Glucose metabolic reprogramming-related parameters for the prediction of 28-day neurological prognosis and all-cause mortality in patients after cardiac arrest:a prospective single-center observational study

BACKGROUND:We aimed to observe the dynamic changes in glucose metabolic reprogrammingrelated parameters and their ability to predict neurological prognosis and all-cause mortality in cardiac arrest patients after the restoration of spontaneous circulation(ROSC).METHODS:Adult cardiac arrest patients after ROSC who were admitted to the emergency or cardiac intensive care unit of the First Aflliated Hospital of Dalian Medical University from August 1,2017,to May 30,2021,were enrolled.According to 28-day survival,the patients were divided into a non-survival group(n=82) and a survival group(n=38).Healthy adult volunteers(n=40) of similar ages and sexes were selected as controls.The serum levels of glucose metabolic reprogrammingrelated parameters(lactate dehydrogenase [LDH],lactate and pyruvate),neuron-specific enolase(NSE) and interleukin 6(IL-6) were measured on days 1,3,and 7 after ROSC.The Acute Physiology and Chronic Health Evaluation II(APACHE II) score and Sequential Organ Failure Assessment(SOFA) score were calculated.The Cerebral Performance Category(CPC) score was recorded on day 28 after ROSC.RESULTS:Following ROSC,the serum LDH(607.0 U/L vs.286.5 U/L),lactate(5.0 mmol/L vs.2.0 mmol/L),pyruvate(178.0 μmol/L vs.70.9 μmol/L),and lactate/pyruvate ratio(34.1 vs.22.1) significantly increased and were higher in the non-survivors than in the survivors on admission(all P<0.05).Moreover,the serum LDH,pyruvate,IL-6,APACHE II score,and SOFA score on days 1,3 and 7 after ROSC were significantly associated with 28-day poor neurological prognosis and 28-day all-cause mortality(all P<0.05).The serum LDH concentration on day 1 after ROSC had an area under the receiver operating characteristic curve(AUC) of 0.904 [95% confidence interval [95% CI]:0.851–0.957]) with 96.8% specificity for predicting 28-day neurological prognosis and an AUC of 0.950(95% CI:0.911–0.989) with 94.7% specificity for predicting 28-day all-cause mortality,which was the highest among the glucose metabolic reprogramming-related parameters tested.CONCLUSION:Serum parameters related to glucose metabolic reprogramming were significantly increased after ROSC.Increased serum LDH and pyruvate levels,and lactate/pyruvate ratio may be associated with 28-day poor neurological prognosis and all-cause mortality after ROSC,and the predictive eflcacy of LDH during the first week was superior to others.

Linking uric acid metabolism to diabetic complications

Hyperuricemia have been thought to be caused by the ingestion of large amounts of purines, and prevention or treatment of hyperuricemia has intended to prevent gout. Xanthine dehydrogenase/xanthine oxidase(XDH/XO) is rate-limiting enzyme of uric acid generation, and allopurinol was developed as a uric acid(UA) generation inhibitor in the 1950 s and has been routinely used for gout prevention since then. Serum UA levels are an important risk factor of disease progression for various diseases, including those related to lifestyle. Recently, other UA generation inhibitors such as febuxostat and topiroxostat were launched. The emergence of these novel medications has promoted new research in the field. Lifestyle-related diseases, such as metabolic syndrome or type 2 diabetes mellitus, often have a common pathological foundation. As such, hyperuricemia is often present among these patients. Many in vitro and animal studies have implicated inflammation and oxidative stress in UA metabolism and vascular injury because XDH/XO act as one of the major source of reactive oxygen species Many studies on UA levels and associated diseases implicate involvement of UA generation in disease onset and/or progression. Interventional studies for UA generation, not UA excretion revealed XDH/XO can be the therapeutic target forvascular injury and renal dysfunction. In this review, the relationship between UA metabolism and diabetic complications is highlighted.

