Reducción de eventos cardiovasculares de Lomitapida versus Estatinas en pacientes con diagnóstico de hipercolesterolemia fa-miliar. Protocolo de revisión sistemática / Reduction of cardiovascular events of Lomitapida versus Statins in patients with a diagnosis of familial hypercholesterolemia: A systematic review protocol

Introducción: La hipercolesterolemia familiar (HF) un trastorno genético autosómico domi-nante que produce un desarrollo prematuro de enfermedades cardiovasculares. Las estati-nas han sido el medicamento de elección en estos pacientes, sin embargo, un buen por-centaje de pacientes no pueden alcanzar sus objetivos terapéuticas con las dosis máximas por lo que la Lomitapida se podría establecer como una nueva alternativa de tratamiento. Objetivo: El objetivo de esta revisión sistemática es determinar si la Lomitapida reduce los eventos cardiovasculares en pacientes con diagnóstico de Hipercolesterolemia familiar comparado con estatinas. Métodos: Se incluirán ensayos controlados aleatorios (ECA) y cuasialeatorios de pacientes con diagnóstico de HF. Las medidas de resultado los niveles de LDL, HDL pos tratamiento y eventos cardiovasculares. Las búsquedas electrónicas se realizarán en PUBMED, The Coch-rane Central Register of Controlled Trials (CENTRAL), EMBASE y Scientific electronic library (Scielo). En la evaluación del riesgo de sesgo se utilizará la herramienta de Cochrane. Las medidas del efecto del tratamiento serán las diferencias de medias (DM) y los intervalos de confianza (IC) del 95%. La evaluación de heterogeneidad se realizará mediante la inspec-ción visual del diagrama de embudo. La evaluación de la calidad de la evidencia se reali-zará usando la evaluación GRADE.

Introduction: Familial hypercholesterolemia (FH) is an autosomal dominant genetic disor-der that produces hypercholesterolemia and premature development of cardiovascular diseas-es. Statins are the drug of choice in these patients; however, a high percentage of patients cannot achieve their therapeutic goals with the maximum recommended doses, so Lo-mitapide may prove to be useful as a new treatment alternative to traditional statins. Objective: The objective of this systematic review is to determine if Lomitapide is better than statins at reducing cardiovascular events in patients with a diagnosis of FH. Methods: Randomized controlled trials (RCTs) and quasi-randomized trials of patients di-agnosed with FH will be included. Primary outcome measures included several parameters: 1. Post-treatment low- and high-density lipoprotein (LDL and HDL, respectively) levels and 2. Presence of cardiovascular events. Electronic searches will be conducted in PUBMED, The Cochrane Central Register of Controlled Trials (CENTRAL), EMBASE, and the scientific elec-tronic library (Scielo). The assessment of the risk of bias will be used by the Cochrane tool. The measures of the treatment effect will be considered the mean differences (MD) and the 95% confidence intervals (CI). The evaluation of heterogeneity will be done by visual inspec-tion of the funnel diagram. The evaluation of the quality of the evidence will be done using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) ap-proach.

Reducción de eventos cardiovasculares de Lomitapida versus Estatinas en pacientes con diagnóstico de hipercolesterolemia fa-miliar. Protocolo de revisión sistemática / Reduction of cardiovascular events of Lomitapida versus Statins in patients with a diagnosis of familial hypercholesterolemia: A systematic review protocol

Introducción: La hipercolesterolemia familiar (HF) un trastorno genético autosómico domi-nante que produce un desarrollo prematuro de enfermedades cardiovasculares. Las estati-nas han sido el medicamento de elección en estos pacientes, sin embargo, un buen por-centaje de pacientes no pueden alcanzar sus objetivos terapéuticas con las dosis máximas por lo que la Lomitapida se podría establecer como una nueva alternativa de tratamiento. Objetivo: El objetivo de esta revisión sistemática es determinar si la Lomitapida reduce los eventos cardiovasculares en pacientes con diagnóstico de Hipercolesterolemia familiar comparado con estatinas. Métodos: Se incluirán ensayos controlados aleatorios (ECA) y cuasialeatorios de pacientes con diagnóstico de HF. Las medidas de resultado los niveles de LDL, HDL pos tratamiento y eventos cardiovasculares. Las búsquedas electrónicas se realizarán en PUBMED, The Coch-rane Central Register of Controlled Trials (CENTRAL), EMBASE y Scientific electronic library (Scielo). En la evaluación del riesgo de sesgo se utilizará la herramienta de Cochrane. Las medidas del efecto del tratamiento serán las diferencias de medias (DM) y los intervalos de confianza (IC) del 95%. La evaluación de heterogeneidad se realizará mediante la inspec-ción visual del diagrama de embudo. La evaluación de la calidad de la evidencia se reali-zará usando la evaluación GRADE.

