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.

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.

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 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 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 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 screen and isolate the radioresistance related genes of IRM-2 mice.Methods cDNA library of IRM-2 mice was constructed by SMART technique.Total RNA was isolated from spleens of IRM-2 male mice.The first-strand cDNA was synthesized by using PowerScript reverse transeriptase,and double-strand cDNA was synthesized and amplified by long PCR.The PCR products were purified,digested with restriction enzyme Sfi I.The ds-cDNA fragment lessthan 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector.The ligation mixture was transformed into E.coil DH5α by electroporution transformation to generate the unamplified cDNA library.The quality of cDNA library was identified by PCR technique.130 clones from cDNA library were sequenced and compared with GenBank database.Results The cDNA library contained 2.25 x 106 independent clones with an average insert size of 1.2 kb.The ratio of recombination and full-length was 95% and 55%,respectively.21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database,with registered number DW474856-DW474876.Conclusions cDNA library of IRM-2 mice has been constructed successfully.21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice,which will lay a foundation for isolating and identifying radioresistance related genes in further study.