Function of Lactate Dehydrogenase in Cardiac and Skeletal Muscle of Phrynocephalus Lizard in Relation to High-Altitude Adaptation

Poikilothermic animals living in high-altitude environments can be greatly affected by the anaerobic metabolism and lactate recycling, which are catalyzed by an enzyme called lactate dehydrogenase(LDH). However, the function and possible regulatory mechanisms of their anaerobic glycolysis remained elusive. We compared the difference in LDH between a native high-altitude(4 353 m) lizard, Phrynocephalus erythrurus, and a closely related species, Phrynocephalus przewalskii that lives in intermediate altitude environment(1 400 m). The activity of LDH, the concentration of lactate, the distribution of isoenzyme, and the mRNA amounts of Ldh-A and Ldh-B were determined. In cardiac muscle, the lactate-forming activity of P. erythrurus in LDH was higher than of P. przewalskii LDH at all three temperatures tested(10 °C, 25 °C and 35 °C), while lactate-oxidation activity of LDH was significantly different between the two species only at 25 °C and 35 °C. In skeletal muscle, both lactate-forming and lactate-oxidation rates of P. erythrurus were lower than that of P. przewalskii. There was a higher proportion of H subunit and a significantly higher expression of Ldh-B, with a concomitant decrease of lactate concentration in P. erythrurus. These results indicate that P. erythrurus may have a strong potential for anaerobic metabolism, which is likely adapted to the hypoxic environment at high altitudes. Furthermore, P. erythrurus is capable of oxidizing more lactate than P. przewalskii. The Ldh-A cDNA of the two species consists of a 999 bp open reading frame(ORF), which encodes 332 amino acids, while Ldh-B cDNA consists of a 1 002 bp ORF encoding 333 amino acids. LDHA has the same amino acid sequence between the two species, but three amino acid substitutions(V12 I, N21S and N318K) were observed in LDHB. Structure analysis of LDH indicated that the substitutions of residues Val12 and Asp21 in P. erythrurus could be responsible for the highaltitude adaptation. The LDH characteristics of LDH in P. erythrurus suggest unique adaptation strategies of anaerobic metabolism in hypoxia and cold environments at high altitudes for poikilothermic animals.

A rice XANTHINE DEHYDROGENASE gene regulates leaf senescence and response to abiotic stresses

Xanthine dehydrogenase, a member of the molybdenum enzyme family, participates in purine metabolism and catalyzes the generation of ureides from xanthine and hypoxanthine. However, the mechanisms by which xanthine dehydrogenase affects rice growth and development are poorly understood. In the present study, we identified a mutant with early leaf senescence and reduced tillering that we named early senescence and less-tillering 1(esl1). Map-based cloning revealed that ESL1 encodes a xanthine dehydrogenase, and it was expressed in all tissues. Chlorophyll content was reduced and chloroplast maldevelopment was severe in the esl1 mutant. Mutation of ESL1 led to decreases in allantoin, allantoate, and ABA contents. Further analysis revealed that the accumulation of reactive oxygen species in esl1 resulted in decreased photosynthesis and impaired chloroplast development, along with increased sensitivity to abscisic acid and abiotic stresses. Ttranscriptome analysis showed that the ESL1 mutation altered the expression of genes involved in the photosynthesis process and reactive oxygen species metabolism.Our results suggest that ESL1 is involved in purine metabolism and the induction of leaf senescence.These findings reveal novel molecular mechanisms of ESL1 gene-mediated plant growth and leaf senescence.

Positive effect of ethanol-induced Lactococcus lactis on alcohol metabolism in mice

It is well known that exposure to environmental stresses could enhance the adaptability of bacteria and up-regulate the expression of a variety of oxidative stress-related genes and antioxidant enzymes.It is unclear whether the adaptability of microorganisms formed naturally in special environments could transfer to other organisms.The study aimed to evaluate the effects of untreated and ethanol-induced Lactococcus lactis intracellular extracts(U-IE and E-IE)on alcohol metabolism in mice.The positive effects of E-IE on alcohol metabolism in mice were revealed by the enhanced latency of loss of righting reflex(LORR),the reduced duration of LORR,the decrease of blood alcohol concentration,as well as the elevation of alcohol dehydrogenase(ADH)activities in the stomach and liver tissues.Furthermore,the potential benefits of E-IE on the liver were evaluated by biochemical parameters including the activities of serum transaminase,the levels of antioxidant enzymes,and the pathological changes of liver tissue.The present work put forward a new point that appropriate ethanol stress could enhance the intracellular ADH activity of L.lactis,and its intracellular extracts could continue to enhance alcohol metabolism in mice.