Introduction: Familial hypercholesterolemia (FH) is an autosomal dominant genetic disor-der that produces hypercholesterolemia and premature development of cardiovascular diseas-es. Statins are the drug of choice in these patients; however, a high percentage of patients cannot achieve their therapeutic goals with the maximum recommended doses, so Lo-mitapide may prove to be useful as a new treatment alternative to traditional statins. Objective: The objective of this systematic review is to determine if Lomitapide is better than statins at reducing cardiovascular events in patients with a diagnosis of FH. Methods: Randomized controlled trials (RCTs) and quasi-randomized trials of patients di-agnosed with FH will be included. Primary outcome measures included several parameters: 1. Post-treatment low- and high-density lipoprotein (LDL and HDL, respectively) levels and 2. Presence of cardiovascular events. Electronic searches will be conducted in PUBMED, The Cochrane Central Register of Controlled Trials (CENTRAL), EMBASE, and the scientific elec-tronic library (Scielo). The assessment of the risk of bias will be used by the Cochrane tool. The measures of the treatment effect will be considered the mean differences (MD) and the 95% confidence intervals (CI). The evaluation of heterogeneity will be done by visual inspec-tion of the funnel diagram. The evaluation of the quality of the evidence will be done using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) ap-proach.

Reducción de eventos cardiovasculares de Lomitapida versus Estatinas en pacientes con diagnóstico de hipercolesterolemia fa-miliar. Protocolo de revisión sistemática / Reduction of cardiovascular events of Lomitapida versus Statins in patients with a diagnosis of familial hypercholesterolemia: A systematic review protocol

Introducción: La hipercolesterolemia familiar (HF) un trastorno genético autosómico domi-nante que produce un desarrollo prematuro de enfermedades cardiovasculares. Las estati-nas han sido el medicamento de elección en estos pacientes, sin embargo, un buen por-centaje de pacientes no pueden alcanzar sus objetivos terapéuticas con las dosis máximas por lo que la Lomitapida se podría establecer como una nueva alternativa de tratamiento. Objetivo: El objetivo de esta revisión sistemática es determinar si la Lomitapida reduce los eventos cardiovasculares en pacientes con diagnóstico de Hipercolesterolemia familiar comparado con estatinas. Métodos: Se incluirán ensayos controlados aleatorios (ECA) y cuasialeatorios de pacientes con diagnóstico de HF. Las medidas de resultado los niveles de LDL, HDL pos tratamiento y eventos cardiovasculares. Las búsquedas electrónicas se realizarán en PUBMED, The Coch-rane Central Register of Controlled Trials (CENTRAL), EMBASE y Scientific electronic library (Scielo). En la evaluación del riesgo de sesgo se utilizará la herramienta de Cochrane. Las medidas del efecto del tratamiento serán las diferencias de medias (DM) y los intervalos de confianza (IC) del 95%. La evaluación de heterogeneidad se realizará mediante la inspec-ción visual del diagrama de embudo. La evaluación de la calidad de la evidencia se reali-zará usando la evaluación GRADE.

Introduction: Familial hypercholesterolemia (FH) is an autosomal dominant genetic disor-der that produces hypercholesterolemia and premature development of cardiovascular diseas-es. Statins are the drug of choice in these patients; however, a high percentage of patients cannot achieve their therapeutic goals with the maximum recommended doses, so Lo-mitapide may prove to be useful as a new treatment alternative to traditional statins. Objective: The objective of this systematic review is to determine if Lomitapide is better than statins at reducing cardiovascular events in patients with a diagnosis of FH. Methods: Randomized controlled trials (RCTs) and quasi-randomized trials of patients di-agnosed with FH will be included. Primary outcome measures included several parameters: 1. Post-treatment low- and high-density lipoprotein (LDL and HDL, respectively) levels and 2. Presence of cardiovascular events. Electronic searches will be conducted in PUBMED, The Cochrane Central Register of Controlled Trials (CENTRAL), EMBASE, and the scientific elec-tronic library (Scielo). The assessment of the risk of bias will be used by the Cochrane tool. The measures of the treatment effect will be considered the mean differences (MD) and the 95% confidence intervals (CI). The evaluation of heterogeneity will be done by visual inspec-tion of the funnel diagram. The evaluation of the quality of the evidence will be done using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) ap-proach.

