OBJECTIVE: To study the feasibility of using biodegradable chitosan nanoparticles as new type carriers for controlled GM-CSF delivery system. METHODS: GM-CSF and BSA loaded chitosan nanoparticles were prepared by ion cross linking method using tripolyphosphate as cross-linking agent with its morphology observed by transmission electron microscope. The encapsulating efficiencies of GM-CSF and BSA were determined respectively by fluorescence scanning and ultraviolet spectrophotometer. The in vitro drug release properties of chitosan nanoparticles were studied as well. RESULTS: Morphologically, the nanoparticles were spherical with mean particle size of 201nm. The encapsulating efficiencies of GM-CSF and BSA were 62.1% and 58.5%, respectively, and their in vitro accumulative drug release rates on the 3rd day were 69% and 82% respectively. CONCLUSION: The ion cross linking method is fit for the preparation of GM-CSF loaded chitosan nanoparticles.

OBJECTIVE: To prepare 5-fluorouracil(5-Fu) microemulsion and to investigate the intestinal absorption of 5-Fu microemulsion in rats. METHODS: The formulation of 5-Fu microemulsion was optimzied using pseudo-ternary phase diagrams with isopropyl myristate (IPM) as oil phase and ODO-L as surfactant; the intestinal absorption ex vivo model was prepared using everted intestinal sac method to investigate the absorption sites and absorption efficiency of 5-Fu microemulsion in rats. RESULTS: IPM/ODO-L/EtOH/H2O (Km=1∶2) was selected as the delivery system of 5-Fu; as compared with 5-Fu solution, the intestinal absorption of 5-Fu microemulsion was significantly improved and the middle and lower segments of rat's intestine were the best sbsorption sites for 5-Fu microemulsion and the accumulative aborption rates of 5-Fu microemulsion at 90 min was 3-fold that of 5-Fu solution. CONCLUSION: The prepared 5-Fu microemulsion is stable in quality and well-absorbed in rats' intestine.

Objective In order to study the application of macroporous resins and so on to the purified active components of Euphorbia humifusa、Leaves of Flos Lonicerae、Chrysanthemum morifolium,adsorption and separation properties for 3 types of macroporous resins and polyamide were investigated.Methods The total flavone was used as the evaluating criteria,we selected suitable macroporous resins and studied optimum technological parameters of the adsorption and elution.Spectrophotometry was used for the determination of total flavone.Results The suitable macroporous resins which were used to the purified active components of traditional Chinese medicine were D101 and DA201 and DM301 for Euphorbia humifusa、DA201 for Leaves of Flos Lonicerae、D101 and DA201 and DM301 for Chrysanthemum morifolium,The concentration of the sample of Euphorbia humifusa for DA201 and D101 were 0.49～1.47 and 0.42～1.31 mg/ml.The concentration of the sample of Leaves of Flos Lonicerae for DA201 was 1.03～2.07 mg/ml.The concentration of the sample of Chrysanthemum morifolium for DA201 and D101 was 0.50～1.00 and 0.71～1.99 mg.ml.In the adsorption course,appeared leaking were 8 and 10、2、2 and 1 BV respectively.In the elution course,when the alcohol concentrations were 20%、30%、40% and 20%、30%、40%;10%、20%、30%;30%、40%、50% and 20%、30%、40%;respectively,the total flavone content in the elution solutions was higher.The influence of temperature to DA201 and D101 adsorpting total flavone for Euphorbia humifusa was not great.But the influence of temperature to DA201 and D101 adsorpting total flavone for Leaves of Flos Lonicerae and Chrysanthemum morifolium were certain degree.Conclusions It is obviously different to refine the total flavone active components of traditional Chinese medicine,while using 3 types of macroporous resins and polyamide.

