EBP50(ERM-binding phosphoprotein-50),a multifunctional adapter protein with 358 amino acids and two PDZ domains, regulates cell growth and migration. Lines of evidences indicate that it is a potential cancer suppressor protein. Loss of heterozygosity (LOH) and intragenic mutation of the ebp50 gene have been found in both primary breast tumors and breast cancer cell lines. EBP50 suppresses the breast cancer cell proliferation via its interaction with many tumor suppressor protein including PTEN, SYK, MERLIN, etc. Here the molecular structure of EBP50, signal pathway regulated by EBP50, and the relationship between breast cancer development and EBP50 are discussed.

Objective To evaluate the effects of curcumin pretreatment on the expression of Nrf2 protein during ventilator-induced lung injury in rabbits.Methods Twenty-four healthy male New Zealand white rabbits,aged 3-6 months,weighing 2.5-3.0 kg,were randomized into 3 groups (n =8 each) using a random number table:two-lung ventilation (TLV) group; one-lung ventilation (OLV) group; and curcumin pretreatment group (group Cur).In group Cur,curcumin 40 mg/kg (dissolved in 2 ml of 1％ sodium carboxymethylcellulose) was given via a gastric tube into the stomach twice a day for 7 consecutive days starting from 7 days before ventilation,while the equal volume of sodium carboxymethylcellulose was given via a gastric tube instead of curcumin in TLV and OLV groups.All the rabbits were tracheostomized,and a tracheal tube was inserted to perform TLV in TLV group,and a tracheal tube was inserted into the right bronchus to establish OLV in OLV and Cur groups.Volumecontrolled ventilation was used in the three groups and the ventilatory parameters were regulated to maintain SpO2 ＞ 90 ％.Immediately before beginning of ventilation (T0) and at 1,2 and 3 h of ventilation (T1-3),arterial blood samples were obtained for blood gas analysis and determination of PaO2.The oxygenation index was calculated.At the end of ventilation,the rabbits were sacrificed and right lungs were removed for determination of wet/dry lung weight ratio (W/D ratio).The right lower lobe was isolated and puhmonary specimens were obtained for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity (using colorimetric method) and the expression of Nrf2 and HO-1 protein (by Western blot) in lung tissues and for microscopic examination of pathological changes of the lung which were scored.Results Compared with group TLV,the W/D ratio,pathological scores,MDA content,and expression of Nrf2 and HO-1 were significantly increased,and the SOD activity and oxygenation index at T2,3 were decreased in OLV and Cur groups (P ＜ 0.05).Compared with group OLV,the W/D ratio,pathological scores,and MDA content were significantly decreased,and the SOD activity,oxygenation index at T3,and expression of Nrf2 and HO-1 were increased in group Cur (P ＜ 0.05).Conclusion Curcumin pretreatment reduces ventilator-induced lung injury through promoting the expression of Nrf2 protein in lung tissues in rabbits.

AIM: To construct the recombinant adenovirus carrying fusion suicide gene CDglyTK with the C promoter(Cp),one of the key factors in controlling Epstein-Barr virus latent gene expression,and to investigate if the Cp mediates the expression of CDglyTK in CNE1 cells and kills the cancer cells specifically.METHODS: The tk,cd,Cp sequences were amplified by PCR and subcloned into corresponding sites of pDC316 vector with directional cloning method to construct the pDC316-CP-CDglyTK.The plasmid was analyzed by DNA sequencing and enzyme digestive method.The recombinant adenovirus of Ad-Cp-CDglyTK was packaged,amplified and purified in 293 cells,and the virus titre was determined by TCID50 method.The CDglyTK gene expression in CNE1 and NP69 were examined by reverse transcription-polymerase chain reaction(RT-PCR) after in vitro transfection in CNE1 and NP69 cells.The killing effect of Ad-Cp-CDglyTK/GCV+5-FC on CNE1 cells was detected by MTT method.RESULTS: The results of restriction enzyme digestion and DNA sequencing showed that the tk,cd,and Cp gene were inserted into the pDC316 plasmid in correct orientations.The titer of the recombinant adenovirus was 5.6?1012 TCID 50/L.The Cp fragment was amplified from the total RNA of the transfected CNE1 cells by RT-PCR.The mRNA of CDglyTK gene expression was not detected in NP69 cells.The MTT results showed that after administration of GCV and 5-FC,the killing effects of fusion gene were much better than those of single gene therapy.CONCLUSION: The C promoter specifically mediates the expression of CDglyTK in CNE1 cells.The Ad-Cp-CDglyTK/GCV+5-FC has much better killing effects on CNE1 cells than single gene.