Zaluzania montagnifolia: essential oil composition and biological properties / Zaluzania montagnifolia: composición del aceite esencial y actividades biológicas

The chemical composition of the seasonal essential oils (2015-2016) from the leaves and flowers of Zaluzania montagnifolia is presented. The chemical content of those oils showed quantitative and qualitative differences. Germacrene D (19.9-29.8%), camphor (12.4- 19.4%) and ß-caryophyllene (13.7-18.5%) were the most abundant volatiles in the leaves. The essential oils from the flowers contained high amounts of camphor (32.7-37.2%) limonene (19.8-24.9%) and germacrene D (3.2-7.3%). All the seasonal essential oils showed a potent in vitro inhibition against HMG-CoA reductase. The essential oils from flowers (IC50, 40.5-55.1 µg mL-1) showed better inhibition properties than those of leaves (IC50, 84.4-123.5 µg mL-1). Camphor (IC50, 72.5 µg mL-1) and borneol (IC50, 84.4 µg mL-1) exerted a non-competitive inhibition on the enzyme. Additionally, the hydrodistillates exhibited antibacterial activity against the phytopathogenic Pseudomonas syringae pv. tabaci TBR2004 (MIC, 62.7-76.5 µg mL-1) P. syringae pv. tomato DC3000 (MIC, 45.4-50.4 µg mL-1) and P. syringae pv. phaseolicola NPS3121 (MIC, 26.7-31.9 µg mL-1). Germacrene D (MIC, 35.4-66.2 µg mL-1) and ß-caryophyllene (MIC, 36.5-54.2 µg mL-1) were the strongest anti-Pseudomonas syringae agents.

Se presenta la composición química de los aceites esenciales estacionales (2015-2016) provenientes de hojas y flores de Zaluzania montagnifolia. El contenido químico de los aceites esenciales mostró diferencias cualitativas y cuantitativas. El germacreno D (19.9-29.8%), alcanfor (12.4-19.4%) y ß-cariofileno (13.7-18.5%) fueron los volátiles más abundantes en las hojas. Los aceites esenciales de las flores contuvieron altas concentraciones de alcanfor (32.7-37.2%), limoneno (19.8-24.9%) y germacreno D (3.2-7.3%). Todos los aceites esenciales estacionales mostraron una potente inhibición in vitro contra la HMG-CoA reductasa. Los aceites esenciales de las flores (IC50, 40.5-55.1 µg mL-1) mostraron mejores propiedades inhibitorias que aquellos de las hojas (IC50, 84.4-123.5 µg mL-1). El alcanfor (IC50, 72.5 µg mL-1) y el borneol (IC50, 84.4 µg mL-1) ejercieron una inhibición no competitiva sobre la enzima. Adicionalmente, los hidrodestilados exhibieron una actividad antibacterial contra los fitopatógenos Pseudomonas syringae pv. tabaci TBR2004 (MIC, 62.7-76.5 µg mL-1) P. syringae pv. tomato DC3000 (MIC, 45.4-50.4 µg mL-1) y P. syringae pv. phaseolicola NPS3121 (MIC, 26.7-31.9 µg mL-1). El germacreno D (MIC, 35.4-66.2 µg mL-1) y ß-cariofileno (MIC, 36.5-54.2 µg mL-1) fueron los agentes más fuertes contra los patovares de Pseudomonas syringae.

Effect of leech on lipid metabolism and liver in hyperlipidemia rats / 中国中药杂志

To explore the effect of leech on lipid metabolism and liver function in hyperlipidemia rats and the possible mechanism, biochemical analyzer was used to examine the regulation of leech on levels of serum triglycerides(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C). The levels of ALT and AST in serum were detected by ELISA. The proteins expression of ACAT-2, Fas and HMGCR in liver tissue was detected by Western blot. The weight of body and liver were weighed, and liver index was calculated. Oil red O staining was used to observe the lipid accumulation in liver tissue of rats by light Microscope. The results showed that leech could decrease the levels of TC, LDL-C obviously, and increase HDL-C, decrease the levels of ALT, AST and the liver index, down-regulate the proteins expression of ACAT-2, Fas and HMGCR. And oil red O staining indicated that the lipid accumulation was less in the liver tissue of the rats intervented by leech. These data indicated that leech may affect the expression of ACAT-2, Fas and HMGCR in liver tissue to reduce the synthesis of cholesterol and fatty acid, and promote the cholesterol transforming, then regulate lipid metabolism to decrease the levels of serum lipid, and reduce lipid accumulation in liver tissue and ease liver injury of rats, then slowing down the process of nonalcoholic fatty liver disease(NAFLD) in hyperlipidemia rats.

