AIM: To study the effects of rhIFN-? and r hI L-2 on human umbilical vein endothelial cell (HUVEC) proliferation, migration, cell cycle and vascular endothelial growth factor (VEGF). METHODS: Cultured HUVECs were used as model to determine the cel l proliferation with MTT method. Cell cycle was analyzed by cytometry. The cell migration was investigated by agarose scraping method and vascular endothelial g rowth factor (VEGF) content in supernatant of cultured HUVEC was determined by e nzyme-linked immunosorbent assay (ELISA). RESULTS: The growth and migrating number of endothelial cells in rhIFN-? group were 0.199?0.009 and 75.750?23.330, in rhIL-2 group was 0 .217?0.005 and 49.250?8.140, and in combined group was 0.183?0.080 and 40.500?17.230, respectively. In comparison with control group (0.248?0.005 and 160.500?13.220), the effects showed more significant (all P0.05). CONCLUSIONS: rhIFN-? and rhIL-2 show inhibitory effect on vasc ular endothelial cell proliferation, migration and DNA synthesis. When used in c ombination, synergistic effect of rhIFN-? and rhIL-2 is observed, suggesting th at these cytokines play an important role in angiogenesis diseases.

Objective To study whether transient overexpression of tumor suppressor gene PTEN could lead to growth suppression and up-regulate the sensitivity to doxorubicin of human breast cancer MCF-7 cells. Methods The eukaryotic expression plasmid pEGFP-C 1-PTEN containing whole cDNA of PTEN was constructed and transfected into MCF-7 cells by Lipofectamine 2000 in vitro. Growth inhibition of the cells was observed by phase contrast microscope and flow cytometry. The clonogenic cell survival ability was studied by clony forming assay. MCF-7 cells′ chemosensitivity to adriamycin was studied with MTT assay. Results PTEN overexpression led to morphological changes characteristic of apoptosis of MCF-7 cells. PTEN overexpression also resulted in a significant increase in G 0/G 1 cell population (14.79%) and apoptosis (10.60%) detected by flow cytometry. The clonogenic survival rate of cells transfected with PTEN was significantly decreased after doxorubicin treatment compared with control. The transfected cells were more sensitive to doxorubicin compared with the control cells ( ? 2=8.59 , P

Objective:To observe the expression of IL-12 family subunit genes by real-time quantitative PCR in mice C6 glioma cells,construct the basis of the brain glioma research on IL-12 family in the future.Methods:Mice C6 glioma RNA was abstracted and reversed transcription cDNA.The mice C6 glioma cells mRNA expression influence of IL-12 family subunit genes was compared and analyzed by real-time quantitative PCR.Results: In mice C6 glioma cells, high expression abundances in IL-23a, IL-12a, midlde expression abundances in EBI3, IL-27, low expression abundance in IL-12b.Conclusion: IL-12 families are closely related to the occurrence and development of glioma,IL-12,IL-23 are regarded as the most potential anti-glioma cytokines among them,research de-velopments will bring a new way of brain glioma immune therapy.

Objective To explore the effects of atorvastatin and valsartan on high glucose-induced human umbilical vein endothelial cells (HUVECs) injury.Methods Cultured HUVECs were divided and assigned to 9 groups:normal control group,mannitol control group,high glucose group,low dose atorvastatin group (0.1 μ mol/L),medium dose atorvastatin group (1 μmol/L),high dose atorvastatin group (10 μ mol/L),low dose valsartan group (0.1 μmol/L),medium dose valsartan group (1 μmol/L),and high dose valsartan group (10 μmol/L).H UVECs were pretreated with or without 30 mmol/L glucose plus various concentrations of atorvastatin and valsartan (0.1,1,10 μmol/L) for 16 hours and then incubated with 5.5 mmol/L glucose for 6 days.The levels of vascular cell adhesion molecule 1 (VCAM-1),monocyte chemotactic protein 1 (MCP-1),and plasminogen activator inhibitor 1 (PAI-1) in the culture supernatant were measured by enzyme-linked immunosorbent assay.Results Compared with normal glucose group,hyperglycemia memory increased the levels of VCAM-1,MCP-1,and PAI-1 (all P＜0.05),which were still maintained at high levels even after withdrawal of high glucose.Atorvastatin and valsartan treatment decreased the levels of VCAM-1,MCP-1,and PAI-1 (all P＜0.05).Conclusion Atorvastatin and valsartan may lower the secretion of VCAM-1,MCP-1,and PAI-1,and prevent high glucose memory-induced injury to endothelial cell.