Objective To evaluate the effect of ixeris sonchifolia on mitochondrial transcription factor A (mtTFA) expression following myocardial injury induced by exhausting exercise in rats.Methods Forty male Wistar rats,weighing 200-220 g,were randomly divided into 3 groups using a random number table:control group (group C,n =8),exhausting exercise group (group ES,n =16) and ixeris sonchifolia group (group IS,n =16).The model of myocardial injury was established by exhausting swimming.In IS group,the rats were subjected to exhausting swimming after intraperitoneal ixeris sonchifolia 20 g/kg.In ES and IS groups,8 rats were chosen at 6 and 24 h after exhaustion,and blood samples were taken from the inferior vena cava for determination of serum cardiac troponin I (cTnI) concentrations by ELISA.The animals were sacrificed after blood sampling and myocardial specimens were obtained for microscopic examination and for determination of mtTFA expression by Western blot.Results Compared with group C,the concentration of serum cTnI was significantly increased,and the expression of mtTFA in myocardial tissues was down-regulated at 6 and 24 h after exhaustion in ES and IS groups.Compared with group ES,the concentration of serum cTnI was significantly decreased,and the expression of mtTFA in myocardial tissues was up-regulated at 6 and 24 h after exhaustion in group IS.Conclusion Ixeris sonchifolia can reduce exhausting exercise-induced myocardial injury through up-regulating myocardial mtTFA expression in rats.

Objective To evaluate the effect of anisodamine on endoplasmic reticulum stress during acute kidney injury (AKI) in rats.Methods Forty-two nale Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 3 groups:control group (group C,n =6),AKI group (n =18) and anisodamine group (group AD,n =18).AKI was induced by intramuscular injection of 50％ (v/v) glycerol 10 ml/kg into bilateral hind limbs in groups AKI and AD.In addition,anisodamine 10 mg/kg was injected intraperitoneally 20 min before intramuscular injection of glycerol in group AD.In group C the rats received intramuscular injection of normal saline 10 ml/kg into bilateral hind limbs.Six rats were chosen immediately after injection of normal saline in group C or at 1,6 and 24 h after glycerol injection in groups AKI and AD and then anethetized with intraperitoneal pentobarbital.The animals were sacrificed and kidney specimens were obtained and cut into sections which were stained with hematoxylin and eosin for pathological examination.The pathological changes of the renal tubules were scored.The expression of glucose-regulated protein 78 (GRP78) and oxygen-regulated protein 150 (ORP150) in renal tissues was determined by immuno-histochemistry and Western blot.Results Compared with group C,the pathological scores were significantly increased and the expression of GRP78 and ORP150 was up-regulated at all time points in groups AKI and AD (P ＜ 0.01).Compared with group AKI,the pathological scores were significantly decreased and the expression of GRP78 and ORP150 was down-regulated at all time points in group AD (P ＜ 0.05 or 0.01).Conclusion Anisodamine can ameliorate AKI through inhibiting endoplasmic reticulum stress in renal tubular epithelial cells and decreasing endoplasmic reticulum stress-induced cell apoptosis in rats.

Objective To investigate the knowledge about risk factors for stroke in stroke patients. Methods 1043 stroke patients in 10 hospitals were investigated with self-designed questionnaire. Results The rate of awareness of risk factors for stroke was: hypertension 61.55%, hyperlipidemia 40.27%, drinking 32.21%, smoking 30.11%, diabetes mellitus 28.67%, heart disease 23.11%. 6.42% knew all the 6 risk factors, and 20.13% did not know any risk factor. The rate of awareness of diabetes mellitus was different among various aged groups (P<0.01), and the rate of awareness of hypertension, hyperlipidemia, and diabetes mellitus increased with the years of education. Conclusion The stroke patients know less about the common risk factors for stroke, who need more health education.

OBJECTIVE: To study the effects of Lycium barbarum polysaccharide (LBP) on tumor microenvironment T-lymphocyte subsets and dendritic cells in H22-bearing mice and the mechanisms for intervention of tumor immune escape by LBP. METHODS: H22-bearing mice were given LBP orally for two weeks. T-lymphocyte subsets and the phenotypes of dendritic cells in tumor-infiltrating lymphocytes (TIL) were detected by flow cytometry (FCM). RESULTS: LBP could significantly increase the numbers of CD4(+) and CD8(+) T cells in TIL as compared with those in model control group (P<0.05). In model control group, the number of dendritic cells in tumor microenvironment decreased markedly, while in LBP-treated group, the increased number of dendritic cells and B7-1 expression were observed, but there were no significant differences between these two groups. CONCLUSION: LBP has anti-tumor effect probably by increasing the numbers of CD4(+) and CD8(+) T cells in TIL to relieve the immunosuppression and enhance the anti-tumor function of the immune system. But whether LBP can recover the phenocyte and function of dendritic cells in H22-bearing mice should be further studied.

