Objective To explore the reversal effect on MDR1 gene-mediated multidrug resistance in human colon carcinoma LOVO/5-Fu cells by tetrandrine ( Tet) and to clarify its molecular mechanism.Methods LOVO/5-Fu cells were treated for 48 h with Tet.Drug sensitivity was measured by MTT.The cell cycle, apoptosis of cells and expression of P-glycoprotein (P-gp) were determined by flow cytometry assay.Expression of MDR1 mRNA was detected by real-time quantitative PCR (real-time PCR).P-gp expression was detected by Western blot.Results After LOVO/5-Fu cells were treated for 48 h with Tet, the IC50 of 5-Fu decreased to ( 4.15 ± 0.31 ) μg/ml ( P ＜ 0.05 ) ; and the apoptotic rate increased to (3.44% ± 0.28% ) ( P ＜ 0.05) ; the expression of MDR1 mRNA reduced to (570 ± 85) (P ＜ 0.05 ).Conclusions Tetrandrine reverses MDR1 gene-mediated multidrug resistance in human colon carcinoma LOVO/5-Fu cells possibly by inhibiting the expression of MDR1, decreasing the expression of P-gp, thus enhancing the sensitivity of LOVO/5-Fu cells to 5-fluorouracil.

Objective:Small bowel IRI models in rats were established to explore the effect of glutamine enriched parenteral nutrition on mucosal barrier,and to discuss the probable mechanisms.Methods:Thirty Wistar rats were randomly assigned into 3 groups:The control group(N group,n=10) ,conducted fictitious operation and fed with common forage,TPN group(n=10) and TPN+Glu group(n=10) .The morphous of mucous,serum and intestinal mucosal Gln concentration,levels of D-lactate,endotoxin,TNF-?,IL-6,HO-1 positive ratio and HO-1 mRNA were detected.Results:Glutamine obviously improved the structure of intestinal mucosal and decreased the expressions of D-lactate,endotoxin,TNF-?and IL-6.And enhanced the expressions of HO-1 mRNA and HO-1.Conclusion:Glutamine enriched parenteral nutrition can alleviate small intestinal IRI and inflammatory reaction and enhance the HO-1 and HO-1 mRNA expressions.HO-1 and its metabolin's anti-oxygen,anti-apoptosis,anti-inflammator action may be the mechanism of the protective action of Gln on mucosal barrier of small bowel.

Colorectal neoplasms always present with thickness of the intestinal wall or a soft tissue mass in the enteric cavity. Multi-slice computed tomography (MSCT) with high spatial resolution and advanced post-processing techniques can demonstrate the above signs of the tumor, and the invasive signs of adjacent structures and lymph node metastasis. Combined with three dimensional reformation images, MSCT shows a higher sensitivity than that of double contrast barium enema and electronic colonoscope. MSCT is promising in the diagnosis and preoperative evaluation of colorectal neoplasms.

Objective To investigate the expression of VEZT and its clinical pathological significance in gastric cancer tissues,and to construct the VEZT over-expression vector.Methods The expression of VEZT was examined in 119 cases of gastric carcinoma and their corresponding normal mucus tissues by SP immunohistochemical staining.The relationships between the VEZT expression levels and its clinicopathological characteristics,prognosis were also investigated.VEZT cDNA was extracted and amplificated from 293 cells by polymerase chain reaction (PCR).Using DNA recombinant technique,the target gene was connected to the pEGFP-N1 vector to construct recombinant plasmid vector after the target gene and pEGFP-N1 were purified.The recombinant plasmid was identificated by 1％ enzyme electrophoresis and DNA sequencing.Results The VEZT-positive expression in gastric carcinoma was 30.3％ and 60.5％ in normal gastric tissues.VEZT expression in high differentiated cancer tissues was higher than that in lower differentiated tissues(x2 =5.002,P ＜ 0.05),expression of VEZT in early gastric cancer was significantly higher than that in patients with advanced gastric cancer (x2 =5.551,P ＜ 0.05),expression of VEZT in patients without lymph node metastasis was significantly higher than that of lymph node metastasis (x2 =5.878,P ＜ 0.05).Patients with positive VEZT expression have better prognosis than the negative patients(x2 =6.908,P ＜ 0.01).The target fragment,consistent with the theoretical value,was verified by gel electrophoresis.DNA sequencing confirmed that the gene sequence had no mutation and the eukaryotic expression plasmid vector pEGFP-N1-hVEZT was successfully constructed.Conclusions VEZT protein was correlated with TNM staging,tumor stage,invasion depth and lymph node metastasis,positive VEZT expression predicting better five year survival.The VEZT over-expression plasmid vector was successfully constructed.

