OBJECTIVE:To optimize the precipitation process of the extract of Folium Isatidis using chitosan and ZTC1+ 1-Ⅱ natural clarify agents.METHODS:Using the remaining rates of polysaccharide and indirubin and the defecation level as indexes,the optimum process of removing impurity was investigated with single-factor test.RESULTS:The optimum process conditions were as follows:chitosan:precipitating the extract solution which was condensed to 1∶10(herb:the herb solution) with the concentration of chitosan clarify agent at 8% at a heating temperature of 50 ℃;ZTC1+1-Ⅱ:precipitating the extract solution which was condensed to 1∶10(herb:the herb solution) with the concentration of chitosan clarify agent at 5% and the heating temperature at 70 ℃,and the adding sequence of ZTC 1+1-Ⅱ natural clarify agents was clarifier B before clarifier A.CONCLUSION:Chitosan and ZTC1+1-Ⅱ natural clarify agents can be used to effectively remove the impunity of extract of Folium Isatidis.

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.

ObjectiveTo investigate influence on adults'visual function and fluid intelligence with N-back working memory training based on Gabor signal.MethodsControlling group including 13 adults had no training,training group including 14 adults received an eight-days training,half an hour a day.The stimulus was N-back training which has on improved Gabor signal with adjustable spatial frequency and contrast sensitivity.The contrast sensitivity and fluid intelligence were record using OPTEC 6500 and Raven's Standard Progressive Matrices before and after training,then the data was analyzed and processed by SPSS.ResultsContrast sensitivity:there was a siguificant different of the contrast sensitivity between pretest and posttest ( ( 1.93 ± 0.17 ) log,( 1.76 ±0.20 ) log) in training group ( t =-4.579,P =0.001 ).Fluid intelligence:there was a significant different of fluid intelligence between pretest and posttest( ( 129.9 ± 9.0 ) scores,( 113.7 4-16.0 ) scores ) in training group ( t =-4.373,P =0.001 ),and superior to controlling group,which also had a statistical significance (F =1.353,P =0.004).ConclusionThe method of N-back working memory training based on Gabor signal not only enhances working memory and fluid intelligence,but also improves the visual function effectively,and more various effect is acquired comparing to traditional training method.

Objective　To evaluate the diagnosis and surgical treatment of hilar cholangiocarcinoma(H-CC). Methods　Retrospective analysis was made on the clinical feature, surgical treatment and the effect on 73 patients with H-CC. Results　Diagnosis was made in all of the patients preoperatively and the correct diagnostic rate of BUS was 69.9%. In the treatment, radical resection was performed on 15 patients with good results in a short-term period. Of the 43 patients who underwent biliary tract internal drainage or exterrnal drainage, 37 patients had good results in a short-term period, while 6 died after operation. Laparotomy or hepatic artery cannulization with chemotherapy was performed on 15 patients and no change occurred in a short-term period after operation. In 15 cases subjected to radical resection, 11 cases were followed up. The 1,3-year survival rates was 90.9%, 20.0% respectively, but none of the patients survived for over 5 years. In patients undergoing other operations, none survived more than 9 months. Conclusions　It's still difficult to mak early diagnosis of H-CC, which mainly depends on imaging technics. The BUS should be choiced first. Radical resection rate is still low nowadays. The lobus quadratus resection is helpful to select the operation.

Objective To study the curative effect of XRCC2 gene silencing mediated by shRNA combined with radiation on human colonic trans?planted carcinoma in nude mice. Methods Colonic carcinoma T84 cells were transfered into BALB/c nude mice to establish a tumor xenograft mod?el in vivo. Mice were divided into three groups:control,shRNA?SC and shRNA?XRCC2 and exposed to X?ray radiation. The change of volume and weight of the xenografts were examined after receiving radiotherapy and the pathological analysis of tumor tissues were conducted. Results Tumor xenografts transfected with shRNA?XRCC2 in nude mice grew slowly. The xenograft volume in the shRNA?XRCC2 group was decreased significant?ly from day 12 to day 28 after radiotherapy compared with the control group(P<0.01). The xenograft weight in the shRNA?XRCC2 group was small?er than in the control group,with statistically significant difference(t=18.843,P<0.01). The inhibited rate of xenografts in the shRNA?XRCC2 group(56.25%),was markedly higher than that in the shRNA?SC group(4.69%). Pathological analysis of colonic transplanted carcinoma showed that nuclear atypia was not obvious,karyokinesis was decreased and small areas of necrosis were present in tumor xenografts treated with shRNA?XRCC2 transfection. Conclusion XRCC2 gene silencing combined with radiation has significant inhibition effect on colonic transplanted carcino?ma in nude mice.

