(2 μmol·L - 1 ) for 12 h, 24 h and 48 h was valued by morphological detection using optical microscope and sub-sphere forming assay. Mo-reover, the related markers of stem cells, such as CD133, were detected in shikonin treated GSCs by western blot assay. Protein expression of PI3K, p-PI3K, Akt and p-Akt was detected by western blot af-ter shikonin treatment alone. Furthermore, by combi-nation with insulin-like growth factor-1 ( IGF-1), we observed the alteration of stemness maintance of shiko-nin-treated GSCs. Results The presence of neural stem cell related markers CD133 and nestin proved the characteristics of GSCs. Shikonin treatment significant-ly inhibited the morphology of GSCs and the sub-sphere forming. Besides, the reduced expression of CD133 was detected in shikonin treated GSCs. Though, the expression of PI3K and Akt did not change compared with the control group, the expression of p-PI3K and p-Akt was reduced. Furthermore, the combination of IGF-1 markedly attenuated the inhibitory effect of shikonin on stemness maintance of GSCs. Conclusion The stemness maintance of GSCs can be significantly inhibited by shikonin treatment, in which PI3K/ Akt pathway is involved.

Aim To investigate the signaling mecha-nisms in endothelial monocyte-activating polypeptide-Ⅱ( EMAP-Ⅱ)-induced increase in blood-tumor barri-er ( BTB ) permeability. Methods Relatively pure cerebral microvessel fragments were obtained from the cortex of 3-5 days old Wistar rats by using careful dis-section, enzyme digestion, and dextran centrifugation. Then, these fragments were seeded on dishes and cul-tured primarily. In vitro BTB models were constructed by co-cultivation of rat brain microvascular endothelial cells ( BMECs) with C6 glioma cells. Confluent mono-layers of co-cultured BMECs were divided randomly in-to 5 groups ( each n=6 ): control, EMAP-Ⅱ, H7 +EMAP-Ⅱ, C3 exoenzyme + EMAP-Ⅱ, and C3 ex-oenzyme + H7 + EMAP-Ⅱ groups. Transendothelial electric resistance values and horseradish peroxidase flux were measured to evaluate changes in the BTB permeability . The expression levels of tight junction-re-lated protein occludin and ZO-1 in BMECs were meas-ured by Western blot. Immunofluorescence was used to identify the expression and distribution of occludin and ZO-1 in BMECs. Also, Western blot were used to de-tect the expression levels of myosin light chain ( MLC) and phosphomyosin light chain ( pMLC ) in BMECs. Results Compared with control group, the BTB per-meability of EMAP-Ⅱ group was increased significant-ly. The expression levels of occludin and ZO-1 in BMECs were significantly decreased, accompanied with marked increase in the expression level of pMLC. These above-mentioned effects of EMAP-Ⅱ were sig-nificantly inhibited by pretreatment with H7 ( an inhib-itor of PKC ) or/and C3 exoenzyme ( an inhibitor of RhoA ) . Conclusion Signaling molecules PKC and RhoA play important roles in EMAP-Ⅱ-induced in-crease in BTB permeability; signaling pathways PKC-pMLC and RhoA-pMLC are involved in this process.

Objective: To design the proficiency testing (PT) project (No. NIFDC-PT-183) for assays of bismuth potassium citrate capsules and organize to assess the proficiency of complexometric titration in laboratories, and provide some technical analyses and advices.
Methods: Two groups of samples with different concentration were prepared. The uniformity was evaluated with one-way analysis of variance and the stability was confirmed with t-test, whose results all conformed the requirements. The samples with three combinations were randomly distributed to 279 laboratories. The determination was performed according to the assays of bismuth potassium citrate capsules in Volume Ⅱ of the Chinese Pharmacopoeia 2015. The median value and normalized interquartile range (NIQR) of robust statistical analysis was adopted and Z-scores were used to evaluate the results from each of laboratories.
Results: Among 279 laboratories, 240 laboratories results were satisfactory, 23 were questionable, and the other 16 were unsatisfied. The satisfaction rate was 86.0%.
Conclusion: The overall capacity of national laboratories for assays of bismuth potassium citrate capsules is good while a portion of participants require further improvement.