Objective To construct the RNA interference(RNAi)recombinant adenovirus vector targeting at human hypoxia-inducible transcription factor 1?(HIF-1?)and to evaluate its effect on human lung adenocarcinoma cell line SPCA-1.Methods The recombinant adenovirus Ad was constructed.HIF-1? inserted with HIF-1? RNAi fragment via AdEasy system.The virus was purifed by CsCl gradient centrifuge.The functional titer of recombinant adenovirus was measured by transfection test in HEK 293 cells.SPCA-1 cells were transducted with 2 multiplicity of infection(MOI)Ad.HIF1? in vitro,the expression rate of green fluorescence protein(GFP)was recorded by flow cytometry,HIF-1? mRNA and protein level was measured by Real-Time RT-PCR and flow cytometry.ResultsThe recombinant shuttle plasmid PAdTrack.HIF-1? and adenovirus plasmid Ad.HIF-1? were all correct shown by enzyme digestion confirmation.The plasmid pAd.HIF-1? was transducted into HEK293 cells,15%GFP expressionwere seen after 3 days.The final titers of recombinant adenovirus were 5.0?1010 TU/mL.SPCA-1 cells was transducted by Ad.HIF-1? in vitro for 48 h,GFP expression rate was 92%,HIF-1? mRNA and protein level decreased 89% and 87%,respectively.Conclusion RNAi adenovirus vector of human HIF-1? gene has been successfully constructed,which could facilitate the research onHIF-1? gene related gene therapy for lung cancer.

AIM:To investigate the regulatory role of hypoxia mimic reagent cobalt chloride ( CoCl2 ) on cave-olin-1 (Cav-1) generation and the influence of Cav-1 on the abilities of migration and invasion of human lung adenocarcino-ma A549 cells.METHODS:The concentrations of Cav-1 and hypoxia-inducible factor ( HIF)-1αin pleural effusion of the patients with lung cancer ( MPE) or tuberculous pleurisy ( TBPE) were detected, and the correlation was also compared. A549 cells were treated with CoCl2 at different concentrations and time in the presence or absence of HIF-1αinhibitor YC-1.The concentrations of Cav-1 and HIF-1αin the cell supernatants were measured by ELISA.The effects of Cav-1 induced by CoCl2 on the migration and invasion of A549 cells were determined by scratch test and Transwell invasion trial, respec-tively.RESULTS:The levels of Cav-1 and HIF-1αin MPE were significantly higher than those in TBPE.There was a highly positive correlation between Cav-1 and HIF-1αlevels in the pleural effusion.CoCl2 induced the generation of Cav-1 and HIF-1αin A549 cells in a concentration-and time-dependent manner, the peak occurred at 200 μmol/L or 24 h, while the concentration over 200 μmol/L or after treated over 24 h, a concentration-or time-dependent inhibition was ob-served.HIF-1αinhibitor YC-1 concentration-dependently inhibited the generation of HIF-1αand Cav-1 induced by CoCl2 in A549 cells.CoCl2 enhanced A549 cells migration and invasion, with 200 μmol/L played the strongest role, which were down-regulated significantly in the presence of YC-1.CONCLUSION:The alteration of hypoxia-induced Cav-1 generation might be involved in the migration and invasion of A549 cells.A possible role for HIF-1αis indicated in Cav-1 generation.

Purpose:To study the relation between regional lymph node metastasis and nm23 H 1 protein expression and DNA ploidy of esophageal squamous cell carcinoma patients. Methods:Flow cytometry was used to analyze nm23 H 1 protein and DNA ploidy in surgical specimens from the patients with esophageal carcinoma. The object tissues included: the primary tumor tissue, the pericancerous mucosa, the incisional margin and the regional lymph node. Results:There were obvious differences between nm23 H 1 protein expression in lymph node metastasis positive and negative groups, primary tumor tissue and pericancerous mucosa, cancer tissue and the incisal margin( P 0.05).The differences of DI were distinct between cancer tissue and the pericancerous mucosa, cancer tissue and the incisal margin, cancer tissue and the lymph node( P 0.05).In corresponding tissues of lymph node metastasis positive and negative groups, nm23 H 1 protein expression was different( P 0.05). As to DI, it was different only among pathological grades( P 0.05). Conclusions:nm23 H 1 protein higher expression can produce certain inhibitory effect on metastasis of esophageal carcinoma. Combining DNA ploidy detection can help to understand the biological behavious and deduce the prognosis of esophageal carcinoma.

