OBJECTIVE:To find an efficient avenue to the solution of problems existed in GSP authentication so as to provide references for the concerned governmental departments in the process of working out further laws and regulations and policies.METHODS:Practical problems existed in the GSP authentication of drug supply enterprises in Shannxi province were analyzed by the on the spot research method.RESULTS&CONCLUSION:Only through multi-sectional cooperation,further improving personnel quality of different kinds of people,clarifying each one's duty and regulating different management sys-tem,can GSP be put into practice successfully.

BACKGROUND:Studies have found that erythropoietin has a protective effect on embryonic development of the retina, but there are rare studies concerning the embryonic development of the lens. OBJECTIVE: To observe the difference in erythropoietin expression in the lens from Wistar rats at different embryonic periods and to investigate the effect and significance of erythropoietin in the embryonic development of the lens. METHODS:Clean Wistar rats with pregnancy for 10 days (n=5), 12 days (n=5), 14 days (n=5), 16 days (n=5), 18 days (n=5), 20 days (n=5) were randomly colected and divided into six groups. Every two embryonic ratsof the different 30 pregnant rats were obtained randomly by the caesarean operation under ketamine-induced anesthesia. The eye tissues of al the embryonic rats were isolated and cut into sections. The expression of erythropoietin protein and mRNA in rat lens was detected by immunohistochemistry and reverse t ranscription-PCR, respectively. RESULTS AND CONCLUSION:Erythropoietin was distinctly expressed at the six different embryo stages, and the expression of erythropoietin protein and mRNA gradualy increased from embryonic day 10 to embryonic day 16, and decreased from embryonic day 10 to embryonic day 20. There were significant differences between the six groups. These findings indicate that the expression of embryonic appears in a low to high to low fashion during the embryonic development of Wistar rats, which may be closely associated with the developing procedure of lens.

Objective To observe the effect of different concentration netrin-1 on retinal vascular permeability in diabetes mellitus (DM) rats.Methods Eighty adult Sprague-Dawley rats were randomly divided into 8 groups,10 rats in each group,including normal control group (group A),normal+balanced salt solution (BSS) group (group B),normal+netrin-1 (500 μg/ml) group (group C) and DM group (50 rats in 5 subgroups).DM rats were induced by intraperitoneal injection of streptozocin.Three months after intraperitoneal injection,10 DM rats in the control group were injected with BSS (group D).Forty DM rats were injected with 5 μl of different concentrate netrin-1,and were divided into DM+netrin-1 10 μg/ml group (group E),DM+netrin-1 50 μg/ml group (group F),DM+netrin-1 100 μg/ml group (group G),DM+netrin-1 500 μg/ml group (group H)according to the different concentration.Non-DM rats in group C were injected with netrin-1 500 μg/ml.The expression of occludin was determined by immunohistochemistry for protein,and by real-time fluorescence quantitative reverse transcription polymerase chain reaction for mRNA level.Retinal vascular permeability was measured by Evans blue infusion.Results The expression of occludin protein and mRNA in group D were less than group A (t=27.71,8.59;P=0.00,0.00).However,the retinal vascular permeability increased in group D (t=-42.72,P=0.00).The expression of occluding protein,occludin mRNA and retinal vascular permeability showed significant differences between group D,E,F,G and H (F=146.31,16.54,67.77;P=0.00,0.00,0.00).Compared the group B with group C,there was no significant differences between the expression of occludin protein,occludin mRNA and the retinal vascular permeability (t=-1.13,0.93,1.04;P=0.27,0.36,0.31).The concentrate of netrin-1 showed a significant positive correlation to the expression level of occludin and occludin mRNA (r=0.73,0.81;P=0.00,0.00),but negative correlation to the vascular permeability (r=-0.61,P=0.00).Conclusion Netrin-1 can reduce the DM rats' retinal vascular permeability,which depended on the concentration of netrin-1.

OBJECTIVE:To systematically review the effectiveness and safety of methotrexate(MTX)in the treatment of rheuma-toid arthritis,and provide evidence-based reference for clinical treatment. METHODS:Retrieved from Medline,EMBase,Cochrane Library,PubMed,CJFD,CBM,VIP and Wanfang Database,randomized controlled trials(RCT)about MTX in the treatment of rheu-matoid arthritis were collected,Meta-analysis was performed after data extraction and quality evaluation. RESULTS & CONCLU-SIONS:33 effectiveness evaluation were included,involving 8 253 patients,and 53 safety evaluation were included,involving 4 803 patients. Results of analysis show,the efficacy of MTX was not higher than leflunomide in the treatment of rheumatoid arthritis,superi-or to cyclosporine A,and similar to sulfasalazine. In addition,MTX shows similar efficacy with etanercept(7.5-20 mg per week),goli-mumab,adalimumab or rituximab,but less effective than trastuzumab. The incidence of adverse reactions of MTX is high,but mainly mild or moderate,and the most common adverse reactions are gastrointestinal symptoms. Oral administration of MTX is more secure than intramuscular injection and subcutaneous injection.

