Cardiac energy metabolism disorder mediated by energy substrate imbalance and mitochondrial damage upon tebuconazole exposure

Tebuconazole exposure has been described as an increasing hazard to human health.An increasing number of recent studies have shown a positive association between tebuconazole exposure and cardiovascular disease risk,which is characterized by the reduction of adenosine triphosphate(ATP)synthesis.However,researches on the damage of tebuconazole exposure to energy metabolism and the related molecular mechanisms are limited.In the present study,male C57BL/6 mice were treated with tebuconazole at different low concentrations for 4 weeks.The results indicated that tebuconazole could accumulate in the heart and further induce the decrease of ATP content in the mouse heart.Importantly,tebuconazole induced an obvious shift in substrate utilization of fatty acid and glucose by disrupting their corresponding transporters(GLUT1,GLUT4,CD36,FABP3 and FATP1)expression,and significantly repressed the expression of mitochondrial biogenesis(Gabpa and Tfam)and oxidative phosphorylation(CS,Ndufa4,Sdhb,Cox5a and Atp5b)related genes in a dosedependent manner.Further investigation revealed that these alterations were related to the IRS1/AKT and PPARγ/RXRαpathways.These findings contribute to a better understanding of triazole fungicide-induced cardiovascular disease by revealing the key indicators associated with this phenomenon.

Using pipette tips to readily generate spheroids comprising single or multiple cell types

Three-dimensional(3D)cell culture methods have been validated that can replicate the tumor environment in vivo to a large extent,providing an effective tool for studying tumors.In this study,we demonstrated the use of standard laboratory pipette tips as micro vessels for generating 3D cell spheroids.No microfabrication or wet-chemistry surface modifications were involved in the procedure.Spheroids consisting of single or multiple cell types were generated within 24 h just by pipetting and incubating a cell suspension in pipette tips.Scanning electron microscope and optical microscope proved that the cells grew together tightly,and suggested that while gravity force might have initiated the sedimentation of cells at the bottom of the tip,the active aggregation of cells to form tight cell-cell interactions drove the formation of spheroids.Using common laboratory micropipettes and pipette tips,the rate of spheroid generation and the generation reproducibility was characterized from five boxes each with 80 tips.The ease of transferring reagents allowed modeling of the growth of microvascular endothelial cells in tumor spheroids.Moreover,the pairing and fusion of tumor spheroids could be manipulated in the pipette tips,suggesting the potential for building and assembling heterogeneous micro-tumor tissues in vitro to mimic solid tumors in vivo.This study demonstrated that spheroids can be readily and cost-effectively generated in standard biological laboratories in a timely manner using pipette tips.

Highly selective kinetic resolution of D/L-syn-p-sulfone phenylserine catalyzed by D-threonine aldolase in two-phase ionic solvent

In the chemical synthesis of L-syn-p-methylsulfoxide phenylserine ethyl ester(D-ethyl ester),L-tartaric acid or enzymatic resolution is employed to resolve the racemate,and thus obtain the target compound,and the remaining isomer can be recycled to obtain the raw material.In this study,high-purity L-syn-p-methylsulfoxide phenylserine(L-syn-MPS)was obtained.The kinetics of the D-threonine aldolase enzymatic hydrolysis reaction reveals that D-syn-p-sulfoxylphenylserine resolves well in[BMIM][BF 4]ionic solvents.The D/L-syn-MPS racemate was resolved using a two-phase ionic solvent[BMIM][NTf 2]to afford L-syn-MPS(ee(enantiomeric excess)>99%)and a white solid in 41.7%yield.Therefore,this system is suitable for the separation of insoluble aldehydes and successfully avoids the condensation of hydroxyl aldehydes to form D-anti-MPS.

Biallelic mutations in UGDH cause congenital microcephaly

Hengel et al recently reported that bi-allelic loss-of-function mutations in UDP-Glucose 6-Dehydrogenase(UGDH)caused a severe epileptic encephalopathy syndrome-Jamuar syndrome(OMIM#618792).1 The functional studies partially recapitulated the clinical phenotypes in the patient-derived cerebral organoid.A reduced number of proliferating neuronal progenitors in cerebral organoids was shown,which is a critical mechanism in congenital microcephaly(CM)whose patients were born with an occipitofrontal circumference(OCF)more than 2 standard deviations below average for age and sex.However,none of the reported patients in the article presented the phenotype as CM.

