Objective: To study optimum inclusion process conditions for volatile oils from Herba Schizonepetae and Pericarpium Citri Reticulatae Methods : The grinding method was compared with the saturated water solution method by determining the ultilization ratio of volatile oil from both herbs. The optimum preparation conditions were investigated by the orthogonal design. The quality of inclusion compound of volatile oil before and after being included were analyzed by TLC. Results : The optimum preparation conditions for inclusion were established as follow ultilization ?-CD and oil was 8∶ 1, The proportion of water and ?-cyclodextrin was 4∶1 (g:mL), and inclusing time up to three hours, The ultilization ratio of oil was 87.1% . Conclusion : The grinding method is better than the saturated water solution method. The method is applicable.

Objective To study the elongation factor 1α(EF-1α) gene functions in prostate cancer cell line DU145 in the aspects of cell proliferation and clone formation by using the RNA interference technique. Methods DU145 cell lines were divided into control group, transfection control group transfected with scramble siRNA and experimental group transfected with EF-1α siRNA. After transfecting EF-1α siRNA into DU145 cell line, the down-regulation of EF-la expression in DU145 cell line was confirmed by Western blotting and immunofluorescence staining. Then, the cell proliferation and clone formation assays were carried on in these 3 groups of DU145 cells. Results Compared with controls, the specific down-regulation of EF-1α expression was achieved in experimental group only. Compared with control group, after the down-regualtion of EF-1α in DU145 cell line, the cell proliferation rate decreased from day 4 to day 7 after transfection by 45. 9%, 53. 5% , 35. 3% and 38. 1% , respectively(P<0. 05). The clone formation number in experimental group decreased by 67.0% (P<0. 01). Conclusions The down-regulation of EF-1α has a negative impact on prostate cancer cell proliferation and clone formation. EF-1α might be an appropiate targeting gene in prostate cancer targeting therapy.

Objective To study the effect of down-rdgulation of EF-1 alpha gene in prostate cancer cell line DU-145 on cancer cell migration and invasion by using RNA interference technique. Methods The prostate cancer cell line DU-145 was divided into three groups: the control group (untransfected with siRNA), randomly control group (randomly transfected with siRNA) and experimental group (transfected with EF-1 alpha siRNA). Localization of EF-1 alpha and its relationship with F-actin in cytoplasm were analyzed by immunofluorescence technique. Cancer cell migration and invasion capability of DU145 cells were studied by transwell technique in these three groups. Results EF-1 alpha expression in DU145 cell line was down-regulated by using RNA interference technique. EF-1 alpha was localized in cytoplasm and co-located with F-actin. The down-regualtion of EF-1 alpha did not change the F-actin distribution in cytoplasm. The cell migration and invasion study showed that after seeding 20×104 DU145 cells into the upper chamber of transwall for 12 hours, the cells collected in the lower chambers were (10.6±1.0)×104 in control group, (11.2±0.8)×104 in randomly control group and (3.9±0.6)×104 in experimental group. Compared with controls, the cancer cell migration and invasion capability was significantly inhibited to only 37.1% (t= 13.9, P<0.05) after the specific down-regulation of EF-1 alpha expression in DU145 cells. Conclusions The down-regulation of EF-1 alpha expression has negative impacts on prostate cancer cell migration and invasion. EF-1 alpha plays important roles in prostate cancer local invasion.

Objective:To construct the Helicobacter pylori infected C57BL/6 mice model to observe the activation of NOD1/NF-κB signaling pathways in the gastric tissues,and study its roles in inflammatory response during Hp infection.Methods:6-8 week-old C57BL/6 mice were randomly divided into two groups,the Hp infection group and the control group,and mice were given by gavage every 48 h for five times with Hp or PBS,respectively.All the animals were sacrificed at different time point and the gastric tissue were stained with hematoxylin-eosin( HE);The mRNA expression of NOD1 and RIP2 in gastric tissues were examined by RT-PCR;Levels of IFN-βand IP-10 in mice serum were assessed by ELISA;Nuclear translocation of p65 in gastric tissue was detected by Western blot.Results:Hp infection elicits an inflammatory cell response,glands in gastric tissue were reduced or atrophic,as compared with that in the control group.The levels of IP-10 and IFN-βincreased in the model group, and peaked at 16 weeks after Hp infection.Hp infection increased the mRNA expression of NOD1 and the p65 content in nuclear between 24-120 h(P<0.05),and the highest level at 48 h,subsequently the expression levels were began to decrease.The mRNA expression level of RIP2 was up-regulated after Hp was administrated, peaked at 48 h and declined after 72 h.However, the expression levels would rise again at 120 h.Conclusion: Hp infection can activate the NOD1/NF-κB signaling pathways and induce the production of IFN-βand IP-10 in gastrics of mice.