Down-regulation of single-stranded DNA-binding protein 1 expression induced by HCMV infection promotes lipid accumulation in cells

The objective of this study was to observe the infection of human cytomegalovirus (HCMV) to human umbilical vein endothelial cells, and its effect on the expression of single-stranded DNA-binding protein (SSBP1) and on lipid metabolism in endothelial cells. We screened the differential expression of mRNAs after HCMV infection by suppression subtractive hybridization and the expression levels of SSBP1 mRNA and protein after HCMV infection by real-time PCR and western blot. After verification of successful infection by indirect immunofluorescent staining and RT-PCR, we found a differential expression of lipid metabolism-related genes including LDLR, SCARB, CETP, HMGCR, ApoB and LPL induced by HCMV infection. The expression levels of SSBP1 mRNA and protein after HCMV infection were significantly down-regulated. Furthermore, we found that upregulation of SSBP1 inhibited the expression of atherosclerosis-associated LDLR, SCARB, HMGCR, CETP as well as the accumulation of lipids in the cells. The results showed that the inhibition of SSBP1 by HCMV infection promotes lipid accumulation in the cells.

Continuum Cardiovascular 25 anos - Evolução de um Modelo Etiofisiopatológico / Cardiovascular Continuum 25 years – The Evolution of an Etiopathophysiology Model

O conceito de continuum cardiovascular foi elaborado por Dzau e Braunwald e difundido entre cardiologistas como modelo etiofisiopatológico que direciona o desenvolvimento de medidas intervencionistas na prevenção das doenças cardiovasculares. Após workshop realizado em 1989, foi possível reunir questões resolvidas e não resolvidas sobre fatores relacionados à terapia e proteção cardiovascular, resultando na primeira publicação por Dzau e Braunwald, em 1991. O avanço nos estudos da biologia molecular e celular permitiu entendimento do papel da disfunção endotelial no estresse oxidativo e do óxido nítrico na doença aterosclerótica coronariana (DAC),concedendo prêmio Nobel aos autores Ferid Murad, Robert Furchgott e Louis Ignarro. Em 2006, uma segunda publicação, também liderada por Dzau, consolidou o modelo clássico do continuum cardiovascular, no qual fatores de risco para DAC se associam e desencadeiam cascata progressiva de eventos levando aos estágios finais da doença cardiovascular. A partir da observação da existência de doenças isquêmicas do miocárdio em populações com baixa incidência de aterosclerose coronariana, estudos comprovaram que a doença isquêmica não está apenas associada à aterosclerose, mas também ao envelhecimento vascular. Este achado levou à publicação do terceiro artigo de O’Rourke em 2010, que apresentou um modelo adicional: continuum do envelhecimento vascular. Aevolução desse modelo possibilitou o enfoque não apenas em tratamentos preventivos dos fatores de risco da DAC, mas também na busca de terapias capazes de prevenir dano endotelial causado pelo envelhecimento vascular e de modular o estresse oxidativo. Esta revisão tem por objetivo reunir os principais estudos que embasam a evolução do modelo continuum cadiovascular em 25 anos...

The concept of cardiovascular continuum was devised by Dzau and Braunwald and spread among cardiologists as na etiopathophysiology model that directs the development of interventionist measures in the prevention of cardiovascular diseases. After a workshop held in 1989, it was possible to gather resolved and unresolved issues about factors related to cardiovascular therapy and protection, resulting in the first publication by Dzau and Braunwald, in 1991. Progress in the studies of molecular and cellular biology allowed understanding the role of endothelial dysfunction in oxidative stress and nitric oxide in coronary artery atherosclerosis (CAA), awarding Nobel Prize to authors Ferid Murad, Robert Furchgott and Louis Ignarro. In 2006, a second publication, also ledby Dzau, consolidated the classic model of cardiovascular continuum, in which risk factors for CAA are associated and trigger a progressive cascade of events leading to the final stages of cardiovascular disease. By observing the existence of ischemic myocardial diseases in populations with low incidence of coronary artery atherosclerosis, studies have shown that ischemic heart disease is not only associated with atherosclerosis, but also to vascular aging. This finding led to the publication of O’Rourke’s third manuscriptin 2010, which presented an additional model: vascular aging continuum. The evolution of this model allowed focusing on preventive treatments for the risk factors of CAA and the search for therapies capable of preventing endothelial damage caused by vascular aging and modulating oxidative stress. This review aims to bring together the leading studies that support the evolution of the cardiovascular continuum model over 25 years...

Inhibition of HMG-CoA reductase by MFS, a purified extract from the fermentation of marine fungus Fusarium solani FG319, and optimization of MFS production using response surface methodology / 中国天然药物

The present study was designed to isolate and characterize a purified extract from Fusarium solani FG319, termed MFS (Metabolite of Fusarium solani FG319) that showed anti-atherosclerosis activity by inhibiting 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. Response surface methodology (RSM) was employed to achieve an improved yield from the fermentation medium. The inhibiting effect of the isolate, MFS, on HMG-CoA reductase was greater than that of the positive control, lovastatin. The average recovery of MFS and the relative standard deviation (RSD) ranged between 99.75% to 101.18%, and 0.31% to 0.74%, respectively. The RSDs intra- and inter-assay of the three samples ranged from 0.288% to 2.438%, and from 0.934% to 2.383%, respectively. From the RSM, the concentration of inducer, cultivation time, and culture temperatures had significant effects on the MFS production, with the effect of inducer concentration being more pronounced that other factors. In conclusion, the optimal conditions for the MFS production were achieved using RSM and that MFS could be explored as an anti-atherosclerosis agent based on its ability to inhibit HMG-CoA reductase.