Objective To investigate the effect of recombinant Vibrio vulnificus hemolysin (rVvhA) on the expression of nitric oxide(NO) and induced nitric oxide synthase(iNOS) in J774A. 1 cells.Methods The inhibitory effect of rVvhA on J774A. 1 proliferation was measured by MTF colorimetry technique. The content of nitrite in culture medium was determined by Griess reagent. The expression of iNOS protein and mRNA were measured by immunofluorescence and RT-PCR, respectively. Results The viability of J774A. 1 cells was apparently inhibited when exposed to 0.8 HU/ml rVvhA and up. With the help of IFN-γ, the expression of NO and the activity of iNOS in J774A. 1 cells were remarkably increased when exposed to 0.4 HU/ml rVvhA. Conclusion rVvhA can increase the expression of NO and iNOS, it may play an important role in the pathogenic mechanism of Vibrio vulnificus.

Objective To investigate the cytolytic activity of extracellular cytolytic toxin rVVC of Vibrio vulnificus on the apoptosis of human ECV304 cells, and to analyze the activities of Caspase-3,-8 and -9. Methods The cytotoxic effect of refolded rVVC on the growth and apoptosis of ECV304 cells was identified by MTT, Hochest33342/PI fluorescent staining, flow cytometry and DNA agarose electrophoresis analysis, respectively. The activities of Caspase-3, -8 and -9 was measured using a colorimetric method. Results The viability of human ECV304 cells exposed to rVVC was inhibited by rVVC after 24 h. 2.0 HU/ml rVVC groups had a better cytotoxic effect to human ECV304 than that of 0.5 HU /ml rVVC groups. The apoptosis of human ECV304 cells in 2.0 HU/ml rVVC+40 μmol/L Z-VAD-FMK groups was relative reduced than that of 2.0 HU/ml of rVVC groups. After 0.5 h treatment with 2.0 HU/ml of rVVC, the Caspase-3 activity in human ECV304 cells increased gradually and reached the peak at 3 h (versus control groups, P<0.01). The activity of Caspase-8 and -9 remained unchanging. Conclusion The rVVC has cytotoxic effect on human ECV304 and the cytolysin is probably correlated with Caspase-3.

Objective To observe the effect of netrin-1 on retinal Müller cells in diabetes mellitus (DM) rats. Methods Fifty Sprague-Dawley rats were randomly divided into the normal control group (group A), normal + balanced salt solution (BSS) group (group B), normal+netrin-1 group (group C), DM+BSS group (group D) and DM+netrin-1 group (group E), with 10 rats in each group. DM rats were induced by intraperitoneal injection of Streptozotocin (60 mg/kg). The expression level of glial fibrillary acidic protein (GFAP) on retinal Müller cells was determined by immunohistochemistry, the level of GFAP mRNA was analyzed by real-time fluorescence quantitative reverse transcription polymerase chain reaction. Results Immunohistochemistry showed that GFAP was distributed in retinal ganglion cells and retinal nerve fiber layer in group A, B and C. Compared to group B, GFAP staining was brighter in the group D. There were significant differences in the expression of GFAP protein and mRNA among groups A-E (F=203.43, 72.91; P=0.00, 0.00), they were higher in group D than group A (t=-26.01, 22.26; P=0.00, 0.00), and group E (t=-10.78, 3.93; P=0.00, 0.00). They were higher in group E than group A (t=7.00, -9.82; P=0.00,0.00). There were no significant differences in between group A and group C (t=-0.29, 0.50; P=0.77, 0.62). Conclusion The expression of GFAP in Müller cells of DM rats could be decreased by injecting netrin-1 into vitreous.

OBJECTIVETo develop a three-dimensional multimedia system for enhancing the efficiency of dental education and chairside communication.

METHODSA set of three-dimensional digital models of normal teeth and jaws related to dental education in prosthodontics were acquired or established under Microsoft Windows. The three-dimensional models were re-edited and rendered with texture attached, producing a large number of three-dimensional pictures and short animated pictures. A software platform was established for displaying all sorts of media, including the three-dimensional models. Finally, all media files produced or gathered before were integrated into the platform, similar to the textbook in chapter adopted in dental education at university.

RESULTSThe prosthodontic three-dimensional multimedia system was successfully developed. The system covered basic information within the current textbook of prosthodontics, three-dimensional pictures, animated pictures, and virtual three-dimensional scenes. The system might serve as an assistant tool in dental education and chairside communication.

CONCLUSIONSIt is technically feasible to establish the prosthodontic three-dimensional multimedia system, according to experiences in this study. The success also anticipates the possibility and feasibility of developing similar systems in other disciplines of dentistry.

