Hierarchical activation of compartmentalized pools of AMPK depends on severity of nutrient or energy stress.

AMPK, a master regulator of metabolic homeostasis, is activated by both AMP-dependent and AMP-independent mechanisms. The conditions under which these different mechanisms operate, and their biological implications are unclear. Here, we show that, depending on the degree of elevation of cellular AMP, distinct compartmentalized pools of AMPK are activated, phosphorylating different sets of targets. Low glucose activates AMPK exclusively through the AMP-independent, AXIN-based pathway in lysosomes to phosphorylate targets such as ACC1 and SREBP1c, exerting early anti-anabolic and pro-catabolic roles. Moderate increases in AMP expand this to activate cytosolic AMPK also in an AXIN-dependent manner. In contrast, high concentrations of AMP, arising from severe nutrient stress, activate all pools of AMPK independently of AXIN. Surprisingly, mitochondrion-localized AMPK is activated to phosphorylate ACC2 and mitochondrial fission factor (MFF) only during severe nutrient stress. Our findings reveal a spatiotemporal basis for hierarchical activation of different pools of AMPK during differing degrees of stress severity.

Clinical values of using platelet rich plasma with autologous fat injection in facial rejuvenation / 中华医学美学美容杂志

Objective To explore the clinical value of autologous fat injection with platelet rich plasma in facial rejuvenation.Methods 35 female beauty seekers were collected in this study.They were divided into two groups:sample autologous fat group (FAT group,n=20) and combined autologous fat and platelet-rich plasma group (PRP group,n =15).The average level of injection was 34 ml.The fat was collected at the abdominal or thigh sites using 18 G needle connecting with negative pressure suction-pump.The fat was centrifugated 2 min to get rid of the lower layer of water and the upper layer of oil.Weigao PRP extract kit was used to obtain the PRP.The ratio of PRP to fat was 1:5.The injection technique was according to Colemen method.The survival rate,satisfaction and complication were compared between the 2 groups.Results There were no infection,hematoma,fat liquefaction necrosis occurred in the 2 groups.Compared with the FAT group,the skin texture and facial contour of the PRP group was improved.Conclusions The platelet rich plasma can improve the survival rate of fat and has a stable effect,which is one of the safe and ideal methods of facial rejuvenation.

Salvianolic acid B as a substrate and weak catechol-O-methyltransferase inhibitor in rats.

1. The aim of this study was to investigate the biotransformation of salvianolic acid B (SAB) by catechol-O-methyltransferase (COMT) and its interaction with levodopa (l-DOPA) methylation in rats. 2. The enzyme kinetics of SAB were studied after incubation with rat COMT. The in vivo SAB and 3-monomethyl-SAB (3-MMS) levels were determined after a single dose of tolcapone with or without SAB administration. For l-DOPA, the effect of SAB inhibition on l-DOPA methylation was studied in vitro. The l-DOPA and 3-O-methyldopa (3-OMD) levels were determined after single and multiple doses of SAB with or without l-DOPA administration. 3. After incubation, we found that SAB was methylated mainly by rat liver and kidney COMT. Tolcapone strongly inhibited the formation of 3-MMS in vitro and in vivo, without any change in the plasma concentration of SAB. Moreover, tolcapone significantly increased the cumulative bile excretion of SAB from 3% to 40% in the rat. SAB inhibited the methylation of l-DOPA with an IC50 value of 2.08 µM in vitro. In vivo, a single intravenous dose of SAB decreased the plasma concentration of 3-OMD, with no obvious effect on the pharmacokinetics of l-DOPA. Multiple doses of SAB given to rats also decreased the plasma concentration of 3-OMD, while SAB increased the plasma concentration of l-DOPA.

Mechanism-based inhibitory and peroxisome proliferator-activated receptor α-dependent modulating effects of silybin on principal hepatic drug-metabolizing enzymes.

