OBJECTIVE:To study inhibitory effect of Lycianthes biflora polysaccharide on the proliferation of hepatic stellate HSCs-T6 cells in rats and its mechanism. METHODS:After treated with 0(blank control),50,100,200μg/ml L. biflora polysac-charide for 24 and 48 h,the activity of hepatic stellate HSCs-T6 cells in rats was determined by MTT assay and inhibitory rate of cell proliferation was calculated;the content of Hyp in supernatant was detected by immunohistochemical assay. After 48 h,the ex-pression of cellular α-smooth muscle actin(α-SMA)was measured by immunohistochemical assay;both transforming growth fac-tor β1(TGF-β1)and Smad3 were measured by Western blot assay. RESULTS:Compared to blank control,50,100 and 200 μg/ml L. biflora polysaccharide could inhibit the proliferation of HSCs-T6 cells;cell inhibitory rates were 39.84%-69.31% and 45.16%-82.93% respectively after treated for 24 and 48 h,which were positively associated with time and concentration. The con-tents of Hyp in supernatant were 178.36-93.25 μg/ml and 131.94-68.74 μg/ml respectively after treated with different concentrations of PRP for 24 and 48 h,which were negatively associated with time and concentration. The protein level of TGF-β1 and Smad3 de-creased after treated with L. biflora polysaccharide for 48 h(P<0.01). CONCLUSIONS:L. biflora polysaccharide can inhibit the proliferation of hepatic stellate HSCs-T6 cells in rats,and its mechanism is associated with the inactivation of endogenous TGF-βpathway for reducing collagen production.

Objective To explore the relationship among the expression level of MMP?13, AGG and Col?II in the chon?drocytes caused by nutritional deficiencies in rabbits. Methods 30 New Zealand white rabbits were randomly divided into autolo?gous chondrocyte transplantation group (control group, n=10), nutrition block group (surgery group, n=10), and peripheral nutrition block group (sham surgery group, n=10). 4 weeks after treatment, the rabbits were sacrificed for undergoing the observations on general and histological level;real?time PCR assay was employed for testing the expression level of MMP?13, AGG and Col?II;cel?lular apoptosis percentage was observed through TUNEL stain. The relationship among the apoptosis level, cartilage cells histologi?cal Mankin score as well as the expression level of MMP?13, AGG and Col?II were analyzed. Results Based on the Mankin score, there was a statistic difference between surgery group and control group. On the other side, there were no statistic differenc?es between sham surgery group and control group. 4 weeks after treatment, surgery group presented a higher apoptotic percentage compared with control group;this value between sham surgery group and control showed no significant differences. There was an increased mRNA expression level of MMP?13 and a decreased mRNA expression level of AGG and Col?II in surgery group com?pared with control group;no statistic differences of all these values was found between sham surgery group and control group. His?tological Mankin score and apoptotic percentage presented positive correlation (r=0.922, P<0.001), the regression equation:Y=-0.548+0.404X, R2=0.844 (F=157.735, P<0.001); the mRNA expression level of MMP?13 and apoptotic percentage presented positive correlation (r=0.942, P<0.001), the regression equation:Y=0.951+0.116X, R2=0.883 (F=219.054, P<0.001). There was a nega?tive correlation between the mRNA expression level of MMP?13 and the mRNA expression level of AGG as well as Col?II (r=-0.956,-0.945, P<0.001). Conclusion Damage of cartilage cells causes the up?regulation of the MMP?13 expression which could ex?acerbate the degeneration of cells. It could induce the down?regulation of AGG and Col?II mRNA expression, which will cause the extracellular matrix synthesis disorder.

Objective:To investigate the biofilm-forming abilities of pathogens associated with contact lens related microbial keratitis and to compare the efficacies of different treatments in eliminating biofilms in contact lens cases (CLCs).Methods:Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans biolm formation in polypropylene CLCs was examined by using a static biofilm formation model, which was incubated at 37 ℃ for 24 hours.According to the CLC treatment methods, the experimental groups were divided into a control group, a room temperature air-drying group, a contact lens care solution soaking group, a heat-drying group and a soaking-heating group.A pathogen colony counting-based serial dilution micro-counting method was applied to evaluate the biofilm elimination efficacies and pathogen killing rates of treatments.Results:All four tested strains formed biofilms on the inner walls of the CLC, and the biomasses of S. aureus, P. aeruginosa, E.coli and C. albicans biofilm were (10.78±2.12), (9.19±0.57), (8.03±0.30), and (7.50±0.07)lg CFU/ml, respectively.The S. aureus biofilm biomass was significantly higher than those of the other strains (P<0.05). The biofilm biomasses of all the tested strains in the heat-drying and the soaking-heating groups were significantly lower than those in the control group (all at P<0.05); and the biofilm biomasses of S. aureus and E. coli in the soaking group were significantly lower than that in the control group (all at P<0.05). The heat-drying treatment resulted in a killing rate of (51.76±16.75)% for S. aureus, (68.63±4.43)% for P. aeruginosa, (83.51±13.97)% for E. coli, and (97.13±5.19)% for C. albicans, respectively (F=31.806, P<0.001). Significant differences were observed between the killing rates for each bacterium (all at P<0.05). The E. coli and C. albicans killing rates of the soak-heating treatment were (100.00±0.00) % and (97.79±7.67)%, respectively, and were significantly higher than (81.13±14.86)% of S. aureus and (74.22±11.91)% of P. aeruginosa (all at P<0.05).Conclusions:Heating alone or combined with the use of contact lens care solution treatment can improve the pathogen killing rates and effectively eliminate the bacterial and fungal contaminations in CLCs.