Silencing of Os XDH reveals the role of purine metabolism in dark tolerance in rice seedlings

Xanthine dehydrogenase(XDH) is a crucial enzyme involved in purine metabolism. To evaluate the effect of XDH deficiency on rice growth during dark treatment, wild type(WT) Nipponbare(Oryza sativa L.) and two independent transgenic lines with severe RNAi suppression(xdh3 and xdh4) were used in the present experiment. Under normal growth conditions, chlorophyll levels and biomass were indistinguishable between WT and the two RNAi transgenic lines, but XDH enzyme activity and ureide levels were suppressed in XDH RNAi transgenic lines. When XDH RNAi transgenic lines were subjected to dark treatment, chlorophyll content and biomass were significantly decreased, while O~–· production rate and malonaldehyde(MDA) were significantly increased compared to WT. The spraying test of exogenous allantoin raised chlorophyll content and biomass and reduced O~–· production rate and MDA in WT and both transgenic lines, and it also simultaneously reduced differences between RNAi and WT plants caused by XDH deficiency in growth potential and anti-oxidative capacity under dark treatment. These results suggested that fully functional purine metabolism plays an important role in reducing the sensitivity of rice seedlings to dark stress.

Aqueous Components of Tomato Accelerate Alcohol Metabolism by Increasing Pyruvate Level

Consumption of food while drinking alcohol has been suggested to play important roles in alleviating the physiological and pharmacological influences of alcohol. Vegetables are believed to provide health benefits, but there is little evidence for their influence on the effects of alcohol consumption. The present study aimed to investigate the effect of a common vegetable, tomato, on alcohol metabolism. In a randomized, controlled, crossover study with12 Japanese healthy men aged between 24 and 56 years, drinking tomato juice containing 5% (v/v) alcohol (TJAlc) significantly attenuated the elevation of blood ethanol level and subsequently increased the level of acetate compared with a water-based alcoholic beverage with an equal dose of alcohol (0.4 g/kg body weight). Significantly higher levels of blood pyruvate and lactate were also observed in subjects who had consumed TJAlc compared with those consuming the water-based beverage. Additionally, a biphasic alcohol effects scale method showed that subjective feelings for alcohol-induced stimulant effects were significantly enhanced by drinking TJAlc. Animal experiments using male Sprague Dawleyrats suggested that the effect on blood biomarkers was attributable to the serum fraction of tomato (TS), which largely consisted of aqueous compounds, but not lipophilic compounds such as the carotenoid lycopene. Furthermore, it was suggested the TS possibly included potent compound(s) in addition to alanine, glutamine, and citric acid, all of which have previously been reported to affect alcohol metabolism. Administration of TS clearly increased the activity of NAD (H)-dependent enzymes such as lactate-(LDH), alcohol-, and aldehyde-dehydrogenase in rat liver cytosols. These findings suggest that aqueous compound(s) in tomato promote alcohol metabolism, probably through increasing pyruvate level, enhancing LDH activity, and improving the ratio of NAD to NADH.

EFFECT OF ACUPUNCTURE ON CARBOHYDRATE METABOLISM IN PATIENTS WITH SIMPLE OBESITY

In an attempt to elucidate the effect ofacupuncture on carbohydrate metabolism inpatients with simple obesity and its mecha-nism in reducing excess body fat,obesity in-dexes,fasting blood sugar,insulin contentand activity of lactic dehydrogenase(LDH)were all observed before and afteracupuncture treatment.

Prognostic performance of an index based on lactic dehydrogenase and transaminases for patients with liver steatosis and COVID-19