Objective To observe the influence of human umbilical cord mesenchymal stem cells (hUCMSC) transplanted into the tail vein of diabetic rats on apoptosis of retinal neurons and the retinal expression level of glial fibrillary acidic protein (GFAP).Methods Seventy clean male Sprague-Dawley rats were randomly divided into the normal control group (group A),diabetes mellitus (DM) only group (group B),DM + balanced salt solution (BSS) group (group C),DM + hUCMSC group (group D),with 10 rats in each group.DM rats were induced by intraperitoneal injection of streptozotocin.Apoptosis of retinal cells was assayed by dUTP nick end labeling.Immunohistochemistry and Western blot was performed to detect the retinal expressions of GFAP in rats.Results Compared with group A,large numbers of apoptotic cells could be found in the retinal ganglion cell layer (GCL) and inner nuclear layer (INL) of group B and group C,however the apoptotic cells in group D were significantly reduced than group B and C.The expression of GFAP was mainly located in the retinal GCL and retinal nerve fibre layer (RNFL) in group A,throughout the inner plexiform layer (IPL) in group B and C,only distributed in RNFL and GCL in group D.It was obvious that the expression of GFAP in group B and C was higher than group A.Compared with group B and C,the expression of GFAP in group D was significantly reduced.The difference of GFAP expression among the 4 groups was significant (F=79.635,P＜0.05).Conclusion hUCMSC could inhibit the apoptosis of retinal cells and activation of glial cells in early DM rats.

Objective To observe the effect of Delta-like ligand 4 (Dll-4) on the pathological structure of retina in early diabetic rats (DM) and its relationship with vascular endothelial growth receptor-2 (VEGFR-2).Methods A total of 70 male Sprague-Dawley rats were randomly divided into normal group and DM group,with 10 and 60 rats in each group,respectively.The rats of DM group was induced by intraperitoneal injection of streptozotocin to established DM model.The rats with blood glucose recovery and death were excluded,and the final 60 rats were included in the statistics.Rats in the normal group were injected with an equal volume of citric acid-sodium citrate buffer.Rats in the DM group were divided into DM 1 month (DM lm) group,DM 2 months (DM 2m) group,DM 3 months (DM 3m) group and DM 3m + Anti group,DM 3m + phosphate buffer solution (PBS) group by random number table method,and 10 rats in each group.In the DM 3m+Anti group,4 μl ofantiDll-4 polyclonal antibody was injected into the vitreous cavity,and the antibody concentration was 0.25 mg/ml.The DM 3m+PBS group was intravitreally injected with an equal volume of PBS.Five days after the injection,the rats were sacrificed.Rats in the DM 3m group and the normal group were not treated,and were sacrificed 3 months after the model was established.The structure and microvascular changes of the retina were observed by hematoxylin-eosin staining,and the total thickness of the retina was measured.The expression of Dll-4 and VEGFR-2 in the retina was detected by immunohistochemistry and fluorescence quantitative polymerase chain reaction (PCR).One-way analysis of variance was used to compare the expression of Dll-4 and VEGFR-2 in the retina of each group.The least significant difference t test was used to compare the two groups.Results Light microscopy showed that the retinal ganglion cells layer in the DM 3m group were obviously edematous,the inner and outer nuclear layers were thinner,the number of cells was reduced,the arrangement was disordered,the edema of outer plexiform layer was obvious,and the microvessels were abnormally dilated.In the DM 3m+Anti group,the edema of outer plexiform layer was lessened than that of the DM 3m group,and the other layers were not significantly different from the DM 3m group.Compared with the normal group,the total retinal thickness of the DM 3m group,the DM 3m+Anti group and the DM 3m+PBS group increased (t=5.596,3.290,4.286;P=0.000,0.008,0.002).Immunohistochemical staining showed that a small amount of Dl14 was positively expressed in the retinal ganglion cell layer of the normal group;a small amount of VEGFR-2 was positively expressed in the ganglion cell layer and the inner and outer nuclear layers.The positive expression of Dll-4 and VEGFR-2 in retinal vascular endothelial cells of DM 3m group increased significantly.The expression of Dll-4 was significantly decreased in the retinal layers and vascular endothelial cells ofDM 3m+Anti group,while the expression of VEGFR-2 was significantly increased.There was no significant difference between the positive expression of Dll4 and VEGFR-2 in the DM 3m+PBS group and the DM 3m group.The results of real-time PCR showed that the relative expression of Dll-4 and VEGFR-2 mRNA in the DM 3m group was significantly higher than that in the normal group (t=6.705,20.871;P＜0.05).Compared with DM 3m group,the relative expression of Dll-4 mRNA in DM 3m+Anti group decreased,and the relative expression of VEGFR-2 mRNA increased (t=2.681,3.639;P＜0.05).The relative expressions of Dll-4 and VEGFR-2 mRNA in the DM 3m+PBS group and DM 3m group were not statistically significant (t=0.513,0.657;P＜0.05).Conclusions The expression of Dll-4 in retinal vascular endothelial cells is gradually increased during the early retinopathy of DM rats.The expression of Dll-4 is inhibited,the expression of VEGFR-2 is up-regulated,and the plexus edema is alleviated.