Objective To examine the sensitivity of human colorectal carcinoma cells to 5-fluorouracil treatment by stable transfection of extrinsic Fas-associated death domain protein(FADD) gene，both in vitro and in vivo，so as to investigate the feasibility of combination therapy of FADD gene and 5-fluorouracil in human colorectal carcinoma.Methods①RT-PCR and Western blotting were used to detect the expressions of both mRNA and protein of FADD gene in SW480／FADD (stably transfected with FADD)，SW480／neo and SW480 cells.②After treatment with 5-fluorouracil as an apoptotic inducer，in vitro cell growth activities were investigated by MTT assay.Cell apoptosis and its rates were determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay and flow cytometry of annexin V-FITC／PI staining.The expressions of caspase-8 and caspase-3 were examined by Western blotting.③To examine the inhibitory effect of FADD gene combined with 5-fluorouracil, tumor xenograft model was prepared for in vivo study.Results ① Compared with SW480 and SW480/neo cells, FADD mRNA and protein levels of SW480/FADD cells were higher (P<0.05). ② Inhibitory rate of SW480/FADD cells was remarkably higher than SW480 and SW480/neo cells (P＜0.05 ). ③ Forty-eight hours after treatment with 5-fluorouracil (10 mg/L), the apoptotic rate of SW480/FADD cells was (33.3 ± 4.5)%, which was higher than SW480 [(13. 9 ± 3. 2)%3 and SW480/neo [(14. 1 ± 3. 4)%], with significant difference (P< 0.05).④ Forty-eight hours after treatment with 5-fluorouracil (10 mg/L),procaspase-8 and procaspase-3 expressions of SW480 and SW480/neo cells were higher than SW480/FADD cells, whereas their cleaved caspase-8 and cleaved caspase-3 expressions were lower than SW480/FADD cells (P＜0. 05).⑤ In in vivo study, SW480/FADD cells increased the efficacy of fluorouracil-induced inhibition of tumor growth in nude mice. ConclusionsStable overexpression of FADD increases sensitivity of the cells to 5-fluorouracil and combination of FADD with 5-fluorouracil will he a promising alternative in colorectal cancer treatment.

Objective To investigate the correlation between expression of human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood and postoperative recurrence and prognosis in the patients with advanced colorectal cancer (CRC).Methods The preoperative and 1 month postoperative quantitative expression of hTERT mRNA in peripheral blood in 85 patients with advanced CRC were detected with real-time fluorescence quantitative real-time fluorescence quantitative polymerase chain reaction (RTPCR).The quantitative expression of hTERT mRNA in peripheral blood in 30 normal controls were also detected.The postoperative prognosis was followed-up,and the quantitative expression of hTERT mRNA in peripheral blood in the patients with recurred CRC and the patients without recurred CRC were detected repeatedly.The factors that affected the preoperative hTERT mRNA expression were analyzed.The relationship was investigated between the hTERT mRNA expression levels and postoperative recurrence and prognosis in the patients with advanced CRC.Results The hTERT mRNA expression levels in peripheral blood in advanced CRC group were significantly higher than those in the normal controls [(6.22 ±5.85)2-△△Ct vs (0.45 ±0.27)2-△△Ct,(P ＜0.01)],and were significantly decreased at the time point of 1 month after the operation (P ＜ 0.01).The hTERT mRNA expression levels in the postoperative recurred CRC group were significantly higher than those in the 1 month postoperation group and no recurrence CRC group [(5.07 ± 2.87)2-△△Ct vs (1.83 ± 1.08)2-△△Ct,(2.15 ± 1.49)2-△△Ct,(P ＜0.01)].Univariate analysis revealed that preoperative high hTERT mRNA expression levels in peripheral blood were related to the differentiated degree of tumor,tumor size,lymph node metastasis ratio,and liver metastasis (P ＜ 0.05 or P ＜ 0.01).Multivariate logistic regression analysis revealed that differentiated degree of the tumor,lymph node metastasis,and liver metastasis were independent risk factors for the elevation of preoperative hTERT mRNA expression in the advanced CRC.Elevated preoperative hTERT mRNA expression was positively related to the postoperative recurrence rate,and was negatively related to postoperative survival rate.Conclusions Preoperative hTERT mRNA expression levels in peripheral blood were related to the important pathologic features in patients with advanced CRC,and might be a valuable prognostic factor.Postoperatively elevated hTERT mRNA expression in peripheral blood was related to recurrence of advanced CRC.