BACKGROUND Metabolic associated fatty liver disease(MAFLD)is associated with complications and mortality in patients with coronavirus disease 2019(COVID-19).However,there are no prognostic scores aimed to evaluate the risk of severe disease specifically in patients with MAFLD,despite its high prevalence.Lactate dehydrogenase,aspartate aminotransferase and alanine aminotransferase have been used as markers of liver damage.Therefore,we propose an index based on lactate dehydrogenase,aspartate aminotransferase and alanine aminotransferase for the prediction of complications and mortality in patients with MAFLD and COVID-19.AIM To evaluate the prognostic performance of an index based on lactate dehydrogenase and transaminases(aspartate aminotransferase/alanine aminotransferase)in patients with COVID-19 and MAFLD[liver fibrosis and nutrition(LNF)-COVID-19 index].METHODS In this retrospective cohort study,two cohorts from two different tertiary centers were included.The first was the derivation cohort to obtain the score cutoffs,and the second was the validation cohort.We included hospitalized patients with severe COVID-19 and MAFLD.Liver steatosis was evaluated by computed tomography scan.Area under the receiver operating characteristic(ROC)curve analysis and survival analysis were used.RESULTS In the derivation cohort,44.6%had MAFLD;ROC curve analysis yielded a LFN-COVID-19 index>1.67 as the best cutoff,with a sensitivity of 78%,specificity of 63%,negative predictive value of 91%and an area under the ROC curve of 0.77.In the multivariate analysis,the LFN-COVID-19 index>1.67 was independently associated with the development of acute kidney injury(odds ratio:1.8,95%confidence interval:1.3-2.5,P<0.001),orotracheal intubation(odds ratio:1.9,95%confidence interval:1.4-2.4,P<0.001),and death(odds ratio:2.86,95%confidence interval:1.6-4.5,P<0.001)in both cohorts.CONCLUSION LFN-COVID-19 index has a good performance to predict prognosis in patients with MAFLD and COVID-19,which could be useful for the MAFLD population.

磷化氢的毒理及害虫对磷化氢的抗性机制研究进展

近60年来,熏蒸剂磷化氢被广泛用于储藏物害虫的防治。但是,长期、不合理地使用磷化氢,导致储藏物害虫抗药性的广泛产生。了解磷化氢的毒理机制可以为磷化氢抗性机制研究提供思路。早期的研究发现,磷化氢通过干扰神经传导、抑制能量代谢和破坏氧化还原系统,引起害虫死亡;但近年的研究表明,磷化氢致死害虫的主要机制是抑制能量生成和通过干扰氧化还原系统来增加氧化损伤。早期的研究发现,害虫对磷化氢的抗性机制主要包括主动排斥磷化氢、保护性昏迷和增强解毒酶活性。近年来,随着基因组学、蛋白组学和代谢组学的应用,相继出现一些新的磷化氢抗性机制,如穿透抗性、磷化氢作用靶标敏感性降低、能量代谢模式调整。越来越多的研究表明,靶标二氢硫辛酰胺脱氢酶突变以及抗氧化酶和解毒酶活性增强是主要的抗性机制,而能量代谢模式调整可能是抗性形成初期抵抗磷化氢不良影响的重要机制。采用基因渐渗的方法研究害虫磷化氢抗性突变的适合度代价可以更精准地预测抗性突变的进化方向。研究害虫的磷化氢抗性机制和抗性突变的进化潜力不仅有助于理解害虫抗药性的形成和生物的进化,同时对害虫的磷化氢抗性监测和治理有重要的意义。

酱香型白酒风味成分对小鼠乙醇代谢及关键酶活性的影响

选取产自山东、四川、贵州的酱香型白酒(编号为SD、SC和GZ),分别灌胃小鼠相同浓度乙醇、酱香型白酒、高醇白酒和高酯白酒,测定小鼠行为学指标、血液中乙醇和乙醛含量以及肝脏中乙醇脱氢酶(ADH)和乙醛脱氢酶(ALDH)的活性。结果表明,分别添加正戊醇、异丁醇、乙酸乙酯均会明显降低小鼠运动能力,而异戊醇、乳酸乙酯均能减缓乙醇对小鼠运动能力的影响,醇、酯并不能显著影响小鼠平衡能力;与同浓度乙醇溶液相比,灌胃酒样SD后,乙醇质量浓度降低33.32%;在酒样GZ中添加正戊醇,使ADH质量浓度增加11.83%,3种酱香型酒样中添加丁酸乙酯均能使ALDH质量浓度增加。

