Objective:To construct a vector that suits the construction of large phage antibody libraries.Methods:Hie phage antibody vector p3MH was modified by insertion of synthesized oligos and PCR mediated site-specific mutations. Hie resultant vector was checked by expression of phage antibody, ELBA and in vivo recombination. Results: Vector pAL was obtained by following modifications of p3MH: insertion of Ioxp511 and loxp sequences, substitution of Lac promoter by Ara promoter, and modification of the cloning site for antibody genes. pAL was proved capable of expressing functional Fab phage antibodies under tighter control. In ere+ bacteria, pAL exhibited loxp-cre mediated recombination. Conclusion: pAL is useful for construction of large phage antibody libraries.

Objective To study the HLA-DR gene frequency in systemic lupus erythematosus (SLE) patients and its relationship with renal involvement and AN As profile. Mefliods HLA-DRB1 genotype frequency assigned by PCR amplification with sequences-specific primers (PCR-SSP) in 89 Chinese patients with SLE and 106 ethnically and geographically matched controls. Results HLA-DR2 and DR9 were significantly more frequent in SLE patient group (0.36/0.20, RR= 2.36, P

To evaluate the clinical value of NMP22 for the diagnosis of urothelial cancer. Urinary NMP22 was determined with enzyme linked immunoaorbent assay (ELISA) in 50 patients in whom 24 patients were suffering from cancer of urothelium, and 20 cases of cancer of other origins, and 6 cases of artificial bladder after total cystectomy for cancer. The median NMP22 value of urothelial cancer was 37 49U/ml, which was significantly higher than those of other patients (4 33U/ml, P

Objective To observe the expressions of senescence-associated biomarkers in fibroblasts after repeated exposures to subtoxic doses of ultraviolet B (UVB), and to study the effect of ginsenoside Rb1 and Rg1 as well as Lycium barbarum polysaccharide on the UVB-induced premature senescence and on the expressions of senescence-associated signals including pl6, p21 and pS3. Methods Skin fibroblasts were classified into 8 groups to receive pretreatment with traditional Chinese medicine (TCM) monomers only, UVB irradiation only, no treatment, or both pretreatment and irradiation. UVB was given successively at a dose of 15 mJ/cm~2 for 10 times, and the concentration of three monomers was 50 mg/L. After 5 days of treatment, light microscopy was used to observe the morphology of fibroblasts, transmission electron microscopy to study the cell ultrastructure,β-galactosidase histochemical staining to detect senescent cells, flow cytometry to analyze cell cycle, and RT-PCR to measure the mRNA expressions of p16, p21 and p53 in these skin fibroblasts. Results None of the 3 monomers had any effect on cell morphology, P-galactosidase activity, cell cycle or the mRNA expression of p53, p21 and pl6 in skin fibroblasts. After UVB irradiation, some changes occurred to cell morphology and ultrastructure; 91.5% of the cells were stained positively for P-galactosidase. The proportion of cells in G1 phase was 88.63% ± 4.67% in irradiated fibroblasts, significantly different from that in untreated controls (49.18% ± 5.53%, P< 0.05) and that in irradiated fibroblasts pretreated with ginsenoside Rbl and Rgl as well as Lycium barbarum polysaccharide (71.04% ± 1.64%, 70.38% ± 2.58%, 80.09% ± 3.46%, all P < 0.05). Compared with untreated fibroblasts, the mRNA expression of p53, p21 and pl6 significantly increased in irradiated fibroblasts (P < 0.05), however, the induced increase in the mRNA expression of pl6 was inhibited by all the three monomers (all P< 0.05), that of p2l by ginsenoside Rb1 and Rg1 (P< 0.05), and that of p53 by ginsenoside Rbl and Lycium barbarum polysaccharide (both P < 0.05). Conclusions Ginsenoside Rbl, Rgl and lycium barbarum polysaccharide can inhibit UVB-induced premature senescence, which may be associated with the down-regulation of mRNA expressions of pl6, p21 and p53.