Toxoplasma gondii GRA15II effector-induced M1 cells ameliorate liver fibrosis in mice infected with Schistosomiasis japonica

Recent studies indicated that type II Toxoplasma gondii(Tg)GRA15II favored the generation of classically activated macrophages(M1),whereas type I/III TgROP16I/III promoted the polarization of alternatively activated macrophages(M2).A number of studies have demonstrated that M2 cells are involved in the pathogenesis of the liver fibrogenesis caused by Schistosoma japonicum.The purpose of the present study was to explore the inhibitory effect of Toxoplasma-derived TgGRA15II on mouse hepatic fibrosis with schistosomiasis.The gra15II and rop16I/III genes were amplified from strains T.gondii PRU and Chinese 1 Wh3,respectively.Lentiviral vectors containing the gra15II or rop16I/III plasmid were constructed and used to infect the RAW264.7 cell line.The polarization of the transfected cells was evaluated,followed by co-culture of the biased macrophages with mouse hepatic stellate JS1 cells.Then,mice were injected with GRA15II-driven macrophages via the tail vein and infected with S.japonicum cercariae.TgGRA15II induced a M1-biased response,whereas TgROP16I/III drove the macrophages to a M2-like phenotype.The in vitro experiments indicated that JS1 cell proliferation and collagen synthesis were decreased following co-culture with TgGRA15II-activated macrophages.Furthermore,mice inoculated with TgGRA15II-biased macrophages displayed a notable alleviation of collagen deposition and granuloma formation in their liver tissues.Our results suggest that TgGRA15II-induced M1 cells may dampen the M2 dominant pathogenesis of hepatic fibrosis and granulomatosis.These results provide insights into the use of parasite-derived immunomodulators as potential anti-fibrosis agents and to re-balance the schistosomiasis-induced immune response.

Qualitative and quantitative analysis of glucosinolates and nucleosides in Radix Isatidis by HPLC and liquid chromatography tandem mass spectrometry

Multi-component fingerprinting and quantitation of the glucosinolates and nucleosides in samples of Radix Isatidis have been carried out using high-performance liquid chromatography with diode-array detection and electrospray ionization tandem mass spectrometry(HPLC–DAD–ESI/MS).Five nucleosides together with one glucosinolate were identified by comparing retention times,ultraviolet spectra,mass spectra and/or empirical molecular formulae of reference compounds.Quantitation of these six compounds was carried out simultaneously by HPLC on a Phenomenex Luna C18 column using gradient elution with methanol and water and detection at 254 nm.All calibration curves were linear(r40.9994)within test ranges.Limits of detection and quantitation were 0.33 ng and 2.50 ng on column,respectively.Intra-and inter-day precision(as relative standard deviation)for all analytes was o2.19%with recoveries in the range 99.6%–101.8%at three concentration levels.The validated method was successfully applied to fingerprinting and assay of 25 batches of Radix Isatidis sourced from different geographical regions of China.The method is simple and reliable and has potential value in the quality control of Radix Isatidis.

Three-dimensional bioprinting sodium alginate/gelatin scaffold combined with neural stem cells and oligodendrocytes markedly promoting nerve regeneration after spinal cord injury

Accumulating research has indicated that the transplantation of combined stem cells and scaffolds is an effective method for spinal cord injury(SCI).The development of three-dimensional(3D)bioprinting technology can make the 3D scaffolds combined with cells more accurate and effective for SCI treatment.However,unmyelinated newborn nerve fibers have no nerve signaling conduction,hampering recovery of motor function.In this study,we designed and printed a type of sodium alginate/gelatin scaffold loaded with neural stem cells and oligodendrocytes,which were involved in the formation of the myelin sheaths of neural cell axons.In order to observe the effectiveness of this 3D bioprinting scaffold,we transplanted it into the completely transected rat spinal cord,and then immunofluorescence staining,hematoxylin–eosin staining and behavioral assessment were performed.The results showed that this 3D bioprinting scaffold markedly improved the hindlimb motor function and promoted nerve regeneration.These findings suggested that this novel 3D bioprinting scaffold was a good carrier for cells transplantation,thereby enhancing spinal cord repair following injury.