Objective To detect p27 expression in rectal carcinoma and serum transforming growth factor ? 1 (TGF ? 1) level in these patients, and to elucidate the modulatory effect of serum TGF ?1 on p27 expression in rectal carcinoma. Methods Expression of p27 was measured in 37 cases of rectal carcinoma, 22 of rectal adenoma and 19 of normal control specimens by immunohistochemical staining using antibodies to p27. Serum level of TGF ?1 was measured in these patients by enzyme linked immunosorbent assay (ELISA) method. Results p27 protein was expressed in normal rectal tissue, rectal adenoma and rectal carcinoma, and the positive rate was 89.47%, 90.91% and 64.87%, respectively. The positive rate of p27 in rectal carcinoma was significantly lower than that of normal rectal tissue and rectal adenoma ( P =0.025). p27 was mainly located in nucleolus of normal rectal tissue and rectal adenoma, and the positive rate of p27 in cytoplasm of rectal carcinoma was higher than that of normal and rectal adenoma. The positives rates of serum TGF ?1 in normal group, rectal adenoma group and rectal carcinoma group were 21.05%, 27.27% and 51.35%( P =0.045),respectively. The expression of p27 related to histological differentiation, lymph node metastasis and infiltration depth. Serum level of TGF ?1 related to lymph node metastasis, infiltrated depth and CEA level. The positive rate of p27 in TGF ?1 negative group and positive group was 88.89% and 42.11%(Mantel Haenszel ? 2=6.755, P =0.009), respectively. Conclusion TGF ?1 may be useful in assessment of malignance and prognosis of rectal carcinoma. TGF ?1 can down regulate p27 expression in rectal carcinoma.

Objective:To explore the diagnostic value of mammography and ultrasonography(US)in diagnosis of breast disease with microcalcifications.Methods:Seventy-eight cases with breast microcalcifications were detected by mammography.The detection rate of breast microcalcification at US was analyzed.And the sensitivity,specificity and the accuracy of US in diagnosis breast disease with microcalcifications was compared with these of mammography.Results:The detection rate of breast microcalcification was 66.7%,And US depicted more malignant(87.5%)than benign microcalcification(33.3%).The sensitivity specificity and accuracy of US was 68.7%、83.3%、 74.3%respectively,and the sensitivity specificity and accuracy of mammography was 72.9%、 73.3%、75.6%respectively,there were no significant difference between the two methods.The sensitivity,specificity and accuracy of the combined the two methods was 89.6%,90.0%,92.3% respectively.Conclusion:US can effectively detect and identify the breast microcalicications.The combination can improve the assessment of breast disease with microcalicications,and has a significant clinical practical value in diagnosis of early breast cancer.

Objective To describe and analyze the clinical characters of patients with FRG from 7 Chinese families.Then analyze and identify their mutations in SGLT2 gene,and explore the association of genotype and phenotype.Methods Quantitative test for 24-hour urine glucose and other laboratory tests were carried out among 7 probands (14 patients in all) and their family members from 7 pedigrees (totaling 23 subjects).All coding regions,including intron-exon boundaries,were analyzed using PCR followed by direct sequence analysis.Results Five novel mutations in SLC5A2 gene were identified in this investigation,including four missense mutations (A Serine to Glycine at position 335 (c.1003A＞G,p.S335G),a Glutamine to Arginine at position 448 (c.1343A ＞ G,p.Q448R),an alanine to proline at position 474 (p.A474P,c.1420G ＞ C) and a glycine to aspartic acid at position 580 (c.1739G ＞ A,p.G580D) and a deletion in intron 7 (c.886(-10_-31)del).By the minigene studies using the pSPL3 plasmids,we confirmed the deletion c.886(-10_-31)del as a splicing mutation.In this study,the mutation c.886(-10_-31)del accounted for about 43％ of the total alleles (12/28).These patients with compound heterozygous or homozygous mutations manifested middle degree or severe glycosuria (Quantitative test for 24-hour urine glucose:10.56-50.68 g/1.73 m2),however those with heterozygous variants presented with mild to moderate glycosuria (Quantitative test for 24-hour urine glucose ≤ 2.45 g/1.73 m2).This fits co-dominant inheritance pattern.Conclusions Five novel mutations which may be related to FRG are found in this study,and c.886(-10-31) del may be a high frequency mutation in Chinese patients.