AIM: To investigate the effects of TNF receptor-associated death domains (TRADD) of latent membrane protein 1 (LMP1) on the proliferation of nasopharyngeal cancer SP18 cells.METHODS: The SP18-LMP1 cells and SP18-LMP1TRADD cells, which expressed LMP1 and LMP1 TRADD proteins, respectively, were established.The proliferation of SP18 cells affected by LMP1TRADD was detected by cell counting to analyze the cell growth curve, and by colony formation assay, soft agar formation assay, and flow cytometry.Moreover, the expression profile of differential genes between SP18-LMP1 cells and SP18-LMP1TRADD cells was analyzed by gene chips.RESULTS: The cell growth curve, and the results of colony formation and soft agar formation displayed that the growth velocity and colony forming ability of SP18-LMP1 cells were stronger than those of SP18-LMP1TRADD cells (P<0.01).The results of flow cytometry analysis showed that the proliferation index of SP18-LMP1 cells was higher than that of SP18-LMP1TRADD cells (P<0.01).Sixty-three differentially expressed genes associated with cell proliferation were screened out, in which 33 genes were up-regulated and 30 were down-regulated in the SP18-LMP1TRADD cells.CONCLUSION: TRADD active region is an important functional site of LMP1 to promote the proliferation of SP18 cells.LMP1 may improve the cell proliferation index and induce the proliferation of SP18 cells through TRADD.

Objective To explore the expression and relationship of PTEN and mTOR in the development of cholangiocarcinoma,determine the effect of the PI3K/PTEN/AKT/mTOR signal transduction on the(development) of cholangiocarcinoma.Methods The expression of mTOR and PTEN in the cholangiocarcinoma was detected by the immunohistochemistry and RT-PCR method.Results Compared to normal tissues,the(expression) of mTOR gene in cholangiocarcinoma significantly increased,but the expression of PTEN gene(decreased).There was a negative correlation between mTOR gene and PTEN gene expression in(cholangiocarcinoma). Conclusions The expression of mTOR gene in cholangiocarcinoma was increased and the expression of PTEN was decreased.It suggested that the mTOR gene and PTEN gene could play an important role in the process of development of cholangiocarcinoma.

Objective To study the differential diagnostic value of soluble epithelial cadherin (sE-cad) in malignant and benign ascites. Methods The concentrations of sE-cad and carcino-embryonic antigen (CEA) in abdominal cavity synovia patients with benign diseases (31 cases), benign ascites(82 cases) and malignant ascites(119 cases) were ineasured by ELISA technique. Results The concentrations of sE-cad and CEA in malignant ascites patients were significantly higher than those in benign ascites patients and abdominal cavity synovia patients (P < 0.01). The sensitivity, negative predictive value and totally accurate rate of sE-cad was 89.9%, 86.8% and 92.5% respectively,which was significantly higher than those of CEA(68.1%, 65.8% and 76.6%)(P < 0.05). sE-cad in malignant ascites showed a certain tissue specificity. Conclusion sE-cad is an useful tumor marker in differential diagnosis of benign and malignant ascites.

Objectives To explore the differences in therapeutic efficacy between decitabine combined with low dose HAG(D+ HAG)and CAG in elderly patients with acute myeloid leukemia (AML).Methods Totally 32 elderly patients with AML in our department from July 2012 to July 2015 were retrospectively analyzed.15 patients were on a therapy of decitabine combined with low dose HAG,and 17 patients with CAG.Efficacy and side effects were compared between the two groups.Results In(D+HAG)versus CAG groups,the complete remission(CR)was 10 cases(66.7％)vs.6 cases (35.3 ％),partial remission(PR) was 2 cases (13.3％) vs.1 case(5.9 ％),non-remission(NR) was 3 cases(20％)vs.10 cases(58.8％),and the total efficacy rate(CR+PR)was 80％vs.41.2％ (P＜0.05).The side effects in two groups mainly included bone marrow depression,respiratory tract infection,gastrointestinal reaction,which were all alleviated after symptomatic treatments.The incidence rate of side effects had no statistical difference (P ＞ 0.05).Conclusions The treatment of decitabine combined with low dose of HAG is prior to CAG,and the side effects are all tolerated.So decitabine combined with low dose of HAG can be served as the first-line therapy for elderly AML patients.

