Objective To establish a HPLC determination method for the effective constituents extracted from Fructus Schisandrae Chinensis by low-temperature water extraction.Methods The assay was conducted on Kromasil C18 column(250 mm? 4.6 mm,5 ? m)using a gradient elution with acetonitrile-water as mobile phase.The flow rate was set at 1.0 mL/min,column temperature was kept at 30 ℃,and the detection wavelength was at 240 nm.Results The linear range for schizandrol A was 5.0～ 200.0 ? g/mL(r=0.999 9),and the average recovery was 101.27 % with RSD being 1.97 %(n=6).Conclusion The method is simple,reliable and reproducible for the determination of schizandrol A.It can be used for quality control of preparations including Fructus Schisandrae Chinensis.

Objective:To evaluate the clinical efficacy and safety of repaglinide and gliclazide in the treatment of elderly type 2 di-abetes mellitus (T2DM). Methods:A total of 225 T2DM patients with the age over 60 were enrolled and randomly divided into repa-glinide group (n=112) and gliclazide group (n=113). The repaglinide group was given repaglinide 0. 5~2 mg, po, tid, while the gliclazide group was given gliclazide modified release tablets 30~120 mg, po, qd,both with the treatment course of 12 weeks. The level of FPG, 2h PG, HbAlC, TC and TG and the side effects were observed and the clinical efficacy was compared between the two groups. Results:The level of FPG, 2h PG, HbAlc and TC in the two groups showed significant decrease after the treatment (P<0. 01 or P<0. 05), and the level of 2h PG was decreased more significantly in the repaglinide group (P<0. 05). There were no significant differ-ences in the total effective rate (91. 8% vs 86. 2%), hypoglycemia incidence (4. 5% vs 8. 2%) and ADR incidence (6. 4% vs 10. 1%) in the two groups (P>0. 05). Conclusion:Both repaglinide and gliclazide show safe and reliable efficacy in the treatment of elderly T2DM.

Objective To observe the expression and mechanism of hypoxia-inducible factor-1α (HIF-1α) and p53 protein at the altitude of 5000 meter plateau hypoxia environment in rats,as well as the effect of Astragalus injection.Methods Sixty Sprague Dawley rats were randomly divided into the Astragalus injection intervention group and normal saline control group,30 rats in each group.Astragalus injection group rats were intraperitoneal injected of Astragalus injection (15 ml/kg) before 30 minutes into the plateau environment simulation cabin,normal saline group rats were intraperitoneal injected with the same volume of saline.30 minutes after injection,rats in each group were reared in the plateau experiment cabin which simulated altitude of 5000 m (oxygen partial pressure 11.3 kPa) for 2,6,8,12,24 hours,each time period of 6 rats.When get out,the rats were executed immediately and eyes were harvested.Retinal sections were studied by hematoxylin eosin stain,and immunohistochemical method for HIF-1α and p53 expression.Results For control rats,after 2 hours in the cabin,there was edema in retinal layers.HIF-1α and p53 were expressed mainly in the cytoplasm of retinal layers.When the periods in cabin extended,there was atrophy of retinal nerve fiber layer,swelling and degeneration of ganglion cells.The expression of HIF-1α and p53 was increased.Compared with the control group,the intervention group rat had similar but less severe retinal changes,and the expression of HIF-1α and p53 was significantly decreased (P ＜0.05).Conclusion Astragalus injection can reduce pathological retinal damage in rats at high altitude environment,and its mechanism may be associated with reduced HIF-1α,p53 expression.

Objective To determine the contents of free ferulic acid and total ferulic acid in different processed products of Rhizoma Chuanxiong.Methods With methanol-formic acid(95∶5) as the extraction solvent for free ferulic acid and with methanol-2% NaHCO3 water solution(95∶5) as the extraction solvent for total ferulic acid,ultrasonic method was used for the extraction.The contents of free ferulic acid and total ferulic acid were determined by HPLC,and the chromatographic conditions were as follows: C18 column(250mm?4.6mm,5?m),the mobile phase consisting of acetonitrile-1%acetic acid solution(20∶80),the detecting wavelength at 320nm,flow rate being 1.0 mL/min and detection at room temperature.Results The average contents of free ferulic acid and total ferulic acid in Rhizoma Chuanxiong roasted by wine were higher than those in raw and other processed Rhizoma Chuanxiong,and free ferulic acid content in raw and different processed Rhizoma Chuanxiong was lower than the total ferulic acid content.Conclusion The method is simple,accurate and can be used for the quality control standard for Rhizoma Chuanxiong,and the chemical assay of total ferulic acid content would be a better choice of assessing the herbal quality of Rhizoma Chuanxiong.