Objective To study the CT and MR characteristic features of the respiratory epithelial adenomatoid hamartoma of olfactory clefts. Methods (1)The CT and MRI findings of 29 patients with histologically proved respiratory epithelial adenomatoid hamartoma in the olfactory clefts were retrospectively reviewed.All patients underwent CT and 8 of them underwent MRI. Location, CT and MRI features, and associated findings of the disease were reviewed;(2)The CT findings, olfactory clefts width, total nasal distance, and the ratio of OC to the total nasal distance of the case patients (29 cases) and the control patients (33 patients with sinusitis) were compared to investigate the correlation of the olfactory clefts distance and the incidence of respiratory epithelial adenomatoid hamartoma in olfactory clefts. Results All patients were associated with sinusitis, and 23 had sinonasal polyps, 1 had papilloma. On nonenhanced CT, the OC lesions with the OC widening were isodense to gray matter in all cases, and the lesions caused the adjacent bony expansion and absorption rather than erosion; 15 cases were bilateral diseases and 14 were unilateral;The olfactory clefts width of the case patients and the control patients were (1.03±0.24) cm, (0.71± 0.17) cm, respectively. There was statistically significant difference (t=4.963, P<0.01) for the olfactory clefts width between the case patients and the control patients, and there was no significant difference (t=1.640, P>0.05) for the total nasal distance, and was significant difference(t=6.029,P<0.01)in the ratio of OC to the total nasal distance between the two groups. On T1WI, the disease appeared isointense in 6 patients and slightly hypointense in 2 patients compared with gray matter. On T2WI, the lesions revealed heterogeneous isointense in all patients. Regular cribriform pattern was found on MR T2WI and enhanced TlWI. Conclusions The unilateral or bilateral olfactory cleft opacification in chronic sinusitis patients with or without sinonasal polyposis, with involved OC widening and the adjacent bony walls compressed and remodeled may highly suggests the presence of REAH in the OC. The lesions showed inhomogeneous isointense signal on T2WI images, regular cribriform pattern enhancement are typical imaging feature of this entity.

Objective To establish a tandem affinity purification(TAP) system of innate immune-regulatory protein PKR and analyze PKR function, for the future screen and identification of novel PKR-interaction proteins. Methods PKR gene was amplified by PCR, and then cloned into a mammalian expression vector pcTAP-A. Recombinant pcTAP-PKR was transfected into PKR knock-down(PKRkd) HeLa cells by LipofectAMINE 2000,and the PKR overexpressed HeLa cells were harvested for mitogen-activated protein kinases(MAPK) activation analysis. Cell extracts of PKR overexpressed cells were purified using TAP kit and examined by Western blot. Results Cal modulin resin(CBP) and streptavidin resin(SBP) tagged PKR was detected in PKRkd HeLa cells as early as 24 h upon transfection with pcTAP-PKR, and its expression decreased at later time points. The overexpression of PKR was autophosphorylated, and thus involved in the regulation of MAPK actviation. After small-scale TAP kit purification, PKR protein was detectable by Western blot. Conclusion We have successfully established a TAP system that over-expresses functional PKR, providing a useful tool for the future study on the identification of PKR interacting proteins.

【Objective】 To study the location effect of the Mc Ab G9 to human esophageal carcinoma in tumor bearing nude mice. 【Methods】 125　I-G9 was prepared by Chloramine-T method. The distribution of 125 　 I-G9 was detected at different time (24, 48, and 72 h) after peritoneal injecti on. The percentage of the injected dose per gram of tissue(%ID/g) and the ratio of Tumor/non-Tumor were calculated. 【Results】 The distribution of 125　 I -G9 in tumor at the third day was showed by autoradiography obviously higher th an in other organ/tissue (except blood) and the highest is at the 48 h. The T/NT values are 2-7. The autoradiography indicated that radioactivity concentrated in tumor tissue. The concentration in tumor edge is more obvious than in the ce nter. 【Conclusion】 125　I-G9 has a considerable targeting activity and can well locate in esophageal carcinoma tissue in tumor bearing nude mice.