OBJECTIVETo study the effects of companion fungus on hyphal growth and polysaccharide content of Polyporus umbellata.

METHODThe mycelia and culture filtrate of companion fungus were added to the liquid culture system, and the biomass yield and polysaccharide of P. umbellatus were measured.

RESULTMycelia and appropriate unsterilized culture filtrate of companion fungus could enhance the biomass yield of P. umbellatus significantly, while sterilized culture filtrate of companion fungus could decrease the biomass yield of P. umbellatus significantly. Either mycelia or culture filtrate of companion fungus could increase the intracellular polysaccharide content of P. umbellatus significantly. At the same time, they also could decrease extracellular polysaccharide content of P. umbellatus evidently.

CONCLUSIONThe mycelia and culture filtrate of companion fungus could be used in further fermentation of P. umbellatus.

Biomass ; Extracellular Space ; metabolism ; Hyphae ; growth & development ; Intracellular Space ; metabolism ; Polyporus ; cytology ; growth & development ; metabolism ; Polysaccharides ; metabolism ; Symbiosis

A stochastic model of intracellular calcium oscillations is put forward by taking into account the random opening-closing of Ca2+ channels in endoplasmic reticulum (ER) membrane. The numerical results of the stochastic model show simple and complex calcium oscillations, which accord with the experiment results.
Calcium Channels/*metabolism ; *Calcium Signaling ; Endoplasmic Reticulum/*metabolism ; Intracellular Space ; Kinetics ; Mathematics ; Models, Biological ; Stochastic Processes

Being an essential component of systematic biology, metabolomics has received attention in recent years. It is a post genomic technology aimed at qualitative and quantitative analysis of all low molecular-mass metabolites present in complex biological samples, and mainly investigates the change of endogenous metabolites of a stimulated or disturbed biological system. Investigations into intracellular endogenous metabolites in metabolomics have great advancement in recent years. This review outlines the progress of metabolomics in cell culture analysis including sample preprocessing methods and metabolite target analysis, metabolic profiling analysis, metabolomics analysis and metabolic footprinting analysis.
Animals ; Cell Culture Techniques ; Chemical Fractionation ; methods ; Humans ; Intracellular Space ; metabolism ; Metabolome ; Metabolomics ; methods

A stochastic model of intracellular calcium oscillations is put forward by taking into account the random opening-closing of Ca2+ channels in endoplasmic reticulum (ER) membrane. The numerical results of the stochastic model show simple and complex calcium oscillations, which accord with the experiment results.
Calcium Channels ; metabolism ; Calcium Signaling ; Endoplasmic Reticulum ; metabolism ; Humans ; Intracellular Space ; Kinetics ; Mathematics ; Models, Biological ; Stochastic Processes

Myocardial infarction afflicts close to three quarters of a million Americans annually, resulting in reduced heart function, arrhythmia, and frequently death. Cardiomyocyte death reduces the heart's pump capacity while the deposition of a non-conductive scar incurs the risk of arrhythmia. Direct cardiac reprogramming emerged as a novel technology to simultaneously reduce scar tissue and generate new cardiomyocytes to restore cardiac function. This technology converts endogenous cardiac fibroblasts directly into induced cardiomyocyte-like cells using a variety of cocktails including transcription factors, microRNAs, and small molecules. Although promising, direct cardiac reprogramming is still in its fledging phase, and numerous barriers have to be overcome prior to its clinical application. This review discusses current findings to optimize reprogramming efficiency, including reprogramming factor cocktails and stoichiometry, epigenetic barriers to cell fate reprogramming, incomplete conversion and residual fibroblast identity, requisite growth factors, and environmental cues. Finally, we address the current challenges and future directions for the field.
Animals ; Cellular Reprogramming ; Epigenesis, Genetic ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Intracellular Space ; metabolism ; Myocardium ; cytology ; Signal Transduction

The commercial application of plant cell cultures is often hindered by the instability of secondary metabolite biosynthesis, where the metabolite yield fluctuates and decline dramatically over subcultures. This study proposed that such instability is due to the fluctuations of culture variables. To validate this hypothesis, the effects of the fluctuations of two culture variables (subculture cycle and inoculum size) on the biomass, anthocyanin biosynthesig, intracellular carbon, nitrogen and phosphate during continuous 10 subculture cycles were investigated. The subculture cycle was fluctuated for 12h in a 7 day cycle (6.5, 7 and 7.5 d), and the inoculum size was fluctuated by 20% on basis of 2.00 g (1.60, 2.00 and 2.40 g). It was found that all the measured culture parameters fluctuated over the 10 subculture cycles. The fluctuation in terms of inoculum sizes had a greater effect on the stability of anthocyanin biosynthesis in suspension cultures of V. vinifera. Among all the subculture conditions investigated, 7d-subculture cycle and 1.60 g-inoculum size was the best one to hold the relatively stable anthocyanin production. The anthocyanin yield presented a negative correlation with intracellular sucrose content or intracellular total phosphate content.
Anthocyanins ; biosynthesis ; Carbohydrate Metabolism ; Cell Culture Techniques ; methods ; Intracellular Space ; metabolism ; Phosphates ; metabolism ; Plant Proteins ; metabolism ; Suspensions ; Vitis ; cytology ; growth & development ; metabolism

