Objective To observe biological character iistcs of cultivated cornea limbal epithelial cells of rabbit.Methods Cultivated cornea limbal epithelial cells with tissue inoculation were observed by inverted microscope every two days; ultrastructure of cell was observated by electron microscope; Proliferative ability of cell was examined by flow cytometry; the cell growth curve was examined by monotetrazolium(MTT) colorimetry.Results Cultivated limbal cornea epithelial cells formed monolayer at six day,the cultivated conea limbal epithelial cells had high metabolism and proliferative ability, growth of cornea limbal epithelial cells reached climax after 4 days by passage culture. Conclusion Cultivated limbal epithelial cells during the first and second growth generation can keep cornea epitheliam property and high proliferative ability.

Objective To review the surgical experience of intrapericardial pneumonectomy for Ⅲ stage lung cancer. Methods 35 patients with Ⅲ stage lung cancer received intrapericardial pneumonectomy from January 1990 to December 2000. Results None of the patients was operative death. The side effect rate was 34.3 %. 1, 3, and 5 years survival rate were 75.2 %, 35.8 %, 21.3 % respectively. Conclusion Intrapericardial pneumonectomy can increase obiousiy resect rate of Ⅲ stage lung cancer which is a kind of safe and dependable surgical operation. It can prolong the life and improve the quality of life. [

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Objective To observe the effect of circadian rhythm on hypnotic median effective dose( ED50) of ketamine. Methods Sixty mice were randomly divided into 4 groups which had 15 mice in each group. They were intraperitoneally injected with ketamine at different times of 2 Am,8 Am,2 Pm and 8 Pm, respectively. Righting reflex was recorded and the value of ED50 was measured with sequential experimental method. Results The hypnotic ED50 of ketamine at 2 Am was(54.57?0.82) mg/kg, with 95% confidence limit of ED50 38.06-78.22 mg/kg;ED50 was(49. 27?0. 12) mg/kg at 8 Am, with 95% confidence limit of ED50 40. 21-60. 37 mg/kg;ED50 at 2 Pm was (42.28?0.21) mg/kg, with 95% confidence limit 37.35 - 47 83 mg/kg;and ED50 at 8 Pm was(57.42?0.14) mg/kg, with 95% confidence limit 37.51-73 72 mg/kg,respectively. The ED50 were significant different at 2 Pm and 8 Pm. However, there were no significant difference in ED50 value among other groups. Conclusion The hypnotic effect of ketamine has circadian rhythm - dependent.

Objective To explore the expression and clinical significance of chemokine receptor 6 (CCR6)/chemokine ligand 20(CCL20) axis in hepatocellular carcinoma (HCC).Methods From March 2003 to December 2005,48 patients with HCC were selected,and one specimen of HCC tissue and one of corresponding adjacent tissue were taken from every patient.Another eight patients with benign liver lesions were selected,and one specimen of surgical sectioned normal liver tissue of each was taken.The relative expression quantity of CCR6 and CCL20 at mRNA level was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR).And the expression of CCR6 and CCL20 at protein level was determined by immunohistochemisty methods.One-way analysis of variance (ANOVA) was performed for comparison among groups of measurement data.Chi-square test was used for rate comparison.The correlation coefficient was calculated by Spearman's method.Survival curves were plotted by Kaplan-Meier method and the survival rate was compared by Log-rank test.Results The relative expression quantity of CCR6/CCL20 at mRNA level in HCC tissues (0.99±0.21 and 0.46± 0.11) were significantly higher than those of para carcinoma tissues (0.33 ± 0.09 and 0.31 ± 0.07) and normal liver tissues (0.22±0.06 and 0.28±0.05),and the differences were statistically significant (F=127.43 and 21.10,both P＜0.05).The positive percentage of CCR6 protein expression in HCC tissues (54.17％,26/48) was significantly higher than that in para carcinoma tissues (16.67％,8/48) and normal liver tissues (0/8),and the difference was statistically significant (x2 =19.55,P＜0.05).There was no statistically significant difference in the positive percentage of CCL20 protein expression among HCC tissues (50.00％,24/48),paracarcinoma tissues (33.33％,16/48),normal liver tissues (2/8) (all P＜0.05).There was correlation between the positive percentage of CCR6 protein expression and that of CCL20 protein expression in HCC tissues (r=0.42,P＜0.05).The positive percentage of CCR6 protein expression was correlated with the pTNM stage of HCC,vascular tumor thrombosis,intrahepatic metastasis and lung metastasis (x2 =5.48,4.02,5.07 and 5.19,all P＜0.05).The positive percentage of CCL20 expression was significantly correlated to tumor maximum diameter and pTNM stage (x2 =4.09 and 4.00,both P＜0.05).Both the disease-free survival (DFS) rate and overall survival (OS) rate of CCR6-positive group were significantly lower than those of negative group (x2 =4.57 and 6.57,both P＜ 0.05).There were no significant differences in DFS rate and OS rate between CCL20-positive group and negative group (both P＞0.05).Conclusion CCR6/CCL20 axis may be related with the malignant behavior and the prognosis of HCC.