AIM: To investigate the effect of sodium ferulate on expression of apoptosis-related genes, bcl-2 and bax , in rat lens epithelial cells (LEC) injured by oxidation.METHODS: Eyes of SD rats were divided randomly into four groups: control group, hydrogen peroxide group (H 2O 2), pirenoxine sodium group (PS) and sodium ferulate group (SF). Eyes were excised and lenses were separated under operating microscope and sterilized condition. Lenses were incubated in CO 2 incubator for 24 h with 300 ?mol?L -1 H 2O 2 and with or rithout 5 mmol?L -1 SF. The expression of Bcl-2 and Bax protein of LEC were measured and compared by tearing the LEC anterior capsule via immunohistochemical analysis.RESULTS: (1) There were Bcl-2 and Bax expression in normal lenses of SD rates, Bcl-2 expression was stronger than Bax. (2) Bcl-2 expression decreased and Bax expression increased markedly ( P

AIM: To investigate the effects of Schisandrin B (Sch B) on apoptosis of lens epithelial cells (LEC) treated with H 2O 2. METHODS: Eyes in SD rats were excised and lenses were separated under operating microscope in sterilized condition. Lenses were divided randomly into four groups with different treatment: control group, hydrogen peroxide group (H 2O 2), pirenoxine sodium group (PS) and schisandrin B group (Sch B). Lenses were incubated in CO 2 incubator for 24 h with 300 ?mol?L -1 H 2O 2 and with or without 0 5 mmol?L -1 Sch B. LEC aoptosis and apoptosis rate were measured by TUNEL method. Ultrastructure changes and apoptosis bodies of LEC were observed via transmitted electron microscope. RESULTS: (1) Apoptosis rate in H 2O 2 group (92.0?2.6) was significantly higher than that in control group (3.5?1.8). Apoptosis rate in Sch B group (13.8?3.27) was remarkably lower than that in H 2O 2 group and PS group. (2) Ultrastructure observation indicated that apoptosis cells occurred in most LEC in H 2O 2 group and the changes were severe presenting different stages. While a few apoptosis cells were observed in Sch B group, the changes were slight and most of them were in early and middle stages. CONCLUSION: These data indicated that Sch B significantly inhibited apoptosis of LEC during experimental oxidative injury, the effects were stronger than PS.

Objective To elucidate the correlation between the dose of ultraviolet B (UVB) irradiation and apoptotic phase of keratinocytes (HaCaT cells). Methods Cultured immortalized human ker-atinocyte cell line, HaCaT cells, was irradiated with ultraviolet B (UVB) in doses of 20, 40, 60, 80, 100 and 120 mJ/cm2. Cellular viability, cell cycles and apoptosis were simultaneously detected at 2, 12, 24, 48 and 72 h after irradiation. The caspase-mediated apoptosis was detected by FITC-labelled VAD-FMK, which could irreversibly bind to activated caspases. FITC-Annexin V and PI double staining detected cell membrane-mediated apoptosis. Cell nucleus-mediated apoptosis was detected by DNA ladder electrophoresis. Results Cell cycle analysis revealed that UVB treated HaCaT cells were blocked predominantly in G2/M phase in a dose-dependent manner. Treatment with UVB decreased the viability of HaCaT cells in dose- and time-dependent manners. The rates of apoptosis were increased with increasing dose and prolongation of time after UVB irradiation. The apoptosis of UVB-treated HaCaT presented in an obvious time-dependent manner, in which caspase- and cell membrane-mediated apoptosis was predominantly in early phase and cell nucleus-mediated apoptosis predominantly in late phase. Conclusions The apoptosis of UVB treated HaCaT cells appears in dose- and time-dependent manners, which warrants further detection with multiple parameters and at different phases.

Objective To establish a method for the determi nation of emodin and chrysophanic acid in Compound Rhubarb Spray by HPLC.Methods The ODS column was applied.methanol∶water(87∶13)served as mobile phase.The detectio n wavelength was at 254nm and the flow r ate was 0.65ml /min.Results The average recovery of emodin was 99.48%with relative standard deviation being 1.75%(n=9)and the average recovery of chrysoph anic acid was 101.46%with rela-tive standard deviation being 2.85%(n=9).Conclusion The method was simple,rapid and reli able.It is suitable for the quality control of Compound Rhubarb Spray.

AIM: To optimize the best prescription of micro-emulsions of volatile oil in rhubarb of compound liquid spray by triangular phase diagram METHODS : Poloxamer-108 was used as emulsifiers,absolute alcohol as assistant emulsifiers,volatile oil from Lonicera dasystyla and Curcuma longa as the oil phase,the proportion between the emulsifier and append-emulsifier was established RESULTS : Optimized ratio of Poloxamer-108 and alcohol was 1∶10,the volatile oil in prescription formed micro-emulsions,distributing stably in the spray CONCLUSION : The study of micro-emulsions of volatile oil in compound liquid preparation of traditional Chinese medicine by triangular phase diagram offered a short-cut for resolving its stability. The experiment also suggested that the micro-emulsions could increase solubility of insoluble volatile oil from TCM.

Medical genetics is one of the important basic courses in medical education. The teaching reform in course content, teaching method and experimental teaching was carried out to arouse their enthusiasm in study, cultivate their capabilities of analyzing of medical practice problem.