Intracellular calcium overload is a key factor for myocardial ischemia reperfusion injury (IR). However, there was no report for interstitial calcium concentration dynamics. We investigated the interstitial calcium dynamics in rat myocardial IR model in vivo. A microdialysis system was involved, and the time delay of the system and recovery time was introduced and tested with a fluids switching method. Twelve SD rats were divided into IR or control group. Myocardial IR was induced by ligating (20 min) then releasing (60 min) the suture underlying left anterior descending branch. Mycrodialyisis probe was implanted into the left ventricular myocardium perfusion area for occlusion. Dialysate samples were collected every 10 min. Dialysate calcium concentration was detected with an atomic absorption spectrophotometer. Recovery time for the microdialysis system was 20 min, and recovery rate was 16%. Dialysate calcium concentration showed no changes during ischemia, descended immediately after reperfusion, reached the lowest level (67% of baseline value) 20 min after reperfusion, then escalated slowly. Recovery time was an important parameter for mycrodialysis technique, and it should not be neglected and needed to be tested. Our data suggest that interstitial calcium concentration in rats with myocardial IR in vivo kept steady in ischemia, descended rapidly at the initial reperfusion, then rebounded slowly. In conclusion, we introduced the concept of recovery time for microdialysis and provided a simple testing method.
Animals ; Calcium ; metabolism ; Dialysis Solutions ; metabolism ; Intracellular Space ; metabolism ; Kinetics ; Male ; Microdialysis ; methods ; Myocardial Reperfusion Injury ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spectrophotometry, Atomic ; Time Factors

OBJECTIVETo found the subcellular location of the human gene 6 transactivated by nonstructural protein 5A of hepatitis C virus (NS5ATP6).

METHODSGreen fluorescent protein (GFP) expression vector pEGFP- NS5ATP6 was established. The pEGFP- NS5ATP6 was transfected into HepG2 cells, and analyze the subcellular location of the proteins expressed by NS5ATP6 through Green fluorescent microscopy after 24 hours.

RESULTSThe pEGFP- NS5ATP6 gene was successful cloned, NS5ATP6 can express protein in cells and subcellularly located in cell plasma.

CONCLUSIONNS5ATP6 can express protein, and the protein expressed by NS5ATP6 subcellularly located in cell plasma.

Cell Line, Tumor ; Genetic Vectors ; genetics ; metabolism ; Hepacivirus ; Humans ; Intracellular Space ; metabolism ; Microscopy, Fluorescence ; Transcriptional Activation ; Transfection ; Viral Nonstructural Proteins ; metabolism

Mammalian carboxylesterases hydrolyze a wide range of xenobiotic and endogenous compounds, including lipid esters. Physiological functions of carboxylesterases in lipid metabolism and energy homeostasis in vivo have been demonstrated by genetic manipulations and chemical inhibition in mice, and in vitro through (over)expression, knockdown of expression, and chemical inhibition in a variety of cells. Recent research advances have revealed the relevance of carboxylesterases to metabolic diseases such as obesity and fatty liver disease, suggesting these enzymes might be potential targets for treatment of metabolic disorders. In order to translate pre-clinical studies in cellular and mouse models to humans, differences and similarities of carboxylesterases between mice and human need to be elucidated. This review presents and discusses the research progress in structure and function of mouse and human carboxylesterases, and the role of these enzymes in lipid metabolism and metabolic disorders.
Amino Acid Sequence ; Animals ; Carboxylic Ester Hydrolases ; chemistry ; genetics ; metabolism ; Humans ; Intracellular Space ; metabolism ; Lipid Metabolism ; Mice ; Polymorphism, Single Nucleotide ; Protein Domains

Na+ -Ca2+ exchanger (NCX) transports Ca2+ coupled with Na+ across the plasma membrane in a bi-directional mode. Ca2+ flux via NCX mediates osteogenic processes, such as formation of extracellular matrix proteins and bone nodules. However, it is not clearly understood how the NCX regulates cellular Ca2+ movements in osteogenic processes. In this study, the role of NCX in modulating Ca2+ content of intracellular stores ([Ca2+](ER)) was investigated by measuring intracellular Ca2+ activity in isolated rat osteoblasts. Removal of extracellular Na+ elicited a transient increase of intracellular Ca2+ concentration ([Ca2+](i)). Pretreatment of antisense oligodeoxynucleotide (AS) against NCX depressed this transient Ca2+ rise and raised the basal level of [Ca2+](i). In AS-pretreated cells, the expression and activity of alkaline phosphatase (ALP), an osteogenic marker, were decreased. However, the cell viability was not affected by AS-pretreatment. Suppression of NCX activity by the AS-pretreatment decreased ATP-activated Ca2+ release from intracellular stores and significantly enhanced Ca2+ influx via store operated calcium influx (SOCI), compared to those of S-pretreated or control cells. These results strongly suggest that NCX has a regulatory role in cellular Ca2+ pathways in osteoblasts by modulating intracellular Ca2+ content.
Alkaline Phosphatase/metabolism ; Animals ; Calcium/*metabolism ; Cell Membrane/metabolism ; Cell Survival ; Cells, Cultured ; Cytoplasm/metabolism ; Endoplasmic Reticulum/metabolism ; Intracellular Space/metabolism ; Oligodeoxyribonucleotides, Antisense/pharmacology ; Osteoblasts/drug effects/*physiology ; Rats ; Signal Transduction ; Sodium/physiology ; Sodium-Calcium Exchanger/*physiology