Purpose:To study COX 2 protein expression and its significance in familial adenomatous polyposis (FAP) and rectal cancer tissues. Methods: Flow cytometry was used to detect COX 2 protein expression in surgical specimens from 12 normal rectal mucosa tissues, 19 FAP tissues, 7 transformed FAP tissues and 36 rectal cancer tissues.Results:The COX 2 protein expression was negative in the 12 normal rectal mucosa tissues ( FI=0.67?0.28). The COX 2 protein expression was positive in 18 (94.74%) of the 19 FAP tissues (FI=1.97?0.36),the 7(100%) transformed FAP tissues(FI=2.02?0.24) and the 36(100%) rectal cancer tissues (FI=2.14?0.31). The COX 2 protein and positive expression rates showed that there were obvious differences between normal rectal mucosa tissues and FAP tissues, transformed FAP tissues, rectal cancer tissues( P 0.05). Conclusions:It is suggested that the detection of COX 2 protein expression can have important significance for predicting adenomatous polyposis carceration and chemo interfering carceration progress. [

Objective:To investigate the relationship of osteoporotic vertebral body compression fracture(OVCF)with multifidus muscle and intervertebral disc degeneration in the lower lumbar spine and the lumbar-pelvic parameters in elderly women.Methods:Clinical data of a total of 108 elderly women with osteoporosis were retrospectively analyzed.They were divided into the fracture group(n=56)and the control group(n=52)according to the presence of vertebral body compression fractures.Age, body mass index(BMI)and bone mineral density(BMD)were collected in two groups.The angle of pelvic incidence(PI), angle of pelvic tilt(PT), low lumbar lordosis(LLL)and sacral slope(SS)were measured by the standing lumbar lateral X-ray photography.The degree of degeneration of intervertebral disc L 3-S 1 and multifidus muscle were detected by using the lumbar MRI.These parameters were compared between the two groups. Results:The age, BMI and BMD had no significant difference between the two groups( P>0.05). The degree of intervertebral disc degeneration of L 3-L 4 and L 4-L 5 was statistically significant higher in the fracture group than in the control group( Z=-4.656 and -2.675, P=0.000 and 0.007), while the degree of intervertebral disc degeneration of L 5-S 1 had no statistically significant difference between the two groups( Z=-1.784, P=0.075). There were statistically significant higher multifidus muscle degeneration of L 3-L 4 and L 4-L 5 in the fracture group than in the control group( Z=-3.248 and -4.073, P=0.001 and 0.000), while no significant difference between the two groups in multifidus muscle degeneration of L 5-S 1( Z=-1.096, P=0.282). No statistically significant differences were found in PI, PT, SS and LLL between the two groups( t=1.162, 0.827, -0.082 and -0.677, P=0.248, 0.410, 0.935 and 0.500). Conclusions:The degeneration of intervertebral disc and multifidus muscle of L 3-L 4 and L 4-L 5 is positively correlated with OVCF in elderly women.

Objective To explore the relationship of chemotherapy sensitivity and expression of multidrug resistance genes and apoptosis regulation genes in human breast cancer cell lines.Methods MTT assay was used to detect the sensitivity to adriamycin(ADM),cisplatinum(DDP),mitomycin C(MMC),fluorouracilum(5-Fu),carmustine(BCNU) in five breast cancer cell lines including Bcap37,MCF-7,T47D,MDA-MB-231and MDA-MB-435.Multidrug resistance genes including P-glycoprotein(P-GP),Glutsthione-s-transferases-?(GST-?),Lung resistance protein(LRP),multidrug resistance related protein(MRP),MGMT and apoptosis regulation genes FAS,BCL-2,P53 and P16 were examined by flow cytometry(FCM).Results The chemotherapy sensitivity was obviously divergent in different breast cancer cell lines.The correlation analysis showed that there were positive correlations between the sensitivity to 5-Fu in breast cancer cell lines and the expression of P16.There was no correlation among the sensitivity to other drugs and expression of other genes.Conclusions The sensitivity to 5-Fu is related to the expression of P16 in breast cancer cell lines.