Study on anti-hyperlipidemia mechanism of high frequency herb pairs by molecular docking method / 中国中药杂志

Traditional Chinese medicine (TCM) has definitely clinical effect in treating hyperlipidemia, but the action mechanism still need to be explored. Based on consulting Chinese Pharmacopoeia (2010), all the lipid-lowering Chinese patent medicines were analyzed by associated rules data mining method to explore high frequency herb pairs. The top three couplet medicines with high support degree were Puerariae Lobatae Radix-Crataegi Fructus, Salviae Miltiorrhizae Radix et Rhizoma-Crataegi Fructus, and Polygoni Multiflori Radix-Crataegi Fructus. The 20 main ingredients were selected from the herb pairs and docked with 3 key hyperlipidemia targets, namely 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), peroxisome proliferator activated receptor-α (PPAR-α ) and niemann-pick C1 like 1 (NPC1L1) to further discuss the molecular mechanism of the high frequency herb pairs, by using the docking program, LibDock. To construct evaluation rules for the ingredients of herb pairs, the root-mean-square deviation (RMSD) value between computed and initial complexes was first calculated to validate the fitness of LibDock models. Then, the key residues were also confirmed by analyzing the interactions of those 3 proteins and corresponding marketed drugs. The docking results showed that hyperin, puerarin, salvianolic acid A and polydatin can interact with two targets, and the other five compounds may be potent for at least one of the three targets. In this study, the multi-target effect of high frequency herb pairs for lipid-lowering was discussed on the molecular level, which can help further researching new multi-target anti-hyperlipidemia drug.

Regulatory effect of coptisine on key genes involved in cholesterol metabolism / 中国中药杂志

To study the effect of cholesterol and 25-OH-cholesterol on cholesterol metabolism in HepG2 cells and the effect of coptisine (Cop) extracted from Coptidis Rhizoma (CR) in reducing and regulating cholesterol. In this study, TC, TG, LDL-c and HDL-c were measured by biochemical analysis; mRNA and protein expressions of LDLR, HMGCR and CYP7A1 were detected by qRT-PCR and Western blot. According to the results, cholesterol and 25-OH-cholesterol inducing could decrease in mRNA and protein expressions of LDLR and CYP7A1, so as to increase TC and LDL-c contents. However, Cop could up-regulate mRNA and protein expressions of LDLR and CYP7A1 and down-regulate that of HMGCR, so as to reduce TC and LDL-c levels. These findings suggested that Cop has potential pharmacological activity for reducing cholesterol, and may reduce cholesterol by regulating mRNA and protein expressions of key genes involved in cholesterol metabolism, such as LDLR, CYP7A1 and HMGCR. This study laid a firm theoretical foundation for developing new natural drugs with the cholesterol-lowering activity.

Inhibition of proprotein convertase subtilisin/kexin type 9: a novel mechanism of berberine and 8-hydroxy dihydroberberine against hyperlipidemia / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect and molecular mechanisms of different doses of 8-hydroxy dihydroberberine (Hdber) for the treatment of hyperlipidemia in rats.</p><p><b>METHODS</b>A rat model of hyperlipidemia was established by feeding rats a high-fat diet for 4 weeks in 70 rats of 80 animals, and 10 rats were randomly selected as control group. The hyperlipidemic rats were then randomly divided into the following groups: a model group (MOD); a berberine group [BBR, 156 mg/(kg day)]; Hdber groups, which were treated with different doses of Hdber [78, 39 and 19.5 mg/(kg day)]; and a simvastatin group [SIM, 4 mg/(kg day)]. The corresponding therapy was administered to the rats of each treatment via gastric tubes. Normal animals were used as a control group. The blood levels of various lipids, including total cholesterol, triglycerides, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, free fatty acid (FFA), apolipoprotein AI(Apo-AI) and apolipoprotein B (Apo-B) were examined. The protein expressions of low-density lipoprotein receptor (LDL-R), sterol regulatory element-binding protein 2 (SREBP-2), 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) and proprotein convertase subtilisin/kexin type 9 (PCSK-9) in liver tissues were determined by Western blot analysis.</p><p><b>RESULTS</b>Compared with the control group of rats, the model group demonstrated a deteriorated blood lipid profile and exhibited increased expression levels of PCSK-9 protein in their liver tissues (P<0.01). In addition, the high-fat diet decreased the expression levels of LDL-R, SREBP-2 and HMGCR proteins in murine liver tissues. However, the addition of berberine or Hdber reversed the blood lipid profile changes (P<0.05 or P<0.01), decreased the expression levels of PCSK-9 proteins (P<0.01), and increased the expression levels of LDL-R proteins in the hyperlipidemic rats (P<0.01). These compounds did not significantly influence the expression levels of SREBP-2 and HMGCR proteins in the hyperlipidemic rats.</p><p><b>CONCLUSIONS</b>Hdber is effective in the treatment of hyperlipidemia in rats. The therapeutic mechanisms of Hdber may be associated with increasing the expression of LDL-R protein and decreasing the expression of PCSK-9 protein in liver tissues.</p>

