Objective To investigate the inhibition effect of silencing Ku80 gene combined with irradiation on growth of esophageal cancer xenografs. Methods shRNA-Ku80 vector was constructed.The expression of Ku80 protein was inhibited by shRNA-Ku80 vector by using Western blotting. 20 BALB/c nude mice were randomly divided into 4 groups, including control group, radiation group, shRNA-Ku80 group and combined group. The growth of esophageal cancer xenografs was observed. The expression of Ku80 was examined in esophageal cancer xenografs by IHC. Results Effective target sequence was selected. The growth of esophageal cancer xenografs was inhibited by shRNA-H2 and radiation, especially in combined group. The inhibition rate of growth in three groups above was 32.0% , 39. 9% and 68. 9% ,respectively. The expression of Ku80 was reduced to 58% by shRNA-H2 in esophageal cancer xenografs ( t = 3.77, P ＜ 0. 05 ). Conclusions Combination of silencing Ku80 and radiation could enhance radiotherapeut effect of esophageal cancer xenografs.

Objective To study the role of KuS0 in the treatment of esophageal cancer through inhibiting the Ku80 expression by shRNA. Methods shRNA-KuS0 vector was constructed. The effectiveness and feasibility of RNA interference were confirmed by Western blot and RT-PCR methods. The cell sensitivity to ganuna-rays was studied by colony formation assay. The effects of shRNA-Ku80 on cell cycle were observed by flow cytometry analysis. Results ShRNA-Ku80 vector was constructed successfully. Inhibition of Ku80 expression by shRNA enhanced the sensitivity of esophageal cancer cells to gamma-rays, shRNA K3 or shRNA H2 showed higher percentage of cells in G2/M phase (61.8% vs 28.6% ;64.3% vs 28.6%). Conclusions Inhibitions of Ku80 expression by shRNA play a role in the treatment of esophageal cancer. Ku80 might be a new target of tumor treatment.

0.05).CONCLUSION:It is feasible to reduce the dose of hormone in weekly docetaxel chemotherapy

Objective To evaluate the blond-saving effect of low central venous pressure(CVP) combined with acute hypervolemic hemedilution(AHHD)in patients undergoing hepatic lobectomy.Methods sixty ASA I orⅡpatients of both sexes aged 32-48 yr weighing 47-72 kg undergoing hepatic lobectomy for primary malignant hepatonm under epidural combined with general anesthesia were randomly divided into 3 groups(n=20 each);group I control(C);group 1I AHHD and group Ⅲ low CVP+AHHD.Group C received crystalloid and coloid in a ratio of 1.5:1 during operation.In groupⅡ4% suecinylated gelatin was infused at 50 ml·kg-1·h-1 for 30 min after tracheal intubation (AHHD);while inⅢ group low CVP was induced and maintained by epidural administration of a mixture of 1.5% lidnoaine +O.2% bupivacaine 6-8 ml combined with intravenous infmion of propofol at 6 mg·kg-1·h-1 until 10 min after hepatic lobectomy was completed.then 4% succinylated gelatin was infused at 50 ml·kg-1·h-1 for 30 min.Blood glucose,Hb,Hct, WBC count,blood coagulation (PT,AVIT,Fib),shtmic-pyruvic transaminase (GPT) and renal function (BUN,Cr) were determined before operation (baseline),immediately before skin incision,immediately before and 10 min after liver lobe was removed,at the end of operation and 7 d after operation.Urine output,intraoperative blood loss and blood transfusion and complications were recorded.Results The glood glucose concentration.WBC count and GPT levd were significantly lower;the amount of fluid infused and urinary output before hepatic lobe resection and the percentage of the patients with allogeneic blood transfusion during operation were less;Hb,Hct and the amounl of fluid infused and urinary output after hepatic lobe resection were uigher in grolp Ⅲ than in group I and ⅡⅡⅡ.There were no significant differences in blood coagulation,renal function,the total amount of fluid infused and urine output among the 3 groups.No patient developed any complication.Conclusion The low CVP hefor combined with AHHD after hepatic resection can decrease intraoperative blood loss and allogeneic blood transfusior and is safe.