Silybin, a major pharmacologically active compound in silymarin, has been widely used in combination with other prescriptions in the clinic to treat hepatitis and a host of other diseases. Previous studies suggested that silybin is a potential inhibitor of multiple drug-metabolizing enzymes (DMEs); however, the in vitro to in vivo translation and the mechanisms involved remain established. The aim of this study was to provide a mechanistic understanding of the regulatory effects of silybin on principal DMEs. Silybin (50 or 150 mg/kg/d) was administered to mice for a consecutive 14 days. The plasma and hepatic exposure of silybin were detected; the mRNA, protein levels, and enzyme activities of principal DMEs were determined. The results demonstrated that the enzyme activities of CYP1A2, CYP2C, CYP3A11, and UGT1A1 were significantly repressed, whereas little alteration of the mRNA and protein levels was observed. Silybin inhibits these DMEs in a mechanism-based and/or substrate-competitive manner. More importantly, silybin was found to be a weak agonist of peroxisome proliferator-activated receptor (PPAR)α, as evidenced from the molecular docking, reporter gene assay, and the targeting gene expression analysis. However, silybin could significantly compromise the activation of PPARα by fenofibrate, characterized with significantly repressed expression of PPARα targeting genes, including L-FABP, ACOX1, and UGT1A6. This study suggests that silybin, despite its low bioavailability, may inhibit enzyme activities of multiple DMEs in a mechanism-based mode, and more importantly, may confer significant drug-drug interaction with PPARα agonists via the repression of PPARα activation in a competitive mode.

Clinical efficacy of dual plane breast augmentation using perinipple incision / 中华医学美学美容杂志

Objective To study the technology and the result of dual plane breast augmentation using nipple margin vertical incision of areola.Methods Totally 60 cases of augmentation mammaplasty were involved in this study.The nipple margin vertical incision of areola was applied obliquely into the breast through the pectoralis major fascia.The rib starting point of pectoralis major were cut off,medial to the side of the sternum.Under the pectoralis major the cavity was peeled according to the preoperative design range.Based on the different situation of the breast types Ⅰ,Ⅱ,Ⅲ,dual plane breast augmentations were stripped respectively.After implanting the breast prosthesis,the upper part of the prosthesis was under the pectoralis major and the lower part was under the mammary gland.Results The 60 patients were all after childbearing,20 of whom underwent type 2 dual plane breast augmentation,4 underwent type 3 double plane and the rest underwent type 1 double plane.After 3 months to 2 years follow-up,all cases got satisfactory results,except 1 case of postoperative hematoma and 1 case appeared capsular contracture.Conclusions The nipple margin vertical incision of areola can complete types Ⅰ,Ⅱ,Ⅲ dual plane breast augmentation operation,at the same time it can correct mild-to-moderate mastoptosis.

Discovery of Imigliptin, a Novel Selective DPP-4 Inhibitor for the Treatment of Type 2 Diabetes.

We report our discovery of a novel series of potent and selective dipeptidyl peptidase IV (DPP-4) inhibitors. Starting from a lead identified by scaffold-hopping approach, our discovery and development efforts were focused on exploring structure-activity relationships, optimizing pharmacokinetic profile, improving in vitro and in vivo efficacy, and evaluating safety profile. The selected candidate, Imigliptin, is now undergoing clinical trial.

Prediction of human pharmacokinetics from preclinical information of rhein, an antidiabetic nephropathy drug, using a physiologically based pharmacokinetic model.

The aim of the study was to develop a physiologically based pharmacokinetic (PBPK) model of rhein to predict human pharmacokinetics before dosing for the first time in human beings. After oral administration of rhein at the doses of 35, 70 and 140 mg/kg in rat, rhein had the following mean plasma pharmacokinetic properties: t1/2 of 3.2, 3.6 and 4.3 hr, AUC∞ of 69.5, 164.3 and 237.8 µg/h/ml and CL/F of 503.4, 426.1 and 588.8 ml/hr/kg, respectively. In vitro, the intrinsic clearance (Clint ) of rhein in cytochrome P450 (CYP450), UDP-glucuronosyltransferase (UGT) and sulfotransferase (SULT) metabolism of rat was 0.6, 7.8, and 5.5 µl/min/mg protein, respectively. The Clint of rhein in CYP450, UGT and SULT of human beings was 0.10, 1.36 and 0.68 µl/min/mg protein. The rat pharmacokinetics and the metabolism data in vitro were used to construct the PBPK model of rhein, and the observed plasma drug concentration profiles of rhein in rat were validated by a PBPK model. Subsequently, the plasma drug concentration profiles of human beings by the present PBPK model were validated by experimental data in human beings accurately.

The identification and pharmacokinetic studies of metabolites of salvianolic acid B after intravenous administration in rats.

AIM: To identify and quantify the major metabolites of salvianolic acid B (SAB) after intravenous injection in rats. METHODS: LC-IT/TOF-MS was used to identify the metabolites in rat bile, plasma, and urine; LC-MS/MS was used to quantify the two major metabolites. RESULTS: In rat bile, plasma, and urine, nine metabolites were identified, including methylated metabolites of SAB, lithospermic acid (LSA), the decarboxylation and methylation metabolites of LSA, salvianolic acid S (SAS), and dehydrated-SAS. The t1/2 of monomethyl-SAB and LSA were both very short, and monomethyl-SAB had a larger AUC than LSA in rats. CONCLUSION: Nine metabolites were found, the metabolic pathway was described, and the pharmacokinetic profiles of LSA and monomethyl-SAB were studied, thereby clarifying that methylation was the dominant metabolic pathway for SAB in rats.