AIM: To evaluate the potential acylation stimulating protein (ASP) resistance in both adipocytes and preadipocytes under the conditions by which insulin resistance is produced by the stimulation of free fatty acids (FFA), and to explore the mechanism of ASP resistance on post-receptor level. METHODS: 3T3-L1 preadipocytes were induced to differentiate. Then the cells were treated with oleate or palmitate at concentration of 0 mmol/L (FFA-free DMEM/F12), 0.125 mmol/L, 0.5 mmol/L or 1.0 mmol/L overnight. Glucose transport was assessed by [~3H] 2-deoxyglucose uptake to evaluate insulin resistance and ASP resistance. Both non-FFA treated and FFA treated 3T3-L1 cells were cultured with ASP at concentration of 5.0 μmol/L for 4 h, then the cell proteins were extracted, and the expressions of guanine nucleotide binding protein beta (Gβ), guanine nucleotide-binding protein alpha-q/11(Gαq/11), phosphorylated-protein kinase Cα (p-PKCα) and phosphorylated-protein kinase Cζ (p-PKCζ) were measured by Western blotting. RESULTS: Both adipocytes and preadipocytes were responsive to ASP. ASP stimulation increased glucose transport by 198% in adipocytes and by 287% in preadipocytes (P<0.01 vs PBS). FFA at concentration of 0.125 mmol/L did not change ASP-stimulated glucose transport significantly, but high dose of oleate or palmitate effectively reduced the ASP response with a significant reduction by 47% (P<0.05 for oleate) and 34% (P<0.05 for palmitate) at 1 mmol/L FFA in adipocytes. Similarly in preadipocytes, glucose uptake rates were decreased by 43% (P<0.05 for oleate) and 62% (P<0.01 for palmitate) at 1 mmol/L FFA. Effects were comparable to those obtained with insulin. After overnight incubation with oleate or palmitate in adipocytes and preadipocytes, Gβ, Gαq/11, p-PKCα and p-PKCζ were downregulated both in the absence of ASP treatment and in the presence of ASP treatment in adipocytes. At concentration of 1.0 mmol/L, oleate inhibited the expressions of ASP-induced Gβ, Gαq/11, p-PKCα and p-PKCζ in adipocytes by 47%, 44%, 39% (P<0.05, P<0.01) and 20% (P>0.05), respectively. Palmitate also effectively blocked the expressions of ASP (at concentration of 1.0 mmol/L)-induced Gβ, Gαq/11, p-PKCα and p-PKCζ by 50%, 43%, 44% and 43% (P<0.05, P<0.01) in adipocytes. In preadipocytes, oleate only inhibited ASP-induced p-PKCα and p-PKCζ significantly by 39% and 19%, respectively (P<0.05). However, overnight exposure of 3T3-L1 preadipocytes to 1 mmol/L palmitate leaded to 45%, 50%, 52% and 21% (P<0.05, P<0.01) inhibition of ASP-induced expressions of Gβ, Gαq/11, p-PKCα and p-PKCζ, respectively. CONCLUSION: Oleate and palmitate inhibit ASP-mediated stimulation of glucose transport both in adipocytes and preadipocytes. The study provides direct evidence of ASP resistance under the condition of insulin resistance induced by FFA in a cellular model. The mechanism of action involves both changes in expression of C5L2 as well as signaling parameters. Fatty acid-induced ASP resistance may contribute to the physiological abnormalities associated with insulin resistance and obesity phenotype.

China ; Humans ; Logistic Models ; Non-alcoholic Fatty Liver Disease ; Risk Factors

AIM: The purpose of the present study was to examine the effect of insulin and different kinds of free fatty acid (FFA) on glucose transport in cultured 3T3-L1 preadipocytes or adipocytes. METHODS: Following the exposure of preadipocytes to insulin at different concentrations and treated times, glucose transport was assessed as [3H]2-deoxy glucose uptake. Furthermore, 3T3-L1 preadipocytes and adipocytes with either the monounsaturated FFA oleate (C18:1) or the saturated FFA palmitate (C16:0)were used and glucose transport was examined as above. RESULTS: Insulin increased specific membrane glucose transport in 3T3-L1 preadipocytes at the time of 15 min to 1 h stimulation. However, after 6 h exposure to insulin, downregulation of glucose transport was observed. Dose response studies demonstrated that 2-DG transport increased by 336% at 50 nmol/L of insulin (P