自杀质粒同源重组介导阴沟肠杆菌adh基因缺失突变体构建及其生物学特性

本研究旨在探究敲除阴沟肠杆菌(Enterobacter cloacae)代谢通路上乙醇脱氢酶基因对乙偶姻代谢合成的影响,进一步提高乙偶姻产量。实验基于E.cloacae SDM中的乙醇脱氢酶adh基因序列设计特异性引物,并构建adh基因自杀质粒pKR6K-∆adh。利用热激法将自杀质粒转入E.cloacae∆budCldh,成功得到多基因缺失菌株E.cloaca∆budC-ldh-adh,并进行发酵性能测试。研究结果显示,与出发菌株相比,新构建的重组菌株在敲除了负责乙醇合成的关键酶乙醇脱氢酶adh基因后,其乙偶姻的生产强度提高了20.6%,产量提高了22.6%,同时乙醇产量提高了92.8%。此外,由于通过基因改造阻断了多条分支代谢路径,流向丁二酸代谢路径的碳通量明显变多,丁二酸产量提高101.3%。本研究成果不仅为构建高效乙偶姻生产菌株提供了有价值的参考,也为乙偶姻的大规模工业生产奠定了基础。

鲟鱼子酶解产物对酒精损伤肝细胞的保护作用及活性肽虚拟筛选

本研究旨在探究鲟鱼子酶解产物对酒精损伤肝细胞的保护作用,并深入阐明其作用机理。以鲟鱼子为研究对象,结合酶解效率和体外活性等关键指标,对其酶解工艺进行优化。借助酒精损伤HepG2细胞模型,系统研究鲟鱼子酶解产物对肝细胞的保护作用。运用计算机虚拟筛选技术,探索潜在的生物活性多肽序列。研究结果表明,在碱性蛋白酶(1343 U/g)及胰酶(27 U/g)1:1复配酶解8 h的条件下获得的鲟鱼子酶解产物最优,蛋白回收率为49.30%±0.57%,水解度为46.05%±0.92%,ABTS+自由基清除率为51.45%±0.66%。不同浓度下均表现出显著的乙醇脱氢酶(Alcohol Dehydrogenase,ADH)激活率(P<0.05),在5 mg/mL时为168.64%±1.42%。在此酶解条件下制备的产物,0.3~2 mg/mL时与模型组相比能显著提升酒精损伤HepG2细胞内超氧化物歧化酶(Superoxide Dismutase,SOD)活性、谷胱甘肽(Glutathione,GSH)含量(P<0.05),显著降低丙二醛(Malondialdehyde,MDA)含量(P<0.05)。此外,酶解产物在0.3~2 mg/mL时显著降低丙氨酸氨基转移酶(Alanine Aminotransaminase,ALT)和天门冬氨酸氨基转移酶(Aspartate Aminotransferase,AST)的活性(P<0.05)。虚拟筛选结果显示,鲟鱼子酶解产物中的LPG和FLPR等5条多肽显示潜在的酒精损伤肝细胞保护作用。因此,本研究为进一步发展鲟鱼子等水产原料的精深加工利用提供了基础支持。

羟酰辅酶A脱氢酶在癌中作用机制的研究进展

脂肪酸代谢与癌的发生发展之间存在密切联系。羟酰辅酶A脱氢酶(HADH)作为脂肪酸β-氧化的关键酶,近年来被发现在多种癌中为抑癌因子,在急性髓系白血病等中为促癌因子。在癌细胞中,HADH除直接催化脂肪酸β-氧化外,还间接作用于PPAR、TNF-α、JAK-STAT3、PI3K/Akt、IFN-γ、MAPK与非经典Wnt等多种信号通路,影响癌细胞增殖与迁移。HADH作为不同种癌诊断、治疗和预后的潜在肿瘤标志物,具备较高临床应用价值。

