OBJECTIVETo observe the effect of pure total flavonoids from Citrus (PTFC) on the hepatic fatty degeneration, inflammation, oxidative stress and SIRT1/PGC-1alpha expressions in mice with non-alcohol steatohepatitis (NASH), and discuss the action mechanism of PTFC on NASH.

METHODMice were given high-fat diet for 16 weeks to induce the NASH model. Since the seventh week after the model establishment, the mice were intervened with 100, 50 and 25 mg x kg(-1) x d(-1) PTFC for 10 weeks. The pathologic changes in hepatic tissues were observed with HE staining. The contents of TG, CHOL in hepatic tissue, as well as the levels of AST, ALT in serum were detected by using the biochemical process. The expression of SIRT1, PGC-1alpha and MnSOD mRNA in hepatic tissues were detected with Real-time PCR assay. SIRT1, PGC-1alpha protein and 8-OHdG expressions were determined with the immunohistochemical method. The SOD level in hepatic tissues was tested by the xanthine oxidase method. The MDA content in hepatic tissues was examined by the thiobarbituric acid method.

RESULTThe contents of TG, CHOL, NAFLD activity scores and ALT level in serum in hepatic tissues of mice in the model induced by fat-rich diet were obviously higher than that of the normal group (P < 0.010. The SIRT1, PGC-1alpha, MnSOD mRNA and protein expression in hepatic tissues were significantly lower than that of the normal group (P < 0.01). The expression of 8-OHdG and the content of MDA in hepatic tissues were obviously higher than that of the normal group (P < 0.01). After the intervention with different doses of PTFC, the NAFLD activity scores, the content of TG and the level of AST in serum were notably lower than that of the normal group (P < 0.01, P < 0.05); whereas the SIRT1, PGC-1alpha, MnSOD mRNA and protein expression were obviously higher than that of the normal group (P < 0.01, P < 0.05), with the significant decrease in the expression of 8-OHdG and the content of MDA (P < 0.01).

CONCLUSIONOxidative stress/lipid peroxidation enhancement in in NASH mice induced by high-fat diet may be related to the changes in SIRT1/PGC-1alpha signal transduction pathway. PTFC could enhance the anti-oxidant capacity in liver, relieve the damage of reactive oxygen during the fatty acid metabolic process, and prevent NASH from the occurrence and development by regulating the SIRT1/PGC-1alpha signal pathway.

Animals ; Citrus ; chemistry ; Fatty Liver ; drug therapy ; genetics ; metabolism ; Flavonoids ; chemistry ; pharmacology ; Inflammation ; drug therapy ; genetics ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; Oxidative Stress ; drug effects ; genetics ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Sirtuin 1 ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

Three candidate antisense target sites of mouse Fas gene were screened by PARASS (poly-A anchored RNA accessible sites screening) technology. They were target at Fas gene 297nt-317nt, 618nt- 638nt and 662nt-682nt. Antisense oligos (A1, A2 and A3) and DNAzymes (D1, D2, and D3) for every target site were designed and synthesized. In vitro, the validation of the sites were judged by antisense oligos included RNase H splicing and the DNAzyme degradation. The results indicated that A1, A2 and A3 introduced RNase H degradation. DNAzymes D1, D2 and D3 cleaved Fas mRNA effectively. Neither degradation observed in antisense oligo RNase H group in non-target site (1211-1231nt) and 2 bases mismatched of A3, nor splicing occurred in DNzyme group in non-target site ( 1211-1231nt) and 2 bases mismatched of D3. Site 2 and 3 were at the same positions with those of ISIS Pharmaceuticals. The effective antisense oligos and DNAzymes for Fas gene could be used for the research subsequently.

Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Diagnosis, Differential ; Histiocytoma, Benign Fibrous ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neprilysin ; metabolism ; Receptors, Cell Surface ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery ; Thigh ; Vimentin ; metabolism

