The 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase activity of Sclerotinia sclerotiorum is one of the multifunctional enzyme AROM activities, which catalyzes a reversible conversion of shikimate 3-phosphate (S3P) and phosphoenolpyruvate (PEP) to EPSP and inorganic phosphate, and is inhibited by the herbicide glyphosate (N-phosphonomethyl glycine). AROM protein has been purified from Sclerotinia sclerotiorum and the EPSP synthase has been analyzed. The results indicated that the optimal pH and temperature of EPSP synthase were 7.2 and 30℃ respectively. The activation energy of the heat-deactivated reaction of the enzyme was found to be 69.62 kJ/mol. Both of the substrates, S3P and PEP, were showed to inhibit the reaction rate when their concentrations exceeded 1 mmol/L and 2 mmol/L respectively. The Km of 140.98 μmol/L for PEP and 139.58 μmol/L for S3P were obtained by Dalziel equation which was a steadystate kinetic equation of the enzymatic reaction with the double substrates. The kinetic pattern of the enzyme was consistent with a sequential mechanism. Inhibition of the EPSP synthase reaction by glyphosate was competitive with respect to PEP, with the Ki 0. 32 μmol/L, and noncompetitive with regard to S3P. Activation by [ K+ ] was observed in the forward reaction. The Km (PEP) was lowered by increasing [ K+ ], while the Km (S3P) changed irregularly and the Ki (PEP) was enhanced.

4 female Wistar rats,weighing 287.5?21.7 gms,about 7～7.5 months old,were used.Using ~3H-TdR and ~(14)C-TdR as labels, double labelling method was employed. The dose of ~3H-TdR used was 1 ?Ci/gm body weight and that of ~(14)C-TdR used was 0.125 ?Ci/gm body weight. Bone marrow smears of both femurs were made and autoradiographs were coated with No. 4 liquid nuclear emulsion.The following results were obtained:In erythropoietic series: Nc/N_H=5.0?1.0, LI=62.3?17.9％, MI=3.4?1.9％. According to the above figures, by calculation, we obtained the cell cycle time and the duration of 4 phases as follows: TG_1≤4.3 hrs,Ts=5.0 hrs, TG_2≥1.0 hr, Tm=0.4 hr and Tc=10.7 hrs.In neutrophilic granulopoietic series: Nc/N_H=3.9?0.9, LI=45.3?5.3％, MI=3.3?1.4％. According to the above figures, the following parameters of the cycle. were obtained as follows: TG_1≤5.6 hrs, Ts=3.9 hrs, TG_2≥1.0 hr, Tm=0.4 hr and Tc=10.9 hrs.There was no obvious difference of Tc between the erythropoietic series, and theneutrophilic granulopoietic series in rats, but significant difference of Tc between them in mice.

By using double labelling methods with ~3H-TdR and ~(14)C-TdR, autoradiographs were prepared in young (40 days old) and in old (16 months old) male LACA mice and in adult (7～7.5 months old) female Wistar rats to observe the cell cycle time and relevant parameters of dorsal skin and both testes.The following results were obtained:1. In young and old mice, duration of cell cycle (Tc) and relevant parameters of epidermis were as follows respectively: labeling index (LI)=1.0 and 1.5％; mitotic index (MI)=0.4 and 0.4%; duration of DNA synthesis (Ts)=3.5 and 4.2 hours; Tc=20.3 and 17.5 days.In adult female rats, they were 3.0%, 0.3%, 4.7 hours and 8.3 days respectively. Tc of rat epidermis was obviously shorter than that of mouse epidermis (P0.50).

Objective:To study the morphologic standard values of craniofacial hard-tissue of the youths in Xi'an.Methods:CBCT scanned cephalometric data of 100 selected volunteers (50 males and 50 females)with individual normal occlusion were collected.31 landmarks and 31 measurements were compared between sexes and between 3D and 2D data with software InvivoDental 5.2,WinCeph 8.0.and SPSS 19.0.Results:1.In the 3D measurements,vertical growth of mandible in the females was more than that in the males. The values of torque of lower incisor,basis length,height of rumi mandibulae and length of corpora mandibulae in the males were bigger than those in the females.2.Compared with 2D measurements,there existed statistically significant differences in most parameters except U1-NA(mm).Conclusion:3D analysis with CBCT may provide more accurite morphologic data for craniofacial hard tissues.