ObjectiveTo evaluate in vivo antithrombosis property of optimized FW-Ⅱ axial blood pump and provides evidence for future clinical use.MethodsA left ventricle-pump-descending aorta bypass model was established in five healthy sheep (60-70 kg) and the circulation of these sheep was assisted by FW-Ⅱ axial blood pump for 2 weeks.In preoperative and postoperative day 1,2,3,7,10 and 14,blood was drawn from the jugular vein to examine platelet activation and leukocyte-platelet aggregation respectively quantified with Annexin V,CD41/61 and CD14-PE by flow cytometry assays.Immediately after termination of the experiment,FW-Ⅱ axial blood pumps were explanted and each part was inspected for thrombus formation.Macroscopic and histological examinations were checked on heart,brain,kidney and spleen,respectively for thrombosis.ResultsCompared with preoperative baseline,the number of platelet activation and leukocyte-platelet aggregation reached a peak at postoperative day 2,it retained a high level within 7 days,then gradually decreased,but was still higher than preoperative level at dayl4.According to rotating speed,the number of platelet activation and platelet-leukocyte aggregation were lowest at the speed of 8000 r/min Minus thrombus were found in the front and rear hub of the pump rotor,and there was no thrombus at other components (flow straighter,impeller and pump housing).There were no ischemia and infarction evidences in macroscopic and histological examination of the heart,brain,kidney and spleen.ConclusionFW-II axial blood pump can be used to assist left ventricular circulation for 2 weeks with a satisfactory antithrombosis property.The level of platelet activation and leukocyte-platelet aggregation can be reduced to a lowest level at an optimized pump rotating speed.

Objective To discuss the role of assistance of intraoperative uitrasonography in ret-roperitoneal laparoscopic nephron-sparing surgery for renal tumors. Methods The intraoperative laparoscopic ultrasonography was applied in retroperitoneal nephron-sparing surgery for 20 patients, of whom 11 were men and 9 were women. The average age was 53(range 33 to 73) years. There were 12 patients with renal cell carcinoma and the mean tumor size was 2.9(range 1.4 to 4.6)cm in diame-ter. All of them were staged as T1 N0 M0. Seven patients had angiomyolipoma and the mean tumor size was 4.5(range 1.8 to 8.0)cm in diameter and 1 patient had a 3.1 cm oncocytoma in diameter. The ul-trasonography was used to locate the tumor, observe the bloodstream and detect whether there were small satellite tumors. The surgical time, time of renal artery occlusion and operative effect were ob-served. Results Laparoscopic surgery was successful in all cases without conversion to open surger-y. Mean operative time was 115 (range 85 to 270) min, mean time of renal artery occlusion was 28 (range 22 to 50) min. During the mean followup of 16(range 4 to 30) months, no patients with renal cell carcinoma had local or port site recurrence or metastatic disease. Conclusion In retroperitoneal laparoscopic nephron-sparing surgery, the intraoperative uhrasonography is helpful to locate the tumor in the surgery, to estimate whether the renal artery is occluded completely and to excise the tumor pre-cisely.

This study aimed at identifying pig-hide gelatin from tortoise shell glue using ultra-performance liquid chromatography-electrospray-time of flight mass spectrometry(UPLC-ESI-QTOF-MS/MS).The samples of tortoise shell glue were digested by trypsin before the analysis on UPLC coupled with QTOF mass spectrometry.The ions at m/z 925.4 were selected as the marker peptides of pig-hide gelatin.It was found that the test results of the ions at m/z 925.4 were positive in the samples with low contents of the marker peptides of tortoise shell glue,indicating the existence of pig-hide gelatin.However,the result of bovine-hide gelatinordonkey-hide gelatin was negative,while negative results were detected in tortoiseglue standard.The finding was in conformity with previous references over the fragmentation regularity of MS/MS.In conclusion,the method can be adopted to identifying the pig-hide gelatin from tortoise shell glue and applied to the quality control of tortoise shell glue with high specificity and sensibility.