Objective To investigate the effect of radiation on the expressions of RANKL and OPG in osteoblasts in order to disclose the molecular mechanism of bone injury induced by ionizing radiation.Methods The osteoblasts were differentiated from MC3T3-E1 cells.After 2 or 4 Gy137 Cs γ-irradiation,the mRNA and protein expression levels of RANKL and OPG of osteoblast precursor and osteoblast were detected by real-time PCR and Western blot.Results The expressions of RANKL mRNA (t=5.41,P＜0.05)and protein(t=68.37,P＜0.01)were up-regulated after 4 Gy irradiation,while the expressions of OPG mRNA(t=5.20,7.02,P＜0.05)and protein(t=7.78,9.45,P＜0.05)were down-regulated after 2 and 4 Gy irradiation.Conclusions 2 and 4 Gy ionizing radiation alters RANKL/RANK/OPG pathway in osteoblasts,which may promote the osteoclast differentiation and maturation and hence promote bone resorption of osteoclasts.

Objective To investigate the effects of pulsed electromagnetic fields (PEMF) on homing and proliferation-related genes of mouse osteoblasts.Methods 9 week-old C57BL/6 mice were treated with PEMF (70 Hz,1 mT) for 4～5 weeks,while mice in control group didn't not receive PEMF.Bone marrow cells of femurs and tibias were flushed out,and the bones were minced and incubated at 37 ℃ with a type Ⅰ collagenase.Bone associated mononuclear cells (MNCs) were isolated via density centrifugation with Lymphoprep.Magnetic cell sorting was used before flow-cytometric sorting,and the ALCAM+Sca-1-cells were collected.The homing and proliferation-related genes expressed in ALCAM+Sca-1-cells were detected with high throughput microarray and RT-PCR.Results The expression of Jag1 and Ang-1 in mouse osteoblasts increased under the effects of PEMF.Conclusions PEMF may have regulation effects on HSC (hematopoietic stem cell) survival through modulating the homing and proliferationrelated genes in ALCAM+Sca-1-osteoblasts.

Objective To study the influence of irradiation on the osteoblast function by the gene expression changes of RANKL and OPG.Methods Bone marrow stromal cells were induced to develop into early and mature osteoblasts in vitro.The characterization of osteoblasts was indentified by ALP staining.The RANKL and OPG mRNA levels in early and mature osteoblasts, which exposed to 0 -4 Gy radiation were determined by RT-PCR.Results Bone marrow stromal cells had been induced to early and mature osteoblasts by osteoblast differentiation medium in vitro.In early stage of osteoblast, RANKL mRNA expression levels treated with 1Gy irradiation was 2.83-fold higher than those other irradiation dosage groups.The RANKL mRNA expression levels of each group in early stage of osteoblasts were significantly higher than those in the mature counterpart ( t = 8.34 - 103.57, P ＜ 0.05 ).The ratio of RANKL/OPG mRNA was obviously greater in early osteoblast compared with the mature cells ( t = 2.84 - 20.99, P ＜0.05 ), and it was the highest in 1Gy irradiation treated early osteoblast.Conclusions Radiation exposure of the early osteoblasts promotes osteoclasts function and results in the bone loss.

objective To assess the DNA damage of radiation workers in different grade hospitals,and to explore the correlation between the types of work or work time and the levels of DNA damage.Methods DNA single strand break were detected by using alkaline single cell gel electrophoresis(SCGE),and the comet was analyzed with CASP(Comet Assay Software Project).TDNA,TL,TM and OTM were calculated.Results The parameters of SCGE in the radiation greup were higher than those of control group(F=3.93,P<0.01).The significant difference was found not only among the different types of work or difierent work time,but also among the different grade hospitals(F=1.83,1.9 1,P<0.05).Conclusions Various levels of DNA damage could be detected in the radiation workers of the two hospitals.DNA damage of radiation workers is less serious in the higher-grade hospital than the lower grade one.Different types of work or work time might affect the DNA damage level.