^(18)F-FDG PET/CT代谢参数辅助评估弥漫大B细胞淋巴瘤化疗疗效

目的探讨^(18)F-FDG PET/CT代谢参数与弥漫大B细胞淋巴瘤(DLBCL)化疗疗效的相关性。方法回顾性总结2021年2月至2023年2月本院确诊DLBCL患者共78例,采用R-CHOP化疗方案。治疗前采用^(18)F-FDG PET/CT测量代谢参数包括标准摄取值的最大值(SUVmax)、峰值(SUVpeak)和平均值(SUVmean)、肿瘤代谢体积(MTV)和糖酵解总量(TLG),解剖学参数包括肿瘤位置、最大直径和AnnArbor分期。治疗4个疗程(21 d为1个疗程)后复查PET/CT,根据Lugano修订淋巴瘤疗效评价标准分为有效组62例和无效组16例,比较两组PET/CT参数。结果与无效组比较,有效组年龄和最大直径较小,改良国际预后指数(NCCN-IPI)和乳酸脱氢酶(LDH)水平较低,Ⅲ~Ⅳ期和骨髓侵犯较少;SUVmax、MTV和TLG值降低(P<0.05)。Spearman检验显示,SUVmax、MTV和TLG值与最大直径、NCCNIPI、LDH和AnnArbor分期呈显著正相关(P<0.05)。ROC计算SUVmax、MTV和TLG值预测DLBCL患者化疗疗效的AUC为0.741、0.886和0.869(P<0.001),MTV和TLG明显优于SUVmax(P<0.001),但MTV和TLG无明显差异(P>0.05)。结论^(18)F-FDG PET/CT作为DLBCL临床分期、疗效评估和预后随访的重要方法,代谢参数MTV和TLG值与化疗疗效密切相关。

6-Phosphogluconate dehydrogenase 2 bridges the OPP and shikimate pathways to enhance aromatic amino acid production in plants

The oxidative pentose phosphate(OPP)pathway provides metabolic intermediates for the shikimate pathway and directs carbon flow to the biosynthesis of aromatic amino acids(AAAs),which serve as basic protein building blocks and precursors of numerous metabolites essential for plant growth.However,genetic evidence linking the two pathways is largely unclear.In this study,we identified 6-phosphogluconate dehydrogenase 2(PGD2),the rate-limiting enzyme of the cytosolic OPP pathway,through suppressor screening of arogenate dehydrogenase 2(adh2)in Arabidopsis.Our data indicated that a single amino acid substitution at position 63(glutamic acid to lysine)of PGD2 enhanced its enzyme activity by facilitating the dissociation of products from the active site of PGD2,thus increasing the accumulation of AAAs and partially restoring the defective phenotype of adh2.Phylogenetic analysis indicated that the point mutation occurred in a well-conserved amino acid residue.Plants with different amino acids at this conserved site of PGDs confer diverse catalytic activities,thus exhibiting distinct AAAs producing capability.These findings uncover the genetic link between the OPP pathway and AAAs biosynthesis through PGD2.The gain-of-function point mutation of PGD2 identified here could be considered as a potential engineering target to alter the metabolic flux for the production of AAAs and downstream compounds.

Decoding 17-Beta-hydroxysteroid Dehydrogenase 13:A Multifaceted Perspective on Its Role in Hepatic Steatosis and Associated Disorders

Chronic liver disease(CLD)represents a significant global health burden,with hepatic steatosis-associated disorders—such as metabolic dysfunction-associated steatohepatitis(MASH),alcoholic liver disease,and hepatitis C virus infection—being major contributors.Recent genome-wide association studies have identified the rs72613567:TA variant in the 17-beta-hydroxysteroid dehydrogenase 13(HSD17B13)gene as a protective factor against the development and progression of these conditions.In this review,we summarized the current evidence surrounding the HSD17B13 rs72613567 variant,aiming to elucidate its impact on CLD risk and outcomes,and to explore the potential mechanisms behind its hepatoprotective effects.The rs72613567:TA variant induces a splice donor site mutation,resulting in a truncated,nonfunctional HSD17B13 protein.Numerous studies have demonstrated that this loss-of-function mutation confers protection against the development of cirrhosis and hepatocellular carcinoma(HCC)in patients with MASH,alcoholic liver disease,and hepatitis C virus infection.Moreover,the rs72613567:TA variant has been associated with reduced liver enzyme levels and improved survival in HCC patients.Integrating this variant into genetic risk scores has shown promise in predicting the progression of fatty liver disease to cirrhosis and HCC.Furthermore,inhibiting HSD17B13 expression through RNA interference and small molecule inhibitors has emerged as a potential therapeutic strategy for MASH.However,the precise molecular mechanisms underlying the hepatoprotective effects of the HSD17B13 rs72613567 variant remain to be fully elucidated.Future research should focus on clarifying the structure-function relationship of HSD17B13 and its role in liver pathophysiology to facilitate the development of targeted therapies for CLD associated with hepatic steatosis.