[Summary] A total of 352 pregnant women were selected in Affiliated Hospital of Qingdao University. Serum levels of TSH and FT4 were determined and pregnancy outcome were observed in all subjects. According to the standard of American Thyroid Association(ATA) published in 2011 and the Chinese Guideline of Gestation Thyroid Disease published in 2012, the subjects were grouped into control(0. 1≤TSH≤2. 5 mIU/ L), observation(2. 55. 17 mIU/ L)during first-trimester(T1)and control(0. 2≤TSH≤3. 0 mIU/ L), observation(3. 05. 22 mIU/ L) during second-trimester(T2). The results showed that the rupture of membranes, preterm labor, and total obstetrical adverse events in the observation group were significantly higher than those in control group during T1(all P<0. 05), no statistical significance between control group and treatment group. The rupture of membrances in the observation group was significantly higher compared with control group during T2 ( P < 0. 05), no significant difference in other adverse pregnancy outcomes between these two groups. Compared with control group during T1, the proportion of adverse pregnancy outcomes in observation group during T1 was significantly increased(P<0. 05). The elevated TSH is one of the risk factors for the increased incidence of adverse pregnancy outcomes at the first half of pregnancy, especially during T1. L-T4 treatment reduces the incidence of adverse pregnancy outcomes.

Objective To evaluate the effect of PUN282987 on global cerebral ischemia-reperfusion (I/R) injury in aged rats.Methods One hundred and twenty pathogen-free healthy male SpragueDawley rats,aged 18-22 months,weighing 450-600 g,were divided into 3 groups (n=40 each) using a random number table:sham operation group (group S),global cerebral I/R group (group I/R) and α7 nicotinic acetylcholine receptor (α7nAChR) agonist PNU282987 group (group PUN).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 0.4 ml/100g,and global cerebral I/R was produced by 4-vessel occlusion technique in I/R and PUN groups.PUN282987 2.4 mg/kg was injected intraperitoneally before ischemia in group PUN.At 1,5,12 and 24 h of reperfusion,10 rats were randomly selected in each group and then sacrificed,and the brains were removed for detection of the neuronal apoptosis and for determination of the expression of α7nAChR,choline acetyltransferase (ChAT),tumor necrosis factor-α (TNF-α) and intedeukin-1β (IL-1β) in the hippocampal CA1 region.Apoptosis rate was calculated.Results Compared with group S,the apoptosis rate was significantly increased,and the expression of α7nAChR,ChAT,TNF-α and IL-1β was up-regulated at each time point in I/R and PUN groups (P＜0.05).Compared with group I/R,the apoptosis rate was significantly decreased,the expression of α7nAChR and ChAT was up-regulated,and the expression of TNF-α and IL-1β was down-regulated at each time point in group PUN (P＜0.05).Conclusion PUN282987 can reduce global cerebral I/R injury in aged rats.

Objective To establish an early cognitive disorder model in rats and investigate the early cognitive functioning after ischemic hypoxic brain injury during the neonatal period. Methods Forty-six newborn Sprague-Dawley rats were randomized into a 21-d-old group and a 31-d-old group. These 2 groups were then subdivided into model and sham-operated subgroups (M21, n=12; SH21, n=11; M31, n=12; SH31, n=11). A model of neonatal early cognitive disorder was established in the rats of the M21 and M31 groups using a modification of Rice's method. Rats in the SH21 and SH31 groups received skin incisions and common carotid artery separation without ligation or hypoxia. Each group was tested with a Morris water maze. The rats were sacrificed after testing, and brain tissue was examined under the electron microscope. Nissl staining allowed Nissl body quantification and neurocyte acin the M21 group was significantly longer than in the SH21 group. The 31-d-old subgroups had shorter average escaping latencies than the corresponding 21-d-old subgroups. (b) Spatial memory: The average platform times, Ⅰ region times and Ⅰ region distances showed no significant differences among groups. ②Brain pathology (a) Gross appearance: Obvious ischemic hemisphere atrophy was observed in the M group, and no abnormality was observed in the SH group. (b) Electron microscopic observation: In the SH group cell ultrastructures in the ischemic hippocampus were normal. Karyopyknosis and dilated endoplasmic reticulums were found in the M group. More mitochondria were found in the presynaptic membranes of the ischemic hippocampus in the M group than that in the SH group. (c) Nissl body quantification and neurocyte activity analysis: Significantly less activity in the ischemic cortex was found in the M21 group compared to the SH21 group. More activity was observed in the 31-d-old subgroups than in the corresponding 21-d-old subgroups. Conclusions ①The neonatal rats with ischemic hypoxic brain injury had prolonged average escaping latency and depressed neuronal activity. ②The 31-d-old rats had better spatial localization learning ability than the 21-d-old rats.