BACKGROUND: Recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) could stimulate the proliferation of fibroblasts, keratinocyte and skin mucosae cells to different degrees.OBJECTIVE: To observe the effect of rhGM-CSF on the healing of drug exosmose induced skin ulcers.DESIGN: A randomized and controlled animal experiment.SETTING: Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out at the Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University of Chinese PLA from June to November 2004. Totally 20 male Kunming white mice, with body mass of 18 to 24 g, were chosen.METHODS: Prepared skin ulcers animal models were randomly divided into control group and treated group with 10 white mice in each group.Mice in the control group were given 1mg phentolamine ,20 mg lidocaine and 1mg dexamethasone diluted by normal saline to 0.5 mL ,then sealed up , once a day for 7 days; 25 μg rhGM-CSF was diluted by normal saline to 0.5 mL , then the solution was injected into the periphery of ulcers of mice in treated group , once every other day, for 7 days. Healing time and histological change of skin tissue at ulcer were observed.MAIN OUTCOME MEASURES: To observe the effect of rhGM-CSF on the healing time of drug exosmose induced skin ulcer and anabrosis and histological changeRESULTS: Totally 20 mice entered the stage of result analysis. ①Healing time: the healing time of ulcer and erosion was significantly longer in control group than in treated group [(20-24,8-12)d,t=2.264,P=0.01];②Histological observation: hyperplasia of granulation tissue was not obviously on 7 days after treatment in control group; Hyperplasia of granulation tissue appeared and the newly born blood vessel was abundant on 7 days after treatment in the treated group.CONCLUSION: rhGM-CSF can promote the wound healing of drug induced anabrosis and ulcer.

AIM: To obtain a vaccine with sLAG-3 as immunoadjuvant and investigate its biologic activity in order to establish the safe and effective way for asthma as one of the specific immunotherapy.METHODS: The coding sequence of LAG-3 was amplified by polymerase chain reaction,the expression vector pcDNA-sLAG-3-Ig was constructed by inserting the PCR products of sLAG-3 and Fc sequence of IgG.With electroporation transfection,pcDNA-sLAG-3-Ig was transfected into COS-7 cells and its biologic activity was investigated by Western blotting analysis.RESULTS: By temperature induction,the LAG-3-Ig was highly expressed in E.coli DH5?.LAG-3-Ig fusion protein was observed by SDS-PAGE and Western blotting,the results showed that the LAG-3-Ig protein was an antagonist of the IL-4-induced synthesis of IgE in B cells.CONCLUSION: A new vaccine with sLAG-3 as immunoadjuvant was obtained.It could inhibit synthesis of IgE in B cells.Thus,LAG-3-Ig would be hopeful to establish the safe and effective way for asthma as one of the specific immunotherapy.

Objectives To analyze prognosis and risk factors of Barcelona Clinical Liver Cancer (BCLC) stage B hepatocellular carcinoma patients treated with hepatectomy.Methods Clinical data of 162 BCLC stage B patients who underwent hepatectomy at Tianjin Medical University Cancer Institute & Hospital and the Second Hospital of Tianjin Medical University from June 2007 to December 2013 were retrospectively studied.The correlations between factors (age,gender) and long-term outcome were analyzed to determine independent risk factors.Subsequently,subgroup analysis of BCLC stage B hepatocellular carcinoma was performed.Results Multiple tumors,maximum tumor diameter ＞ 10 cm and AFP ＞ 100 μg/L were con firmed as independent risk factors of overall survival in postoperative BCLC B patients.Based on the risk factors,patients were divided into two groups,namely low-risk subgroup (≤ 1 risk factor) and high-risk subgroup (≥ 2 risk factors).In low-risk subgroup,1,3 and 5-year overall survival (OS) rates were 91.6％,65.5％,61.9％ respectively,and mean OS was 72 months.By contrast,1,3 and 5-year OS rates in high-risk subgroup were 67.4％,25.6％,10.8％ respectively,and mean OS was 29 months.Further more,1,3 and 5-year OS rates of all patients were 85.2％,54.9％,48.0％ respectively,and mean OS was 61 months.Conclusions Relatively favorable long-term outcomes could be yielded in BCLC stage B hepatocellular carcinoma patients treated with liver resection.The independent risk factors including multiple tumors,maximum tumor diameter ＞ 10 cm and AFP ＞ 100 μg/L were closely correlated with overall survival.BCLC stage B hepatocellular carcinoma patients could be divided into low-risk and high-risk subgroups based on the risk factors mentioned above.Survival rates in low-risk subgroup are remarkably superior to those in high-risk subgroup.

Three-dimensional structure model of azoreductase AZR of Rhodobacter sphaeroides was con- structed using homology modeling method. It is a flavodoxin adopting ?/? structure. Structure alignment of two different types of flavin-dependent azoreductases revealed that they possessed high similarity. Based on sequence and structure analysis, site-directed mutagenesis of K109H and K109A were performed. The opti- mal pH values are pH 6 and pH 9 for K109H and K109A mutant protein, respectively. The optimal tempera- ture (30℃) is not affected by mutagenesis. Positively charged residues at position 109 is necessary for the binding of methyl red, while K109H is not a conserved mutagenesis for the binding of NADPH. K109 may only be involved in the binding of the 2’-phosphate group of NADPH and have no effect on the binding of NADH.

Adolescent ; Adult ; China ; epidemiology ; Female ; Hepatitis B ; epidemiology ; prevention & control ; Hepatitis B Vaccines ; immunology ; Humans ; Incidence ; Male ; Vaccination