Combined Modality Therapy ; Congresses as Topic ; Head and Neck Neoplasms ; therapy ; Humans

Objective To isolate,cultivate and identify the human embryonic fibroblasts(hEFs) derived from the gonadal ridges and dorsal mesenteries of human embryos,and to detect the expression by hEFs of cytokines crucial for the growth of human embryonic germ cells(hEG) in vitro.Methods The hEFs were isolated by enzyme digestion from gonadal ridges and dorsal mesenteries of 5-to 9-week old human embryos.The cells were then cultivated.The biologic characteristics(morphology,growth characteristics and cell cycle) of these cells were also studied.The reverse transcription-polymerase chain reaction(RT-PCR) was used to seek the expressions of a specific fibroblast marker,prolyl 4-hydroxylase ?,and a specific marker of epithelial cells,cytokeratin-4,in the cells.The expressions of cytokines essential for the growth of hEG,namely basic fibroblast growth factor(bFGF) and leukemia inhibitory factor(LIF),were also examined by RT-PCR.Results The hEFs were successfully isolated and cultivated from gonadal ridges and dorsal mesenteries of human embryos.They could be passaged beyond the 25th generation.The biologic characteristics of the cells did not change,even in high-passage cells or frozen-thawed cells.The cells expressed prolyl 4-hydroxylase ?, but not cytokeratin-4,which was similar to the fibroblasts.The cultured cells expressed bFGF and LIF.Conclusion The hEFs derived from gonadal ridges and dorsal mesenteries of human embryos are successfully isolated and cultivated,and the cells express the cytokines essential for the growth of hEG in vitro.

Objective:To explore the effects of different types of music on Caspase-3 and Livin expression levels of the hippocampus of brain tissue of aged rats.Methods: Healthly elderly male Wistar rats 40 were randomly divided into :control group , Mozart music group ,Liangzhu music group and rock music group ,n=10;water maze training every morning while music intervention 2 h,afternoon pure music intervention 2 h,continuous 7 d,the eighth days ,the rats were sacrificed and taking blood and brain tissue ,by ELISA in serum and brain tissue Caspase-3 and Livin protein contents change;using immunohistochemistry assay in rat brain tissue Caspase-3 and Livin protein levels;real-time PCR detection of rat brain tissue changes in gene level of Caspase-3 and Livin.Results:Compared with the control group ,Mozart music group ,Liangzhu music rats the protein content ,protein level and gene level of Caspase-3 of serum and brain tissue were lower statistically significant ( P<0.01 ) , Livin was increased there was statistically significant ( P<0.01 ) ,Mozart music group and Liangzhu music group were no significant difference ( P>0.05 );while rock music group the protein content,protein level and gene level of Caspase-3 and Livin of brain tissue were not significantly difference (P>0.05).Conclusion:Mozart music group and Liangzhu music group by reducing serum and brain tissue Caspase -3 and increased Livin , to inhibit the apoptosis of the body′s nerve cells,which play a neuroprotective effect;the rock music group in the present study ineffective.

Objective To establish a HPLC method for the determination of kaempferol-7-O-?-D-glucopyranoside in Hosta ventricosa stearn.Methods HPLC ana1ysis was carried out on a Phenomenex Luna C18(250 mm?4.6 mm,5 ?m) column,gradient elution with the mobile phase of 0.5 % phosphate acid in water-methanol.The detective wavelength was set at 364 nm.Results calibration curve was linear within the range of 0.113 ～ 2.260 ?g for kaempferol-7-O-?-D-glucopyranoside,coefficient correlation being 1.The average recovery was 101.1 %(RSD= 2.58 %,n=6).Conclusion The content of kaempferol-7-O-?-D-glucopyranoside in Hosta ventricosa stearn is 0.2 mg/g(0.02 %) on average.This method is simple,accurate and practical for the determination of kaempferol-7O-?-D-glucopyranoside.