Aim To observe the effects of the ethyl ace-tate extract from Coreopsis tinctoria on glucose and lipid metabolism and liver, kidney function in diabetic rats. Method By high-sugar, high-fat diet combined with intraperitoneal injection of streptozotocin ( streptozoto-cin, STZ ) Type 2 diabetes SD rat model was estab-lished. Model rats were randomly divided into six groups ( control group, model group, three dose groups Coreopsis tinctoria extract:low, middle,high 0. 15 g· kg-1;0. 3 g·kg-1;0. 6 g·kg-1 , positive drug met-formin 0. 16 g · kg-1 group ) . The control group and the model group were given physiological saline and the remaining groups intragastric administration coreofosis tinctoria extrat. Random blood glucose and body weight of rats were measured weekly. After 4 weeks of admin-istration, The rats were killed and rat serum was col-lected to detect serum lipids ( TC/TG/HDL/LDL ) , liver and renal function indicators, serum insulin, and glycated hemoglobin levels. Result Coreopsis tincto-ria ethyl acetate extract effectively reduced the diabetic rats random blood glucose, glycated hemoglobin,serum triglycerides, LDL, total serum protein, serum creati-nine and uric acid levels, and increased serum white protein content in diabetic rats. Conclusion Coreop-sis tinctoria ethyl acetate extract can reduce blood glu-cose and lipid in diabetic SD rats and protect their liver and kidney function.

Objective To explore the influence of fluorine on mRNA and protein expression of the insulin-like growth factor-1 (IGF-1) and its receptor of rat osteoblasts.Methods Osteoblasts were isolated from rat bone by enzyme digestion.Different fluorine concentration [0 (control),10-7,10-6,10-5,10-4,10-3 mol/L] was add to the second generation osteoblasts.The IGF-1 in the culture medium was determined by radioimmunoassay (RIA) at different fluorine concentration and different time (24,48 h).The expression of IGF-1 receptor was measured by the method of fluorescent quantitation PCR and the expression of protein IGF-1 receptor was measured by Western blotting.Results ①With increased dose of fluoride exposure,IGF-1 concentration in the osteoblastic culture medium increased first and then decreased at 24,48 h,respectively.Compared to the control group [(38.83 ± 3.48)ng/L],IGF-1 concentration of the 24 h 10-6 mol/L group[(65.45 ± 4.84)ng/L] was higher,and the difference was statistically significant(P ＜ 0.05).The same result was also shown in the 48 h 10-5 mol/L group [(59.14 ± 1.53)ng/L] to its corresponding control group [(33.79 ± 1.84)ng/L,P ＜ 0.05].②The mRNA expression of IGF-1 receptor of the 24,48 h 10-5 mol/L groups (0.0055 ± 0.0004,0.0262 ± 0.0040) was significantly higher than their corresponding control groups (0.0022 ± 0.0001,0.0073 ± 0.0008,all P ＜ 0.05).③With increased dose of fluoride exposure,the protein expression of IGF-1 receptor increased first and then decreased ;the expression of 24 h 10-5 mol/L group (1.39 ± 0.16) was compared with the corresponding control group (0.86 ±0.12),and the difference was statistically significant (P ＜ 0.05) ; the expression of 48 h every fluorine group was also compared with the corresponding control group,and the difference was not statistically significant(all P＞ 0.05).Conclusions Fluorine can affect the mRNA and protein expression of osteoblastic IGF-1 and its receptor.It indicates that IGFS signal transduction pathways play an important role in fluorine regulation of bone metabolism.

6-11 years old were 9.67%, 6.81%, 3.49% and 0.80%, respectively.Furthermore, the infection rates between each two age stages were significantly different(Pa0.05).4.Infection rates in 2005,2006 and 2007 were 4.0%, 8.92%, 8.85%,respectively.Infection rates between 2005 and 2006,2007 were significantly different(Pa