Objective To analyze and verify the expression profiles of long non-coding RNAs (lncRNAs) in gastric cancer (GC) metastatic lymphonodus.Methods Microarray analysis was performed in 3 GC positive lymphonodus and 1 normal lymph node with Agilent Array platform to measure the expression levels of lncRNAs and mRNAs and to investigate the expression differences of lncRNAs in GC metastatic lymphonodus and normal lymphonodus, and hierarchical clustering used to screen out the differently expressed lncRNAs.15 up-regulated lncRNAs and 15 down-regulated lncRNAs were randomly chosen and RT-PCR was used to verify the expression differences.Results Comparing with normal lymphonodus, 353 lncRNAs and 547 mRNAs are up-regulated, but 464 lncRNAs and 562 mRNAs are down-regulated in GC metastatic lymphanodus as 6 times or more variation was found.The expressions of lncRNA OR3A4, LOC84740, FCGR1C and C21orf 96 were increased in GC metastatic lymphonodus, but lncRNA MSTO2P, LOC344595, TUG1, TYW3 and KRT8P10 decreased.Conclusions LncRNAs are aberrantly expressed in GC metastatic lymphonodus.

Objective To investigate the effect of astaxanthin on the peripheral blood endothelial progenitor cells (EPCs) apop-tosis induced by oxidative stress in vitro and to explore its underlying mechanism .Methods Human peripheral blood EPCs were in vitro cultured and divided into the control group ,model group with 100 μmol/L tert-butyl hydroperoxide(tBHP) and the astaxan-thin plus tBHP group(with 0 .10 ,1 .00 ,10 .00 nmol/L astaxanthin pretreatment for 24 h ,then adding the final concentration of 100μmol/L tBHP for 6 h continuous culture) .The cell viability was measured by the MTT method .The level of reactive oxygen spe-cies (ROS) was determined by the DCFH-DA method .The changes of mitochondrial membrane potential(MMP) and the apoptosis ratio were detected by the JC-1 method and the DAPI method ,respectively .Results Compared with control group ,100μmol/L tB-HP could obviously caused the apoptosis of EPCs(P<0 .05) ,while astaxanthin could decrease tBHP induced apoptosis ,which man-ifested by the decrease of the apoptosis ratio (P<0 .05) and MMP increase .Conclusion Astaxanthin has the protective effect on the apoptosis of EPCs ,its mechanism may be related with the protection of the mitochondrial function .

Objective To establish a model of pancreatic cancer induced by 7,12-dimethylbenzathracene (DMBA) in SD rats, and to detect the expression levels of RAD51 and Myc-associated factor X (MAX) and their effect on carcinogenesis of rat pancreas. Methods Ninety SD rats were randomly divided into 3 groups: a model group, an intervention group, and a control group. DMBA was directly implanted into the parenchyma of rat pancreas (the model group and the intervention group). Rats in the intervention group were treated with 1 mL trichostatin A (TSA) saline solution (1 μg/mL) via ip weekly. Rats within 3～5 months in the model group and the intervention group were executed and observed by macrograph and under microscope. Meanwhile, the rats in the control group were executed at 5th month. The EnVision~(TM) immunohistochemistry to assay the expression levels of RAD51 and MAX was used in conventional paraffin-embedded sections from the above pancreatic specimens.Results The incidence of pancreatic cancer in the model group within 3-5 months was 48.7% (18/37), including 17 ductal adenocarcinomas and 1 fibrosarcoma. The incidence of pancreatic cancer in the intervention group within 3-5 months was 33.3%(12/36), including 11 ductal adenocarcinomas and 1 fibrosarcoma. The maximal diameter of mass in the model group was significantly higher than that in the intervention group (P＜0.05). No pathological changes were found in pancreas of the control group and other extra-pancreatic main organs of the model group and the intervention group (such as the liver, biliary tract, gastrointestine tract, kidney, and lung). The positive rate of RAD51 was significantly higher in ductal adenocarcinoma in the model group, the intervention group, and the model group +the intervention group than those in corresponding groups of non-cancerous pancreatic tissues (P＜0.01), but the positive rate of MAX expression was opposite to RAD51 expression(P＜0.01). The positive tissues of RAD51 expression and/or negative tissues of MAX expression in non-cancerous tissues showed atypical-hyperplasia of ductal epitheli. Pacncreas of the control group showed the negative expression of RAD51 and positive expression of MAX. Two cases of fibrosarcoma showed the negative expression of RAD51 and MAX.Conclusion DMBA directly implanted into the parenchyma of pancreas can obtain an ideal pancreatic cancer model with high incidence in a short time. The TSA might have an inhibitive effect on carcinogenesis and growth of rat pancreas. The over-expression of RAD51 and/or lose-expression might have important effect on carcinogenesis induced DMBA in rat pancreas.