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.

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: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.

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 compare the reversal effects of short hairpin RNA (shRNA) interference and tetrandrine on multidrug resistance (MDR) of human colorectal cancer cell line LoVo/5-fluorouracil(5-FU ).Methods An eukaryotic expression plasmid of shRNA targeting MDR1 was constructed and transfected into human colorectal cancer cell line LoVo/5-FU (transfection group).LoVo/5-FU was also pretreated with tetrandrine (tetrandrine group).Drug sensitivity was detected by methyl thiazolyltetrazolium colorimetric method.Cell cycle,apoptosis of cells and positive expression rate of P-glycoprotein (P-gp) were determined by flow cytometry assay.The expressions of MDR1 mRNA and P-gp were detected by real-time polymerase chain reaction and Western blot,respectively.All data were analyzed by analysis of variance and SNK-q test.Results (1)Drug sensitivity:the 50％ concentration of inhibition(IC50)of the control group,tetrandrine group and transfection group were (7.3 ± 0.3),(4.4 ±0.7) and (2.4 ±0.4) mmol/L,respectively,a significant difference between the 3 groups was found(F =65.27,P ＜ 0.05 ).There was a significant difference in the IC50 between the tetrandrine group and the transfection group (q =6.67,P ＜ 0.05 ).(2) Changes of cell cycle:the proportion of cells in the G1 phase and S phase of the control group,tetrandrine group and transfection group were 38.13％ ± 3.75％,51.36％ ± 2.76％,59.24％±4.31％ and 20.46％±2.23％,14.32％± 1.91％,9.40％± 1.65％,respectively,a significant difference between the 3 groups was found(F =25.23,24.37,P ＜ 0.05 ).There were significant differences in the proportion of cells in the G1 phase and S phase between the tetrandrine group and the transfection group(q =3.67,4.35,P ＜ 0.05 ).(3) Cell apoptosis:the cell apoptotic rates of the control group,tetrandrine group and transfection group were 1.32％ ± 0.47％,3.24％ ± 0.26％,5.88％ ±- 0.44％,respectively,a significant difference between the 3 groups was found(F =99.26,P ＜ 0.05 ).There was a significant difference in the cell apoptotic rate between the tetrandrine group and transfection group(q =11.48,P ＜ 0.05 ).(4)The expression of P-gp:the positive expression rates of P-gp of the control group,tetrandrine group and transfection group were 96.9％ ± 2.3％,61.6％ ± 4.9％,76.6％ ± 3.6％,respectively,a significant difference between the 3 groups was found(F =67.83,P ＜ 0.05 ).There was a significant difference in the positive expression rate of P-gp between the tetrandrine group and transfection group (q =6.97,P ＜ 0.05 ).(5)The mRNA expression of MDR1:the mRNA expressions of MDR1 of the control group,tetrandrine group and transfection group were 1462 ±161,570 ±85,233 ± 81,respectively,a significant difference between the 3 groups was found(F =90.59,P ＜ 0.05 ).There was a significant difference in the mRNA expression of MDR1 between the tetrandrine group and transfection group (q =5.12,P ＜ 0.05 ).Conclusions MDR1 shRNA and tetrandrine could reverse M DR1 gene-mediated m.ultidrug resistance in human colon cancer cell line LoVo/5-FU,but the effect of MDR1 shRNA is better than that of tetrandrine.MDR1 shRNA and tetrandrine might take effect by inhibiting P-gp expression and down-regulating mRNA expression of MDR1.

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.

Objective To compare the efficacy of XELOX and FOLFOX4 in the treatment of locally advanced unresectable gastric cancer. Methods The clinical data of 72 patients with gastric cancer who were admitted to the Shandong Provincial Hospital from July 2006 to October 2009 were prospectively analyzed. Of all the patients, 3 lost follow-up, and 69 patients with locally advanced unresectable gastric cancer were randomly divided into XELOX group ( n = 36 ) and FOLFOX4 group ( n = 33 ) according to the random number table.All patients received chemotherapy for six weeks. The efficacy of the two regimens were evaluated by the multidiscipline team six weeks later. The cell cycle of patients with complete or partial remission and received surgical treatment was detected by flow cytometry. All data were analyzed using the Pearson chi-square test, Levene test or t test. Results The curative rates of XELOX and FOLFOX4 were 53% (19/36) and 52% (17/33), respectively,with no significant difference between the two groups ( x2= 0. 01 , P ＞ 0. 05 ). The incidences of nausea and vomiting, phlebitis and hand-foot syndrome were 25% (9/36), 6% (2/36) and 19% (7/36) in the xELOX group, and 55% ( 18/33), 39% (13/33) and 3% (1/33) in the FOLFOX4 group, respectively, with significant difference between the two groups ( x2 = 6.31, 11.59, 4.53, P ＜ 0.05 ). Nineteen patients in the XELOX group and 17 patients in the FOLFOX4 group received surgical resection of the gastric cancer, and no complications such as anastomotic leakage and hemorrhage occurred postoperatively. In the XELOX group, the s-phase fraction (SPF),proliferation index (PI) and G2/M of the gastric cancer cells were 5.89% ± 0.79%, 9.22% ± 1.99% and 5.19% ± 1. 66% after neoadjuvant chemotherapy, which were significantly lower than 6.76% ± 1.21%, 10.44% ±2.12% and 6. 04% ± 0. 57% before neoadjuvant chemotherapy, while the ratio of gastric cancer cells in the G0/G1 phase after neoadjuvant chemotherapy was 90.39% ±4.78%, which was significantly higher than 87.54%±6.34% before neoadjuvant chemotherapy (x2 =3.61, 2.52, 2. 15, 2.91, P ＜0.05). In the FOLFOX4group, the SPF, PI and G2/M of the gastric cancer cells were 6.09% ± 0.96%, 10.65 % ± 2.47% and 4.88% ±0.87% after neoadjuvant chemotherapy, which were significantly lower than 7.15% ± 1.45%, 11.87% ± 2.33%and 5.67% ± 1.03% before neoadjuvant chemotherapy, while the ratio of gastric cancer cells in the G0/G1 phase after neoadjuvant chemotherapy was 91.45% ± 5.22%, which was significantly higher than 88.01% ± 4.23%before neoadjuvant chemotherapy ( x2 = 3.50, 2.06, 3.37, 2.94, P ＜ 0.05 ). There was a significant difference in PI between XELOX group and FOLFOX4 group after neoadjuvant chemotherapy ( x2 = 2.66, P ＜ 0.05 ).Conclusions XELOX and FOLFOX4 are safe and effective in the treatment of locally advanced unresectable gastric cancer, and they can significantly restrain the proliferation of gastric cancer cells. XELOX regimen is more effective than FOLFOX4 regimen.

Two patients with coexistence of thyroid disease and suspected Gitelman's syndrome underwent SLC12A3 gene analysis. The results confirmed that both patients were compound heterozygotes of SLC12A3 gene mutation. Three novel variants of SLC12A3 were found in this study. This report suggests that Gitelman's syndrome may coexist with other disorders associated with hypokalemia, such as Graves' disease.

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.