This study aimed at developing an ultra-performance liquid chromatography electrospray time of flight-mass spectrometry (UPLC-ESI-QTOF-MS) method for the identification of bovine hide gelatine from the shell glue of the red-eared slider.Tortoise shell glue was made from the shell of red-eared slider or Reeves' turtle (Chinemys reevesii).The samples were analyzed on UPLC-ESI-QTOF-MS after being digested by trypsin,and the ions at m/z 604.8,m/z 641.3,m/z 790.8 of the marker peptides of bovine hide gelatin were detected.As a result,it was found that the test results of the ions made from the shell of red-eared slider at m/z 641.3 were positive,taking the same fragmentation regularity of MS/MS as the standard of bovine-hide gelatine,while negative results were detected from the ions made from the shell of Reeves' turtle.In conclusion,the method was simple and accurate provided a basis for distinguishing the tortoise shell glue from bovine hide,which could be also used for the quality control of tortoise shell glue.

Objective To investigate the different radioprotective effects of total flavonoids of Astragalus (TFA) on human normal mesenchymal stem cells(hMSCs) and hepatoma cells injured by 60 Coγ-ray radiation.Methods hMSCs and HepG-2 cells were cultured and randomly divided into TFA-treated and untreated groups.The cells of different groups were irradiated with 60 Co γ-rays at the dose of 6 Gy.MTT method was utilized to detect the survival rates of the hMSCs and HepG-2 cells pretreated or untreated with TFA before irradiation.Cell clone formation test was used to measure the cellular radiosensitivity.The apoptosis rates of different groups were determined by flow cytometer assay.The expression rates of the apoptosis-promoting proteins Fas and Bax and the apoptosis-inhibiting protein Bcl-2 were analyzed by Western blotting.Results MTT showed that the survival rates of hMSCs pretreated by TFA were 1.15-1.95 times higher than that of the pure irradiation group.On the contrary,the survival rates of the TFA pretreated HepG-2 cells were only 0.53-0.23 times that of the pure irradiation group.There was a good dose-effect relationship between the cell survival rate and the TFA concentration.Cell clone formation rate indicated that combined treatment of TFA and radiation inhibited the cell proliferation more effectively than single TFA or pure radiation.Flow cytometry showed that 6,24 and,48 h post-irradiation to 6 Gy,the apoptosis rates of the hMSCs were 23.3% ,11.2% ,and 2.9% ,respectively in the TFA pretreated group and were 29.3% ,24.9% ,and 13.6% in the pure radiation group.However,the apoptosis rates of the HepG-2 cells at 6,24,and 48 h post-irradiation to 6 Gy were 11.6% ,17.3% ,and 20.1% ,respectively in the TFA pretreated group and were 6.9% ,9.3% ,and 15.8% ,respectively in the direct radiation group.Western blotting showed that the expression levels of Fas and Bax proteins in the HepG-2 cells were significantly higher in the TFA pretreated group than in the pure radiation group.On the contrary,the expression level of the apoptosis inhibiting protein Bcl-2 was significantly lower in the TFA pretreated group than in the pure radiation group.Conclusions TFA has obvious effects of radiological protection on human hMSCs and has no effects of radiological protection but effects of apoptosis enhancement on hepatoma cells.The promotion of apoptosis of TFA on hepatoma cells is primarily through increasing the expression of apoptotic proteins such as Fas and Bax and reducing the expression of anti-apoptotic protein Bcl-2.

Objective:To determine the equilibrium solubility and the apparent oil/water partition coefficients of nicotinate-curcu-min ester,so as to provide basis for the new formula design. Methods:Nicotinate-curcumin ester was dissolved in buffer solution with pH of 1.2-7.8. HPLC was used to detect the equilibrium solubility of nicotinate-curcumin ester in various solutions. The apparent oil/water partition coefficients in octanol-water and octanol-buffer solution system were measured by a shaking-flask method. Results:The solubility of nicotinate-curcumin ester in pH 6.8 was the highest. The apparent oil-water partition coefficients of nicotinate-curcumin ester in pH 1.2,5.8,6.5 and 7.8 were lgPapvalues within the range(lgPap=1.69-1.98) beneficial to the absorption in vivo. But in pH 2,5 and 6.8,the lgPapvalues were greater than 2 with strong lipophilicity. Conclusion:The equilibrium solubility as well as ap-parent oil/water partition coefficients of nicotinate-curcumin ester is greatly influenced by the pH value of media. In the pH value with relatively high solubility,lipophilicity is stronger,suggesting it is difficult to be absorbed by the body and needs to be further studied on dosage forms.