Effect of Puerariae Lobatae Radix on Expression of Endoplasmic Reticulum Stress Related Proteins GRP78 and ATF6 in T2DM Rats / 中国实验方剂学杂志

Objective:To observe the effect of Puerariae Lobatae Radix on type 2 diabetes mellitus (T2DM) rats and related mechanism. Method:Ninety SD rats were divided into normal group, model group, metformin group, Puerariae Lobatae Radix high dose group, Puerariae Lobatae Radix medium dose group and Puerariae Lobatae Radix low dose group, 15 rats in each group. The rats in abnormal group were fed with high-fat and high sugar diet for 4 weeks, and then T2DM model was established by intraperitoneal injection of 40 mg·kg-1 streptozotocin (STZ). Puerariae Lobatae Radix high-dose group was intragastrically administered with 2.1 g·kg-1 of Puerariae Lobatae Radix extract powder, Puerariae Lobatae Radix medium-dose group was intragastrically administered with 1.4 g·kg-1 of Puerariae Lobatae Radix extract powder, Puerariae Lobatae Radix low-dose group was intragastrically administered with 0.7 g·kg-1 of Puerariae Lobatae Radix extract powder, 0.2 g·kg-1 of metformin hydrochloride in metformin group, distilled water once a day in normal group and model group. After 8 weeks, fasting blood glucose (FBG), glycosylated serum protein (GSP), insulin (FINS), triglyceride (TG), cholesterol (TC) were measured, and insulin resistance index （HOMA-IR） was calculated. The expression of tumor necrosis factor -α(TNF-α) was observed by immunohistochemistry. Western blot was used to detect the protein expression of glucose regulatory protein 78 （GRP78）, activated transcription factor 6 （ATF6） in pancreatic tissue. Result:Compared with normal group, the contents of FBG, GSP, TG, TC, FINS, TNF-α in model group were significantly increased (P<0.05), the HOMA-IR was significantly increased (P<0.05), the protein expressions of GRP78 and ATF6 in pancreatic tissue were significantly increased (P<0.05). Compared with model group, the contents of FBG, GSP, TG, TC, FINS and TNF-α in the metformin group and Puerariae Lobatae Radix high, medium and low dose groups were significantly decreased (P<0.05), HOMA-IR decreased significantly (P<0.05), and the expression of GRP78 and ATF6 protein in pancreatic tissue decreased significantly (P<0.05). Conclusion:Puerariae Lobatae Radix can significantly improve insulin resistance in T2DM rats, inhibit the expression of inflammatory factor TNF-α in pancreatic tissue, reduce the protein expression of GRP78 and ATF6 in pancreatic tissue.

Clinical observation on acupuncture for intervening the response in gastroscopy / 中国针灸

<p><b>OBJECTIVE</b>To observe the clinical effect of acupuncture for controlling the adverse response in gastroscopy.</p><p><b>METHODS</b>Ninety-seven cases of gastroscopy were randomly divided into an observation group of 52 cases and a control group of 45 cases. The observation group were treated by acupuncture at Hegu (LI 4), Neiguan (PC 6) and Zusanli (ST 36), in combined with oral administration of Lidocaine; the control group were treated by simple administration of Lidocaine. Changes of the adverse response, blood pressure and heart rate, and satisfactory degrees and the willing re-examination rate were investigated in the two groups. Results In the observation group, the nausea and vomiting, salivation, restlessness, breath holding and other adverse responses in gastroscopy were significantly decreased as compared with those in the control group (P < 0.01), and the blood pressure and heart rate were more stable than in the control group, and the satisfactory degree and willing re-examination rate were higher than the control group (P < 0.01).</p><p><b>CONCLUSION</b>Acupuncture can effectively control the adverse response in gastroscopy.</p>

A Pichia pastoris with alpha-1, 6-mannosyltransferases deletion and its use in expression of HSA/GM-CSF chimera / 生物工程学报

Yeast is a widely used host for recombinant protein expression. However, glycoproteins derived from yeast contain N-glycan of high mannose type and are usually hyperglycosylated. alpha-1,6-mannosyltransferases gene (och1) encodes the enzyme that initiates the first step of out-chain elongation of high mannose type N-glycan in yeast, which is different from that in human. So, a high efficient method to knockout target gene by two-step recombination was established and was used to delete och1. In the first recombinant, a plasmid with och1::ADE1 and ura3 gene was linearized in the downstream of och1 and inserted to the och1 site of P. pastoris genome, where the upstream and downstream of och1 were duplicated. In the second recombinant, the duplicated fragments of och1 were exchanged and the och1 deletion strains were selected on the plates containing 5-FOA, but no adenine. Then the och1 deletion strain was applied to express an human serum albumin (HSA) granulocyte-macrophage colony-stimulating factor (GM-CSF) chimera. Different with the hyperglycosylated HSA/GM-CSF chimera expressed in wild type P. pastoris, the chimera expressed in the och1 deletion strain, contained smaller N-glycan. The results suggested that the och1 mutant yeast may be more suitable for production of recombinant glycoproteins. And the och 1 deletion strain could be used for further re-engineering to produce complex human glycoproteins.

The inhibition of CYP2C9 isoenzyme in Cunninghamella blakesleeana AS 3. 910 / 药学学报

<p><b>AIM</b>To investigate the variation of CYP2C9 isoenzyme activity in the microbial model in response to inhibitors of CYP2C9.</p><p><b>METHODS</b>Using C. blakesleeana AS 3. 910 as a model strain, the impact of CYP2C9 inhibitors on the metabolites yields of CYP2C9 substrates was determined and the drug-drug interactions among CYP2C9 substrates were evaluated. Liquid chromatography-mass spectrometry was used to analyze biotransformation products.</p><p><b>RESULTS</b>Benzbromarone decreased the yield of 4'-hydroxytolbutamide from 100% to 14.5%; sulfaphenazole decreased the yield of O-demethylindomethacin from 75.2% to 9.9%; valproic acid decreased the yield of 4'-hydroxydiclofenac from 98.6% to 2.7%, separately. Tolbutamide, indomethacin and diclofenac interacted with each other, resulting in the decreased formation of metabolites catalyzed by CYP2C9.</p><p><b>CONCLUSION</b>Three CYP2C9 inhibitors inhibit the activity of CYP2C9 isoenzyme in C. blakesleeana AS 3. 910 differently, and there are drug-drug interactions among CYP2C9 substrates.</p>

Investigation on the hydroxylation metabolism of imrecoxib in vitro by using recombinant human CYPs / 药学学报

<p><b>AIM</b>To identify the drug-metabolizing enzymes involved in the hydroxylation of the new anti-inflammatory and anodyne imrecoxib.</p><p><b>METHODS</b>Imrecoxib was incubated with heterologous expression human cytochrome P450 (rCYPs) in vitro, and metabolites and remained parent drug were detected with liquid chromatography-multistage mass spectrometry. The contribution of 4 CYPs in the hydroxylation metabolism of imrecoxib was evaluated by total normalized rate (TNR) method.</p><p><b>RESULTS</b>Imrecoxib is metabolized by CYP2C9, CYP2D6 and CYP3A4, with the rate of 62.5%, 21.1% and 16.4%, respectively.</p><p><b>CONCLUSION</b>CYP2C9 is the major enzyme involved in imrecoxib hydroxylation metabolism.</p>