Objective To evaluate the effects of hepatitis B virus on liver function,liver fibrosis,and liver pathological staging at different immune stages.Methods We made a retrospective analysis of 657 patients with chronic hepatitis B diagnosed in the First Hospital of Lanzhou University.Their liver function parameters,liver fibrosis parameters,and hepatitis B virus load were measured by automatic biochemical analyzer,automatic gammaradiation immunity analyzer,and quantitative PCR analyzer,respectively.Effects of hepatitis B virus on liver function,liver fibrosis in different immune stages were analyzed by variance analysis.Effects of hepatitis B virus on liver pathological staging at different immune stages were analyzed by linear trend chi square test analysis.Results In ALT normal chronic hepatitis B patients group,viral load had mild effects on liver function and liver fibrosis parameters.However,in ALT abnormal chronic hepatitis B patients group,viral load had a significant effect on liver function and liver fibrosis parameters,and the effect was most obvious in ALT＞double upper limit of normal group.The specific manifestation was that with viral load increasing,liver function parameters including ALT,AST,TBiL,DBiL,and IBiL increased,while TP and ALB decreased.Liver fibrosis parameters HA,LN,PcⅢ,and CIV all increased (P＜0.05).In ALT normal chronic hepatitis B patients group,viral load had no relationship with liver pathological staging.However,in ALT abnormal chronic hepatitis B patients group,especially ALT≥double upper limit of normal group,viral load was significantly related to liver pathological staging.Conclusion The effects of hepatitis B virus on patients' liver function at different immune stages were different,thus providing evidence-based medicine support for clinical antiviral treatment.

Objective To explore the value of comprehensive biochemical and serological test in Salmonella clinical test .Methods A total of 95 cases of suspected salmonella infection with enteric fever were selected as subjects in this study from April 2013 to A‐pril 2014 ,and 48 cases randomly divided in research group ,47 cases in control group .Regular inspection was conducted in the con‐trol group ,comprehensive biochemical and serological test were conducted in the control group .Compared the results in the two groups .Results The salmonella detection rate in the research group was 93 .75% ,which was significant higher than 68 .09% in the control group (P< 0 .05) .Conclusion Combined detection of comprehensive biochemical and serology test could detect Salmonella effectively ,it is worthy of application and popularization in clinic .

Objective To evaluate the changes in cholinergic anti-inflammatory pathway in hippocampi global in aged rats with cerebral ischemia/reperfusion (I/R ) injury .Methods One hundred and twenty male Sprague-Dawley rats , aged 18-22 months ,weighing 450-600 g ,were randomly divided into 2 groups ( n= 60 each):sham operation group (group S) and global cerebral I/R group (group I/R) .The animals were anesthetized with intraperitoneal 10% chloral hydrate 0.4 ml/100 g .Global cerebral I/R was induced by 4-vessel occlusion method described by Pulsinelli .Fifteen rats were sacrificed at 1 ,3 ,5 and 7 days of reperfusion ,and brains were removed for determination of neuronal apoptosis and expression of α7 nicotinic acetylcholine receptor (α7nAChR ) , choline acetyltransferase (ChAT ) ,tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) in the hippocampal CA1 region .The apoptosis rate was calculated .Results Compared with group S ,the apoptosis rate was increased and the expression of α7nAChR ,ChAT ,TNF-αand IL-1βwas up-regulated in group I/R ( P<0.05 or 0.01 ) . The expression of α7nAChR and ChAT was up-regulated gradually during reperfusion and peaked at 5 day of reperfusion ( P< 0.05 ) .Conclusion Global cerebral I/R injury can activate cholinergic anti-inflammatory pathway in aged rat hippocampi ,and the activation of this pathway is the endogenous mechanism of inhibition of excessive inflammatory responses in brain tissues .