BACKGROUND: Ankle fracture combined with deltoid ligament rupture and distal tibiofibular syndesmosis injury occurs occasionally. Its treatment with distal tibiofibular syndesmosis screw fixation or deltoid ligament repair remains controversial. The former appears with poor reduction, broken nails, secondary surgery and other problems.OBJECTIVE: To observe the clinical effectiveness of suture anchor repair for ankle fracture combined with deltoid ligament injury.METHODS: Twelve patients with ankle fracture combined with deltoid ligament injury were selected from the First Affiliated Hospital of Guangzhou University of Chinese Medicine between January 2013 and December 2015. All patients were treated with open reduction, internal fixation, and anchor repair, but without distal tibiofibular syndesmosis screw fixation. The curative efficacy and joint stability were observed.RESULTS AND CONCLUSION: (1) All patients were followed-up for more than 12 months. (2) The modified Baird-Jackson scoring system showed excellent in nine cases, good in two cases, average in one case, poor in none case, and the excellent and good rate was 92%. (3) At 1 year after internal fixation, the X-ray examination showed the malleolus gap and talus slope angle of the affect side were (3.47±0.12) mm and (0.45±0.18)°, and the malleolus gap and talus slope angle of the healthy side were (3.44±0.05) mm and (0.43±0.14)°, and there was no significant difference between two sides (P > 0.05). (4) These results indicate that the suture anchor can repair the anatomy and biomechanics of deltoid ligament with stable ankle joint, and secondary surgery is unnecessary.

D-amino acid oxidase (DAAO) is one of important industrial enzymes. To increase the solubility and activity of the TvDAAO from Trignoposis variabilis expressed in recombinant Escherichia coli (E. coli), a maltose binding protein (MBP) and Vitreoscilla hemoglobin (VHb) was introduced to fuse with N-terminal of the TvDAAO, respectively. Fusion protein of MBP-TvDAAO was constitutively expressed in JM105/pMKC-DAAO and inductively expressed in JM105/pMKL-DAAO. With respect to the control strain of BL21 (DE3)/pET-DAAO without MBP fusion, the constitutive fusion expression obtained 28% of soluble protein with 3.7 folds of solubility improvement. As for the inductive fusion expression, corresponding results changed to 17% and 1.8 folds, respectively. However, the DAAO activity significantly decreased in the MBP-fusing expression. Fusion protein of VHb-TvDAAO was constructed and inductively expressed in BL21 (DE3)/pET-VDAAO. Its DAAO activity highly reached 3.24 u/mL in flask culture, about 90% increase in contrast to the control without VHb.
Bacterial Proteins ; biosynthesis ; genetics ; Carrier Proteins ; biosynthesis ; genetics ; D-Amino-Acid Oxidase ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Maltose-Binding Proteins ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Truncated Hemoglobins ; biosynthesis ; genetics ; Yeasts ; enzymology ; genetics

OBJECTIVETo characterize the effect of omeprazole on the spectrum of gene expression in the cultured human umbilical vein endothelial cell (HUVEC) line (EA.hy926), and explore the underlying molecular mechanism.

METHODSAffymetrix U133 plus2.0 oligonucleotide microarray was used to detect the alteration in the gene expression profiles induced by 1×10(-5) mol/L omeprazole in HUVECs. Real-time PCR was employed to verify the results of selected differentially expressed genes, and Western blotting was performed to test the expression levels of the related proteins.

RESULTSA total of 282 genes were found to show at least 1.5-fold changes in EA.hy926 cells after treatment with omeprazole for 48 h, including 236 up-regulated and 46 down-regulated ones. These genes were involved in the regulation of transcription, inflammatory response, immune response, cell adherence, anti-apoptosis, and signal transduction.

CONCLUSIONOmeprazole modulates the function of endothelial cells by regulating the gene expression profiles of multiple pathways.

Cell Line ; Computational Biology ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Oligonucleotide Array Sequence Analysis ; Omeprazole ; pharmacology ; Transcriptome ; drug effects

Objective To report our first clinical experience with a novel modified culotte technique for the treatment of true coronary bifurcation lesions. Methods The novel modified culotte technique (the mono-ring culotte) stenting was done in which the side branch (SB) stent was deployed firstly followed by ex vivo wiring of a most proximal cell of SB stent with the hard end of main branch (MB) wire. Secondly, the MB stent was deployed through the most proximal cell of SB stent. The procedure was ended with kissing balloon dilation. From June 2014 to March 2015, 15 patients with true coronary bifurcation lesion were treated with mono-ring culotte stenting in our center. Results The procedures were successful in all cases without procedural complication and in-hospital major adverse cardiovascular events. The procedural time was (34. 3 ± 9. 6) min, fluoroscopic time was (18. 1 ± 3. 8) min, and contrast volume was (112. 0 ± 24. 5) ml, respectively. Post-procedurally, the residual stenosis of the main and the side branch were (10. 0 ± 2. 5)% and (10. 2 ± 5. 3)% , respectively. Conclusions The mono-ring culotte stenting is safe and feasible for treatment of true coronary bifurcation lesions, and may be superior to the conventional culotte stenting.