Cloning and characterization of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) gene from Paris fargesii Franch.

3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) plays an important role in catalyzing the first committed step of isoprenoids biosynthesis in mevalonic acid (MVA) pathway. Here, we cloned a full-length transcript of Paris fargesii Franch. The full-length cDNA of P. fargesii HMGR (Pf-HMGR, GenBank accession no. JX508638) was 1,973 bp and contained a 1,728 bp ORF encoding 576 amino acids. Sequence analysis revealed that the deduced Pf-HMGR had high similarity with HMGRs from other plants, including Ricinus communis (77%), Litchi chinensis (76%), Michelia chapensis (75%) and Panax quinquefolius (72%). It had a calculated molecular mass of about 62.13 kDa and an isoelectric point (pI) of 8.47. It contained two transmembrane domains, two putative HMGR binding sites and two NADP(H)-binding sites. The predicted 3-D structure revealed that Pf-HMGR had a similar spatial structure with other plant HMGRs. Three catalytic regions, including L-domain, N-domain and S-domain were detected by structural modeling of HMGR. Tissue expression analysis revealed that Pf-HMGR was strongly expressed in roots and stems than in leaves. Taken together, our data laid a foundation for further investigation of HMGR's functions and regulatory mechanisms in plants.

Efeitos pleiotrópicos das estatinas na sepse / Pleiotropic effects of statins in sepsis

O objetivo do presente estudo foi revisar os efeitos da terapia com estatinas em pacientes com quadro de sepse. Foi realizada uma busca bibliográfica de artigos na base de dados MEDLINE/PubMed, SciELO e LILACS, no período de publicação delimitado entre 2009 e 2013. Foram incluídos artigos referentes a ensaios clínicos randomizados controlados e estudos de coorte observacionais clínicos. Foram encontrados diversos estudos clínicos e observacionais que investigaram o efeito do uso de estatinas em infecções e em sepse, tanto de uso contínuo pré-hospitalar (com ou sem interrupção durante internação) ou com início imediato pós-internação. Alguns estudos descrevem prováveis efeitos positivos e benéficos da terapia com estatinas no quadro de sepse, dentre eles a melhora dos parâmetros inflamatórios e da taxa de mortalidade, porém, esses resultados não se sustentam quando são aplicados métodos estatísticos que levamem conta diferentes variáveis, tais como idade, sexo, comorbidades e gravidade da doença. Até o momento nenhum estudo demonstrou evidências sólidas e significativas quanto à redução de mortalidadee morbidade no quadro séptico associado ao uso de estatinas, indicando que seu efeito benéfico e protetor ainda não foi plenamente delimitado, sendo necessários mais estudos que confirmem os resultados até agora encontrados.

The objective of this study was to review the effects of statin therapy in patients with signs of sepsis. A bibliographic search of articles published between 2009-2013 was performed in the MEDLINE/PubMed, SciELO and LILACS databases. Randomized controlled trials and observational clinical cohort studies were included. The results show that several clinical and observational studies have investigated the effect of statin in infection and sepsis, both pre-hospital continuous use (with or without interruption during hospitalization) or starting immediately post-hospitalization. Some studies describe positive and beneficial effects of statin therapy in the contextof sepsis, including improvement on inflammatory parameters and mortality rates. However, these results do not hold on when statistic methods, which take into account different variables such as age, sex, comorbidities and severity disease, are applied. To date, no study has demonstrated strong and significant evidence regarding the reduction of morbidity and mortality in sepsis associated with the use of statin. This indicates that beneficial and protective effects have not been fully defined yet. More researches are required to confirm the results found so far.

