Objective To screen microRNA (miRNA) that inhibit expression of the metastasisrelated gene ezrin in ovarian cancer cells and explore their correlation to the invasion and metastasis of ovarian cancer. Methods The differential expression of ezrin in two paired high-metastatic and lowmetastatic cell lines were examined by real time reverse transcription (RT)-PCR and western blot. A functional screen with microarray was employed to identify miRNA that were differentially expressed between SKOV3 and SKOV3ip cell lines. Three programs, TARGETSCAN ( http://www. targetscan. org ),MICROCOSM ( http ://www. ebi. ac. uk/enright-srv/microcosm/htdocs/targets/v5/) and PICTAR (http://www. pictar. mdc-berlin. de), were employed to identify all miRNA, which may inhibit the expression of ezrin and were differentially expressed between SKOV3 and SKOV3ip cells. To test the repressive potential of these miRNA, synthetic mimetics were transfected individually into SKOV3ip cells and endogenous ezrin expression levels monitored by western blot and real-time RT-PCR. Results ( 1 ) The mRNA average level of ezrin were (81.74 ± 5.34) -fold higher expression level in SKOV3ip versus SKOV3 cells ( P ＜ 0. 01 ), while (2. 61 ±0. 14)-fold in HO-8910PM versus HO-8910 cells (P ＜0. 01 ). Elevated protein level of ezrin were observed in SKOV3ip cells compared with that in SKOV3 cells, and the same that in HO-8910PM cells compared with HO-8910 cells. Paired SKOV3 and SKOV3ip cells were employed to study the more significant difference in ezrin expression between them. (2) By a functional screen using miRNA microarray combined with bioinformatics analysis,the miR-183 and miR-22 were indentified as two candidate miRNA,which may have the potential regulatory role in ezrin expression. Real time RT-PCR assays revealed that miR-183 and miR-22 were, respectively, an average of (5.84 ± 0.66)-fold and(6.67 ± 0.67)-fold higher expression level in SKOV3ip versus SKOV3 cells (P ＜0. 01 ), which were in agreement with the microarray data. A subsequent validation by western blot and real time RT-PCR revealed that over-expression of miR-183 and miR-22 could both lead to an obvious decrease in ezrin protein level,while there were not signicant difference in the level of ezrin mRNA( P ＞0. 05 ). Conclusion Increased expression of miR-183 and miR-22 may both repress the protein level of ezrin,indicating that miR-183 and miR-22 may bear a potential role in inhibiting ovarian cancer metastasis in a ezrin-mediated way.

Objective To investigate the biological functions of hepatitis B virus e antigen (HBeAg) protein. Methods Yeast-two hybrid technique was performed to look for proteins interacting with HBeAg from hepatocytes. HBeAg bait plasmid was constructed by cloning the HBeAg gene into carrier plasmid pGBKT7, then the yeast AH109 (a type) was transformed. The transformed yeast cells was amplified and mated with yeast cells Y187 (?type) containing liver cDNA library plasmid pCAT2 in 2?YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium and were selected for two times. Results Plasmids of positive colonies were extracted and analyzed by DNA sequencing and BLAST in GenBank. Conclusions After the full sequence of new gene E-36 was amplified from the mRNA of HepG2 cells by reverse transcription polymerase chain reaction (RT-PCR) and cloned into pGADT7 vector, the recombinant plasmid was translated by reticulocyte lysate and analyzed by immuno-coprecipitation technique in vitro together with HBeAg. The result provided some new clues for the study of the biological functions of HBeAg.