An integral strategy toward the rapid identification of analogous nontarget compounds from complex mixtures.

Identification of nontarget compounds in complex mixtures is of significant importance in various scientific fields. On the basis of the universal property that the compounds in complex mixtures can be classified to various analogous families, this study presents a general strategy for the rapid identification of nontarget compounds from complex matrixes using herbal medicine as an example. The proposed strategy consists of three sequential steps. First, a blank control sample is prepared for the purpose of removing interferences in the complex matrixes via automatic chromatographic and mass spectrometric data comparisons. Second, the diagnostic ions guided bridging network strategy is developed for the rapid classification of analogous compounds and structural characterizations. Finally, a quantitative structure retention relationship (QSRR) is built to validate the identifications and to differentiate isomers. Using this strategy, we have successfully identified a total of 45 organic acids from Mai-Luo-Ning and Flos Lonicerae injection, and 46 ginsenosides from Shen-Mai injection samples. The QSRR approach enabled a successful differentiation of most isomers. The proposed strategy will be expected to be applicable to the identification of nontarget compounds in complex mixtures.

Dysregulations of intestinal and colonic UDP-glucuronosyltransferases in rats with type 2 diabetes.

Diabetes mellitus is a chronic disease of complex metabolic disorder associated with various types of complications. UDP-glucuronosyltransferases (UGTs), the major phase II conjugation enzymes, mediate the metabolism of both drugs and endogenous metabolites that may raise great concerns in the condition of diabetes. The aim of this study was to determine whether diabetes could affect UGTs in the intestinal and colonic tract. A high-fat diet combined with low-dose streptozotocin was used to induce a type 2 diabetic model in rats. The mRNA levels and enzymatic activities of UGT1A1, -1A6, and -1A7 in the diabetic intestine and colon were higher than those in nondiabetic rats. In contrast, both the activity and mRNA level of UGT2B1 in diabetic rats were lower than those in nondiabetic rats. Notably, the diabetic intestine and colon exhibited an inflammatory state with increased pro-inflammatory cytokines. Various transcriptional factors involved in UGT regulation were unanimously upregulated in the diabetic intestine and colon. These findings strongly suggest that the regulating pathways of the UGT1 family are adaptively upregulated in the diabetic gastrointestinal tract. Given the essential regulatory role of the gastrointestinal site in drug disposition, such changes in UGTs may have a dynamic and complex impact on therapeutic drugs and endogenous metabolomes.

The pharmacokinetic-pharmacodynamic model of azithromycin for lipopolysaccharide-induced depressive-like behavior in mice.

A mechanism-based model was developed to describe the time course of lipopolysaccharide-induced depressive-like behavior and azithromycin pharmacodynamics in mice. The lipopolysaccharide-induced disease progression was monitored by lipopolysaccharide, proinflammatory cytokines, and kynrenine concentration in plasma. The depressive-like behavior was investigated by forced swimming test and tail suspension test. Azithromycin was selected to inhibit the surge of proinflammatory cytokines induced by lipopolysaccharide. Disease progression model and azithromycin pharmacodynamics were constructed from transduction and indirect response models. A delay in the onset of increased proinflammatory cytokines, kynrenine, and behavior test compared to lipopolysaccharide was successfully characterized by series transduction models. The inhibition of azithromycin on proinflammatory cytokines was described by an indirect response model. After lipopolysaccharide challenging, the proinflammatory cytokines, kynrenine and behavior tests would peak approximately at 3, 12, and 24 h respectively, and then the time courses slowly declined toward a baseline state after peak response. During azithromycin administration, the peak levels of proinflammatory cytokines, kynrenine and behavior indexes decreased. Model parameters indicated that azithromycin significantly inhibited the proinflammatory cytokines level in plasma and improved the depressive-like behavior induced by inflammation. The integrated model for disease progression and drug intervention captures turnovers of proinflammatory cytokines, kynrenine and the behavior results in the different time phases and conditions.

Disturbance of hepatic and intestinal UDP-glucuronosyltransferase in rats with trinitrobenzene sulfonic acid-induced colitis.