Objective To investigate effect of Bailing Capsule combined with Xihuangwan in stage III breast cancer patients of ER, MMP-9, tumor markers and estrogen.Methods 76 Patients with stage III breast cancer were selected and divided into control group anf experiment group.Control group were treated by clinical routine method.Experiment group were treated on the base of the control group with Bailing Capsule combined with Xihuangwan.The ER, MMP-9, tumor markers, estrogen, survival time and quality of life were observed and compared.ResuIts Compared with control group, the survival period of the experiment group were longer(P<0.05),the quality of life score were higher(P<0.05),the ER, MMP-9, E2, E1 level and the serum levels of CA15-3, CA125 and CEA were lower(P<0.05).ConcIusion Bailing capsule combined with Xihuang Pill can effectively reduce stage III breast cancer tumor marker levels, decrease the ER, the expression of MMP-9 and estrogen level and prolong survival time, improve the quality of life, has the vital significance to the clinical therapy for stage III breast cancer.

Objective Studies show that the ERCC1 gene may be involved in secondary cisplatin resistance.This article aims to investigate the effects of shRNA targeting silencing excision repair cross-complementation group 1 (ERCC1-shRNA) on the proliferation and apoptosis of lung cancer A549/DDP cells treated with different concentrations of cisplatin.Methods Lung cancer A549/DDP cells were divided into a negative control, a blank control, an ERCC1-shRNA1, and an ERCC1-shRNA2 group.Human interfering RNA (RNAi) targeting the human ERCC1 gene was constructed and transfected into the A549/DDP cells using Lipofectamine 2000.The mRNA and protein expressions of ERCC1 in the A549/DDP cells were detected by real-time PCR and Western blot respectively, the proliferation-inhibition rate was assessed by MTT, and their cell cycle and apoptosis were determined by flow cytometry.Results ERCC1-shRNA was successfully constructed and transfected into the A549/DDP cells.Both the mRNA and protein expressions of ERCC1 were significantly lower in the ERCC1-shRNA1 (0.20±0.04 and 0.24±0.10) and ERCC1-shRNA2 (0.47±0.28 and 0.37±0.11) than in the negative control (0.96±0.12 and 1.32±0.13) and blank control groups (0.84±0.07 and 1.45±0.23) (P<0.01).Compared with the negative and blank control groups, the ERCC1-shRNA1 group showed a significantly decreased IC50 value (16.71±2.33 and 16.69±1.69 vs 7.78±0.54, P<0.01) and an increased proportion of G0/G1 phase cells ([72.87±3.23] and [71.75±4.56] vs [82.99±4.23]%, P<0.05), with the cell cycle arrested in the G0/G1 phase.The apoptosis rate of the cells in the ERCC1-shRNA1 group was remarkably lower after treated with cisplatin at the concentrations of 6.25 and 12.5 μg/mL than at 0 μg/mL ([8.17±0.65] and [11.91±1.41] vs [29.97±3.14]%, P<0.05).Conclusion ERCC1-shRNA can inhibit the proliferation and enhance the apoptosis of A549/DDP cells by silencing the expression of the ERCC1 gene.

Objective To investigate the effect of S-1 on serum thymidine kinase-1 ( TK1 ) , vascular endothelial growth factor ( VEGF ) and insulin-like growth factor-1 (IGF-1) and tumor biomarkers, estrgen levels and quality of life in patients with advanced breast cancer.Methods 69 cases with advanced breast cancer from January 2014 to May 2015 in the hospital were selected and randomly divided into two groups.34 cases in control group were treated by conventional therapy, and 34 cases in experimental group were treated with S-1.Serum TK1, VEGF and IGF-1, tumor biomarkers, estrogen level and quality of life score were compared pre-and post-treatment.ResuIts Compared with control group, the levels of VEGF, IGF-1 and TK1 were lower (P<0.05), serum CA125, CEA and CA15-3 concentrations were lower (P<0.05),the LH, E2 and E1 levels were lower (P<0.05) and the survival quality score was higher in experimental group ( P<0.05 ) .ConcIusion S-1 has better clinical curative effect in treatment of patients with advanced breast cancer, and effectively reduce serum TK1, VEGF, IGF-1, tumor biomarker levels, improve the quality of survival, which has important significance.