Objective:To investigate the effects of the anti-TLR2 antibody blocking TLR2 signaling pathway on inflammatory response in Staphylococcus aureus pneumonia murine models.Methods: Sixty C57BL/6J mice were divided randomly into normal control,SA pneumonia,and anti-TLR2 antibody group,killed 3 and 8 days after inoculation respectively.Normal control mice inoculated sterile PBS intranasally ,SA pneumonia mice inoculated SA ,anti-TLR2 antibody group of mice injected with anti-TLR2 antibody by tail vein and then inoculated SA intranasally.At the predetermined point , the colony-forming units ( CFU ) of bacteria were higher , leukocytes and neutrophil percentage were counted in bronchoalveolar lavage fluid ( BALF ) , the concentrations of KC and IL-10 in BALF and serum were assayed by ELISA ,changes in pulmonary histopathology were observed with HE staining and TLR 2 expression was detected by immunohistochemical.Results:3 days after intranasal inoculation ,the concentrations of KC and IL-10 in BALF and serum was increased in SA pneumonia mice , pulmonary histopathology changes significantly in HE staining.Compared with SA pneumonia mice,the CFU of bacteria were higher,leukocytes count and neutrophil percentage ,the concentrations of KC in BALF and serum,as well as HE pathological scores were reduced significantly in anti-TLR2 antibody group mice ,while no significant difference in IL-10.8 days after intranasal inoculation , HE pathological scores of anti-TLR2 antibody group mice were significantly lower than SA pneumonia group mice ,the CFU of bacteria in BALF were not statistically different between those two groups.Conclusion:Anti-TLR2 antibody attenuates the production of inflammatory mediators and inflammatory cell infiltration in SA pneumonia mice .

Objective To investigate the effect of taurine transporter in the process of protection of brain edema in rats with severe traumatic head injury. Methods A total of 24 Male Sprague-Dawley rats were randomly divided into 4 groups. Except the control rats (Group Sham), all other three groups were subjected to lateral fluid percussion head injury. The TBI (Traumatic brain injury) models (Group TBI) and surgical control rats (Group Sham) were injected with saline through caudal vein after surgery, while the Taurine prevention and Taurine treatment models (Group Pre Tau and Group Tau) were injected with 120 g/L taurine solution before or after surgeries respectively. Water content in each brain, mRNA and protein expres?sion of aquaporin 4 and taurine transporter in the injured rat brain hemispheres were all evaluated over the time course of the study (7 d) in each group. Results Compared with rats in Group Sham, water content in each brain increase, mRNA tran?scription and protein expression of AQP4 were both up regulated but the mRNA transcription and protein expression of TauT were both down-regulated in rats in TBI group. Compared with rats in TBI group, brain water content, mRNA transcription and protein expression of AQP4 all decrease while mRNA transcription and protein expression of TauT all increase in rats in Pre tau and Tau groups. There is no statistical difference of TauT expression between rats in pre-tau group and Tau group. Conclusion Taurine exert its neuron protection role through draining water content from brain and down regulating expres?sion of AQP4 but rising expression of TauT after TBI.

Objective: To investigate the effects of taurine on blood glucose, number of platelets in plasma, pletelet aggregation and metabolism of oxygen free radicals in diabetic rats. Methods: Diabetes was induced by a single injection (ip) of streptozotocin (STZ,70 mg/kg body weight). The rats were maintained on drinking water with or without taurine (1.5 g/kg body weight per day). The concentration of blood glucose, maximal aggregation rate of platelets induced by adenosine 5 diphosphate(ADP) or thrombin(Thr) and the number of platelets in plasma were measured. Metabolism of oxygen free radicals in platelets was assessed as superoxide dismutase(SOD) activity and content of malondialdehyde(MDA) in platelets. [WT5FZ]Results: [WT5BZ]The concentration of blood glucose and maximal aggregation rate of platelets induced by ADP or Thr were significantly higher in diabetic rats than in non diabetic rats. The diabetic rats showed decreased platelet number and SOD activity with higher MDA level. In taurine treated diabetic rats concentration of blood glucose was obviously decreased, but no significant change was found in platelet number, SOD activity, MDA content and aggregation of platelets induced by ADP or Thr. [WT5FZ] Conclusion: [WT5BZ]There is a correlation between the increase in platelet aggregability and the abnormal metabolism of oxygen free radicals in platelets and taurine can decrease the blood glucose level but has no significant effect on platelet aggregability and metabolism of oxygen free radicals in diabetic rats.