UDP-glucuronosyltransferase (UGT) is an important class of phase II metabolizing enzymes, playing a pivotal role in detoxifying various substances and in the pathological procedures of some diseases. The present study aims to uncover the potential dysregulation pattern of UGTs in trinitrobenzene sulfonic acid (TNBS)-induced colitis. Colitis was induced by intra-rectally administering a single dose of TNBS (100 mg/kg). The expression and enzyme activity of hepatic UGTs of colitis rats were all down-regulated significantly except UGT1A7, for which the mRNA level was up-regulated. In contrast, UGT isoforms in the small intestine were relatively unaffected. In the colon, where the inflammation occurs, the mRNA level and enzyme activity of UGT1A1 and 1A6 were down-regulated, but those of UGT1A7 and 2B1 up-regulated. The mRNA levels of various transcription factors, including AhR, CAR, PXR, PPARγ, and FXR were all decreased, except for AhR and CAR in the small intestine and colon. Our data suggests that colitis induces an isoform-dependent and tissue-specific dysregulation of UGTs and their related transcription factors.

Preliminary study of the role of canonical Wnt signalling pathway in non-Hodgkin's lymphoma cell lines / 肿瘤

Objective: To determine whether the canonical Wnt signalling pathway is abnormally activated in NHL (non-Hodgkin's lymphoma), and to explore its association with pathogenesis of NHL. Methods: The expression levels of β-catenin, GSK-3β (glycogen synthase kinase-3β) and its inactive form p-GSK-3β (Ser9) (phosphorylation of GSK-3β at Ser9 residue) in NHL cell lines including SUDHL-4, Raji and Namalwa were detected by Western blotting, and the normal human lymphocytes were served as the controls. The mRNA levels of CTNNB 1 gene (encoding β-catenin), canonical Wnt signaling pathwayrelevant gene LRP 5 (low-density lipoprotein receptor-related protein 5) and target gene c -Myc were detected by real-time fluorogenic quantitative PCR. Results: The expressions of total cellular proteins and β-catenin protein in cell nuclei were significantly up-regulated in three NHL cell lines as compared with in those normal human lymphocytes. The elevated expression levels of p-GSK-3β (Ser9) were observed in lymphoma cell lines, but the expression level of GSK-3β was not significantly different between the lymphoma cell lines and the normal human lymphocytes. The mRNA expression levels of CTNNB1, LRP5 and c-Myc were significantly higher in lymphoma cell lines than those in the normal human lymphocytes. Conclusion: Given the important role of β-catenin as an activation marker in canonical Wnt signaling pathway, the evidence of up-regulated expression of β-catenin and the changes of other relevant molecules in canonical Wnt signaling pathway suggests that the constitutional activation of canonical Wnt signaling pathway may contribute to the pathogenesis of NHL. Copyright © 2013 by TUMOR.

Clinical application of intraoperative sustained skin and soft-tissue expansion in augmentation mammaplasty using transaxillary approach with endoscopic assistance / 中华医学美学美容杂志

Objective To evaluate the results of prosthesis augmentation,relieve suffering patient experience,improve shape and touch of breast.Methods According to design,in dual-plane mammaplasty transaxillary approach with endoscopic assistance,we used soft-tissue expansion before prosthesis,saline volume in expander was the same as prosthesis size at beginning; adjusting expander volume and position,then determined prosthesis size according to expander volume lastly.after intraoperative sustained expansion for 30 to 60 minutes twice prosthes volume,replace expander with prosthesis.Results This method was applied in 120 patients,showing that pain significantly reduced after expansive without use of analgesia pump; complications included that 3 patients appeared seroma and absorbed soon; 2 patients had poor prosthese location,which was adjusted in reoperation; one capsular contracture was repaired later.After 6-months to 2-years follow-up,the results showed that breast was nature and dynamic; satisfactory rate reached to 95 ％.Conclusions In augmentation mammaplastic operation,pre-adoption of an expander can make it easy to adjust the location of prosthesis,to avoid being injured and to determine prosthesis volume by expander injection of saline volume,especially for patient with different size of both breasts.

Advances in research on canonical Wnt signaling pathways in malignant lymphomas / 肿瘤

Wnt signaling pathway is an important pathway for regulating the development of embryo and organs. It has been shown to play a key role in proliferation, differentiation, polarity, adhesion and motility of cells. The constitutive activation of canonical Wnt pathway is involved in the pathogenesis of many carcinomas. In recent years, accumulating studies have suggested its association with the pathogenesis of malignant lymphomas. This review summarizes the latest progress in research on canonical Wnt pathway and its role in lymphocyte development and the occurrence of malignant lymphomas, in order to provide the new clues of mechanism for pathogenesis of lymphomas and then disclose new potential therapeutic targets which are based on this pathway. Copyright © 2012 by TUMOR.