Objective To investigate the protective effect and mechanism of oleanolic acid(OA)on kidneys in rats with type 2 diabetes mellitus(T2DM).Methods A total of 35 Sprague-Dawley(SD)rats were enrolled in this study.25 SD rats were randomly selected to establish T2DM model,after modeling,20 rats remained and divided into T2DM group(n=6),low-dose oleanolic acid group(LOA,n=6)and high-dose oleanolic acid group(HOA,n=8).And ten rats were selected as normal control group(NC,n=10).The biochemical indicators,24 h urine volume and 24 h urinary microalbumin(UAlb)were compared among the four groups.Renal lipid deposition was evaluated by Oil red O staining.The protein expressions of Adenosine 5'-monophosphate-activated protein kinase(AMPK),p-AMPK and peroxisome proliferator-activated receptor γ coactivator-1 α(PGC-1 α)were detected by Western blot.Results Compared with the NC group,the levels of 24 h urine volume,fasting blood glucose(FPG),serum total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),serum creatinine(Scr),serum uric acid(SUA)and 24 hUAlb were increased(P<0.05),while the body weight,high density lipoprotein cholesterol(HDL-C),p-AMPK and PGC-1α were decrease in the T2DM group(P<0.05).Compared with the T2DM group,the expressions of HDL-C,p-AMPK and PGC-1α were increased(P<0.05),while the levels of 24 h urine volume,FPG,TG,TC and LDL-C,Scr,SUA and 24 hUAlb were decreased in the LOA and HOA groups(P<0.05).Compared with LOA group,the expressions of HDL-C,p-AMPK and PGC-1α were increased(P<0.05),while the levels of 24 h urine volume,FPG,TG,TC,LDL-C,Scr,SUA and 24 h UAlb were decreased in the HOA group(P<0.05).Compared with the NC group,a large number of red-stained lipid droplets were deposited in the renal tubular epithelial cells in the T2DM group.Compared with the T2DM group,the lipid droplet deposition was reduced in the LOA and HOA groups,and the improvement was more significant in the HOA group.Conclusion OA can alleviate renal injury in T2DM rats,which may be linked to activation of AMPK/PGC-1α pathway.

AIM:To observe the anti-scarring effects and safety of triamcinolone acetonide(TA)-loaded hydrogel sustained-release sheeting on stab incision glaucoma surgery(SIGS)with “one-step tunnel method” in rabbit eyes.METHODS:A total of 48 healthy New Zealand white rabbits were randomly selected and divided into 4 groups(12 rabbits in each group), trabeculectomy(Trab)group, SIGS group, polyvinyl alcohol hydrogel(PVAH)sheeting was implanted under the conjunctiva flap during SIGS(PVAH group), and hydrogel sustained-release sheeting loaded with TA was implanted under the conjunctiva flap during SIGS(TA/PVAH group). On the 1, 2, 3, and 4 wk after surgery, the intraocular pressure, filtering bubble morphology, anterior chamber reaction, and other complications were observed and recorded in each group. Then animals were euthanized, and the surgery area tissues of right eye were taken for pathological tissue paraffin section. Masson staining, picric acid-Sirius rose red staining, as well as α-smooth muscle actin(α-SMA)and fibroblast growth factor 2(FGF2)immunohistochemistry staining was performed on every section. The infiltration of inflammatory cells, proliferation of fibroblasts and synthesis of type I and type III collagen fibers in local tissues were observed. The average positive area ratio of α-SMA and FGF2 antibody immunohistochemical staining in each group was calculated and compared.RESULTS: The TA/PVAH group maintained diffuse and elevated functional filtering blebs, while flat filtering blebs appeared in Trab, SIGS and PVAH groups at 2 wk after surgery. Functional filtering blebs were present in 1 eye(33%), 2 eyes(67%)in the PVAH and TA/PVAH group at 4 wk after surgery, respectively, while the other filtering blebs were flattened. Masson staining showed that the hydrogels in PVAH and TA/PVAH groups did not degrade at 4 wk after surgery. Compared with the Trab and SIGS groups, the filtration passages were more obvious, with less collagen fiber proliferation. Sirius red staining showed that the expression of type I collagen and type III collagen in the TA/PVAH group was less than that in the Trab group, SIGS group and PVAH group at 4 wk after surgery. Immunohistochemical staining showed that the α-SMA expression in the TA/PVAH group was significantly lower than that in the Trab and SIGS groups at 1 wk after surgery(P<0.01). The α-SMA expression was the highest in the Trab and SIGS groups at 2 wk after surgery, while the α-SMA expression in the PHAP and TA/PVAH groups was significantly lower than that in the first two groups(P<0.01). Compared with the Trab group, the expression of FGF2 in the PVAH and TA/PVAH group was significantly increased at 1, 2, 3 and 4 wk after surgery(P<0.05). Compared with the SIGS group, FGF2 expression in the TA/PVAH group was significantly increased at 4 wk after surgery(P<0.05).CONCLUSION:In SIGS surgery of rabbit eyes, implanting hydrogel sustained-release sheeting loaded with TA under conjunctival flap can effectively inhibit the scarring of the filtering bleb, which may be the interaction of the anti-scar effect of TA and the stent function of hydrogel.