When we try to establish the gene recombinant yeast cell to screen the androgenic endocrine disruptors, the key procedure is the androgen receptor (AR) expression in the yeast cell. For this purpose, we obtained the GPD (glyceraldehyde-3-phosphote dehydrogenase) promoter from the yeast genosome of W303-1A using PCR system and inserting it into Swa I and BamH I sites of pYestrp2. The new constructed vector was named pGPD. The V5 epitope tag DNA with a 5'-BamH I and a 3'-EcoR I sticky end was cloned into the corresponding site of the pGPD vector to yield the vector of pGPDV5. The 2 723 bp full length AR ORF amplified by PCR from pcDNA3.1/AR was fused to V5 epitope tag DNA in pGPDV5 to give the AR yeast expression vector of pGPDV5/AR. This fused vector was transformed into the yeast cell (W303-1A). Western blot was used to detect the V5 fused protein of AR, in the protocol of which the primary monoclonal antibody (IgG(2a)) of mouse anti-V5 and the polyclonal secondary antibody of goat anti-mouse (IgG) linked to horseradish peroxidase (HRP) were used to detect the specific protein in the given sample of the transformed yeast extract. The result showed that the fused protein of AR was expressed successfully in the yeast cell.
Base Sequence ; Endocrine Disruptors ; analysis ; Epitopes ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases ; genetics ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Receptors, Androgen ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Yeasts ; genetics ; metabolism

Objective:To establish an in vitro capsular bag model and compare the inhibitory effects of different 360° square-edge intraocular lens (IOL) on lens epithelial cells (LECs) migration. Methods:In vitro capsular bag model with posterior capsule opacification (PCO) was established using Transwell compartment, cell climbing slices, human collagen type Ⅳ, and IOL.The models were divided into Plate-loop HydroSmart group, C-loop HydroSmart group, and C-compensation-loop Hydrophobic group according to the different square-edge IOL implanted.A blank control group was set using the Transwell compartment without IOL.The early PCO pathological manifestations in lens epithelial cell line SRA01/04 cultured in the Transwell compartment were observed with an inverted microscope.The cell morphology in different groups was observed by hematoxylin and eosin staining.The cell counting and cell migration inhibition rate of anterior capsule and posterior capsule were calculated by Transwell assay and cell-exclusion zone assay, respectively. Results:The early pathological characteristics of PCO, such as early Soemmering ring and small Elschnig pearl, could be found in cells in the in vitro capsular bag model after 48-hour culture.The migrating cells in model groups were fibrous.No changes mentioned above were found in blank control group.The number of migrating cells in the anterior capsule of Plate-loop HydroSmart group, C-loop HydroSmart group, C-compensation-loop Hydrophobic group was 18.80±5.53, 24.67±9.80, and 34.47±10.80, respectively, and the number of migrating cells in the optical area of the posterior capsule of the three groups was 56.43±9.00, 162.20±16.38, and 121.30±12.01, respectively.The cell migration inhibition rate in the anterior capsule of Plate-loop HydroSmart group, C-loop HydroSmart group, C-compensation-loop Hydrophobic group was (92.02±1.94)%, (89.76±3.10)%, (86.27±4.54)%, respectively, and the cell migration inhibition rate in optical area of the posterior capsule of the three groups was (91.60±3.65)%, (70.14±5.35)%, (78.43±3.48)%, respectively.The number of migrating cells in the anterior capsule was lower and the cell migration rate inhibition was higher in Plate-loop HydroSmart group than C-compensation-loop Hydrophobic group, with significant differences (both at P<0.05). The number of migrating cells in the optical area of the posterior capsule and the cell migration inhibition rate was greater than those of C-loop HydroSmart group and C-compensation-loop Hydrophobic group, showing statistically significant differences (all at P<0.001). Conclusions:The in vitro capsular bag model can be used in PCO research.Compared with C-loop HydroSmart IOL and C-compensation-loop Hydrophobic IOL, Plate-loop HydroSmart IOL can more effectively inhibit the migration of LECs to the optical area of the posterior capsule.