Epigallocatechin-3-gallate [EGCG] inhibits 3-hydroxy-3-methylglutaryl-CoA reductase in the presence of glycerol

The inhibition of 3-hydroxy-3-methylglutaryl CoA reductase [HMGCR] is considered able to decrease serum cholesterol levels and dramatically reduce the risk for cardiovascular and cerebrovascular diseases. The statins, competitive inhibitors of HMGCR, have been employed to control hypercholesterolemia. But their side effects, especially their safety of long-term administration have attracted great attention. Therefore, there is still an urgent requirement for the development of safer inhibitors of HMGCR with less serious side effects. In this study, we cloned and purified the catalytic domain of human HMGCR [delta HMGCR], and applied the method of Ultra Performance Liquid Chromatography [UPLC] to assay delta HMGCR activity and screen its inhibitors from natural products. The results indicated that EGCG can inhibit delta HMGCR in the presence of some glycerol in vitro and can decrease cellular total cholesterol in HepG2 cells. As a consequence, it is promising to put EGCG into the development of hypolipidemic health product

Molecular characterization of a HMG-CoA reductase gene from a rare and endangered medicinal plant, Dendrobium officinale / 药学学报

The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate in mavalonic acid pathway, which is the first committed step for isoprenoid biosynthesis in plants. However, it still remains unclear whether HGMR gene plays a role in the isoprenoid biosynthesis in Dendrobium officinale, an endangered epiphytic orchid species. In the present study, a HMGR encoding gene, designed as DoHMGR1 (GenBank accession JX272632), was identified from D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, for the first time. The full length cDNA of DoHMGR1 was 2 071 bp in length and encoded a 562-aa protein with a molecular weight of 59.73 kD and an isoelectric point (pI) of 6.18. The deduced DoHMGR1 protein, like other HMGR proteins, constituted four conserved domains (63-561, 147-551, 268-383 and 124-541) and two transmembrane motifs (42-64 and 85-107). Multiple sequence alignment and phylogenetic analyses demonstrated that DoHMGR1 had high identity (67%-89%) to a number of HMGR genes from various plants and was closely related to Vanda hybrid cultivar, rice and maize monocots. Real time quantitative PCR (qPCR) analysis revealed that DoHMGR1 was expressed in the three included organs. The transcripts were the most abundant in the roots with 2.13 fold over that in the leaves, followed by that in the stems with 1.98 fold. Molecular characterization of DoHMGR1 will be useful for further functional elucidation of the gene involving in isoprenoid biosynthesis pathway in D. officinale.

Match of functional module with chassis in 7-dehydrocholesterol synthesis / 生物工程学报

The key challenge to generate engineered cells by synthetic biology for producing 7-dehydrocholesterol (7-DHC) in a high titer is the match between functional module and chassis. Our study focused on solving this problem by combining different promoters and yeast chassis to increase 7-DHC production. To optimize the chassis in order to accumulate zymosterol, the substrate for 7-DHC synthesis, we overexpressed truncated HMG-CoA reductase (tHmglp) and squalene epoxidase (Erglp), both are key genes of yeast endogenous zymosterol biosynthetic pathway. In addition, we knocked out C-24 methyl transferase (Erg6p) and C-22 dehydrogenase (Erg5p) to inhibit the conversion of zymosterol to ergosterol. By introducing heterologous C-24 reductase under three promoters with different strengths, namely TDH3p, PGK1p and TDH1p, we constructed functional modules of diverse activities. Nine engineeredcells were generated based on the combination of these three modules and three chassis. The result shows that the engineered cell composed of functional module regulated by TDH3p and chassis SyBE_000956 had the highest 7-DHC production, indicating a better match than others. This study provides evidences for importance of match and empirical support for rational design of subsequent researches.

Production of β-carotene by metabolically engineered Saccharomyces cerevisiae / 生物工程学报

β-carotene has a wide range of application in food, pharmaceutical and cosmetic industries. For microbial production of β-carotene in Saccharomyces cerevisiae, the supply of geranylgeranyl diphosphate (GGPP) was firstly increased in S. cerevisiae BY4742 to obtain strain BY4742-T2 through over-expressing truncated 3-hydroxy-3-methylglutaryl-CoA reductase (tHMGR), which is the major rate-limiting enzyme in the mevalonate (MVA) pathway, and GGPP synthase (GGPS), which is a key enzyme in the diterpenoid synthetic pathway. The β-carotene synthetic genes of Pantoea agglomerans and Xanthophyllomyces dendrorhous were further integrated into strain BY4742-T2 for comparing β-carotene production. Over-expression of tHMGR and GGPS genes led to 26.0-fold increase of β-carotene production. In addition, genes from X. dendrorhous was more efficient than those from P. agglomerans for β-carotene production in S. cerevisiae. Strain BW02 was obtained which produced 1.56 mg/g (dry cell weight) β-carotene, which could be used further for constructing cell factories for β-carotene production.