Objective To study the different ultrasonic features in patients of polycystic ovary syndrome (PCOS) with or without obesity based on body mass index (BMI), and to investigate whether certain hormonal factors correlate with ovarian morphology and blood flow, and to discuss the role of ultrasound combined with hormone test in the diagnosis of obese PCOS. Methods One hundred and five women with PCOS were recruited. Patients were divided into two groups according to BMI;obese PCOS group (OB-PCOS, n=32, BMI≥25 kg/m2) and non-obese PCOS (NOB-PCOS, n=73, BMI<25 kg/m2). The ultrasonic parameters of follicle number (FN), ovarian volume (Vol), resistance index (RI) of ovarian stromal blood, RI of uterine artery and serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), the ratio of luteinizing hormone and follicle-stimulating hormone (LH/FSH), progesterone (P), estradiol (E2), free testosterone (FT), prolactin (PRL), sex hormoe binding globulin (SHBG), fasting plasma glucose (FPG), fasting insulin (FINS), the extent of insulin resistance and hyperandrogenism (HOMA-IR) were measured and compared. The correlation of the ultrasonic parameters and hormonal factors were analyzed. Results The Vol of OB-PCOS group was significantly higher than NOB-PCOS group [(12.25±4.89) ml vs (10.73±2.30) ml, t=2.20, P < 0.05]. FN and uterine artery RI of OB-PCOS group had a rising trend and RI of ovarian interstitial was on a reducing trend compared with NOB-PCOS group. But the differences were not statistically significant. The levels of FINS and HOMA-IR in OB-PCOS group [(14.82±6.45) mU/L and (3.91±3.30)] were significantly higher than those in NOB-PCOS group [(8.04±4.57) mU/L and (1.64±1.20)] (t=4.87, 3.47, respectively, both P < 0.01). And FSH in NOB-PCOS group was significantly higher than OB-PCOS group [(5.95±1.91) U/L vs (4.65±1.88) U/L, t=-2.77, P<0.01]. In POCS patients, FN was significantly associated with LH/FSH (r=0.35, P<0.01), and FT (r=0.38, P<0.01). Vol was significantly associated with LH/FSH, BMI, HOMA-IR and FPG (r=0.27, P<0.05;r=0.25, P<0.05;r=0.40, P<0.01;r=0.32, P<0.01). RI of ovarian stromal blood flow was significantly associated with SHBG (r=0.28, P<0.05). In OB-POCS group, RI of uterine artery was significantly associated with PRL (r=-0.58, P < 0.05). Vol was significantly associated with HOMA-IR (r=0.47, P < 0.05). In NOB-POCS group, FN was significantly associated with LH/FSH (r=0.33, P<0.05), and FT (r=0. 56, P<0.05). Vol was significantly associated with FT (r=0.31, P < 0.05). Conclusion There are some differences in the ultrasound and endocrine parameters between obese and non-obese PCOS patients, and some correlations exist between them.

Aim To establish a model of the vascular endothelial cell (VEC) injury in SDrats.Methods SD rats were randomly divided into the control and the modelgroups. The model rats were injected with adrenaline diluted to 2. 5 times 0. 05 mg?100 g-1 (tid) for 5 d continously. From the 4th d, they were irritated for 5 min in the0℃ cold-water in the middle between adrenaline injections.The control rats weregiven 0. 9% NS as above. At 6th d, blood samples were taken from carotid arteries ofthe rats and the CEC counts, t - PA、PAI activities, 6-keto-PGF1? concentrations andthe platelet aggregation rate(max) were detected respectively. Results In the modelgroup, as compared with those in the control group, t - PA activity and 6-keto-PGF1?concentration decreased significantly(P

Mycoepoxydiene is a novel antitumor agent extracted from marine lignicolous fungi HLY-2, which is Diaporthe phaseolorum by molecule identification. The medium optimization for mycoepoxydiene by orthogonal design and the comparison of submerged fermentation and solid state fermentation were studied. The rusult is that the maximal yield of the compound is 543mg/L, which is 43 times compared to the customary half-seawater PD medium and 15 times to the best submerged condition. This optimum culture medium included potato 250g/L, seawater 300mL/L, glucose 30g/L, lactose 50g/L, KH_ 2 PO_ 4 0.65mmol/L and (NH_ 4 )_ 2 SO_ 4 1g/L in the solid state condition. Differentiation analysis between submerged and solid state fermentation, and antitumor activity of these ferment products were also studied. The antitumor activity of products of the optimum medium approached the pure compound.

In the reductive branch, glycerol is first dehydrated to 3-hydroxypropionaldehyde that is then reduced to 1,3-PD under the consumption of reducing power (NADH). If over-expression of the gene dhaB encoding glycerol dehydrase is achieved,the reducing power will be scarce and 3-hydroxypropionaldehyde will be accumulated,which is disadvantage to produce 1,3-propanediol.The structure gene yqhD from E.coli encoding 1,3-propanediol oxidoreductase isoenzyme[under the consumption of reducing power (NADPH)]and the gene dhaB encoding glycerol dehydrase from Klebsiella pneumoniae was amplified using PCR method.The two gene were transferred into expression vector pEtac to construct a novel recombinant Klebsiella pneumoniae (pEtac-dhaB-tac-yqhD).Over-expression of yqhD and dhaB in Klebsiella pneumoniae was achieved with pEtac-dhaB-tac-yqhD.The fermentation result on aerobic conversion showed the increase of 20% of 1,3-propanediol yield by Klebsiella pneumoniae(pEtac-dhaB-tac-yqhD) was obtained compared with Klebsiella pneumoniae.The main by-products,acetic acid and butanediol decreased estrogen receptors 30% obviously.