G6PT-H6PDH-11βHSD1 triad in the liver and its implication in the pathomechanism of the metabolic syndrome

The metabolic syndrome, one of the most common clinical conditions in recent times, represents a combination of cardiometabolic risk determinants, including central obesity, glucose intolerance, insulin resistance, dyslipidemia, non-alcoholic fatty liver disease and hypertension. Prevalence of the metabolic syndrome is rapidly increasing worldwide as a consequence of common overnutrition and consequent obesity. Although a unifying picture of the pathomechanism is still missing, the key role of the pre-receptor glucocorticoid activation has emerged recently. Local glucocorticoid activation is catalyzed by a triad composed of glucose-6-phosphate-transporter, hexose-6-phosphate dehydrogenase and 11β-hydroxysteroid dehydrogenase type 1 in the endoplasmic reticulum. The elements of this system can be found in various cell types, including adipocytes and hepatocytes. While the contribution of glucocorticoid activation in adipose tissue to the pathomechanism of the metabolic syndrome has been well established, the relative importance of the hepatic process is less understood. This review summarizes the available data on the role of the hepatic triad and its role in the metabolic syndrome, by confronting experimental findings with clinical observations.

Lipid nanoparticle-mediated CRISPR/Cas9 gene editing and metabolic engineering for anticancer immunotherapy

Metabolic engineering of the tumor microenvironment has emerged as a new strategy.Lactate dehydrogenase A(LDHA)is a prominent target for metabolic engineering.Here,we designed a cationic lipid nanoparticle formulation for LDHA gene editing.The plasmid DNA delivery efficiency of our lipid nanoparticle formulations was screened by testing the fluorescence of lipid nanoparticles complexed to plasmid DNA encoding green fluorescence protein(GFP).The delivery efficiency was affected by the ratios of three components:a cationic lipid,cholesterol or its derivative,and a fusogenic lipid.The lipid nanoparticle designated formulation F3 was complexed to plasmid DNA co-encoding CRISPR-associated protein 9 and LDHA-specific sgRNA,yielding the lipoplex,pCas9-sgLDHA/F3.The lipoplex including GFP-encoding plasmid DNA provided gene editing in HeLa-GFP cells.Treatment of B16F10 tumor cells with pCas9-sgLDHA/F3 yielded editing of the LDHA gene and increased the pH of the culture medium.pCas9-sgLDHA/F3 treatment activated the interferon-gamma and granzyme production of T cells in culture.In vivo,combining pCas9-sgLDHA/F3 with immune checkpoint-inhibiting anti-PD-L1 antibody provided a synergistic antitumor effect and prolonged the survival of tumor model mice.This study suggests that combining metabolic engineering of the tumor microenvironment with immune checkpoint inhibition could be a valuable antitumor strategy.

Molecular Regulation of the Metabolic Pathways of the Medicinal Plants: <i>Phyla dulcis</i>

Phyla (Lippia) dulcis contains hernundulcin sesquiterpene zero-caloric sweetener that is about a thousand times sweeter than sucrose, and also bitter constituents including camphor and limonene. There is yet no simple method to remove the undesirable constituents. The yield of sweetener hernundulcin is very low, and there is no simple method to maximize its composition. The aim of the project was to characterize the mRNA targets that regulate the primary and terpenoid metabolic enzymes of P. dulcis. Restriction fragment differential display polymerase chain reaction of P. dulcis glutamate dehydrogenase-synthesized RNA showed that many mRNAs encoding β-caryophyllene, (+)-epi-α-bisabolol, bicyclogermacrene, bifunctional sesquiterpene, and geraniol synthases shared sequence homologies with ribulose-1,5-bisphophatase carboxylase, granule-bound starch synthase, pyruvate kinase, glucose-6-phosphate dehydrogenase, and phosphoenol pyruvate carboxylase. Sequence similarities between mRNAs encoding primary metabolic enzymes and terpene synthases suggested that photosynthesis could regulate terpenoid metabolism in order to increase the yield of sweetener hernundulcin.