OBJECTIVE:To study the pharmacokinetics and pharmacodynamics of single dose of intramuscularly injected methotrexate(MTX)in the treatment of ectopic pregnancy so as to provide theoretical bases for the choice dosage regimen.METHODS:Serum concentration of MTX in 8 cases with ectopic pregnancy were determined by fluorescence polarization immunoassay after injected i.m with single dose MTX(50mg/m2);The ?-HCG was monitored both before and after treatment and the lump sizes of the ectopic pregnancy were determined by B ultrasound.RESULTS:The pharmacokinetics of single dose of intramuscularly injected MTX conformed to two-compartment open model;Cmax occurred at 1.9h after drug taking with 2.31 ?mol/L;The time for the recovery of ?-HCG to normal was(24.8?10)days,the patency rate of oviducts of those who demanding procreation was 75%.CONCLUSION:Injection intramuscularly with single dose of MTX 50mg/m2 is effective in the treatment of ectopic pregnancy.

Early enteral nutrition (EN) is one of important methods for severe acute pancreatitis (SAP), and it has been used over the world. In this article, different choices of nutrition opportunity, location of intubation and merits and defects of EN were reviewed. The aim of this review is to provide the evidence for EN being applied widely in clinic.

AIM:Scopolamine blocks cholinergic transmission and impairs learning and memory in mice．The purpose of this study was to evaluate the memory-improving properties of curcumin on scopolamine-induced memory im-pairment in mice．METHODS：The mice of memory impairment were induced by scopolamine．Step down test and Morris water maze test were used to observe the earning and memory ability in curcumin-treated ice．Biochemical assessments of AChE，MDA，and GSH-Px levels in brains were performed．RESULTS：Oral administration of curcumin ignificantly reduced the numbers of step-down rrors(P<0．05)and prolonged the step-down latency induced by scopolamine(P<0．05)．The escape latency time in mice treated with curcumin Was remarkably educed compared to that in scopolamine group by Morris water maze test(P<0．05)．After the platform was removed，the total time that the mice swam in the tar-get quadrant Was also longer in curcumin group than that in model group(P<0．05)．The data also indicated that curcumin significantly inhibited AChE ctivity(P<0．01)and prevented oxidative stress characterized by the significant reduction in MDA content and the significant increase in GSH-Px activities in the brain(P<0．01)．CONCLUSl0N：Curcumin in-duces cognitive improvement by enhancing the function of cholinergic system and its antioxidant activity．

ObjectiveTo observe the expression of Caspase-3 mRNA in prefrontal cortex and hippocampus of chronic stress-induced depression rats,and to detect the machnisms of antidepression by electro-acupuncture.MethodsSprague-Dawley rats were randomly divided into four groups:control group,model group,model +electro-acupuncture group and model + paroxetine group,12 rats in each group.Open-field test was used to observe the changes of movements,and real-time fluorescence quantitative PCR (RT-PCR) method was used to detect Caspase-3 mRNA levels in prefrontal cortex and hippocampus.Results ①Open-field test:after stress,compared with control group rats,the model group rats' crossing numbers (29 ± 7),rearing times (6 ± 2) were apparently less than those of control group( (66 ± 13),( 10 ±2) ; P＜0.05,P＞0.05).In comparison with model group,the crossing times and rearing times being increasing in degree in electro-acupuncture group( (61 ±9),( 13 ±1 ) ) and paroxetine group( (39 ± 10),(8 ± 1 ),P＜0.01,P＞0.05).② Compared with the control group,Caspase-3 mRNA in prefrontal cortex and hippocampus significantly increased in model group(P ＜0.05 ) ;and compared with model group,Caspase-3 mRNA in prefrontal cortex in electro-acupuncture group and paroxetine group significantly decreased(P ＜ 0.05 ),and both expression of Caspase-3 mRNA in hippocampus in electro-acupuncture group and expression of Caspase-3 mRNA in hippocampus in paroxetine group decreased (P ＞ 0.05 ).ConclusionChronic stress can increase the expression of Caspase-3 mRNA in prefrontal cortex and hippocampus of chronic stress-induced depression rats,while electro-acupuncture can decrease the expression of caspase-3 mRNA,which may be an important way to anti-depression by electro-acupuncture.