Objective To evaluate the correlation between homologous recombination repair protein RAD51 and methotrexate-enhanced radiosensitivity.Methods Western blot and RT-PCR assays were used to detect RAD51 expression in HOS osteosarcoma cells exposed to γ-ray irradiation alone and in combination with methotrexate.Colony formation assay was used to test the survival fraction of HOS cells exposed to γ-rays and methotrexate.Results Methotrexate inhibited both protein and RNA expressions of RAD51,and the combination of radiation and methotrexate enhanced the inhibition of RAD51 expression.Moreover,transfection of cells with RAD51 gene decreased cellular sensitivity to methotrexate and γ-rays.The sensitizer enhancerment ratios after irradiation in combination with methotrexate were 1.51 and 0.99,respectively.Methotrenate was a preferred radiosensitizer to HOS cell.Conclusions RAD51 might be involved in the methotrexate-enhanced radiosensitivity.

AIM: To study the effects of ultraviolet (UV) on mitochondrial functions and apoptosis in HaCaT cells.METHODS: After irradiation by UV at low dose(UVA 2 J/cm~2,UVB 10 mJ/cm~2) and high dose(UVA 6 J/cm~2,UVB(30 mJ/cm~2),) HaCaT cells were cultured for 15 hours.Flow cytometry was used to measure mitochondrial membrane potential,mitochondrial mass and apoptotic rate.Annexin V-FITC/PI staining of apoptotic cells was analyzed by laser confocal microscopy.RESULTS: After UV irradiation,cell proportion with low mitochondrial membrane potential increased with irradiation doses.The proportion of control group,low dose group and high dose group were 7.94%?1.02%,25.87%?4.55% and 39.27%?5.32%,respectively.Cells proportion with low mitochondrial mass increased with irradiation doses.The proportion of control group,low dose group and high dose group were 15.19%?1.58%,40.36%?4.41% and 68.79%?5.46%,respectively.The hypodiploid peaks of DNA content analysis represented the apoptotic rate of HaCaT cells.The apoptotic rate of control group,low dose group and high dose group were 1.82%?0.51%,30.16%?5.47% and 58.49%?5.98%,respectively.To analyze the cells apoptosis by staining with annexin V-FITC and PI,the results were consistent with those of DNA content analysis.Cells in control group showed almost no positive staining cells.Single annexin V-FITC positive cells in low dose group and double positive cells in high dose group were predominant,respectively.CONCLUSION: UV irradiation induces HaCaT cell mitochondrial depolarization,as well as mitochondrial mass loss.These changes are related to cell apoptosis.

AIM: To study the changes of mitochondrial membrane potential(△?m) and mitochondrial mass in apoptosis of Jurkat cells induced by camptothecin(CPT).METHODS: Jurkat cells were treated with CPT.Annexin V-FITC/propidium iodine(PI) double stainig was used to detected early stage of apoptosis and PI staining for analyzing the cell cycle.Jurkat cells were stained by annexin V-PE/DiOC_6(3) to detect changes of △?m.The mitochondrial mass was measured by cytometry with NAO staining.RESULTS: 6 h after treated with 10 ?mol/L CPT,the rate of early apoptotic cells(22.59?1.04)% had significantly difference compared with control group(3.93?0.73)%(P0.05).Apoptotic peak appeared obviously after treated with CPT,the percentage of late apoptotic cells(13.58?0.97)% had distinctly difference compared with control group(3.18?0.51)%(P

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.

AIM: To observe the effects of puerarin on blood lipid and expression of aorta laminin B_1 mRNA in streptozotocin-induced diabetic rats. METHODS: Diabetic nephropathy rats were induced by intraperitoneal injection of STZ, and the experimental rats were divided into normal control group, model group, and puerarin group. During and after the treatment for 12 weeks, the general state, blood suger(BS), triglyceride(TC), cholesterol, low density lipoprotein(LDL-C), high density lipoprotein(HDL-C), glycosylated hemoglobin(HbA1c) and glycosylated low density lipoprotein(G-LDL) were detected. Aorta alteration of tissue morphology was observed by H.E staining, and the expressions of laminin B1 mRNA were determined by in situ hybridization analysis. RESULTS: Diabetes mellitus and aorta lesion occurred in the two model groups. Puerarin could improve the general state, decrease the level of triglyceride(P