The impact of lipid-metabolizing genetic polymorphisms on body mass index and their interactions with soybean food intake: a study in a Chinese population / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate the association of known polymorphisms in the lipid metabolic pathway with body mass index (BMI), and estimate their interactions with soybean food intake.</p><p><b>METHODS</b>A community-based cross-sectional survey was conducted in a Chinese Han population. BMI, soybean food intake, and single nucleotide polymorphisms of rs599839, rs3846662, rs3846663, rs12916, rs174547, rs174570, rs4938303, and rs1558861 were measured in 944 subjects. A multivariate logistic regression was used to analyze the association of the studied polymorphisms with BMIs. The expectation-maximization algorithm was employed to evaluate the extent of linkage disequilibrium between pairwise polymorphisms. The gene-environment interaction was assessed in the general multifactor dimensionality reduction model.</p><p><b>RESULTS</b>The polymorphisms of rs3846662 and rs3846663 were associated with 10% highest BMIs when comparing to the 10% lowest values both in individuals and haplotype-based association tests. Although no statistically significant gene-environment interactions were found, people with the haplotype composed of C allele in rs3846662 and T allele in rs3846663 and low frequency of soybean intake had significantly higher risk to overweight and obesity as compared with those with the haplotype consisting of T allele in rs3846662 and C allele in rs3846663 and highly frequent soybean food intake, with an odds ratio of 1.64 (95% confidence interval: 1.15-2.34, P<0.01) after adjusting for the common confounders.</p><p><b>CONCLUSION</b>Our study has suggested that rs3846662 and rs3846663 may be the potential candidate polymorphisms for obesity, and their effect on the pathogenesis could be mediated by the frequency of soybean food intake.</p>

Ethylene treatment improves diosgenin accumulation in in vitro cultures of Dioscorea zingiberensis via up-regulation of CAS and HMGR gene expression

Background: The perennial medicinal herb Dioscorea zingiberensis is a very important plant used for steroid drug manufacturing for its high level of diosgenin in rhizome. Although the stimulation of diosgenin accumulation by ethylene has been reported in a few of plant species, its regulation is not yet characterized at the molecular level, the underlying molecular mechanism remains elusive. Results: In this study, the effects of ethylene on diosgenin biosynthesis in in vitro cultures of D. zingiberensis were described. The results showed that, in samples treated with ethylene at concentration E3 (10(4) dilution of 40% ethephon), the diosgenin biosynthesis was significantly promoted in comparison with the control samples. Treatment with high concentrations of ethylene had inhibitory effect, whereas with low concentration of the gas elicitor brought about no detectable deleterious effect on the growth rate and diosgenin content of the cultures. The considerable increase of diosgenin level in in vitro cultured Dioscorea zingiberensis by ethylene application is accompanied by the concomitant increase of soluble proteins and chlorophyll content. The gene expressions of cycloartenol synthase and 3-hydroxy-3-methylglutaryl-CoA reductase but not of squalene synthase or farnesyl pyrophosphate synthase were up-regulated by applied ethylene. Conclusions: Our results suggest that ethylene treatment enhanced diosgenin accumulation via up-regulation of the gene expressions of cycloartenol synthase and 3-hydroxy-3-methylglutaryl-CoA reductase.

Over - expression effect of gene encoding 3-hydroxy-3-methylglutaryl-CoA reductase on production of taxol in Iranian hazel [Corylus avellana L.]

Sustainable and commercial production of taxol as an anti cancer drug is a critical point to its clinical application. Nowadays, hazel because of rapid growth and wide range distribution is considered as an alternative source of Taxol. To increase taxol production the cDNA encoding 3-hydroxy-3-methylglutaryl-CoA reductase [HMGR] from Iranian hazel [GeneBank accession number KF306244, showed by CIHMGR] was isolated and over-expressed in pCAMBIA1304 binary vector. The effect of transient over-expression of HMGR in callus and leaf were evaluated on Taxol production. The calli was established through the culture of immature cotyledon on Murashige and Skoog basal medium supplemented with 2, 4-D and BA. The first strand cDNA of CiHMGR was synthesized by specific primers. Enzymatic assay of recombinant CiHMGR in E. coli were done by western blott and His-tag affinity techniques. Also production of taxol in transformed callus and leaf were evaluated by HPLC analysis. An Open Reading Frame [ORF] with 1698 bp length and a deduced polypeptide with 566 amino acid residues were amplified. The highest and lowest amount of taxol was 0.016 mg/g. DW and 0.004 mg/gDW in transformed calli and untransformed leaves respectively. Generally the over-expression of HMGR increase the total isoprenoids yield, therefore to have high production of target secondary metabolites [taxol] we need both of network of transformed genes and elicited cell culture