AIM and METHODS: To investigate the effect of electrical stimulation of ventlal septal area (VSA) on discharge of pyrogen - treated thermosensitive neurons in preoptic anterior hypothalamus (POAH) region, the discharging rate of thermosensitive neurons in the POAH region of 32 New Zealand white rabbits were recorded by using extracellular microelectrode techinque. RESULTS: (1) Intraceretroventricularly (i c v) injection of interleukin - 1?(IL - 1?) caused decrease of discharging rate of warm - sensitive neurons and increased in discharging rate of cold - sensitive neurons in POAH regions. (2) These effects could be reversed by electrical stimulation of VSA. CONCLUSION: VSA may play a central role of negative regulation in thermoregulation of pyrogen - treated animal.

Objective To investigate levels of IL-17 and IL-22 mRNA in the blood of type 2 diabetes(T2DM)patients with the different stages of kidney injury and explore the relationship between the gene expression levels of IL-17,IL-22 and renal lesions in patients with diabetic nephropathy(DM).Methods Subjects included 60 T2DM patients with or without kidney injury and 20 normal controls(NC,n=20).Diabetes patients were divided into 3 groups by level of urinary albumin to creati-nine ratio (ACR):no proteinuria group (NA,ACR<30 mg/g,n=22),microalb uminuria group (MA,30 mg/g>ACR>300 mg/g,n=18)and diabetic nephropathy group (DN,ACR>300 mg/g,n=20).Quantitative Real-Time RT-PCR was used to detect IL-17 and IL-22 mRNA levels.Analysis differences of IL-17 and IL-22 mRNA levels among NA,MA,DN and NC groups.Results The levels of IL-17 and IL-22 mRNA were significantly higher in DN group than that in MA,NA and NC group (P <0.01,respectively),However,there were not significant difference among MA,NA and NC group (P >0.05,re-spectively).Conclusion Levels IL-17 and IL-22 mRNA were increased in blood of T2DM patients with nephropathy.IL-17 and IL-22 may play role in the pathogenicity of diabetic nephropathy.

The interactions between genetic variations and dietary factors in type 2 diabetes mellitus have attracted some attention. Several studies revealed that dietary carbohydrate quality and quantity and increased dietary fat intake might interact with genetic variations of type 2 diabetes mellitus and increase risk of this disease. Genome-wide association studies suggest that genetic variance may modulate the association between dietary pattern and type 2 diabetes mellitus.

Objective To analyze the ecological environmental differences of hibernation hole and temporary hole of Himalayan marmot based on global positioning system (GPS),remote sensing (RS) and geographical information system (GIS) in Yushu City,Qinghai Province.Methods Hibernation holes and temporary holes were located based on GPS.Ecological environment factors related with Himalayan marmot,such as land surface temperature (LST),digital elevation model (DEM),normalized different vegetation index (NDVI),were extracted based on RS and GIS.The ecological environment differences between hibernation hole and temporary hole were explored based on epidemiological statistical method.Results Monthly LST (degree Kelvin,K) of hibernation hole was slightly higher than that of temporary hole in the same period.Yearly day LST of hibernation hole was 1.1 K higher than that of temporary hole in the same period.Yearly night LST of hibernation hole was 0.3 K higher than that of temporary hole in the same period.Monthly NDVI of temporary hole was slightly higher than that of hibernation hole in the same period.Yearly NDVI of temporary hole was 0.001 higher than that of hibernation hole in the same period.DEM of temporary hole was about 40 m higher than that of hibernation hole.The top two ratios of soil type factors were both alpine meadow soil and dark felty soil for hibernation hole and temporary hole,which were 62.1％ and 29.3％,64.4％ and 25.6％.The top two ratios of land cover factors were both rainfed cropland and mosaic cropland for hibernation hole and temporary hole,which were 43.3％ and 25.8％,40.0％ and 26.2％.Conclusions Soil type factors and land cover factors for hibernation hole and temporary hole are similar.Lower DEM and higher LST are chose for hibernation hole,and better NDVI is chose for temporary hole,which is good for food intake.