Objective To investigate the Reprimo and hMLH1 gene promoter methylation detection value in the diagnosis of early gastric cancer.Methods Chose patints in Shaanxi Provincial People’s Hospital from September 2013 to April 2014,50 cases of patients with chronic atrophicgastritis with intestinal metaplasia,50 cases of patients with gastricmucosal atypical hyperplasia,50 patients with gastric cancer,endoscopic gastric biopsy samples,and 30 cases of normal gastric mucosa biopsy tissues as control group.Analysis abnormal expression in Reprimo gene and hMLH1 genes promoter methylation,compared the differences.between groups of patients.Results The patients with gastric mucosatissues Reprimo and hMLH1 genes promoter methylation positive rate was significantly higher than that of normal group,the difference wasstatistically signifi-cant (P<0.05).Reprimo gene promoter methylation were:patients with chronic atrophic gastritis with intestinal metaplasia 28% (χ2 =10.18,P < 0.05).Patients with gastric mucosalatypical hyperplasia 56% (χ2 = 25.84,P < 0.05)and patients with gastric cancer 62% (χ2 = 30.36,P < 0.05).hMLH1 gene promoter methylation were:patients with chronic atrophic gastritis with intestinal metaplasia 20% (χ2 =4.39,P <0.05),patients with gastric mucosal atypical hyperplasia 44% (χ2 =15.13,P <0.05)and patients with gastric cancer 48% (χ2 = 17.41,P <0.05),high specificity of detection.Conclusion Reprimo and hMLH1 gene’s detect value in the diagnosis of early gastric cancer is very high,high specificity,it is an effec-tive way of diagnosis,treatment in clinical diagnosis of patients with broad prospects.

Objective To detect the infection of human papillomavirus (HPV)in patients with non-small cell lung cancer (NSCLC)and explore the relationship between HPV infection and clinicopathological features.Methods HPV detection and genotyping in 156 cases of NSCLC were performed using a new liquid chip based on Luminex.Patient clinical characteristics were also recorded,and the relationship between HPV infection and clinicopathological features was studied.Results Of the 156 tumor-DNA samples tested,40 (25.6%)cases showed presence of HPV-DNA,of which 37 cases were of a high-risk HPV type (16,18,33, 58).The differences were statistically significant between the HPV-positive and HPV-negative groups in gender (χ2 =4.387,P =0.036),smoking (χ2 =8.130,P =0.004),histologic type (χ2 =4.075,P =0.044)and lymph node metastasis (χ2 =7.082,P =0.008).The differences were not statistically significant between the HPV-positive and HPV-negative groups in age (χ2 =0.013,P =0.910),differentiated degree (χ2 =1.727, P =0.189),clinical stages (χ2 =0.179,P =0.672),distant metastasis (χ2 =3.012,P =0.083).Logistic regression analysis revealed that lymph node metastasis alone was an independent predictive factor of HPV infection in NSCLC (OR =0.384,95%CI:0.153-0.967,P =0.042),and gender (OR =1.402,95%CI:0.522-3.769,P =0.503),smoking (OR =0.506,95%CI:0.194-1.322,P =0.506),histologic type (OR =0.393,95%CI:0.133-0.161,P =0.091)were not independent predictive factors of HPV infection. Conclusion The infection of HPV presents in part of Chinese NSCLC patients,and HPV infection may be connected with occurrence and development of lung cancer.

Objective Network emulation system constructs a virtual network environment which has the characteristics of controllable and repeatable network conditions. This makes it possible to predict the availability and performance of new protocols and algorithms before deploying to Internet. Methods PARNEM, a parallel discrete event network emulation system described in this paper has the following characteristics: ① BREEN - a BSP based real-time event scheduling engine; ② application transparent flexible interactive mechanism; ③ legacy network model reuse. Conclusion PARNEM allows detailed and accurate study of application behavior. Comprehensive case studies covering bottleneck bandwidth measurement and distributed cooperative web caching system demonstrate that network emulation technology opens a wide range of new opportunities for examining the behavior of applications.